US20210247372A1 - Method and apparatus for applying aggregating sampling to food items - Google Patents
Method and apparatus for applying aggregating sampling to food items Download PDFInfo
- Publication number
- US20210247372A1 US20210247372A1 US17/152,566 US202117152566A US2021247372A1 US 20210247372 A1 US20210247372 A1 US 20210247372A1 US 202117152566 A US202117152566 A US 202117152566A US 2021247372 A1 US2021247372 A1 US 2021247372A1
- Authority
- US
- United States
- Prior art keywords
- sampling
- microorganisms
- microbial
- covering
- aggregating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000005070 sampling Methods 0.000 title claims abstract description 219
- 238000000034 method Methods 0.000 title claims abstract description 101
- 230000004931 aggregating effect Effects 0.000 title claims abstract description 68
- 235000013305 food Nutrition 0.000 title claims abstract description 46
- 230000000813 microbial effect Effects 0.000 claims abstract description 82
- 239000000463 material Substances 0.000 claims abstract description 56
- 235000013372 meat Nutrition 0.000 claims abstract description 14
- 238000004519 manufacturing process Methods 0.000 claims abstract description 13
- 244000005700 microbiome Species 0.000 claims description 93
- 244000052769 pathogen Species 0.000 claims description 31
- 238000012864 cross contamination Methods 0.000 claims description 24
- 238000004140 cleaning Methods 0.000 claims description 22
- 238000012216 screening Methods 0.000 claims description 21
- 230000008569 process Effects 0.000 claims description 19
- 238000009739 binding Methods 0.000 claims description 14
- 230000027455 binding Effects 0.000 claims description 13
- 238000000605 extraction Methods 0.000 claims description 13
- 239000012530 fluid Substances 0.000 claims description 12
- 238000012790 confirmation Methods 0.000 claims description 11
- 238000003306 harvesting Methods 0.000 claims description 11
- 238000005520 cutting process Methods 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 238000003070 Statistical process control Methods 0.000 claims description 6
- 239000003446 ligand Substances 0.000 claims description 6
- 108020004707 nucleic acids Proteins 0.000 claims description 5
- 150000007523 nucleic acids Chemical class 0.000 claims description 5
- 102000039446 nucleic acids Human genes 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 230000003993 interaction Effects 0.000 claims description 4
- 108091023037 Aptamer Proteins 0.000 claims description 3
- 239000000427 antigen Substances 0.000 claims description 3
- 108091007433 antigens Proteins 0.000 claims description 3
- 102000036639 antigens Human genes 0.000 claims description 3
- 230000009870 specific binding Effects 0.000 claims description 3
- 239000000047 product Substances 0.000 description 56
- 239000000523 sample Substances 0.000 description 45
- 238000001514 detection method Methods 0.000 description 29
- 238000012360 testing method Methods 0.000 description 26
- 230000008901 benefit Effects 0.000 description 18
- 238000004458 analytical method Methods 0.000 description 16
- 239000004744 fabric Substances 0.000 description 15
- 238000003860 storage Methods 0.000 description 15
- 238000013459 approach Methods 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 12
- 238000005516 engineering process Methods 0.000 description 12
- 238000012545 processing Methods 0.000 description 12
- 238000011021 bench scale process Methods 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 230000009471 action Effects 0.000 description 8
- 241000208822 Lactuca Species 0.000 description 7
- 235000003228 Lactuca sativa Nutrition 0.000 description 7
- 238000013461 design Methods 0.000 description 7
- 230000006872 improvement Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 230000001717 pathogenic effect Effects 0.000 description 6
- 238000012546 transfer Methods 0.000 description 6
- 238000011282 treatment Methods 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 5
- 238000011109 contamination Methods 0.000 description 5
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 239000002953 phosphate buffered saline Substances 0.000 description 5
- 239000000872 buffer Substances 0.000 description 4
- 238000004590 computer program Methods 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 239000013615 primer Substances 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 238000011529 RT qPCR Methods 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000000356 contaminant Substances 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 238000000265 homogenisation Methods 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 238000007747 plating Methods 0.000 description 3
- 235000013547 stew Nutrition 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 238000012952 Resampling Methods 0.000 description 2
- 241000607142 Salmonella Species 0.000 description 2
- 235000009337 Spinacia oleracea Nutrition 0.000 description 2
- 244000300264 Spinacia oleracea Species 0.000 description 2
- 239000002250 absorbent Substances 0.000 description 2
- 230000002745 absorbent Effects 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000012468 concentrated sample Substances 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000013213 extrapolation Methods 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 235000021384 green leafy vegetables Nutrition 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000002906 microbiologic effect Effects 0.000 description 2
- 238000010606 normalization Methods 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 230000000737 periodic effect Effects 0.000 description 2
- -1 polypropylene Polymers 0.000 description 2
- 238000011176 pooling Methods 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 108020004418 ribosomal RNA Proteins 0.000 description 2
- 239000011435 rock Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 210000003813 thumb Anatomy 0.000 description 2
- 229930195730 Aflatoxin Natural products 0.000 description 1
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000589876 Campylobacter Species 0.000 description 1
- 108020001019 DNA Primers Proteins 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000186781 Listeria Species 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 229920001410 Microfiber Polymers 0.000 description 1
- 108091093105 Nuclear DNA Proteins 0.000 description 1
- 241000238413 Octopus Species 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000242583 Scyphozoa Species 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 239000005409 aflatoxin Substances 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 239000013566 allergen Substances 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 238000003491 array Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000008275 binding mechanism Effects 0.000 description 1
- 238000002306 biochemical method Methods 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008364 bulk solution Substances 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000009260 cross reactivity Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000012632 extractable Substances 0.000 description 1
- 239000011536 extraction buffer Substances 0.000 description 1
- 230000005294 ferromagnetic effect Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 210000003811 finger Anatomy 0.000 description 1
- 238000005243 fluidization Methods 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 230000002949 hemolytic effect Effects 0.000 description 1
- 244000052637 human pathogen Species 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000002032 lab-on-a-chip Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000005291 magnetic effect Effects 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000003658 microfiber Substances 0.000 description 1
- 244000005706 microflora Species 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002121 nanofiber Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 239000004745 nonwoven fabric Substances 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 230000008447 perception Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 244000000003 plant pathogen Species 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920000098 polyolefin Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004886 process control Methods 0.000 description 1
- 238000011028 process validation Methods 0.000 description 1
- 230000000135 prohibitive effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000000275 quality assurance Methods 0.000 description 1
- 238000007430 reference method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000001223 reverse osmosis Methods 0.000 description 1
- 210000003705 ribosome Anatomy 0.000 description 1
- 238000004826 seaming Methods 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 239000004758 synthetic textile Substances 0.000 description 1
- 239000013077 target material Substances 0.000 description 1
- 238000004154 testing of material Methods 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000017105 transposition Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/02—Food
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D21/00—Separation of suspended solid particles from liquids by sedimentation
- B01D21/26—Separation of sediment aided by centrifugal force or centripetal force
- B01D21/262—Separation of sediment aided by centrifugal force or centripetal force by using a centrifuge
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/02—Reverse osmosis; Hyperfiltration ; Nanofiltration
- B01D61/025—Reverse osmosis; Hyperfiltration
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/14—Ultrafiltration; Microfiltration
- B01D61/145—Ultrafiltration
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/10—Devices for withdrawing samples in the liquid or fluent state
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N2001/028—Sampling from a surface, swabbing, vaporising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
- G01N2001/2826—Collecting by adsorption or absorption
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
- G01N2001/4088—Concentrating samples by other techniques involving separation of suspended solids filtration
Definitions
- the present disclosure relates generally to improving the food safety of ready-to-eat produce and other food items and providing process validation, and more particularly, to methods and apparatus for microbial sampling of food items and other materials.
- Certain aspects provide a method for microbial sampling of foods and other products.
- the method generally includes gathering a microbial sampling from one or more items, extracting microorganisms from the microbial sampling, concentrating the microorganisms, cleaning the microorganisms, tallying the relative presence of the microorganisms and any potential pathogens, aggregating this information of a microorganism tally from the tallying of microorganisms into a microorganism report, confirming the microorganism tally, and reporting the microorganism report of the microorganism tally.
- Certain aspects provide a method for microbial sampling of foods.
- the method generally includes providing at least one aggregating sampler at one or more sampling locations, and sampling, using the at least one aggregating sampler, a production lot of produce creating one or more samples that makes up a microbial sampling.
- the one or more aspects comprise the features hereinafter fully described and particularly pointed out in the claims.
- the following description and the annexed drawings set forth in detail certain illustrative features of the one or more aspects. These features are indicative, however, of but a few of the various ways in which the principles of various aspects may be employed, and this description is intended to include all such aspects and their equivalents.
- FIG. 1 illustrates example operations for microbial sampling, in accordance with aspects of the present disclosure.
- FIG. 2 illustrates example operations for applying aggregating sampling, in accordance with aspects of the present disclosure.
- FIG. 3 illustrates a Table 1 that includes example treatments and results for meat sampling, in accordance with aspects of the present disclosure.
- FIG. 4 illustrates a Table 2 that includes example treatments and results for leafy sampling, in accordance with aspects of the present disclosure.
- FIG. 5 illustrates an aggregated sampler that includes a pocket in accordance with aspects of the present disclosure.
- FIG. 6 illustrates an aggregated sampler that includes a pocket that extends through a covering in accordance with aspects of the present disclosure.
- FIG. 7 illustrates an aggregated sampler with at least one convex surface in accordance with aspects of the present disclosure.
- FIG. 8 illustrates an aggregated sampler that includes at least one concave surface in accordance with aspects of the present disclosure.
- FIG. 9 illustrates an aggregated sampler that includes a curved shape in accordance with aspects of the present disclosure.
- FIG. 10 illustrates an aggregated sampler that includes a hole or loop in accordance with aspects of the present disclosure.
- FIG. 11 illustrates an aggregated sampler that is shaped as a mitten or glove for a hand in accordance with aspects of the present disclosure.
- aspects of the present disclosure provide apparatus, methods, and/or systems for automated and semi-automated microbial sampling of foods and other materials.
- Other materials can be as diverse as water or air streams. More commonly, it will include kindred products of food such as pet food, medical materials or dietary supplements where microbial testing is needed to confirm hygienic operation.
- Sampling can be active as mediated by material flow or operator mediated by applying the sampler to a surface. Sampling can also be more passive and depend on passive contact or gravity sedimentation.
- devices are assembled and linked to a rapid reporting system to provide a more representative sampling and faster analysis of a food product or other material.
- a similar system provides for robotically sampling a field crop and delivering results with a mobile laboratory. Both embodiments can include a two-stage screening system for speed and for economy, but a conventional testing approach can be considered to take advantage of the sampling improvements.
- Some microbial testing may assume that the test organisms are evenly distributed allowing a grab sample to be representative of the whole. This flawed assumption only accounts for inhomogeneous distributions in aggregate and requires many samples to characterize the microbial population of a lot. Test and release inspection based on grab samples may be flawed to the extent that grab sampling can inherently miss significant contamination. Furthermore, the size the sample limits the detection limit to levels that may be order of magnitude above the background rate and the level where risk becomes imminent. On the other hand, microbial proliferation during enrichment culture occurs under conditions selected to favor the growth of the potential pathogens which often have no relationship with the commercial conditions of storage and therefore generates extorted perceptions of the microbial growth risk.
- One or more embodiments as disclosed herein may address these short comings of the current sampling and testing practice and provides a system to generate more meaningful, real time or near real time, onsite assessment of microbial contamination risk.
- automated and semi-automated microbial sampling of foods and other materials is provided.
- devices may be assembled and linked to a rapid reporting system to provide a more representative sampling and faster analysis of a food product.
- a system may provide robotic sampling of a field crop and may deliver results with a mobile laboratory. Both examples may include a two-stage screening system that may provide speed and for economy. Although less advantageous, it is reasonable to take advantage of improved sampling with conventional enrichment and detection systems.
- a method and system of microbial sampling includes providing a sampling sheet, such as a MicroTally Sheet, which is then used with a continuous sampling device (CSD) to collect biological agents.
- the CSD may change the sampling sheet automatically between samples.
- the sampling sheet is then used for analysis.
- target bacteria may be removed from the sampling medium.
- the samples of target bacteria are cleaned up and concentrated using a treater and made suitable for analysis.
- the treated sample is then analyzed using molecular or biochemical methods and target agents are detected with acceptable accuracy and sensitivity.
- a cloud based data reporting system with user appropriate dashboards to report actionable information and facilitate timely decision making may also be provided.
- FIG. 1 illustrates specific operations 100 for microbial sampling, in accordance with aspects of the present disclosure.
- operations 100 begin, at block 102 , with gathering a microbial sampling from one or more food items which may include, for example, produce, meat, and other food products or other materials.
- operations 100 include extracting microorganisms from the microbial sampling.
- the operations 100 include concentrating the microorganisms.
- the operations 100 include cleaning the microorganisms.
- the operations 100 include, at block 110 , tallying the relative presence of the microorganisms and any potential pathogens.
- the operations 100 also include, at block 112 , aggregating this information of a microorganism tally from the tallying of microorganisms into a microorganism report.
- the operations include confirming the microorganism tally.
- the operations may also include, at block 116 , reporting the microorganism report of the microorganism tally.
- sampling may include using a sampling sheet or swab that collects a sample and is stomached in a 300 micron partitioned bag with 200 mls of eluting buffer and stomached.
- Concentration and cleanup of the sample may include siphoning an eluting buffer and entrained organisms through proprietary sequential filter and the targets may be deposited on a 0.22 micron PC filter.
- the targets are analyzed from the filter which may include for example, DNA purified and qPCR being run for Index elements including pathogen intensity, Enteric status, and/or positive control.
- Cloud based reporting based on, for example Ignition and SQL database, may be provided with user appropriate dashboards to provide timely actionable information
- pathogens may be confirmed by resampling and enrichment procedures, by resampling and doing definitive pathogen tests, or by doing confirmation tests on the original DNA depending on the regulatory guidance.
- a cassette and cartridge system may be implemented to streamline the sampling process.
- eliminating the use of a stomacher and partitioned bag may be provided in one or more cases.
- a binding collector may be provided to replace the PC membrane filter which may help streamline the sample delivery to the detector and potentially eliminate the need for DNA purification.
- Use of Ribosomal RNA may become a new standard for one or more such cases.
- a detector may use flow amplification and laboratory on a chip type technology to further reduce detection times and costs.
- the timeframe from sampling to information may be driven by the needs of the business class but is not limited thereto. Short shelf-life product can justify greater speed. Valuable commodities such as meat products will want more testing to limit exposure when a problem is expected or anticipated. These factors will impact the degree to which a complete automated solution is implemented or conversely when a system more akin to current practices is used to gain some of the benefits of improved sampling.
- the surface area of an aggregating sampler affords advantages beyond material sampling when greater sensitive is desired for surface and fluid flow sampling. Water and air stream sampling are two examples of where flow sampling is advantageous. The surface area is also applicable to the sampling of surfaces where topical contamination is of concern.
- the geometry can be altered to allow sampling of a powder flow through a pipe with a circular geometry where a pipe segment is exchanged between lots.
- the pipe section would either be lined with sampling material or better include baffles maximizing product contact with the sampling surfaces.
- baffles maximizing product contact with the sampling surfaces.
- vertical chutes below pocket fillers to sample product just prior to bagging in a form fill and seal machine.
- a microbial sampler may be included, for example, non-woven fabric, various micro fiber materials, sponges, and/or any absorbent sheet material.
- a non-woven polypropylene or polyethylene fabric are of particular note as these materials are allowed for food contact and therefore have very low extractables which might otherwise contaminant the material stream under examination.
- a feed cartridge can be used to deliver multiple sheets to the sampling location at one time. This cartridge would be placed on the line after sanitation has completed all preparations protecting the drive mechanisms from the harsh cleaning process.
- a magazine of cassettes can be loaded to collect individual pieces of sampling material.
- Both the cartridge and cassette are engineered to advance the sampling material when appropriate, (e.g. When a lot is completed, when a tote is moved, etc.).
- the motive force to advance sampling sheets can be provided by a motor, or supplied by a manual crank or handle, or by an operator depending on the specifics of the operation.
- Both cassette and cartridge may be designed to protect the microbial integrity of the contents. The cartridges may prevent microbial growth after sampling, external contamination, and cross contamination between lots.
- sampling material can be mounted on an inert backing material to facilitate the sanitary placement of sampling sheets.
- sheets of sampling material can be separated by short spaces of inert material to ensure that used sampling materials do not contaminate other sheets.
- the usual design parameters for process equipment may be applied to these devices including, for example, an aggregating sampler.
- an aggregating sampler For example, heavy gauge 316 stainless is an appropriate material.
- the location When the sampling device is not in place, the location may be passive such as a dead plate where product passes without damage or hindrance. It may be sanitary design from the beginning such that it is easily cleaned.
- the cartridges may carry the information regarding the sample they contain. If the sample is used in a manual mode, an electronic transfer of this information along with the sample is also advantageous to avoid human error and speed the flow of information. This transfer of information can occur through the cloud using barcodes, a database and location information.
- sampling has utility beyond testing for human pathogens in that it can be used for testing for plant pathogens that can decrease the productivity of a crop.
- pathogens For example, early detection of mildew spores prompt early harvest of a spinach field. Detection of blight in a wheat or corn field might prompt the use of a disease control measure on the affected field before the blight destroys the crop.
- an octopus tentacle configuration may be the appropriate geometry where strands of the sampling material are slide across the surface of the crop. These strands can be fuzzy cords or strips of material depending on what provide the greatest effective contact. These tentacles can be contacted to the crop by various mechanisms including robots, tractors, hand carrying or drones. It is most important that the altitude be held constant to allow contact while minimizing damage to the crop. To increase the effectiveness of the sampling, it can be advantageous to wick moisture down these tentacles or install vacuuming or sucking mechanism.
- forming the sampling material into pockets, mittens or gloves can facilitate use. Ease of use will generate greater compliance with the sampling protocol.
- the duration of contact is a factor in determining the effectiveness of sampling. Typical durations are minutes. 2-5 minutes durations will work for most applications.
- FIG. 2 illustrates example operations 200 for applying aggregating sampling, in accordance with aspects of the present disclosure.
- operations 200 begin, at block 202 , with providing at least one aggregating sampler at one or more sampling locations.
- operations 200 include, at block 204 , sampling, using the at least one aggregating sampler, a production lot of produce creating one or more samples that makes up a microbial sampling.
- the one or more samples may be configured to be processed to indicate if pathogens are present at no greater than a normal background.
- the one or more sampling locations may include at least one of in a field, at harvest, just after dumping or cutting, in a wash system, or after the wash system.
- additional operations may be include such as, for example, assessing, using the aggregating sampler, a level of cross contamination control to validate or verify a wash process.
- an aggregating sampler may be provided that sufficiently samples a production lot of ready-to-eat produce to confirm that pathogens are present at no greater than the normal background. In one or more cases, an aggregating sampler may assess the level of cross contamination control to validate or verify a wash process.
- an aggregating sampler and a sampling location of the aggregating sampler may be provided. Additionally, sampling by the aggregating sampler may be provided to generate one or more desired samples. Analysis of the one or more samples and interpretation of the analysis results may also be provided. These elements can be practiced individually or together to enhance food safety.
- the aggregating sampler may include a collection surface and an apparatus for holding and positioning the collection surface such that the collection surface contacts product that is to be sampled for micro-organisms or other targets.
- a surface with a sampling efficiency that allows an increased effective sampling size when a production lot of the product is sampled may be provided. For example, two hours of production of a leafy green product may be sampled with such a device. During the two hours a large amount, for example 10,000 to 30,000 pounds, of product will have crossed the sampling surface. If the sampling device is at least 25% efficient as shown in bench scale studies, the effective sample size may be 2500 to 7500 pounds. These sample sizes are enormous when compared to the few hundred grams of a normal grab sample.
- a sampling surface that is compatible with the product is provided.
- a non-woven polyolefin cloth has proven to be effective.
- Another factor may include a design that allows safe and rapid exchange of the sampling surface.
- Another factor that may be included relates to any apparatus left on the line being easily cleaned and may incorporate a sanitary design.
- food grade stainless steel may be a material selected when implementing an aggregating sampler.
- aggregating sampling may yield improvements in food safety. For example, some locations include: 1) In the field; 2) At harvest; 3) Just after dumping or cutting; 4) In the wash system; or 5) After the wash system. In each area, there are particular locations that may be selected for aggregate sampling, but these will vary with the configuration of the specific line and the product to be sampled. Not all areas will be appropriate for all products. The selections may be guided by the desired information. Currently, grab samples are often taken in all of these areas but these grab samples are unable to represent the population under examination and such sampling is generally destructive.
- an aggregating sampler may be provide that can replace the current practice of taking a grab sample in a field by cutting leaves.
- the aggregating sampler can be configured such that it can be carried by hand or mounted on a device designed for traveling through the field.
- the collection surface for field sampling may be divided to allow more conformity to the crop surface.
- an aggregating sampler may be placed on the harvester such that product is sampled during the harvesting process. This approach would allow pairing of harvested product with specific information.
- placement of the aggregating sampler may be just after any sorting is done in the field. For example, if rocks are sorted from the product by density classification, these rocks need not cross the sampler surface.
- the product may still have the flora found in the field.
- An aggregating sampler placed early in the process can sample these organisms. Just after cutting or chopping, the interior of some products will be exposed for the first time allowing a more representative sample to be taken.
- an aggregating sampler can collect a different type of sample in accordance with one or more examples. This sampler may test the cross contamination control of the wash system. The organisms collected will reflect the two most probable mechanisms of cross contamination, water mediated cross contamination and product to product cross contamination.
- An aggregating sampler, placed in the flow of the conveyed product will be impinged by both the water and the product. To avoid overly hindering product flow depending on the line design, the sampler can be placed at any angle from parallel to the product flow to complete perpendicular to product flow.
- the sampler can also be a comb-like device with multiple collecting probes among product and in the wash flow. The angle of attachment may affect the balance between water mediated cross contamination and product to product cross contamination observed. In either case, this type of sampling may be used to validate cross contamination control and effectiveness of wash solution.
- Sampling in the wash system is a case where the sampling surface may be active on multiple surfaces, for example, on both sides or around in the case of the comb like structure mentioned above.
- further advantages may be provided in other ways such as by increasing the thickness of the sampling material.
- designing samplers in devices such as filter housing may be provided.
- placement on a sampling surface in an active area of the wash system may be provided for getting a full measure of the cross contamination potential.
- a sample may be taken after the wash system and will reflect a residual population.
- this area there are a number of specific locations that can be considered depending on the specifics of the line.
- these specific locations include just prior to loading dryers, in a conveyer that might be used to lift the product for packaging, just before a pocket scale, or in the throat of a form fill and seal machine. This in-line continuous sampling may significantly increase sampling efficiency and provide more meaningful data than grab sampling of finished product testing.
- the samples taken in the field, at harvest, or just prior to washing can be used to assess the microbiological status of the raw material. From a food safety perspective the focus heretofore has been on the presence or absence of pathogens. Unfortunately, such analyzes based on grab samples are unable to truly answer the question as to whether pathogens are present due to their lack of sensitivity. Generally, it is known that pathogens are present at very low numbers. These are ubiquitous organisms. A more appropriate question is whether these organisms are present at abnormal concentrations or without the usual competing organism.
- samples taken just prior to washing may be compared to samples taken in the wash system to directly measure cross contamination using wild type flora.
- Water samples may tend to have very low microbial loads in properly managed wash systems even when cross contamination is occurring.
- the wild type flora may also be highly variable.
- a number of metrics for cross contamination can be considered based on the ratio of the results for the samples from prewash to those from in the wash.
- By using more sophisticated analytical procedures one can overcome the flaws in such metrics as aerobic plate count (APC) which would include many organisms that are not relevant to cross contamination control of pathogens.
- APC aerobic plate count
- samples from after a wash process may provide information about microbial populations on the finished product. These samples may provide a much more accurate assessment of the pathogen risk of the product and better detection deviations. These samples can also be used to check for deviations in the normal flora.
- a ratio between the before wash samples and the after wash samples may be used to assess the impact of the wash process.
- the aggregating samplers may reduce the noise and as with the cross contaminations metrics, these ratios can be handled with statistical process control to look for deviations.
- One or more of these examples may be delivered in almost real time because the aggregating samplers collected enough cells for concentration and direct analysis without enrichment.
- an example of a bench scale study demonstrates a sampling efficiency of 15-20% relative to stomaching. It also shows that transfer is essentially instantaneous and that repeated contact collects more organisms. These observations confirm the intuitive assertion that continuous sampling will yield better information than grab sampling.
- E. coli is selected as benign and easy to enumerate.
- the stew meat is allowed to rest at room temperature for 30 minutes to allow adherence.
- the six treatments listed in the table shown in FIG. 3 may be executed in 5 replicates. All sampling cloths may be cut in half, 12 in by 8 in, to reduce the sample requirement and facilitated execution of the experiment.
- the sampling cloth may be, for example, a MicroTally cloth but is not limited thereto.
- the surface areas of the meat cubes are measured directly. For the sampling cloth treatments, meat is arranged in a 10 cm by 10 cm block and the sampling cloth is applied to the upper surface.
- PBS phosphate buffered saline
- a logical extrapolation of this exercise is to estimate the effective sample size of a sampling. This may not truly be possible given the differences in geometry. However, in one or more use cases when using one side only, a sampling cloth is about 6 times larger than those used in the bench scale study experiment and the intended use is to sample almost 2000 pounds of product. Thus, it is reasonable to assert that the effective sample is expected to be 300 to 400 pounds. Larger scale experiments may be implemented that may further confirm this estimate.
- a benefit and advantage of the above method and apparatus of sampling may include providing an improvement over traditional grab samples for meat sampling by providing larger effective samples.
- an example of a bench scale study demonstrates a sampling efficiency of about 30% relative to stomaching. It also shows that transfer is essentially instantaneous and that repeated contact collects more organisms. These observations confirm the intuitive assertion that continuous sampling will yield better information than grab sampling for lettuce.
- E. coli is selected as benign and easy to enumerate.
- the lettuce is allowed to rest at 4° C. for 30 minutes to allow adherence. This short time may explain the higher efficiency observed when compared to a meat study where adhesion maybe faster.
- the six treatments listed in Table 2 shown in FIG. 4 may be executed in 5 replicates. All sampling cloths may be cut in half, 12 in by 8 in, to reduce the sample requirement and facilitated execution of the experiment.
- the sampling cloth may be, for example, a MicroTally cloth but is not limited thereto.
- the surface areas of the lettuce leaves may be measured directly.
- lettuce is arranged in a 10 cm by 10 cm block and the sampling cloth applied to the upper surface. Each mini sampling cloth is extracted in 200 ml of PBS.
- lettuce may be stomached for 60 seconds in 200 mL of PBS. All stomached leaves are measured to estimate surface area. The counts are normalized for surface area and averaged. This normalization provides an apple to apple comparison.
- a logical extrapolation of this exercise is to estimate the effective sample size of a sampling. This may not truly be possible given the differences in geometry. However, in one or more use cases when using one side only, a sampling cloth is about 6 times larger than those used in the bench scale study experiment and the intended use is to sample almost 2000 pounds of product. Thus, it is reasonable to assert that the effective sample is expected to be 400 to 600 pounds. Larger scale experiments may be implemented that may further confirm this estimate.
- a benefit and advantage of the above method and apparatus of sampling may include providing an improvement over traditional grab samples for leafy sampling by providing larger effective samples.
- swabs and/or sheets may be used in accordance with one or more embodiments of the present disclosure.
- a sheet may include a microbial sampling material, such as sterile woven and/or non-woven synthetic fabrics and non-woven cloth for sampling and testing in the field of food safety. These sheets can be folded and curved to allow better conformation to product (e.g., food) streams that are being sampled or when used in a manual mode as driven by the product and the container.
- configurations for sampling raw products or materials may include a sampling device that moves across the stationary product effectively yielding the equivalent of a product stream when sampling needs to occur prior to harvest.
- configurations may include tail shaped sheets similar to, for example, the tentacles of a jellyfish.
- a microbial aggregating sampler may include a covering including a microbial sampling material with a pocket formed in the covering to receive an appendage or a tool for handling of the covering.
- the covering may further include an attachment feature formed in the pocket to receive the tool.
- the attachment feature may include at least one of a hole formed through the covering; a loop positioned within the pocket to receive an end of the tool there through; or a tab positioned within the pocket for an end of the tool to attach thereto.
- the covering may include a sheath formed in the pocket to receive a digit of an appendage.
- the pocket is formed through the covering such that the appendage or the tool for handling the covering extends through the covering.
- the covering may be completely formed from the microbial sampling material.
- the covering includes two sheets attached to each other to form the pocket.
- the covering may include a single sheet folded and attached to itself to form the pocket.
- a sheet may undergo folding and seaming to form a bag or pocket that can be worn as a mitten, glove, sock, or other covering, to facilitate manual sampling.
- a covering may encase an appendage, such as a hand, to facilitate pushing and pulling of the sampler through the product to be sampled.
- aggregating samplers may be created that are more hand like configurations as mitts with thumbs and/or gloves with fingers, allowing the sampler to better conform to the hand. Such configurations may allow easier use when the product is more viscous or more prone to adhering to the sampler.
- a benefit to this aggregating sampler with a pocket may include the ability of the sampler to, when working the sampler, increase or maximize product contact.
- the addition of one or more attachment features, such loops or tabs, to assist with controlling the product contact may be included.
- the aggregating sampler may be used in an automated machine setting to sample a product stream.
- the aggregating sampler may include a number of modifications in accordance with one or more cases.
- the aggregating sampler may include one or more bends and curves that may improve the utility and facilitate use.
- modifications may include, but are not limited to, forming a tube that can slide over one or more shafts for positioning in the product flow. This configuration may remove the need to slide the sampler within any holding device.
- adding tabs or holes can be provided which may allow positioning and facilitate attachment.
- the active sampling surface can be attached to a support material or web that has one or more of these features. One or more of these cases and modifications may help facility contact with the product stream with minimal manual intervention.
- FIG. 5 illustrates an aggregated sampler 500 that includes a pocket in accordance with aspects of the present disclosure.
- the aggregated sampler 500 includes a covering 504 made of a microbial sampling material.
- the covering is formed such that it includes a pocket formed in the covering to receive an appendage or tool 502 within the pocket for handling of the covering.
- FIG. 6 illustrates an aggregated sampler 600 that includes a pocket that extends or is formed through a covering 604 in accordance with aspects of the present disclosure.
- the pocket is formed such that the appendage or tool 602 for handling the covering 604 extends through the covering.
- FIG. 7 illustrates an aggregated sampler 700 with at least one convex surface of the covering 704 in accordance with aspects of the present disclosure.
- the covering forms a pocket for a tool or appendage 702 .
- FIG. 8 illustrates an aggregated sampler 800 that includes at least one concave surface of the covering 804 in accordance with aspects of the present disclosure.
- the covering 804 forms a pocket for a tool or appendage 802 .
- FIG. 9 illustrates an aggregated sampler 900 that includes a curved shape covering 904 in accordance with aspects of the present disclosure.
- the covering 904 is formed to include a pocket for a tool or appendage 902 .
- FIG. 10 illustrates an aggregated sampler 1000 that includes a hole or loop 1008 in accordance with aspects of the present disclosure.
- the covering 1004 may include the hole or loop 1008 , such as positioned within the pocket of the covering 1004 and attached to an inner surface of the covering 1004 .
- This hole or loop 1008 may be used by a hook 1006 of a tool 1002 to attach to the covering 1004 .
- FIG. 11 illustrates an aggregated sampler 1100 that is shaped as a mitten or glove for a hand 1102 in accordance with aspects of the present disclosure.
- the covering 1104 of the aggregated sampler 1100 is formed to contour to a hand 1102 .
- a sheath is formed in the covering 1104 in this embodiment to receive a thumb of the hand 1102 , though one or more sheaths may be used to receive any digit of the hand 1102 .
- a hand 1102 is shown as an example appendage in this embodiment, the present disclosure is not so limited, as other appendages (i.e., a foot) may be contemplated for other embodiments without departing from the scope of the present disclosure.
- the aggregated sampling sheet may be hung into an active zone of a wash line with a cord or cable.
- This line may be attached in number of ways to the sampling material such as, for example, a grommet and clasp.
- a ball and slide clamp may be used.
- a punched hole may be used but may pull out.
- various other clamps may be used.
- an optional float such as a fishing float with or without a weight may be included that will add drag and bouncy which may improve surface exposure. This approach may not include a fixed appliance for holding the sampling material.
- the reference method for extracting microorganisms from grab samples is homogenization by mixing or stomaching with an appropriate amount of fluid, usually a buffer.
- fluid usually a buffer.
- the purpose of adding fluid is to neutralize any antimicrobial or other properties of the sample that may be unfavorable for microbial growth such as low pH and suspend microorganism in liquid to facilitate downstream testing.
- adding fluid also dilutes the concentration of the microorganisms in the sample. Because most microbiological testing only take a portion of the homogenate (e.g., 0.1 mL or 1 mL for plating; 2.5 ⁇ L for direct PCR), the homogenization by dilution method decreases the detectability of the organisms of interest by as much as 1,500,000 times.
- the percentage of negative samples exceeds 99%. Based on simple modeling, one can easily conclude that the number of cells of the organism of interest is seldom more than 1 and probably rarely more than 5 based on a Poisson distribution. With so few cells present, the enrichment is often done in the extraction buffer. This is critical with the small sampling but does not make this type of testing any more representative of the lot under test.
- a volume of 100 to 200 mls of extraction solution is generally appropriate.
- the composition of this solution is driven by the planned detection system and is generally defined in the associated method.
- the concentration two classes of materials may need to be removed, the small, ⁇ 100,000 molecular weight, and the very large, >50 microns.
- the sample may need to be concentrated to about 1 mL to be compatible with the screening detection system. Given that the extraction generally starts at about 200 mL, there is a large amount of water to remove.
- the surface activation may require numerous active binding sites in close proximity.
- the mixed DNA primer arrays of SnapDNA are one class of materials. Another class is a cocktail of antibodies for all the organisms of interest.
- the binding mechanism may bind all the organism of interest. These organisms may be sufficiently bound that other organisms and materials are selectively removed from the area of binding as clean fluid is passed through the channel.
- the motive force to move the cells through the channel can be the mechanical action of fluid flow, electrostatic as the surface of most bacteria is negative, or a size pumping action such as practiced with ferromagnetic particles.
- the screening metric may need to be an index suitable for statistical process control. This implies a measurement with many states as opposed to binary 0 and 1. The magnitude of this metric may relate to the extent of deviation from normal operation and therefore the likelihood that an outbreak could occur. Under these conditions, the detection of deviation can be based on the classic rules for control charting. Furthermore, trend detecting rules have the potential to detect problems before significant deviations have occurred. The statistically based Westcard rules provide one basis for trend analysis.
- an index may be needed that tallies the relative presence of beneficial organisms and potential pathogens and aggregates this information in a useful way. As more information is acquired about specific products, the power of big data will come into play. However, at the simplest levels, the aggregate level of potential pathogen is a useful screening tool. The relative balance between potential pathogens and beneficial organisms is a more sophisticated analysis to compensate for seasonal variations that are inherent in many products. It is reasonable to expect to develop indices that are product specific.
- index data The information behind indices is evolving rapidly.
- the simplest useful index is a ratio of pathogen intensity to a benign organism. These can be generated by many means including classical enumeration with plating, but the classical methods are two slow to meet the functional requirements outlined above. However, given the concentration of the organisms on the cleaning substrate or the concentrated extraction, it is possible to go directly to qPCR in some cases to generate index data.
- the first scheme would be a using a mixture of conjugated antibodies to bind to all types of cells of interest.
- hemolytic E. coli, Salmonella and Listeria are of greatest interest. Poultry focuses on Campylobacter and Salmonella.
- Other industries have other and additional interests.
- These antibodies can be bound to the organisms bound to the cleaning substrate.
- the retained tags with either a fluorescent probe or an enzyme provide signal amplification and detection.
- the cleaning region as a small enough cross section, the signal from the hundreds to thousands of cells on the cleaning substrate are detectable. Micro and nano fluidization are necessary.
- specific detection protocols can be evaluated at a macro scale using an appropriately instrumented microscope that can be used to measure the signal from a small area where cells have been collected. For speed to market, multiple detectors can be run in parallel or series to generate similar multiple channels of data.
- the alternative scheme invokes the lab on a chip concept. By building an array of binding sites, the composition of the samples can be queried. PCR can amplify the contents selectively with a collection of primers. Such detectors can evolve to providing both the screening detection of this step and the ultimate secondary screening. However, it is likely to remain a two-step process due to the economics.
- a number of technologies can meet these requirements including but not limited to, for example: 1) Sensors where the vibrational frequencies (which may for example include optical waveguide), impedance, or other properties of a transistor are modified by the binding of the organisms of interest to the sensor. This approach may require that the sensor be built into the surface of the channel.
- Another technology may include, 2) qPCR where the cells are laid in place and the number of copies of the organisms of interest, or the number of ribosomes of the organisms of interest are estimated.
- Another technology may include, 3) Use tagged antibodies to light up the organisms of interest so that they can detected on the absorbent surface spectrophotometrically. Enzyme, fluorescent probes, and other materials to amplify the signal can be used.
- sampling approach is compatible with various compositing strategies. Samples can be composited anywhere along the processing path before detection. In various situations, one position will be more advantageous than others. Pooling samples prior to extraction reduces work but reduces the resolution of the results if a positive result is found. However, if the result is negative tremendous savings are achieved. After concentrating and cleaning would allow sampled to be retested if only a portion of the extracted sample is used in a wet pooling approach which can avoid detections costs. Each system needs to be evaluated to determine how to best control costs.
- the confirmation step will be applied when there is reason to suspect that a pathogen is present.
- the technology in this area is evolving rapidly with many new approaches and efficiencies being developed and introduced. Some will use the techniques suggested for screening with more specific reagents for the confirmation. At present, all the common procedures rely on either molecular biology or ligand binding reactions. Various strategies have been developed to amplify the signal noise ratio and to identify the contaminant to the desired specificity. The desired specificity ranges from simple speciation to identify specific serotypes.
- a concentrated sample such as afforded the screening detection above may be included.
- the concentration may be included due to the small volumes that are compatible with these types of procedures.
- the organisms, the surface antigens from the organisms, or the nucleic acid from the organisms may need to be extracted from the screening system to the extent that these materials can interact with the reagents of the confirmation procedure.
- the detection module could not be engineered for a second round of chemistries. Any one of these materials may contain the information necessary to characterize the contaminant. There are a number of these processes and they are being improved. The choice of approach will be driven by cost, desired specificity and the desired speed.
- nucleic acid can be nuclear DNA, ribosomal RNA, or messenger RNA.
- sequences of nucleic acid can be nuclear DNA, ribosomal RNA, or messenger RNA.
- RNA messenger RNA
- the molecular assays are built around the use of enzymes to replicate sequences of nucleic acid to generate a large enough signal for detection.
- Various primers are used to select which portions are copied; up to and including the entire genome.
- various fluorescent tags are used for measurement.
- One of the newest techniques utilizes the melting and binding of target material to known sequences to generate a complex matrix of information that can be used chemometrically in lieu of complete sequence data.
- Both detectors should be part of the “Internet of Things”. This connectivity allows results to be pushed to operators on the floor allowing for the rapid release of product or for the redisposition of product if a potential issue is identified that may need to be addressed.
- results Once the results are in an appropriate database, various users can have customized interfaces providing the information. Some may need to track individual results. Others may be more interested in trends and averages. There may be provided a class of users that may want to aggregate even larger data sets to compare across locations.
- Ignition has proven useful in this record as it is open source allowing customization.
- a method for microbial sampling food may include gathering a microbial sampling from one or more food items, extracting microorganisms from the microbial sampling, concentrating the microorganisms, cleaning the microorganisms, tallying a relative presence of the microorganisms and any potential pathogens, aggregating information of a microorganism tally from the tallying of microorganisms into a microorganism report, confirming the microorganism tally, and reporting the microorganism report of the microorganism tally.
- gathering the microbial sampling from the one or more food items includes sampling, using an aggregating sampler, the one or more food items that include a production lot of produce or meat creating one or more samples that makes up the microbial sampling.
- the one or more samples may be configured to be processed to indicate if pathogens are present at no greater than a normal background.
- the method may further include assessing, using an aggregating sampler, a level of cross contamination control to validate or verify a wash process. Gathering the microbial sampling from the one or more food items may include providing an aggregating sampler at a sampling location.
- the sampling location may be at least one of in a field, at harvest, just after dumping or cutting, in a wash system, or after the wash system.
- extracting includes enriching the microbial sampling, and adding fluid to the microbial sample.
- concentrating includes filtering extraction fluid of the microbial sampling using at least one of centrifugation filtering or osmotically filtering.
- cleaning includes binding the microorganisms in a small area including one or more of a microfluidized or nanofluidized channel.
- tallying includes using a collection of ligands that bind and tag all the microorganisms of potential interest yielding a collection of signals that are multiplexed into a family of useful channels.
- ligands include one or more of antibodies, primers, and aptamers.
- tallying includes generating an array of specific binding interactions that are analyzed chemometrically to yield a metric.
- the method may further include building an array of binding sites, wherein the composition of the samples can be queried, and amplifying, using a PCR, the contents selectively with a collection of primers.
- confirming includes extracting surface antigens from the organisms or nucleic acid from the organisms from a screening system to the extent that these materials can interact with the reagents of a confirmation procedure. In some cases confirming further includes amplifying a base signal of the microorganism tally.
- the method may further include use of an index as a surrogate for direct results regarding presence or absence of organisms of interest. In some cases, the method may further include use of a statistical process control for detecting deviations in microbial flora.
- a method of applying aggregating sampling to food items including providing at least one aggregating sampler at one or more sampling locations, and sampling, using the at least one aggregating sampler, a production lot of produce or meat creating one or more samples that makes up a microbial sampling.
- the one or more samples are configured to be processed to indicate if pathogens are present at no greater than a normal background.
- the one or more sampling locations includes at least one of in a field, at harvest, just after dumping or cutting, in a wash system, or after the wash system.
- the method may further include assessing, using the aggregating sampler, a level of cross contamination control to validate or verify a wash process.
- an apparatus for microbial sampling including means for gathering a microbial sampling from one or more food items, means for extracting microorganisms from the microbial sampling, and means for concentrating the microorganisms, means for cleaning the microorganisms, means for tallying a relative presence of the microorganisms and any potential pathogens, means for aggregating information of the microorganism tally into a microorganism report, means for confirming the microorganism tally, and means for reporting the microorganism report of the microorganism tally.
- an apparatus for microbial sampling includes at least one processor configured to generate control signals for controlling gathering a microbial sampling from one or more food items, extracting microorganisms from the microbial sampling, concentrating the microorganisms, cleaning the microorganisms, tallying a relative presence of the microorganisms and any potential pathogens, aggregating information of the microorganism tally into a microorganism report, and confirming the microorganism tally, and a transmitter configured to transmit the microorganism report of the microorganism tally.
- the apparatus may further include an aggregating sampler configured to gather the microbial sampling.
- a non-transitory computer readable medium for microbial sampling having instructions stored thereon for gathering a microbial sampling from one or more food items, extracting microorganisms from the microbial sampling, concentrating the microorganisms, cleaning the microorganisms, tallying a relative presence of the microorganisms and any potential pathogens, aggregating information of the microorganism tally into a microorganism report, confirming the microorganism tally, and reporting the microorganism report of the microorganism tally.
- a method for sampling food including concentrating microorganisms and removing interference, tallying a relative presence of the microorganisms and any potential pathogens, and aggregating information of the microorganism tally into a microorganism report.
- a system capable of implementing one or more of the novel aspects discussed in this application disclosure.
- a microbial aggregating sampler including a covering including a microbial sampling material with a pocket formed in the covering to receive an appendage or a tool for handling of the covering.
- the covering includes an attachment feature formed in the pocket to receive the tool.
- the attachment feature includes one of a hole formed through the covering, a loop positioned within the pocket to receive an end of the tool there through, and a tab positioned within the pocket for an end of the tool to attach thereto.
- the covering includes a sheath formed in the pocket to receive a digit of an appendage.
- the pocket is formed through the covering such that the appendage or the tool for handling the covering extends through the covering.
- the covering is completely formed from the microbial sampling material.
- the covering includes two sheets attached to each other to form the pocket.
- the covering includes a single sheet folded and attached to itself to form the pocket.
- the aggregating sampler can be used to sample for additional analytes beyond bacteria including yeast, molds, viruses, allergens, toxins such as aflatoxin, or particulates such as dust.
- the commonality is the surface presence of the analyte at low levels that can be concentrated.
- the basic process is the same with the same key steps. The biggest differences will be in detection strategy, but these strategies are well known by those who study these analytes.
- the methods disclosed herein comprise one or more steps or actions for achieving the described method.
- the method steps and/or actions may be interchanged with one another without departing from the scope of the claims.
- the order and/or use of specific steps and/or actions may be modified without departing from the scope of the claims.
- a phrase referring to “at least one of” a list of items refers to any combination of those items, including single members.
- “at least one of: a, b, or c” is intended to cover a, b, c, a-b, a-c, b-c, and a-b-c, as well as any combination with multiples of the same element (e.g., a-a, a-a-a, a-a-b, a-a-c, a-b-b, a-c-c, b-b, b-b-b, b-b-c, c-c, and c-c-c or any other ordering of a, b, and c).
- the term “and/or,” when used in a list of two or more items, means that any one of the listed items can be employed by itself or any combination of two or more of the listed items can be employed. For example, if a composition is described as containing components A, B, and/or C, the composition can contain A alone; B alone; C alone; A and B in combination; A and C in combination; B and C in combination; or A, B, and C in combination.
- determining encompasses a wide variety of actions. For example, “determining” may include calculating, computing, processing, deriving, investigating, looking up (e.g., looking up in a table, a database or another data structure), ascertaining and the like. Also, “determining” may include receiving (e.g., receiving information), accessing (e.g., accessing data in a memory) and the like. Also, “determining” may include resolving, selecting, choosing, establishing and the like.
- the term “or” is intended to mean an inclusive “or” rather than an exclusive “or.” That is, unless specified otherwise or clear from the context, the phrase, for example, “X employs A or B” is intended to mean any of the natural inclusive permutations. That is, for example the phrase “X employs A or B” is satisfied by any of the following instances: X employs A; X employs B; or X employs both A and B. All structural and functional equivalents to the elements of the various aspects described throughout this disclosure that are known or later come to be known to those of ordinary skill in the art are expressly incorporated herein by reference and are intended to be encompassed by the claims.
- the various operations of methods described above may be performed by any suitable means capable of performing the corresponding functions.
- the means may include various hardware and/or software component(s) and/or module(s), including, but not limited to a circuit, an application specific integrated circuit (ASIC), or processor.
- ASIC application specific integrated circuit
- DSP digital signal processor
- ASIC application specific integrated circuit
- FPGA field programmable gate array
- PLD programmable logic device
- a general-purpose processor may be a microprocessor, but in the alternative, the processor may be any commercially available processor, controller, microcontroller, or state machine.
- a processor may also be implemented as a combination of computing devices, e.g., a combination of a DSP and a microprocessor, a plurality of microprocessors, one or more microprocessors in conjunction with a DSP core, or any other such configuration.
- an example hardware configuration may comprise a processing system in a wireless node.
- the processing system may be implemented with bus architecture.
- the bus may include any number of interconnecting buses and bridges depending on the specific application of the processing system and the overall design constraints.
- the bus may link together various circuits including a processor, machine-readable media, and a bus interface.
- the bus interface may be used to connect a network adapter, among other things, to the processing system via the bus.
- the network adapter may be used to implement the signal processing functions of the PHY layer.
- a user interface e.g., keypad, display, mouse, joystick, etc.
- a user interface e.g., keypad, display, mouse, joystick, etc.
- the bus may also link various other circuits such as timing sources, peripherals, voltage regulators, power management circuits, and the like, which are well known in the art, and therefore, will not be described any further.
- the processor may be implemented with one or more general-purpose and/or special-purpose processors. Examples include microprocessors, microcontrollers, DSP processors, and other circuitry that can execute software. Those skilled in the art will recognize how best to implement the described functionality for the processing system depending on the particular application and the overall design constraints imposed on the overall system.
- the functions may be stored or transmitted over as one or more instructions or code on a computer readable medium.
- Software shall be construed broadly to mean instructions, data, or any combination thereof, whether referred to as software, firmware, middleware, microcode, hardware description language, or otherwise.
- Computer-readable media include both computer storage media and communication media including any medium that facilitates transfer of a computer program from one place to another.
- the processor may be responsible for managing the bus and general processing, including the execution of software modules stored on the machine-readable storage media.
- a computer-readable storage medium may be coupled to a processor such that the processor can read information from, and write information to, the storage medium. In the alternative, the storage medium may be integral to the processor.
- the machine-readable media may include a transmission line, a carrier wave modulated by data, and/or a computer readable storage medium with instructions stored thereon separate from the wireless node, all of which may be accessed by the processor through the bus interface.
- the machine-readable media, or any portion thereof may be integrated into the processor, such as the case may be with cache and/or general register files.
- machine-readable storage media may include, by way of example, RAM (Random Access Memory), flash memory, phase change memory, ROM (Read Only Memory), PROM (Programmable Read-Only Memory), EPROM (Erasable Programmable Read-Only Memory), EEPROM (Electrically Erasable Programmable Read-Only Memory), registers, magnetic disks, optical disks, hard drives, or any other suitable storage medium, or any combination thereof.
- RAM Random Access Memory
- flash memory Phase change memory
- ROM Read Only Memory
- PROM Programmable Read-Only Memory
- EPROM Erasable Programmable Read-Only Memory
- EEPROM Electrically Erasable Programmable Read-Only Memory
- registers magnetic disks, optical disks, hard drives, or any other suitable storage medium, or any combination thereof.
- the machine-readable media may be embodied in a computer-program product.
- a software module may comprise a single instruction, or many instructions, and may be distributed over several different code segments, among different programs, and across multiple storage media.
- the computer-readable media may comprise a number of software modules.
- the software modules include instructions that, when executed by an apparatus such as a processor, cause the processing system to perform various functions.
- the software modules may include a transmission module and a receiving module. Each software module may reside in a single storage device or be distributed across multiple storage devices.
- a software module may be loaded into RAM from a hard drive when a triggering event occurs.
- the processor may load some of the instructions into cache to increase access speed.
- One or more cache lines may then be loaded into a general register file for execution by the processor.
- any connection is properly termed a computer-readable medium.
- the software is transmitted from a website, server, or other remote source using a coaxial cable, fiber optic cable, twisted pair, digital subscriber line (DSL), or wireless technologies such as infrared (IR), radio, and microwave
- the coaxial cable, fiber optic cable, twisted pair, DSL, or wireless technologies such as infrared, radio, and microwave are included in the definition of medium.
- Disk and disc include compact disc (CD), laser disc, optical disc, digital versatile disc (DVD), floppy disk, and Blu-ray® disc where disks usually reproduce data magnetically, while discs reproduce data optically with lasers.
- computer-readable media may comprise non-transitory computer-readable media (e.g., tangible media).
- computer-readable media may comprise transitory computer-readable media (e.g., a signal). Combinations of the above should also be included within the scope of computer-readable media.
- certain aspects may comprise a computer program product for performing the operations presented herein.
- a computer program product may comprise a computer-readable medium having instructions stored (and/or encoded) thereon, the instructions being executable by one or more processors to perform the operations described herein.
- instructions for performing the operations described herein and illustrated in the appended figures may comprise a computer-readable medium having instructions stored (and/or encoded) thereon, the instructions being executable by one or more processors to perform the operations described herein.
- modules and/or other appropriate means for performing the methods and techniques described herein can be downloaded and/or otherwise obtained by a user terminal and/or base station as applicable.
- a user terminal and/or base station can be coupled to a server to facilitate the transfer of means for performing the methods described herein.
- various methods described herein can be provided via storage means (e.g., RAM, ROM, a physical storage medium such as a compact disc (CD) or floppy disk, etc.), such that a user terminal and/or base station can obtain the various methods upon coupling or providing the storage means to the device.
- storage means e.g., RAM, ROM, a physical storage medium such as a compact disc (CD) or floppy disk, etc.
- CD compact disc
- floppy disk etc.
- any other suitable technique for providing the methods and techniques described herein to a device can be utilized.
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Water Supply & Treatment (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Hydrology & Water Resources (AREA)
- Toxicology (AREA)
- Cell Biology (AREA)
- Nanotechnology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Sampling And Sample Adjustment (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Meat, Egg Or Seafood Products (AREA)
Abstract
Certain aspects of the present disclosure relate to methods and apparatus for microbial sampling of foods. For example, a method may include providing at least one aggregating sampler at one or more sampling locations, and sampling a production lot of produce or other food items such as meat using the at least one aggregating sampler to create one or more samples that makes up a microbial sampling. Certain aspects of the present disclosure relate to methods and apparatus for microbial sampling of foods. For example, an apparatus, such as a microbial aggregating sampler, may include a covering having a microbial sampling material with a pocket formed in the covering to receive an appendage or a tool for handling of the covering.
Description
- The present Application for Patent claims benefit of U.S. Provisional Patent Application Ser. No. 62/656,164, filed Apr. 11, 2018, U.S. Provisional Patent Application Ser. No. 62/589,755, filed Nov. 22, 2017, and U.S. Provisional Patent Application Ser. No. 62/543,220, filed Aug. 9, 2017, assigned to the assignee hereof and hereby expressly incorporated by reference herein.
- The present disclosure relates generally to improving the food safety of ready-to-eat produce and other food items and providing process validation, and more particularly, to methods and apparatus for microbial sampling of food items and other materials.
- The microbial testing process has undergone tremendous change in recent years. Traditional plating techniques for enumeration and detection have given way to faster and more specific antibody and molecular biology based techniques. These newer techniques may not require time for colonies to form but they may generally require enrichment culture to collect enough of the target organism and remove interference.
- In the ready-to-eat produce industry, millions of dollars are spent collecting grab samples attempting to demonstrate the safety of products in an effort to meet demands by customers for an ever increasing numbers of tests. These efforts may be technically and statistically flawed and may not meet the expectations of assuring food safety. Particularly, grab samples are too small to represent the production lots of material. Production lots of material are too heterogeneous for grab sampling to be descriptive of the production lot. Further, results arrive too slowly to make decisions without sacrificing quality. Pathogens levels are generally so low that the occasional positive sample reflects the background that is always present rather than a deviation from the norm. The ready-to-eat produce industry may benefit from an effective assay of cross contamination and cross contamination control. It may also benefit from an effective measure of process efficacy and deviation in processing. Increasing the effectiveness of raw material testing may help improve food safety practice.
- Thus, as the demand for microbial sampling continues to increase, there exists a desire for further improvements in sampling techniques and technology. Preferably, these improvements should be applicable to other related technologies and the methods and devices that employ these technologies.
- The systems, methods, and devices of the disclosure each have several aspects, no single one of which is solely responsible for its desirable attributes. Without limiting the scope of this disclosure as expressed by the claims which follow, some features will now be discussed briefly. After considering this discussion, and particularly after reading the section entitled “Detailed Description” one will understand how the features of this disclosure provide advantages that include improved microbial sampling of foods and other products.
- Certain aspects provide a method for microbial sampling of foods and other products. The method generally includes gathering a microbial sampling from one or more items, extracting microorganisms from the microbial sampling, concentrating the microorganisms, cleaning the microorganisms, tallying the relative presence of the microorganisms and any potential pathogens, aggregating this information of a microorganism tally from the tallying of microorganisms into a microorganism report, confirming the microorganism tally, and reporting the microorganism report of the microorganism tally.
- Certain aspects provide a method for microbial sampling of foods. The method generally includes providing at least one aggregating sampler at one or more sampling locations, and sampling, using the at least one aggregating sampler, a production lot of produce creating one or more samples that makes up a microbial sampling.
- Aspects generally include methods, apparatus, systems, computer readable mediums, and processing systems, as substantially described herein with reference to and as illustrated by the accompanying drawings.
- To the accomplishment of the foregoing and related ends, the one or more aspects comprise the features hereinafter fully described and particularly pointed out in the claims. The following description and the annexed drawings set forth in detail certain illustrative features of the one or more aspects. These features are indicative, however, of but a few of the various ways in which the principles of various aspects may be employed, and this description is intended to include all such aspects and their equivalents.
- So that the manner in which the above-recited features of the present disclosure can be understood in detail, a more particular description, briefly summarized above, may be had by reference to aspects, some of which are illustrated in the appended drawings. It is to be noted, however, that the appended drawings illustrate only certain typical aspects of this disclosure and are therefore not to be considered limiting of its scope, for the description may admit to other equally effective aspects.
-
FIG. 1 illustrates example operations for microbial sampling, in accordance with aspects of the present disclosure. -
FIG. 2 illustrates example operations for applying aggregating sampling, in accordance with aspects of the present disclosure. -
FIG. 3 illustrates a Table 1 that includes example treatments and results for meat sampling, in accordance with aspects of the present disclosure. -
FIG. 4 illustrates a Table 2 that includes example treatments and results for leafy sampling, in accordance with aspects of the present disclosure. -
FIG. 5 illustrates an aggregated sampler that includes a pocket in accordance with aspects of the present disclosure. -
FIG. 6 illustrates an aggregated sampler that includes a pocket that extends through a covering in accordance with aspects of the present disclosure. -
FIG. 7 illustrates an aggregated sampler with at least one convex surface in accordance with aspects of the present disclosure. -
FIG. 8 illustrates an aggregated sampler that includes at least one concave surface in accordance with aspects of the present disclosure. -
FIG. 9 illustrates an aggregated sampler that includes a curved shape in accordance with aspects of the present disclosure. -
FIG. 10 illustrates an aggregated sampler that includes a hole or loop in accordance with aspects of the present disclosure. -
FIG. 11 illustrates an aggregated sampler that is shaped as a mitten or glove for a hand in accordance with aspects of the present disclosure. - To facilitate understanding, identical reference numerals have been used, where possible, to designate identical elements that are common to the figures. It is contemplated that elements disclosed in one aspect may be beneficially utilized on other aspects without specific recitation.
- Aspects of the present disclosure provide apparatus, methods, and/or systems for automated and semi-automated microbial sampling of foods and other materials. Other materials can be as diverse as water or air streams. More commonly, it will include kindred products of food such as pet food, medical materials or dietary supplements where microbial testing is needed to confirm hygienic operation. Sampling can be active as mediated by material flow or operator mediated by applying the sampler to a surface. Sampling can also be more passive and depend on passive contact or gravity sedimentation.
- The following description provides examples, and is not limiting of the scope, applicability, or examples set forth in the claims. Changes may be made in the function and arrangement of elements discussed without departing from the scope of the disclosure. Various examples may omit, substitute, or add various procedures or components as appropriate. For instance, the methods described may be performed in an order different from that described, and various steps may be added, omitted, or combined. Also, features described with respect to some examples may be combined in some other examples. For example, an apparatus may be implemented or a method may be practiced using any number of the aspects set forth herein. In addition, the scope of the disclosure is intended to cover such an apparatus or method which is practiced using other structure, functionality, or structure and functionality in addition to or other than the various aspects of the disclosure set forth herein. It should be understood that any aspect of the disclosure disclosed herein may be embodied by one or more elements of a claim. The word “exemplary” is used herein to mean “serving as an example, instance, or illustration.” Any aspect described herein as “exemplary” is not necessarily to be construed as preferred or advantageous over other aspects.
- In one embodiment, devices are assembled and linked to a rapid reporting system to provide a more representative sampling and faster analysis of a food product or other material. In another embodiment, a similar system provides for robotically sampling a field crop and delivering results with a mobile laboratory. Both embodiments can include a two-stage screening system for speed and for economy, but a conventional testing approach can be considered to take advantage of the sampling improvements.
- There has been little evolution in sampling and sample preparation for submission to these advanced and rapid analytical techniques. One technique is to collect periodic samples or random grabs from a lot. This sample is extracted by homogenization or stomaching and then either a portion analyzed directly when high populations are expected or enriched prior to analysis. Liquid samples and particularly water samples can be filtered to allow analysis of larger samples. For special purposes but generally not for routine analysis liquid samples can be concentrated by centrifugation to pellet microorganisms. There are statistically based sampling plans as recommended by academics or international organizations, but these sampling plans are rarely practical and mostly cost prohibitive. Such sampling plans in use based on periodic samples or random grabs have a very low data density. Furthermore, these sampling plans intrinsically have sampling biases due to lot geometry where portions of the lot are essentially not sampled and the heterogeneous nature of microorganism distribution.
- Some microbial testing may assume that the test organisms are evenly distributed allowing a grab sample to be representative of the whole. This flawed assumption only accounts for inhomogeneous distributions in aggregate and requires many samples to characterize the microbial population of a lot. Test and release inspection based on grab samples may be flawed to the extent that grab sampling can inherently miss significant contamination. Furthermore, the size the sample limits the detection limit to levels that may be order of magnitude above the background rate and the level where risk becomes imminent. On the other hand, microbial proliferation during enrichment culture occurs under conditions selected to favor the growth of the potential pathogens which often have no relationship with the commercial conditions of storage and therefore generates extorted perceptions of the microbial growth risk.
- One or more embodiments as disclosed herein may address these short comings of the current sampling and testing practice and provides a system to generate more meaningful, real time or near real time, onsite assessment of microbial contamination risk.
- In accordance with one or more aspects of embodiments disclosed herein, automated and semi-automated microbial sampling of foods and other materials is provided. For example, devices may be assembled and linked to a rapid reporting system to provide a more representative sampling and faster analysis of a food product. In another case, a system may provide robotic sampling of a field crop and may deliver results with a mobile laboratory. Both examples may include a two-stage screening system that may provide speed and for economy. Although less advantageous, it is reasonable to take advantage of improved sampling with conventional enrichment and detection systems.
- In one or more cases, a method and system of microbial sampling includes providing a sampling sheet, such as a MicroTally Sheet, which is then used with a continuous sampling device (CSD) to collect biological agents. The CSD may change the sampling sheet automatically between samples. The sampling sheet is then used for analysis. Particularly, target bacteria may be removed from the sampling medium. The samples of target bacteria are cleaned up and concentrated using a treater and made suitable for analysis. The treated sample is then analyzed using molecular or biochemical methods and target agents are detected with acceptable accuracy and sensitivity. A cloud based data reporting system with user appropriate dashboards to report actionable information and facilitate timely decision making may also be provided.
- For example,
FIG. 1 illustratesspecific operations 100 for microbial sampling, in accordance with aspects of the present disclosure. - Specifically,
operations 100 begin, atblock 102, with gathering a microbial sampling from one or more food items which may include, for example, produce, meat, and other food products or other materials. Atblock 104,operations 100 include extracting microorganisms from the microbial sampling. Atblock 106, theoperations 100 include concentrating the microorganisms. Theoperations 100, atblock 108, include cleaning the microorganisms. Further, theoperations 100 include, atblock 110, tallying the relative presence of the microorganisms and any potential pathogens. Theoperations 100 also include, atblock 112, aggregating this information of a microorganism tally from the tallying of microorganisms into a microorganism report. Atblock 114, the operations include confirming the microorganism tally. The operations may also include, atblock 116, reporting the microorganism report of the microorganism tally. - In one or more cases, sampling may include using a sampling sheet or swab that collects a sample and is stomached in a 300 micron partitioned bag with 200 mls of eluting buffer and stomached. Concentration and cleanup of the sample may include siphoning an eluting buffer and entrained organisms through proprietary sequential filter and the targets may be deposited on a 0.22 micron PC filter. The targets are analyzed from the filter which may include for example, DNA purified and qPCR being run for Index elements including pathogen intensity, Enteric status, and/or positive control. Cloud based reporting based on, for example Ignition and SQL database, may be provided with user appropriate dashboards to provide timely actionable information
- In accordance with one or more cases, pathogens may be confirmed by resampling and enrichment procedures, by resampling and doing definitive pathogen tests, or by doing confirmation tests on the original DNA depending on the regulatory guidance. In one or more cases a cassette and cartridge system may be implemented to streamline the sampling process. Although not required, eliminating the use of a stomacher and partitioned bag may be provided in one or more cases. A binding collector may be provided to replace the PC membrane filter which may help streamline the sample delivery to the detector and potentially eliminate the need for DNA purification. Use of Ribosomal RNA may become a new standard for one or more such cases. In one or more cases, a detector may use flow amplification and laboratory on a chip type technology to further reduce detection times and costs.
- To best address all the short comings in current practice numerous improved elements may come together. Taken individually each improvement addresses some of the short comings and yields some advantages. Leaping all the way to a complete solution may beyond the sophistication of some industry classes so intermediate steps are considered for each element. For the initial discussion, the elements under consideration include Sampling, Extraction, Concentration, Cleaning, Screening Detection, Second Stage Sampling of Suspected Lots, Reporting, and Information Roll Up. Although not as desirable, a more conventional enrichment can be used instead of Concentration. After such enrichment, any number of detection systems can be used to detect the presence or absence of a target organism. Each of these elements is discussed below.
- Thus, the timeframe from sampling to information may be driven by the needs of the business class but is not limited thereto. Short shelf-life product can justify greater speed. Valuable commodities such as meat products will want more testing to limit exposure when a problem is expected or anticipated. These factors will impact the degree to which a complete automated solution is implemented or conversely when a system more akin to current practices is used to gain some of the benefits of improved sampling.
- There are practical limits to the amount of product that can be sampled by conventional means. Without heroic efforts samples are limited to small fraction of a pound (generally 150 grams or less but some labs are routinely testing as much as 300 grams) which lead to operating curves for c=0 acceptance that have an inflection at about 1 CFU/pound. The net effect of such sampling and testing is the erroneous belief that the worst lots are detected when 1 positive is found in many hundreds or thousands of samples. Unfortunately, this testing is so far removed from the range of interest; it is little more than a random selection of lots to be rejected.
- A manual sheet based sampling can increase the effective sampling weight 20 or 30 fold which is enough to move the operating curve about an order of magnitude to the left to about 0.1 CFU/pound if the same c=0 inspection criteria are used. Similarly, if a continuous sampling method is used the effective sample can be increased 200 to 300 fold yielding an additional order of magnitude in LOD to about 0.01 CFU/pound.
- The surface area of an aggregating sampler affords advantages beyond material sampling when greater sensitive is desired for surface and fluid flow sampling. Water and air stream sampling are two examples of where flow sampling is advantageous. The surface area is also applicable to the sampling of surfaces where topical contamination is of concern.
- The use of the sampler has the advantages of being nondestructive and can yield executional efficiencies. However, the real advantage comes when this LOD is traded off for statistical process control with a two-stage acceptance criterion where deviation from normal are detected as opposed to randomly selecting lots for reallocation. This concept is discussed more fully below when this discussion returns to screening detection. This is an important distinction when the goal is to improve the microbial safety of a product or material. This line of argument also permits the more rapid detection of many cells rather than waiting for one cell to grow into many cells. With the recognition of the power of the larger and effective sampling procedure, there may be a need to expand the range of tooling to apply this technology to a broader array of products with alternative geometries and increased levels of automation. For example, the geometry can be altered to allow sampling of a powder flow through a pipe with a circular geometry where a pipe segment is exchanged between lots. The pipe section would either be lined with sampling material or better include baffles maximizing product contact with the sampling surfaces. Alternatively, one can envision vertical chutes below pocket fillers to sample product just prior to bagging in a form fill and seal machine.
- A microbial sampler may be included, for example, non-woven fabric, various micro fiber materials, sponges, and/or any absorbent sheet material. A non-woven polypropylene or polyethylene fabric are of particular note as these materials are allowed for food contact and therefore have very low extractables which might otherwise contaminant the material stream under examination.
- Additionally, the utility of these sampling approaches can be increased with automation. A feed cartridge can be used to deliver multiple sheets to the sampling location at one time. This cartridge would be placed on the line after sanitation has completed all preparations protecting the drive mechanisms from the harsh cleaning process.
- Similarly, a magazine of cassettes can be loaded to collect individual pieces of sampling material. Both the cartridge and cassette are engineered to advance the sampling material when appropriate, (e.g. When a lot is completed, when a tote is moved, etc.). The motive force to advance sampling sheets can be provided by a motor, or supplied by a manual crank or handle, or by an operator depending on the specifics of the operation. Both cassette and cartridge may be designed to protect the microbial integrity of the contents. The cartridges may prevent microbial growth after sampling, external contamination, and cross contamination between lots.
- The sampling material can be mounted on an inert backing material to facilitate the sanitary placement of sampling sheets. Alternatively, sheets of sampling material can be separated by short spaces of inert material to ensure that used sampling materials do not contaminate other sheets.
- The usual design parameters for process equipment may be applied to these devices including, for example, an aggregating sampler. For example, heavy gauge 316 stainless is an appropriate material. When the sampling device is not in place, the location may be passive such as a dead plate where product passes without damage or hindrance. It may be sanitary design from the beginning such that it is easily cleaned.
- In one case, for this automation to have maximal benefit, the cartridges may carry the information regarding the sample they contain. If the sample is used in a manual mode, an electronic transfer of this information along with the sample is also advantageous to avoid human error and speed the flow of information. This transfer of information can occur through the cloud using barcodes, a database and location information.
- Another class of geometries is necessary to extend the power of this sampling to agricultural commodities in the field. Such sampling has utility beyond testing for human pathogens in that it can be used for testing for plant pathogens that can decrease the productivity of a crop. For example, early detection of mildew spores prompt early harvest of a spinach field. Detection of blight in a wheat or corn field might prompt the use of a disease control measure on the affected field before the blight destroys the crop.
- Depending on the field crop, an octopus tentacle configuration may be the appropriate geometry where strands of the sampling material are slide across the surface of the crop. These strands can be fuzzy cords or strips of material depending on what provide the greatest effective contact. These tentacles can be contacted to the crop by various mechanisms including robots, tractors, hand carrying or drones. It is most important that the altitude be held constant to allow contact while minimizing damage to the crop. To increase the effectiveness of the sampling, it can be advantageous to wick moisture down these tentacles or install vacuuming or sucking mechanism.
- For crops that present a more uniform top surface such as baby greens or spinach, sheet materials can be more effective as new upturned leave surfaces are missed. For these crops, air based sampling with suction or electrostatics that increases microbial sampling efficiency presents an interesting alternative.
- For manual sampling, forming the sampling material into pockets, mittens or gloves can facilitate use. Ease of use will generate greater compliance with the sampling protocol. In a manual mode, the duration of contact is a factor in determining the effectiveness of sampling. Typical durations are minutes. 2-5 minutes durations will work for most applications.
-
FIG. 2 illustratesexample operations 200 for applying aggregating sampling, in accordance with aspects of the present disclosure. Specifically,operations 200 begin, atblock 202, with providing at least one aggregating sampler at one or more sampling locations. Additionally,operations 200 include, atblock 204, sampling, using the at least one aggregating sampler, a production lot of produce creating one or more samples that makes up a microbial sampling. The one or more samples may be configured to be processed to indicate if pathogens are present at no greater than a normal background. The one or more sampling locations may include at least one of in a field, at harvest, just after dumping or cutting, in a wash system, or after the wash system. In one or more cases, additional operations may be include such as, for example, assessing, using the aggregating sampler, a level of cross contamination control to validate or verify a wash process. - In one or more cases, an aggregating sampler may be provided that sufficiently samples a production lot of ready-to-eat produce to confirm that pathogens are present at no greater than the normal background. In one or more cases, an aggregating sampler may assess the level of cross contamination control to validate or verify a wash process.
- In one or more cases, an aggregating sampler and a sampling location of the aggregating sampler may be provided. Additionally, sampling by the aggregating sampler may be provided to generate one or more desired samples. Analysis of the one or more samples and interpretation of the analysis results may also be provided. These elements can be practiced individually or together to enhance food safety.
- The aggregating sampler may include a collection surface and an apparatus for holding and positioning the collection surface such that the collection surface contacts product that is to be sampled for micro-organisms or other targets. In one or more cases, a surface with a sampling efficiency that allows an increased effective sampling size when a production lot of the product is sampled may be provided. For example, two hours of production of a leafy green product may be sampled with such a device. During the two hours a large amount, for example 10,000 to 30,000 pounds, of product will have crossed the sampling surface. If the sampling device is at least 25% efficient as shown in bench scale studies, the effective sample size may be 2500 to 7500 pounds. These sample sizes are enormous when compared to the few hundred grams of a normal grab sample.
- In one or more cases, there are a number of factors to consider in selecting or designing an aggregating sampler. Initially, a sampling surface that is compatible with the product is provided. This typically means that the sampling surface is a food grade material. For example, in one or more cases, a non-woven polyolefin cloth has proven to be effective. Another factor may include a design that allows safe and rapid exchange of the sampling surface. Another factor that may be included relates to any apparatus left on the line being easily cleaned and may incorporate a sanitary design. For example, food grade stainless steel may be a material selected when implementing an aggregating sampler.
- For produce testing, there are a number of areas where aggregating sampling may yield improvements in food safety. For example, some locations include: 1) In the field; 2) At harvest; 3) Just after dumping or cutting; 4) In the wash system; or 5) After the wash system. In each area, there are particular locations that may be selected for aggregate sampling, but these will vary with the configuration of the specific line and the product to be sampled. Not all areas will be appropriate for all products. The selections may be guided by the desired information. Currently, grab samples are often taken in all of these areas but these grab samples are unable to represent the population under examination and such sampling is generally destructive.
- According to an example, an aggregating sampler may be provide that can replace the current practice of taking a grab sample in a field by cutting leaves. Particularly, the aggregating sampler can be configured such that it can be carried by hand or mounted on a device designed for traveling through the field. The collection surface for field sampling may be divided to allow more conformity to the crop surface.
- According to another example, at harvest, an aggregating sampler may be placed on the harvester such that product is sampled during the harvesting process. This approach would allow pairing of harvested product with specific information. Although not required, placement of the aggregating sampler may be just after any sorting is done in the field. For example, if rocks are sorted from the product by density classification, these rocks need not cross the sampler surface.
- In another example, just after dumping or cutting and before washing, the product may still have the flora found in the field. An aggregating sampler placed early in the process can sample these organisms. Just after cutting or chopping, the interior of some products will be exposed for the first time allowing a more representative sample to be taken.
- In a wash system, an aggregating sampler can collect a different type of sample in accordance with one or more examples. This sampler may test the cross contamination control of the wash system. The organisms collected will reflect the two most probable mechanisms of cross contamination, water mediated cross contamination and product to product cross contamination. An aggregating sampler, placed in the flow of the conveyed product will be impinged by both the water and the product. To avoid overly hindering product flow depending on the line design, the sampler can be placed at any angle from parallel to the product flow to complete perpendicular to product flow. The sampler can also be a comb-like device with multiple collecting probes among product and in the wash flow. The angle of attachment may affect the balance between water mediated cross contamination and product to product cross contamination observed. In either case, this type of sampling may be used to validate cross contamination control and effectiveness of wash solution.
- Sampling in the wash system is a case where the sampling surface may be active on multiple surfaces, for example, on both sides or around in the case of the comb like structure mentioned above. In one or more cases, it can prove advantageous to laminate two sheets together with an impermeable tie layer to increase the binding potential relative to the detachment potential by avoiding flow through the sampling surface. Additionally, further advantages may be provided in other ways such as by increasing the thickness of the sampling material. In some cases, designing samplers in devices such as filter housing may be provided. In other cases, placement on a sampling surface in an active area of the wash system may be provided for getting a full measure of the cross contamination potential.
- A sample may be taken after the wash system and will reflect a residual population. In this area there are a number of specific locations that can be considered depending on the specifics of the line. For example, these specific locations include just prior to loading dryers, in a conveyer that might be used to lift the product for packaging, just before a pocket scale, or in the throat of a form fill and seal machine. This in-line continuous sampling may significantly increase sampling efficiency and provide more meaningful data than grab sampling of finished product testing.
- With samples taken, attention may turn to the analysis and interpretation of results as discussed herein. For example, in the specific category of produce, there are specific opportunities to be considered that may be provided with the aggregated sampling using the aggregated sampler. The opportunities are afforded in part due to the more representative nature of the aggregated samples and the greatly increase numbers of organisms available in the samples relative to the typical grab sample. These samples may be analyzed to give multiple channels of data depending on the detector technology employed. In one or more cases, the sample may be analyzed with metagenomics, allowing for the whole population to be studied yielding a large and in some cases a maximum amount of data which can be mined in various ways to gain knowledge and understanding of positive and negative deviations. This range of possibilities can be illustrated with a number of examples but are not limited thereto.
- In one or more examples, the samples taken in the field, at harvest, or just prior to washing can be used to assess the microbiological status of the raw material. From a food safety perspective the focus heretofore has been on the presence or absence of pathogens. Unfortunately, such analyzes based on grab samples are unable to truly answer the question as to whether pathogens are present due to their lack of sensitivity. Generally, it is known that pathogens are present at very low numbers. These are ubiquitous organisms. A more appropriate question is whether these organisms are present at abnormal concentrations or without the usual competing organism.
- In one or more examples, samples taken just prior to washing may be compared to samples taken in the wash system to directly measure cross contamination using wild type flora. Water samples may tend to have very low microbial loads in properly managed wash systems even when cross contamination is occurring. The wild type flora may also be highly variable. However, by using aggregating samplers at both locations the noise can be dampened and cross contamination measured. A number of metrics for cross contamination can be considered based on the ratio of the results for the samples from prewash to those from in the wash. By using more sophisticated analytical procedures one can overcome the flaws in such metrics as aerobic plate count (APC) which would include many organisms that are not relevant to cross contamination control of pathogens. For example, spore forming bacteria such as Bacillus will be unaffected by the wash and just cloud any metric of cross contamination based on APC. However, with more focused channels of data as afforded by modern molecular techniques better information can be obtained. Another aspect of this tool that may be provided is that statistical process control can be applied to detect deviations.
- In one or more cases, samples from after a wash process may provide information about microbial populations on the finished product. These samples may provide a much more accurate assessment of the pathogen risk of the product and better detection deviations. These samples can also be used to check for deviations in the normal flora.
- In some cases, a ratio between the before wash samples and the after wash samples may be used to assess the impact of the wash process. The aggregating samplers may reduce the noise and as with the cross contaminations metrics, these ratios can be handled with statistical process control to look for deviations.
- One or more of these examples may be delivered in almost real time because the aggregating samplers collected enough cells for concentration and direct analysis without enrichment.
- In accordance with one or more cases, an example of a bench scale study demonstrates a sampling efficiency of 15-20% relative to stomaching. It also shows that transfer is essentially instantaneous and that repeated contact collects more organisms. These observations confirm the intuitive assertion that continuous sampling will yield better information than grab sampling.
- For example, for one bench scale study experiment, purchased stew meat is inoculated by immersion in a mixed culture of generic E. coli at ˜105 CFU/ml. E. coli is selected as benign and easy to enumerate. The stew meat is allowed to rest at room temperature for 30 minutes to allow adherence. Then the six treatments listed in the table shown in
FIG. 3 may be executed in 5 replicates. All sampling cloths may be cut in half, 12 in by 8 in, to reduce the sample requirement and facilitated execution of the experiment. The sampling cloth may be, for example, a MicroTally cloth but is not limited thereto. The surface areas of the meat cubes are measured directly. For the sampling cloth treatments, meat is arranged in a 10 cm by 10 cm block and the sampling cloth is applied to the upper surface. Each mini sampling cloth is extracted in 200 ml of phosphate buffered saline (PBS). As a control, cubes of stew meat are stomached for 60 seconds in 200 mL of PBS. All stomached cubes are measured to estimate surface area. The counts are normalized for surface area and averaged. This normalization provides an apple to apple comparison. - This bench scale study experiment has been implemented and the average results are tabulated and shown in Table 1 of
FIG. 3 . These were analyzed with a General Linear Model (GLM) model which indicates that time of contact was not a factor. Multiple contacts yielded about the expected increase and are truly additive. - A logical extrapolation of this exercise is to estimate the effective sample size of a sampling. This may not truly be possible given the differences in geometry. However, in one or more use cases when using one side only, a sampling cloth is about 6 times larger than those used in the bench scale study experiment and the intended use is to sample almost 2000 pounds of product. Thus, it is reasonable to assert that the effective sample is expected to be 300 to 400 pounds. Larger scale experiments may be implemented that may further confirm this estimate. In summary, a benefit and advantage of the above method and apparatus of sampling may include providing an improvement over traditional grab samples for meat sampling by providing larger effective samples.
- In accordance with one or more cases, an example of a bench scale study demonstrates a sampling efficiency of about 30% relative to stomaching. It also shows that transfer is essentially instantaneous and that repeated contact collects more organisms. These observations confirm the intuitive assertion that continuous sampling will yield better information than grab sampling for lettuce.
- For example, in accordance with a bench scale study experiment, purchased lettuce is inoculated by immersion in a mixed culture of generic E. coli at ˜105 CFU/ml. E. coli is selected as benign and easy to enumerate. The lettuce is allowed to rest at 4° C. for 30 minutes to allow adherence. This short time may explain the higher efficiency observed when compared to a meat study where adhesion maybe faster. Then the six treatments listed in Table 2 shown in
FIG. 4 may be executed in 5 replicates. All sampling cloths may be cut in half, 12 in by 8 in, to reduce the sample requirement and facilitated execution of the experiment. The sampling cloth may be, for example, a MicroTally cloth but is not limited thereto. The surface areas of the lettuce leaves may be measured directly. For the sampling cloth treatments, lettuce is arranged in a 10 cm by 10 cm block and the sampling cloth applied to the upper surface. Each mini sampling cloth is extracted in 200 ml of PBS. As a control, lettuce may be stomached for 60 seconds in 200 mL of PBS. All stomached leaves are measured to estimate surface area. The counts are normalized for surface area and averaged. This normalization provides an apple to apple comparison. - This bench scale study experiment has been implemented and the average results are tabulated and shown in Table 2 of
FIG. 4 . These were analyzed with a GLM model which indicates that time of contact was not a factor. Multiple contacts yielded about the expected increase and are truly additive. - A logical extrapolation of this exercise is to estimate the effective sample size of a sampling. This may not truly be possible given the differences in geometry. However, in one or more use cases when using one side only, a sampling cloth is about 6 times larger than those used in the bench scale study experiment and the intended use is to sample almost 2000 pounds of product. Thus, it is reasonable to assert that the effective sample is expected to be 400 to 600 pounds. Larger scale experiments may be implemented that may further confirm this estimate. In summary, a benefit and advantage of the above method and apparatus of sampling may include providing an improvement over traditional grab samples for leafy sampling by providing larger effective samples.
- In one or more cases, swabs and/or sheets, referred to generally as a sheet below, may be used in accordance with one or more embodiments of the present disclosure. A sheet may include a microbial sampling material, such as sterile woven and/or non-woven synthetic fabrics and non-woven cloth for sampling and testing in the field of food safety. These sheets can be folded and curved to allow better conformation to product (e.g., food) streams that are being sampled or when used in a manual mode as driven by the product and the container. In some cases, configurations for sampling raw products or materials may include a sampling device that moves across the stationary product effectively yielding the equivalent of a product stream when sampling needs to occur prior to harvest. In some cases, configurations may include tail shaped sheets similar to, for example, the tentacles of a jellyfish.
- In one or more cases, configurations may be provided that may provide easier use under some conditions. In one case, a microbial aggregating sampler, may include a covering including a microbial sampling material with a pocket formed in the covering to receive an appendage or a tool for handling of the covering.
- In some cases, the covering may further include an attachment feature formed in the pocket to receive the tool. The attachment feature may include at least one of a hole formed through the covering; a loop positioned within the pocket to receive an end of the tool there through; or a tab positioned within the pocket for an end of the tool to attach thereto.
- In some cases, the covering may include a sheath formed in the pocket to receive a digit of an appendage. In some cases, the pocket is formed through the covering such that the appendage or the tool for handling the covering extends through the covering. The covering may be completely formed from the microbial sampling material. In some cases, the covering includes two sheets attached to each other to form the pocket. In some cases, the covering may include a single sheet folded and attached to itself to form the pocket.
- For example, a sheet may undergo folding and seaming to form a bag or pocket that can be worn as a mitten, glove, sock, or other covering, to facilitate manual sampling. Such a covering may encase an appendage, such as a hand, to facilitate pushing and pulling of the sampler through the product to be sampled. In some cases, aggregating samplers may be created that are more hand like configurations as mitts with thumbs and/or gloves with fingers, allowing the sampler to better conform to the hand. Such configurations may allow easier use when the product is more viscous or more prone to adhering to the sampler. A benefit to this aggregating sampler with a pocket may include the ability of the sampler to, when working the sampler, increase or maximize product contact. Further, in some cases, the addition of one or more attachment features, such loops or tabs, to assist with controlling the product contact may be included. These are representative examples and are not meant to limit other configuration embodiments of an aggregating sampler.
- In some cases, the aggregating sampler may be used in an automated machine setting to sample a product stream. The aggregating sampler may include a number of modifications in accordance with one or more cases. In some cases, the aggregating sampler may include one or more bends and curves that may improve the utility and facilitate use. In some cases, modifications may include, but are not limited to, forming a tube that can slide over one or more shafts for positioning in the product flow. This configuration may remove the need to slide the sampler within any holding device. In some cases, adding tabs or holes can be provided which may allow positioning and facilitate attachment. In some cases, the active sampling surface can be attached to a support material or web that has one or more of these features. One or more of these cases and modifications may help facility contact with the product stream with minimal manual intervention.
-
FIG. 5 illustrates an aggregatedsampler 500 that includes a pocket in accordance with aspects of the present disclosure. As shown the aggregatedsampler 500 includes a covering 504 made of a microbial sampling material. The covering is formed such that it includes a pocket formed in the covering to receive an appendage ortool 502 within the pocket for handling of the covering. -
FIG. 6 illustrates an aggregatedsampler 600 that includes a pocket that extends or is formed through a covering 604 in accordance with aspects of the present disclosure. The pocket is formed such that the appendage ortool 602 for handling the covering 604 extends through the covering. -
FIG. 7 illustrates an aggregatedsampler 700 with at least one convex surface of the covering 704 in accordance with aspects of the present disclosure. The covering forms a pocket for a tool orappendage 702.FIG. 8 illustrates an aggregatedsampler 800 that includes at least one concave surface of the covering 804 in accordance with aspects of the present disclosure. The covering 804 forms a pocket for a tool orappendage 802.FIG. 9 illustrates an aggregatedsampler 900 that includes a curved shape covering 904 in accordance with aspects of the present disclosure. The covering 904 is formed to include a pocket for a tool orappendage 902. -
FIG. 10 illustrates an aggregatedsampler 1000 that includes a hole orloop 1008 in accordance with aspects of the present disclosure. The covering 1004 may include the hole orloop 1008, such as positioned within the pocket of thecovering 1004 and attached to an inner surface of thecovering 1004. This hole orloop 1008 may be used by ahook 1006 of atool 1002 to attach to thecovering 1004. -
FIG. 11 illustrates an aggregatedsampler 1100 that is shaped as a mitten or glove for ahand 1102 in accordance with aspects of the present disclosure. As shown, the covering 1104 of the aggregatedsampler 1100 is formed to contour to ahand 1102. In particular, a sheath is formed in the covering 1104 in this embodiment to receive a thumb of thehand 1102, though one or more sheaths may be used to receive any digit of thehand 1102. Further, though only ahand 1102 is shown as an example appendage in this embodiment, the present disclosure is not so limited, as other appendages (i.e., a foot) may be contemplated for other embodiments without departing from the scope of the present disclosure. - In accordance with one or more cases, the aggregated sampling sheet may be hung into an active zone of a wash line with a cord or cable. This line may be attached in number of ways to the sampling material such as, for example, a grommet and clasp. In some cases, a ball and slide clamp may be used. In some cases, a punched hole may be used but may pull out. In other cases, various other clamps may be used. In one or more case, an optional float such as a fishing float with or without a weight may be included that will add drag and bouncy which may improve surface exposure. This approach may not include a fixed appliance for holding the sampling material.
- The reference method for extracting microorganisms from grab samples is homogenization by mixing or stomaching with an appropriate amount of fluid, usually a buffer. The purpose of adding fluid is to neutralize any antimicrobial or other properties of the sample that may be unfavorable for microbial growth such as low pH and suspend microorganism in liquid to facilitate downstream testing. However, adding fluid also dilutes the concentration of the microorganisms in the sample. Because most microbiological testing only take a portion of the homogenate (e.g., 0.1 mL or 1 mL for plating; 2.5 μL for direct PCR), the homogenization by dilution method decreases the detectability of the organisms of interest by as much as 1,500,000 times. To increase the detectability, a lengthy enrichment procedure is incorporated to allow a single viable cell to proliferate to millions so that it can be detected. This procedure costs 24 to 48 hours delay in obtaining detection data and more days in decision making time due to subsequent confirmation steps. It is also important that the samples be preserved properly and extraction be done as close to sampling time as possible. If the sample has changed before extraction, the results do not represent the tested lot.
- For most conventional sampling programs, the percentage of negative samples exceeds 99%. Based on simple modeling, one can easily conclude that the number of cells of the organism of interest is seldom more than 1 and probably rarely more than 5 based on a Poisson distribution. With so few cells present, the enrichment is often done in the extraction buffer. This is critical with the small sampling but does not make this type of testing any more representative of the lot under test.
- However, this conventional enrichment and detection can be applied to extract from an aggregating sampler when presence or absence of the target organism is the desired goal. Modes of detection are considered herein.
- When extracting the sampling materials these same considerations apply. However, the impact on the results of failing to extract one organism is an order of magnitude or smaller due to the larger effective sample. In addition, the cassettes afford the opportunity to begin the extraction faster by adding the fluid immediately after sampling providing maximal time for the extraction to occur while avoiding human interaction.
- When extracting a 24×8 inch sampling sheet of a poly-olefin sampler, a volume of 100 to 200 mls of extraction solution is generally appropriate. The composition of this solution is driven by the planned detection system and is generally defined in the associated method.
- During the concentration, two classes of materials may need to be removed, the small, <100,000 molecular weight, and the very large, >50 microns. In addition, the sample may need to be concentrated to about 1 mL to be compatible with the screening detection system. Given that the extraction generally starts at about 200 mL, there is a large amount of water to remove.
- Two schemes are practical. Traditionally, one can filter the extraction fluid through an inert 50 microns cut off filter and then sediment the organisms of interest by centrifugation. The resulting pellets can be re-suspended in an appropriate buffer and taken on to cleaning. Alternatively, after filtration, the small molecules and water can be removed osmotically with adsorbents or pressure and a semi-permeable membrane such as used for reverse osmosis or ultrafiltration. The latter is more amenable to automation as the resultant concentrated sample remains in solution. However, this option may require a for purpose module to be executed.
- It is important to maintain the connection to the initial Meta data through this process. If enough of interfering material is removed and the sample is sufficiently concentrated, the cleaning step examined next can be skipped and moving directly to the screening determination. This decision determination may be made of a case by case basis.
- It is also possible to use non-specific binding such as a cation exchange surface to collect that target organisms and remove them from the bulk solution. Such as an approach would partially combine concentration and cleaning. This approach is most practical when the there are few larger particles to interfere.
- At this point in the process, samples have been greatly reduced in volume but the organisms of interest have not been segregated from other organism so the signal to noise ratio is till problematic. In addition, further concentration may be needed for detection without enrichment during which one organism is converted to many at the cost of time and delay in decision making.
- Several schemes are practical but all involve binding the organisms of interest in a small area such as a microfluidized or nanofluidized channel. This channel or area may or may not be filled with surface activated nanofiber. Utilizing elasto-inertial microfluidics, the viscoelastic flow enables size based migration of larger particles into a non-Newtonian solution, while smaller bacteria remain in the streamline of the blood sample entrance and can be separated. It is tempting to consider surface activated magnetic particles; however, the mechanical manipulation of these particles to achieve the desired small volume is a larger engineering challenge than activating the small surface area. However, any binding geometry that fixes the organisms of interest and any other organisms selected to represent the other microflora in an appropriate small volume can be used.
- The surface activation may require numerous active binding sites in close proximity. The mixed DNA primer arrays of SnapDNA are one class of materials. Another class is a cocktail of antibodies for all the organisms of interest.
- The binding mechanism may bind all the organism of interest. These organisms may be sufficiently bound that other organisms and materials are selectively removed from the area of binding as clean fluid is passed through the channel.
- The motive force to move the cells through the channel can be the mechanical action of fluid flow, electrostatic as the surface of most bacteria is negative, or a size pumping action such as practiced with ferromagnetic particles.
- With the partially purified organism or organisms of interest bound in a small area or small volume if the cleaning step proves unnecessary, many approaches are available for screening detection for process control. The functional requirements are clearer. First and foremost is that enrichment culture takes substantial amounts of time, and such should be eliminated or reduced in one or more cases. Second, the screening metric may need to be an index suitable for statistical process control. This implies a measurement with many states as opposed to
binary 0 and 1. The magnitude of this metric may relate to the extent of deviation from normal operation and therefore the likelihood that an outbreak could occur. Under these conditions, the detection of deviation can be based on the classic rules for control charting. Furthermore, trend detecting rules have the potential to detect problems before significant deviations have occurred. The statistically based Westcard rules provide one basis for trend analysis. - Expressed differently, an index may be needed that tallies the relative presence of beneficial organisms and potential pathogens and aggregates this information in a useful way. As more information is acquired about specific products, the power of big data will come into play. However, at the simplest levels, the aggregate level of potential pathogen is a useful screening tool. The relative balance between potential pathogens and beneficial organisms is a more sophisticated analysis to compensate for seasonal variations that are inherent in many products. It is reasonable to expect to develop indices that are product specific.
- The information behind indices is evolving rapidly. The simplest useful index is a ratio of pathogen intensity to a benign organism. These can be generated by many means including classical enumeration with plating, but the classical methods are two slow to meet the functional requirements outlined above. However, given the concentration of the organisms on the cleaning substrate or the concentrated extraction, it is possible to go directly to qPCR in some cases to generate index data.
- There are two schemes for generating this type of data at its most sophisticated level. First, one can use a collection of ligands (antibodies, aptamers, or others) that bind and tag all the organisms of potential interest yielding a collection of signals that are multiplexed into a family of useful channels. Alternatively, one can generate an array of specific binding interactions that are analyzed chemometrically to yield a metric. The latter approach will be faster and probably less expensive after the research and analysis is done.
- As an example of the first scheme would be a using a mixture of conjugated antibodies to bind to all types of cells of interest. For produce, hemolytic E. coli, Salmonella and Listeria are of greatest interest. Poultry focuses on Campylobacter and Salmonella. Other industries have other and additional interests. These antibodies can be bound to the organisms bound to the cleaning substrate. The retained tags with either a fluorescent probe or an enzyme provide signal amplification and detection. When the cleaning region as a small enough cross section, the signal from the hundreds to thousands of cells on the cleaning substrate are detectable. Micro and nano fluidization are necessary. However, specific detection protocols can be evaluated at a macro scale using an appropriately instrumented microscope that can be used to measure the signal from a small area where cells have been collected. For speed to market, multiple detectors can be run in parallel or series to generate similar multiple channels of data.
- The alternative scheme invokes the lab on a chip concept. By building an array of binding sites, the composition of the samples can be queried. PCR can amplify the contents selectively with a collection of primers. Such detectors can evolve to providing both the screening detection of this step and the ultimate secondary screening. However, it is likely to remain a two-step process due to the economics.
- A number of technologies can meet these requirements including but not limited to, for example: 1) Sensors where the vibrational frequencies (which may for example include optical waveguide), impedance, or other properties of a transistor are modified by the binding of the organisms of interest to the sensor. This approach may require that the sensor be built into the surface of the channel. Another technology may include, 2) qPCR where the cells are laid in place and the number of copies of the organisms of interest, or the number of ribosomes of the organisms of interest are estimated. Another technology may include, 3) Use tagged antibodies to light up the organisms of interest so that they can detected on the absorbent surface spectrophotometrically. Enzyme, fluorescent probes, and other materials to amplify the signal can be used.
- It may be desirable to avoid confirmation of the presence of one or more specific pathogens during this screening. This is where the trade between LOD and speed to useful information. If the signal is not about three times background, no further action is warranted.
- Given that economics may play a role in testing, it should be noted that the sampling approach is compatible with various compositing strategies. Samples can be composited anywhere along the processing path before detection. In various situations, one position will be more advantageous than others. Pooling samples prior to extraction reduces work but reduces the resolution of the results if a positive result is found. However, if the result is negative tremendous savings are achieved. After concentrating and cleaning would allow sampled to be retested if only a portion of the extracted sample is used in a wet pooling approach which can avoid detections costs. Each system needs to be evaluated to determine how to best control costs.
- The confirmation step will be applied when there is reason to suspect that a pathogen is present. The technology in this area is evolving rapidly with many new approaches and efficiencies being developed and introduced. Some will use the techniques suggested for screening with more specific reagents for the confirmation. At present, all the common procedures rely on either molecular biology or ligand binding reactions. Various strategies have been developed to amplify the signal noise ratio and to identify the contaminant to the desired specificity. The desired specificity ranges from simple speciation to identify specific serotypes.
- For confirmation, a concentrated sample such as afforded the screening detection above may be included. The concentration may be included due to the small volumes that are compatible with these types of procedures. The organisms, the surface antigens from the organisms, or the nucleic acid from the organisms may need to be extracted from the screening system to the extent that these materials can interact with the reagents of the confirmation procedure. In other words, there is not a priory reason that the detection module could not be engineered for a second round of chemistries. Any one of these materials may contain the information necessary to characterize the contaminant. There are a number of these processes and they are being improved. The choice of approach will be driven by cost, desired specificity and the desired speed.
- Many strategies are available to amplify the base signal from these materials that will still be present at only modest concentration in the typical sample. The yield of material from the hundreds to thousands of organisms of interest bound to the screening detector platform is still very small. Amplification based on radio isotopes are largely out of favor but still possible. However, enzyme systems are still in use and new enzymes strategies are still being developed such as those used for ELISA methods or with a luciferase. Fluorescent tags on specific antibodies such as those proposed for the screening determination are less useful for this purpose due the high potential for cross reactivity. However, antibodies of this type are the basis for the serotyping classifications that up until recently has been the standard for characterization. These older serotyping assays may require isolation and growing the organisms.
- Increasingly characterization is based on the presence of sequences of nucleic acid. These can be nuclear DNA, ribosomal RNA, or messenger RNA. At the extreme, it is now practical to sequence the entire genome of the organism. Increasing specificity is useful for identification of the source a contamination. However, it also presents a potential liability in the case of an outbreak of illness.
- The molecular assays are built around the use of enzymes to replicate sequences of nucleic acid to generate a large enough signal for detection. Various primers are used to select which portions are copied; up to and including the entire genome. For measurement, various fluorescent tags are used. One of the newest techniques utilizes the melting and binding of target material to known sequences to generate a complex matrix of information that can be used chemometrically in lieu of complete sequence data.
- Both the screening detection and the confirmation results are reported directly to a sequel database. This reporting may not require human intervention if the included quality assurance standards (positive and negative controls) fall within normal ranges. This avoids transcription and transposition errors. Digital records are more reliable and accurate than manual records.
- Both detectors should be part of the “Internet of Things”. This connectivity allows results to be pushed to operators on the floor allowing for the rapid release of product or for the redisposition of product if a potential issue is identified that may need to be addressed. Once the results are in an appropriate database, various users can have customized interfaces providing the information. Some may need to track individual results. Others may be more interested in trends and averages. There may be provided a class of users that may want to aggregate even larger data sets to compare across locations.
- There are many platforms available for extracting information from the database. For example, but not limited thereto, Ignition has proven useful in this record as it is open source allowing customization.
- In accordance with an aspect of the disclosure, a method for microbial sampling food may include gathering a microbial sampling from one or more food items, extracting microorganisms from the microbial sampling, concentrating the microorganisms, cleaning the microorganisms, tallying a relative presence of the microorganisms and any potential pathogens, aggregating information of a microorganism tally from the tallying of microorganisms into a microorganism report, confirming the microorganism tally, and reporting the microorganism report of the microorganism tally.
- In some cases, gathering the microbial sampling from the one or more food items includes sampling, using an aggregating sampler, the one or more food items that include a production lot of produce or meat creating one or more samples that makes up the microbial sampling. The one or more samples may be configured to be processed to indicate if pathogens are present at no greater than a normal background.
- In some cases, the method may further include assessing, using an aggregating sampler, a level of cross contamination control to validate or verify a wash process. Gathering the microbial sampling from the one or more food items may include providing an aggregating sampler at a sampling location. The sampling location may be at least one of in a field, at harvest, just after dumping or cutting, in a wash system, or after the wash system.
- In some cases extracting includes enriching the microbial sampling, and adding fluid to the microbial sample. In some cases concentrating includes filtering extraction fluid of the microbial sampling using at least one of centrifugation filtering or osmotically filtering. In some cases, cleaning includes binding the microorganisms in a small area including one or more of a microfluidized or nanofluidized channel. In some cases tallying includes using a collection of ligands that bind and tag all the microorganisms of potential interest yielding a collection of signals that are multiplexed into a family of useful channels. In some cases ligands include one or more of antibodies, primers, and aptamers.
- In some cases tallying includes generating an array of specific binding interactions that are analyzed chemometrically to yield a metric. The method may further include building an array of binding sites, wherein the composition of the samples can be queried, and amplifying, using a PCR, the contents selectively with a collection of primers.
- In some cases confirming includes extracting surface antigens from the organisms or nucleic acid from the organisms from a screening system to the extent that these materials can interact with the reagents of a confirmation procedure. In some cases confirming further includes amplifying a base signal of the microorganism tally.
- In some cases, the method may further include use of an index as a surrogate for direct results regarding presence or absence of organisms of interest. In some cases, the method may further include use of a statistical process control for detecting deviations in microbial flora.
- In accordance with an aspect of the disclosure, a method of applying aggregating sampling to food items including providing at least one aggregating sampler at one or more sampling locations, and sampling, using the at least one aggregating sampler, a production lot of produce or meat creating one or more samples that makes up a microbial sampling.
- In some cases the one or more samples are configured to be processed to indicate if pathogens are present at no greater than a normal background. In some cases, the one or more sampling locations includes at least one of in a field, at harvest, just after dumping or cutting, in a wash system, or after the wash system. The method may further include assessing, using the aggregating sampler, a level of cross contamination control to validate or verify a wash process.
- In accordance with an aspect of the disclosure, an apparatus for microbial sampling, including means for gathering a microbial sampling from one or more food items, means for extracting microorganisms from the microbial sampling, and means for concentrating the microorganisms, means for cleaning the microorganisms, means for tallying a relative presence of the microorganisms and any potential pathogens, means for aggregating information of the microorganism tally into a microorganism report, means for confirming the microorganism tally, and means for reporting the microorganism report of the microorganism tally.
- In accordance with an aspect of the disclosure, an apparatus for microbial sampling, includes at least one processor configured to generate control signals for controlling gathering a microbial sampling from one or more food items, extracting microorganisms from the microbial sampling, concentrating the microorganisms, cleaning the microorganisms, tallying a relative presence of the microorganisms and any potential pathogens, aggregating information of the microorganism tally into a microorganism report, and confirming the microorganism tally, and a transmitter configured to transmit the microorganism report of the microorganism tally. In some cases the apparatus may further include an aggregating sampler configured to gather the microbial sampling.
- In accordance with an aspect of the disclosure, a non-transitory computer readable medium for microbial sampling having instructions stored thereon for gathering a microbial sampling from one or more food items, extracting microorganisms from the microbial sampling, concentrating the microorganisms, cleaning the microorganisms, tallying a relative presence of the microorganisms and any potential pathogens, aggregating information of the microorganism tally into a microorganism report, confirming the microorganism tally, and reporting the microorganism report of the microorganism tally.
- In accordance with an aspect of the disclosure, a method for sampling food including concentrating microorganisms and removing interference, tallying a relative presence of the microorganisms and any potential pathogens, and aggregating information of the microorganism tally into a microorganism report. In accordance with an aspect of the disclosure, a system capable of implementing one or more of the novel aspects discussed in this application disclosure.
- In accordance with an aspect of the disclosure, a microbial aggregating sampler, including a covering including a microbial sampling material with a pocket formed in the covering to receive an appendage or a tool for handling of the covering.
- In some cases, the covering includes an attachment feature formed in the pocket to receive the tool. In some cases, the attachment feature includes one of a hole formed through the covering, a loop positioned within the pocket to receive an end of the tool there through, and a tab positioned within the pocket for an end of the tool to attach thereto.
- In some cases the covering includes a sheath formed in the pocket to receive a digit of an appendage. In some cases the pocket is formed through the covering such that the appendage or the tool for handling the covering extends through the covering. In some cases the covering is completely formed from the microbial sampling material. In some cases the covering includes two sheets attached to each other to form the pocket. In some cases the covering includes a single sheet folded and attached to itself to form the pocket.
- The aggregating sampler can be used to sample for additional analytes beyond bacteria including yeast, molds, viruses, allergens, toxins such as aflatoxin, or particulates such as dust. The commonality is the surface presence of the analyte at low levels that can be concentrated. The basic process is the same with the same key steps. The biggest differences will be in detection strategy, but these strategies are well known by those who study these analytes.
- The methods disclosed herein comprise one or more steps or actions for achieving the described method. The method steps and/or actions may be interchanged with one another without departing from the scope of the claims. In other words, unless a specific order of steps or actions is specified, the order and/or use of specific steps and/or actions may be modified without departing from the scope of the claims.
- As used herein, a phrase referring to “at least one of” a list of items refers to any combination of those items, including single members. As an example, “at least one of: a, b, or c” is intended to cover a, b, c, a-b, a-c, b-c, and a-b-c, as well as any combination with multiples of the same element (e.g., a-a, a-a-a, a-a-b, a-a-c, a-b-b, a-c-c, b-b, b-b-b, b-b-c, c-c, and c-c-c or any other ordering of a, b, and c). As used herein, including in the claims, the term “and/or,” when used in a list of two or more items, means that any one of the listed items can be employed by itself or any combination of two or more of the listed items can be employed. For example, if a composition is described as containing components A, B, and/or C, the composition can contain A alone; B alone; C alone; A and B in combination; A and C in combination; B and C in combination; or A, B, and C in combination.
- As used herein, the term “determining” encompasses a wide variety of actions. For example, “determining” may include calculating, computing, processing, deriving, investigating, looking up (e.g., looking up in a table, a database or another data structure), ascertaining and the like. Also, “determining” may include receiving (e.g., receiving information), accessing (e.g., accessing data in a memory) and the like. Also, “determining” may include resolving, selecting, choosing, establishing and the like.
- The previous description is provided to enable any person skilled in the art to practice the various aspects described herein. Various modifications to these aspects will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other aspects. Thus, the claims are not intended to be limited to the aspects shown herein, but is to be accorded the full scope consistent with the language claims, wherein reference to an element in the singular is not intended to mean “one and only one” unless specifically so stated, but rather “one or more.” For example, the articles “a” and “an” as used in this application and the appended claims should generally be construed to mean “one or more” unless specified otherwise or clear from the context to be directed to a singular form. Unless specifically stated otherwise, the term “some” refers to one or more. Moreover, the term “or” is intended to mean an inclusive “or” rather than an exclusive “or.” That is, unless specified otherwise or clear from the context, the phrase, for example, “X employs A or B” is intended to mean any of the natural inclusive permutations. That is, for example the phrase “X employs A or B” is satisfied by any of the following instances: X employs A; X employs B; or X employs both A and B. All structural and functional equivalents to the elements of the various aspects described throughout this disclosure that are known or later come to be known to those of ordinary skill in the art are expressly incorporated herein by reference and are intended to be encompassed by the claims. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the claims. No claim element is to be construed under the provisions of 35 U.S.C. § 112, sixth paragraph, unless the element is expressly recited using the phrase “means for” or, in the case of a method claim, the element is recited using the phrase “step for.”
- The various operations of methods described above may be performed by any suitable means capable of performing the corresponding functions. The means may include various hardware and/or software component(s) and/or module(s), including, but not limited to a circuit, an application specific integrated circuit (ASIC), or processor. Generally, where there are operations illustrated in figures, those operations may have corresponding counterpart means-plus-function components with similar numbering.
- The various illustrative logical blocks, modules and circuits described in connection with the present disclosure may be implemented or performed with a general purpose processor, a digital signal processor (DSP), an application specific integrated circuit (ASIC), a field programmable gate array (FPGA) or other programmable logic device (PLD), discrete gate or transistor logic, discrete hardware components, or any combination thereof designed to perform the functions described herein. A general-purpose processor may be a microprocessor, but in the alternative, the processor may be any commercially available processor, controller, microcontroller, or state machine. A processor may also be implemented as a combination of computing devices, e.g., a combination of a DSP and a microprocessor, a plurality of microprocessors, one or more microprocessors in conjunction with a DSP core, or any other such configuration.
- If implemented in hardware, an example hardware configuration may comprise a processing system in a wireless node. The processing system may be implemented with bus architecture. The bus may include any number of interconnecting buses and bridges depending on the specific application of the processing system and the overall design constraints. The bus may link together various circuits including a processor, machine-readable media, and a bus interface. The bus interface may be used to connect a network adapter, among other things, to the processing system via the bus. The network adapter may be used to implement the signal processing functions of the PHY layer. In the case of a user terminal 120 (see
FIG. 1 ); a user interface (e.g., keypad, display, mouse, joystick, etc.) may also be connected to the bus. The bus may also link various other circuits such as timing sources, peripherals, voltage regulators, power management circuits, and the like, which are well known in the art, and therefore, will not be described any further. The processor may be implemented with one or more general-purpose and/or special-purpose processors. Examples include microprocessors, microcontrollers, DSP processors, and other circuitry that can execute software. Those skilled in the art will recognize how best to implement the described functionality for the processing system depending on the particular application and the overall design constraints imposed on the overall system. - If implemented in software, the functions may be stored or transmitted over as one or more instructions or code on a computer readable medium. Software shall be construed broadly to mean instructions, data, or any combination thereof, whether referred to as software, firmware, middleware, microcode, hardware description language, or otherwise. Computer-readable media include both computer storage media and communication media including any medium that facilitates transfer of a computer program from one place to another. The processor may be responsible for managing the bus and general processing, including the execution of software modules stored on the machine-readable storage media. A computer-readable storage medium may be coupled to a processor such that the processor can read information from, and write information to, the storage medium. In the alternative, the storage medium may be integral to the processor. By way of example, the machine-readable media may include a transmission line, a carrier wave modulated by data, and/or a computer readable storage medium with instructions stored thereon separate from the wireless node, all of which may be accessed by the processor through the bus interface. Alternatively, or in addition, the machine-readable media, or any portion thereof, may be integrated into the processor, such as the case may be with cache and/or general register files. Examples of machine-readable storage media may include, by way of example, RAM (Random Access Memory), flash memory, phase change memory, ROM (Read Only Memory), PROM (Programmable Read-Only Memory), EPROM (Erasable Programmable Read-Only Memory), EEPROM (Electrically Erasable Programmable Read-Only Memory), registers, magnetic disks, optical disks, hard drives, or any other suitable storage medium, or any combination thereof. The machine-readable media may be embodied in a computer-program product.
- A software module may comprise a single instruction, or many instructions, and may be distributed over several different code segments, among different programs, and across multiple storage media. The computer-readable media may comprise a number of software modules. The software modules include instructions that, when executed by an apparatus such as a processor, cause the processing system to perform various functions. The software modules may include a transmission module and a receiving module. Each software module may reside in a single storage device or be distributed across multiple storage devices. By way of example, a software module may be loaded into RAM from a hard drive when a triggering event occurs. During execution of the software module, the processor may load some of the instructions into cache to increase access speed. One or more cache lines may then be loaded into a general register file for execution by the processor. When referring to the functionality of a software module below, it will be understood that such functionality is implemented by the processor when executing instructions from that software module.
- Also, any connection is properly termed a computer-readable medium. For example, if the software is transmitted from a website, server, or other remote source using a coaxial cable, fiber optic cable, twisted pair, digital subscriber line (DSL), or wireless technologies such as infrared (IR), radio, and microwave, then the coaxial cable, fiber optic cable, twisted pair, DSL, or wireless technologies such as infrared, radio, and microwave are included in the definition of medium. Disk and disc, as used herein, include compact disc (CD), laser disc, optical disc, digital versatile disc (DVD), floppy disk, and Blu-ray® disc where disks usually reproduce data magnetically, while discs reproduce data optically with lasers. Thus, in some aspects computer-readable media may comprise non-transitory computer-readable media (e.g., tangible media). In addition, for other aspects computer-readable media may comprise transitory computer-readable media (e.g., a signal). Combinations of the above should also be included within the scope of computer-readable media.
- Thus, certain aspects may comprise a computer program product for performing the operations presented herein. For example, such a computer program product may comprise a computer-readable medium having instructions stored (and/or encoded) thereon, the instructions being executable by one or more processors to perform the operations described herein. For example, instructions for performing the operations described herein and illustrated in the appended figures.
- Further, it should be appreciated that modules and/or other appropriate means for performing the methods and techniques described herein can be downloaded and/or otherwise obtained by a user terminal and/or base station as applicable. For example, such a device can be coupled to a server to facilitate the transfer of means for performing the methods described herein. Alternatively, various methods described herein can be provided via storage means (e.g., RAM, ROM, a physical storage medium such as a compact disc (CD) or floppy disk, etc.), such that a user terminal and/or base station can obtain the various methods upon coupling or providing the storage means to the device. Moreover, any other suitable technique for providing the methods and techniques described herein to a device can be utilized.
- It is to be understood that the claims are not limited to the precise configuration and components illustrated above. Various modifications, changes and variations may be made in the arrangement, operation and details of the methods and apparatus described above without departing from the scope of the claims.
Claims (20)
1. A method for microbial sampling food comprising:
gathering a microbial sampling from one or more food items;
extracting microorganisms from the microbial sampling;
concentrating the microorganisms;
cleaning the microorganisms;
tallying a relative presence of the microorganisms and any potential pathogens;
aggregating information from the tallying of microorganisms into a microorganism report;
confirming the tallying of microorganisms; and
reporting the microorganism report of the tallying of microorganisms.
2. The method of claim 1 , wherein gathering the microbial sampling from the one or more food items comprises:
sampling, using an aggregating sampler, the one or more food items that include a production lot of produce or meat creating one or more samples that makes up the microbial sampling,
wherein the one or more samples are configured to be processed to indicate if pathogens are present at no greater than a normal background.
3. The method of claim 2 , wherein the aggregating sampler comprises:
a covering comprising a microbial sampling material with a pocket formed in the covering to receive an appendage or a tool for handling of the covering.
4. The method of claim 3 , wherein the covering comprises an attachment feature formed in the pocket to receive the tool, and
wherein the attachment feature comprises one of:
a hole formed through the covering;
a loop positioned within the pocket to receive an end of the tool therethrough; and
a tab positioned within the pocket for an end of the tool to attach thereto.
5. The method of claim 3 , wherein the pocket is formed through the covering such that the appendage or the tool for handling the covering extends through the covering.
6. The method of claim 3 , wherein the covering comprises:
two sheets attached to each other to form the pocket, or
a single sheet folded and attached to itself to form the pocket.
7. The method of claim 1 , further comprising:
assessing, using an aggregating sampler, a level of cross contamination control to validate or verify a wash process.
8. The method of claim 1 , wherein gathering the microbial sampling from the one or more food items comprises:
providing an aggregating sampler at a sampling location,
wherein the sampling location is at least one of in a field, at harvest, just after dumping or cutting, in a wash system, or after the wash system.
9. The method of claim 1 , wherein extracting comprises:
enriching the microbial sampling; and
adding fluid to the microbial sample.
10. The method of claim 1 , wherein concentrating comprises:
filtering extraction fluid of the microbial sampling using at least one of centrifugation filtering or osmotically filtering.
11. The method of claim 1 , wherein cleaning comprises:
binding the microorganisms in a small area including one or more of a microfluidized or nanofluidized channel.
12. The method of claim 1 , wherein tallying comprises:
using a collection of ligands that bind and tag all microorganisms of potential interest yielding a collection of signals that are multiplexed into a family of useful channels, wherein ligands include one or more of antibodies, primers, and aptamers.
13. The method of claim 1 , wherein tallying comprises:
generating an array of specific binding interactions that are analyzed chemometrically to yield a metric comprising:
building an array of binding sites,
wherein a composition of the samples can be queried; and
amplifying, using a PCR, the samples selectively with a collection of primers.
14. The method of claim 1 , wherein confirming comprises:
extracting surface antigens from the microorganisms or nucleic acid from the microorganisms from a screening system to an extent that these materials can interact with reagents of a confirmation procedure; and
amplifying a base signal of the tallying of microorganisms.
15. The method of claim 1 , further including use of one or more of an index as a surrogate for direct results regarding presence or absence of organisms of interest, or a statistical process control for detecting deviations in microbial flora.
16. A method of applying aggregating sampling to food items comprising:
providing at least one aggregating sampler at one or more sampling locations; and
sampling, using the at least one aggregating sampler, a production lot of produce or meat creating one or more samples that makes up a microbial sampling.
17. The method of claim 16 , wherein the one or more samples are configured to be processed to indicate if pathogens are present at no greater than a normal background.
18. The method of claim 16 , wherein the one or more sampling locations includes at least one of in a field, at harvest, just after dumping or cutting, in a wash system, or after the wash system.
19. The method of claim 16 , further comprising:
assessing, using the aggregating sampler, a level of cross contamination control to validate or verify a wash process.
20. An apparatus for microbial sampling, comprising:
at least one processor configured to generate control signals for controlling:
gathering a microbial sampling from one or more food items;
extracting microorganisms from the microbial sampling;
concentrating the microorganisms;
cleaning the microorganisms;
tallying a relative presence of the microorganisms and any potential pathogens;
aggregating information from the tallying of microorganisms into a microorganism report; and
confirming the tallying of microorganisms;
an aggregating sampler configured to gather the microbial sampling; and
a transmitter configured to transmit the microorganism report of the tallying of microorganisms.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/152,566 US20210247372A1 (en) | 2017-08-09 | 2021-01-19 | Method and apparatus for applying aggregating sampling to food items |
US17/747,928 US20220276212A1 (en) | 2017-08-09 | 2022-05-18 | Method and Apparatus for Applying Aggregating Sampling to Food Items |
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201762543220P | 2017-08-09 | 2017-08-09 | |
US201762589755P | 2017-11-22 | 2017-11-22 | |
US201862656164P | 2018-04-11 | 2018-04-11 | |
US16/057,137 US10935535B2 (en) | 2017-08-09 | 2018-08-07 | Method and apparatus for applying aggregating sampling to food items |
US17/152,566 US20210247372A1 (en) | 2017-08-09 | 2021-01-19 | Method and apparatus for applying aggregating sampling to food items |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/057,137 Division US10935535B2 (en) | 2017-08-09 | 2018-08-07 | Method and apparatus for applying aggregating sampling to food items |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US17/747,928 Continuation US20220276212A1 (en) | 2017-08-09 | 2022-05-18 | Method and Apparatus for Applying Aggregating Sampling to Food Items |
Publications (1)
Publication Number | Publication Date |
---|---|
US20210247372A1 true US20210247372A1 (en) | 2021-08-12 |
Family
ID=63557670
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/057,137 Active 2038-11-24 US10935535B2 (en) | 2017-08-09 | 2018-08-07 | Method and apparatus for applying aggregating sampling to food items |
US17/152,566 Pending US20210247372A1 (en) | 2017-08-09 | 2021-01-19 | Method and apparatus for applying aggregating sampling to food items |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/057,137 Active 2038-11-24 US10935535B2 (en) | 2017-08-09 | 2018-08-07 | Method and apparatus for applying aggregating sampling to food items |
Country Status (8)
Country | Link |
---|---|
US (2) | US10935535B2 (en) |
EP (1) | EP3665300A1 (en) |
CN (1) | CN111194354A (en) |
AU (1) | AU2018313794A1 (en) |
BR (1) | BR112020002557A2 (en) |
CA (1) | CA3071584A1 (en) |
MX (1) | MX2020001228A (en) |
WO (1) | WO2019032638A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023239933A1 (en) * | 2022-06-09 | 2023-12-14 | Fremonta Corporation | Manual sampling device and methods |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20210017574A1 (en) * | 2019-07-19 | 2021-01-21 | Fremonta Corporation | Method for assessing the lethality and the level of cross contamination control of a process non-invasively |
CN110499243A (en) * | 2019-08-29 | 2019-11-26 | 贵州大学 | A kind of microorganism spreading rod |
CN113804503B (en) * | 2021-09-26 | 2024-06-07 | 自然资源部第二海洋研究所 | Seabed substrate and water body timing sampling system and method |
CN114418270B (en) * | 2021-12-01 | 2023-05-23 | 江苏权正检验检测有限公司 | Sample intelligent screening method and system for food detection |
CN116797053B (en) * | 2023-08-25 | 2023-11-10 | 深圳普菲特信息科技股份有限公司 | Chemical production data analysis method, system and medium based on neural network |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4316462A (en) * | 1980-05-21 | 1982-02-23 | Siloam, Inc. | Filtering device for an injection device |
US20130344478A1 (en) * | 2011-03-09 | 2013-12-26 | 3M Innovative Properties Company | Apparatus and method for processing a sample |
US20160209305A1 (en) * | 2013-09-30 | 2016-07-21 | 3M Innovative Properties Company | Guanidine-functionalized metal silicate particles and methods of making and using such particles |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5868928A (en) * | 1997-06-10 | 1999-02-09 | Bradley; Bruce J. | Microbial sampler and concentrator |
US20030023149A1 (en) | 2001-06-29 | 2003-01-30 | Montemagno Carlo D. | Microfluidic microorganism detection system |
EP1782320A2 (en) * | 2004-07-29 | 2007-05-09 | Kim Laboratories, Inc. | Ultrasensitive sensor and rapid detection of analytes |
CA2536698A1 (en) * | 2006-02-15 | 2007-08-15 | Fio Systems Corporation | System and method of detecting, identifying and characterizing pathogensand characterizing hosts |
EP2183383B1 (en) * | 2007-08-01 | 2013-01-16 | Hitachi Chemical Co., Ltd. | Pathogen detection in large-volume particulate samples |
CN102296103A (en) * | 2010-06-28 | 2011-12-28 | 唐锋 | Solid-phase pour enrichment method and matched device thereof |
FR2962445B1 (en) * | 2010-07-08 | 2013-06-28 | Biomerieux Sa | METHOD FOR DIRECT DETECTION AND IDENTIFICATION OF MICROORGANISM IN A DILUTED BIOLOGICAL SAMPLE IN AN ENRICHMENT BROTH |
TWI582238B (en) * | 2012-07-17 | 2017-05-11 | 奈寇公司 | A method for constructing a diversity index and viability index of microorganisms in process samples |
US20140046722A1 (en) * | 2012-08-10 | 2014-02-13 | Sample6 Technologies, Inc. | System for on-site environment monitoring |
-
2018
- 2018-08-07 US US16/057,137 patent/US10935535B2/en active Active
- 2018-08-08 WO PCT/US2018/045699 patent/WO2019032638A1/en unknown
- 2018-08-08 AU AU2018313794A patent/AU2018313794A1/en active Pending
- 2018-08-08 MX MX2020001228A patent/MX2020001228A/en unknown
- 2018-08-08 EP EP18769247.0A patent/EP3665300A1/en active Pending
- 2018-08-08 CA CA3071584A patent/CA3071584A1/en active Pending
- 2018-08-08 CN CN201880065377.1A patent/CN111194354A/en active Pending
- 2018-08-08 BR BR112020002557-7A patent/BR112020002557A2/en unknown
-
2021
- 2021-01-19 US US17/152,566 patent/US20210247372A1/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4316462A (en) * | 1980-05-21 | 1982-02-23 | Siloam, Inc. | Filtering device for an injection device |
US20130344478A1 (en) * | 2011-03-09 | 2013-12-26 | 3M Innovative Properties Company | Apparatus and method for processing a sample |
US20160209305A1 (en) * | 2013-09-30 | 2016-07-21 | 3M Innovative Properties Company | Guanidine-functionalized metal silicate particles and methods of making and using such particles |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023239933A1 (en) * | 2022-06-09 | 2023-12-14 | Fremonta Corporation | Manual sampling device and methods |
Also Published As
Publication number | Publication date |
---|---|
US10935535B2 (en) | 2021-03-02 |
BR112020002557A2 (en) | 2020-08-04 |
AU2018313794A1 (en) | 2020-03-19 |
EP3665300A1 (en) | 2020-06-17 |
US20190049419A1 (en) | 2019-02-14 |
CA3071584A1 (en) | 2019-02-14 |
WO2019032638A1 (en) | 2019-02-14 |
CN111194354A (en) | 2020-05-22 |
MX2020001228A (en) | 2020-09-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20210247372A1 (en) | Method and apparatus for applying aggregating sampling to food items | |
Martins et al. | Biosensors for on-farm diagnosis of mastitis | |
Bhunia | One day to one hour: how quickly can foodborne pathogens be detected? | |
US9932620B2 (en) | Methods, devices, and systems of detecting microorganisms | |
O’Neil et al. | Comparison of wet-mount, Wright-Giemsa and Gram-stained urine sediment for predicting bacteriuria in dogs and cats | |
KR20120044197A (en) | Automatic realtime pcr system for the various analysis of biological sample, method for automatic nucleic acid purification and realtime quantification of gene amplification, method for automatic viable cell count of pathogenic bacteria using realtime quantitative pcr, method for automatically getting antigene density using quantitative immunity pcr | |
WO2019133756A1 (en) | Automated priming and library loading device | |
US20230166256A1 (en) | Methods and systems for sample processing utilizing filter aid materials and aggregating samplers for equipment | |
JP2023164475A (en) | Application development environment for biological sample assessment processing | |
US20220276212A1 (en) | Method and Apparatus for Applying Aggregating Sampling to Food Items | |
Fung | Predictions for rapid methods and automation in food microbiology | |
Kumar et al. | Foodborne disease symptoms, diagnostics, and predictions using artificial intelligence-based learning approaches: a systematic review | |
US11911717B2 (en) | Method and apparatus for applying aggregating sampling | |
in't Veld et al. | Impact of animal husbandry and slaughter technologies on microbial contamination of meat: monitoring and control | |
Logue et al. | Rapid microbiological methods in food diagnostics | |
Rathnayaka et al. | Rapid Detection of Food Pathogens Using Molecular Methods | |
CN101649316B (en) | Method for quickly extracting pathogen PCR nucleic acid template in foods | |
CN103698530A (en) | Application of mycobacterium tuberculosis protein in preparation of products used for diagnosis of active tuberculosis | |
US20240240264A1 (en) | Rapid milk sample preparation method compatible with molecular tests | |
Pancholi et al. | Rapid screening of urine specimens for bacteriuria by the cellenium system | |
Huegel et al. | Automatic Preparation of Specimens for Diagnostic Testing from a Diverse Range of Animals with the 2A Automatic Homogenization Processor | |
In't Veld et al. | Changing perspectives in food microbiology: implementation of rapid microbiological analyses in modern food processing | |
Jones | To the Window, To the Wall: Improving Environmental Monitoring Protocols | |
Narayana Iyengar et al. | Identifying antibiotic-resistant strains via cell sorting and elastic-light-scatter phenotyping | |
Fung | Methods for Evaluating Microbial Flora in Muscle Foods |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |