ITMI20010555A1 - REDUCED STRAW OF SALMONELLA GALLINARUM-PULLORUM AND VACCINE THAT CONTAINS IT - Google Patents
REDUCED STRAW OF SALMONELLA GALLINARUM-PULLORUM AND VACCINE THAT CONTAINS IT Download PDFInfo
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- ITMI20010555A1 ITMI20010555A1 ITMI20010555A ITMI20010555A1 IT MI20010555 A1 ITMI20010555 A1 IT MI20010555A1 IT MI20010555 A ITMI20010555 A IT MI20010555A IT MI20010555 A1 ITMI20010555 A1 IT MI20010555A1
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- pullorum
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- salmonella
- vaccine
- salmonella gallinarum
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- 241000607142 Salmonella Species 0.000 title claims description 16
- 229960005486 vaccine Drugs 0.000 title claims description 16
- 238000000034 method Methods 0.000 claims description 10
- 241000271566 Aves Species 0.000 claims description 8
- 206010057249 Phagocytosis Diseases 0.000 claims description 8
- 230000008782 phagocytosis Effects 0.000 claims description 8
- 238000002255 vaccination Methods 0.000 claims description 7
- 210000003714 granulocyte Anatomy 0.000 claims description 5
- 241000287828 Gallus gallus Species 0.000 claims description 4
- 206010039438 Salmonella Infections Diseases 0.000 claims description 4
- 235000013330 chicken meat Nutrition 0.000 claims description 4
- 206010039447 salmonellosis Diseases 0.000 claims description 4
- 230000001018 virulence Effects 0.000 claims description 3
- 229940031551 inactivated vaccine Drugs 0.000 claims 1
- 230000006698 induction Effects 0.000 claims 1
- 208000015181 infectious disease Diseases 0.000 description 6
- 230000002238 attenuated effect Effects 0.000 description 5
- 239000013049 sediment Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 229940124590 live attenuated vaccine Drugs 0.000 description 4
- 229940023012 live-attenuated vaccine Drugs 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 206010040047 Sepsis Diseases 0.000 description 3
- 230000000680 avirulence Effects 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 208000013223 septicemia Diseases 0.000 description 3
- 230000002459 sustained effect Effects 0.000 description 3
- 208000004232 Enteritis Diseases 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 241001354013 Salmonella enterica subsp. enterica serovar Enteritidis Species 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000006150 trypticase soy agar Substances 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 241001138501 Salmonella enterica Species 0.000 description 1
- 241000596091 Salmonella enterica subsp. enterica serovar Anatum Species 0.000 description 1
- 241000392514 Salmonella enterica subsp. enterica serovar Dublin Species 0.000 description 1
- 241000607726 Salmonella enterica subsp. enterica serovar Heidelberg Species 0.000 description 1
- 241000210647 Salmonella enterica subsp. enterica serovar Montevideo Species 0.000 description 1
- 241000293871 Salmonella enterica subsp. enterica serovar Typhi Species 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 208000037386 Typhoid Diseases 0.000 description 1
- 206010000210 abortion Diseases 0.000 description 1
- 231100000176 abortion Toxicity 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 230000005571 horizontal transmission Effects 0.000 description 1
- 230000007124 immune defense Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 208000004396 mastitis Diseases 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 201000008297 typhoid fever Diseases 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
Descrizione dell'invenzione industriale avente per titolo: Description of the industrial invention entitled:
“CEPPO ATTENUATO DI SALMONELLA GALLINARUM-PULLORUM E VACCINO CHE LO CONTIENE” "ATTENUATED STRAIN OF SALMONELLA GALLINARUM-PULLORUM AND VACCINE THAT CONTAINS IT"
Campo dell’ invenzione: Field of invention:
La presente invenzione si riferisce a un ceppo attenuato di Salmonella gallinarum-pullorum, al procedimento per la preparazione del nuovo ceppo e ai vaccini contenenti detto ceppo contro le salmonellosi animali ed umane. The present invention relates to an attenuated strain of Salmonella gallinarum-pullorum, to the process for the preparation of the new strain and to vaccines containing said strain against animal and human salmonellosis.
Le Salmonellosi umane ed animali si presentano generalmente in una o due forme: Human and animal Salmonellosis generally occurs in one or two forms:
1. una forma tipicamente enterica, che è causata spesso da salmonelle ubiquitarie (non ospite specifiche), tra le quali S. enteritidis, S. typhimurium, S. heidelberg, S. anatum, S. montevideo, etc. Questi ceppi ubiquitari si trasmettono facilmente da un ospite ad un altro compreso l’uomo. 1. a typically enteric form, which is often caused by ubiquitous (non-host specific) salmonellae, including S. enteritidis, S. typhimurium, S. heidelberg, S. anatum, S. montevideo, etc. These ubiquitous strains are easily transmitted from one host to another including humans.
2. Le salmonelle specie specifiche causano una setticemia nell’ospite correlato. Raramente si trasmettono ad altri ospiti e causano enteriti con diarrea ad altri ospiti. Queste salmonelle sono: S. typhi (o typhosa ) causa della febbre tifoide nella specie umana; S. dublin causa di enterite, setticemia, aborti e mastiti nella specie bovina (ed occasionalmente ovina); S. choleraesuìs, causa della setticemia nella specie suina; S. gallinarum-pullorum, causa della pullurosi e della tifosi aviare. Salmonella gallinarum-pullorum in particolare appartiene al sierogruppo 1,9,12. 2. Species-specific salmonellae cause septicemia in the related host. They rarely pass on to other hosts and cause enteritis with diarrhea in other hosts. These salmonellae are: S. typhi (or typhosa) cause of typhoid fever in humans; S. dublin cause of enteritis, septicemia, abortions and mastitis in bovine (and occasionally ovine) species; S. choleraesuìs, cause of septicemia in the swine species; S. gallinarum-pullorum, cause of pullurosis and avian fans. Salmonella gallinarum-pullorum in particular belongs to the serogroup 1,9,12.
Stato dell’arte State of the art
I vaccini vivi attenuati disponibili contro Salmonella gallinarumpullorum noti e disponibili sul mercato contengono microrganismi attenuati con procedimenti vari. The live attenuated vaccines available against Salmonella gallinarumpullorum known and available on the market contain microorganisms attenuated by various procedures.
In EP-555366 è descritto un procedimento per la preparazione di un vaccino vivo attenuato contro la Salmonella choleraesuis mentre la domanda EP 876476 descrive un vaccino vivo attenuato contro Γ infezione aviare sostenuta da Salmonella enteritidis. Questi procedimenti hanno lo svantaggio di non eliminare completamente la virulenza; inoltre permane il rischio, sebbene molto limitato, di infettare i soggetti. EP-555366 describes a procedure for the preparation of a live attenuated vaccine against Salmonella choleraesuis while the application EP 876476 describes a live attenuated vaccine against Γ avian infection sustained by Salmonella enteritidis. These processes have the disadvantage of not completely eliminating virulence; moreover, the risk remains, although very limited, of infecting subjects.
In nessun prodotto disponibile il microrganismo è stato attenuato con il metodo della fagocitosi da parte dei granulociti eterofili. In no available product the microorganism was attenuated by the phagocytosis method by heterophilic granulocytes.
Si è ora sorprendentemente trovato, ed è oggetto della presente invenzione, un nuovo ceppo di Salmonella gallinarum-pullorum completamente avirulento. A new strain of completely avirulent Salmonella gallinarum-pullorum has now surprisingly been found and is the subject of the present invention.
Il ceppo dell’invenzione è stato depositato in data 27 dicembre 2000 presso l’ATCC (American Type Culture Collection) con numero di deposito PTA 2589 ed è stato designato SGP695AV. The strain of the invention was deposited on 27 December 2000 at the ATCC (American Type Culture Collection) with filing number PTA 2589 and was designated SGP695AV.
Compresi nella presente invenzione sono anche i vaccini vivi attenuati ottenuti dal nuovo ceppo e il metodo di preparazione del ceppo stesso. Also included in the present invention are the live attenuated vaccines obtained from the new strain and the method of preparation of the strain itself.
I vaccini possono essere preparati e commercializzati in forma di liofilizzato, di sospensione o di soluzione per somministrazione orale o parenterale. Vaccines can be prepared and marketed in the form of lyophilisate, suspension or solution for oral or parenteral administration.
Le forme farmaceutiche contengono il ceppo Salmonella gallinarumpulloum oggetto dell’invenzione e adatti eccipienti, adiuvanti e conservanti. The pharmaceutical forms contain the Salmonella gallinarumpulloum strain object of the invention and suitable excipients, adjuvants and preservatives.
Il vaccino secondo l’invenzione è vantaggioso perché rende possibile la profilassi indiretta dell’infezione sostenuta da Salmonella gallinarumpullòrum. In particolare, il vantaggio tipico dei vaccini vivi di stimolare le difese immunitarie cellulo-mediate, importanti nei confronti di infezioni sostenute da batteri come le Salmonelle che possono comportarsi da patogeni intracellulari facoltativi, è associato ad un livello di sicurezza paragonabile a quello dei vaccini spenti garantito dalla esclusiva modalità di attenuazione. The vaccine according to the invention is advantageous because it makes possible indirect prophylaxis of the infection sustained by Salmonella gallinarumpullòrum. In particular, the typical advantage of live vaccines to stimulate the cell-mediated immune defenses, important against infections sustained by bacteria such as Salmonella that can behave as facultative intracellular pathogens, is associated with a level of safety comparable to that of deactivated vaccines. guaranteed by the exclusive attenuation mode.
Descrizione dell’invenzione Description of the invention
L’attenuazione del batterio secondo l’invenzione è stata ottenuta con il metodo della fagocitosi da parte di granulociti eterofili. The attenuation of the bacterium according to the invention was obtained with the phagocytosis method by heterophile granulocytes.
Il processo di fagocitosi operato dagli eterofili sulle Salmonelle modifica le loro caratteristiche di virulenza portandone ad un’attenuazione. I cloni dei batteri fagocitati vengono esaminati per verificarne l’avirulenza, in quanto non tutti i batteri fagocitati sono sufficientemente avirulenti. Il ceppo completamente e sicuramente avirulento è stato ottenuto applicando la fagocitosi in modo ripetuto sulla selezione dei ceppi più avirulenti. The phagocytosis process operated by heterophiles on Salmonellae modifies their virulence characteristics leading to an attenuation. The clones of phagocytosed bacteria are examined to verify their avirulence, as not all phagocytosed bacteria are sufficiently avirulent. The completely and surely avirulent strain was obtained by applying phagocytosis repeatedly on the selection of the most avirulent strains.
I vaccini secondo l’invenzione saranno somministrati secondo modalità (dosi, tempi e vie di somministrazione) del tutto analoghe a quelle di vaccini noti. I vaccini dell’invenzione sono particolarmente utili per la vaccinazione di uccelli, in particolare polli, contro la tifosi aviare e contro la pullorosi aviare. The vaccines according to the invention will be administered according to methods (doses, times and routes of administration) completely similar to those of known vaccines. The vaccines of the invention are particularly useful for the vaccination of birds, in particular chickens, against avian fans and against avian pullorosis.
I vaccini dell’invenzione sono inoltre particolarmente utili per la vaccinazione contro salmonellosi causate da salmonelle appartenenti ai sierogruppi 1,9,12 o 9,12. The vaccines of the invention are also particularly useful for vaccination against salmonellosis caused by salmonella belonging to serogroups 1,9,12 or 9,12.
La presente invenzione viene descritta nel seguente esempio in maggior dettaglio. The present invention is described in the following example in greater detail.
ESEMPIO EXAMPLE
1. Un ceppo patogeno e virulento di Salmonella gallinarum-pullorum (SGP) è stato fatto crescere in brodo di coltura fino ad una concentrazione di 1x10 CFU (unità formanti placca) per mi. 3 mi di granulociti eterofili ad una concentrazione di 6.5xl0<6>/ml, isolati da sangue fresco di pollo, sono stati aggiunti ad 1 ml di SGP ad una concentrazione di 1x1 0<8 >CFU ed incubati per 20 minuti al fine di permettere la fagocitosi. 1. A pathogenic and virulent strain of Salmonella gallinarum-pullorum (SGP) was grown in culture broth to a concentration of 1x10 CFU (plaque forming units) per ml. 3 ml of heterophile granulocytes at a concentration of 6.5xl0 <6> / ml, isolated from fresh chicken blood, were added to 1 ml of SGP at a concentration of 1x1 0 <8> CFU and incubated for 20 minutes in order to allow phagocytosis.
2. Dopo aver centrifugato la miscela SGP-granulociti eterofili a 250g, il sedimento è stato risospeso in 4 mi di HBSS (Hank’s Balanced Salt Suspension) contenente 100 μl/ml di gentamicina ed incubato per 10 minuti. 2. After centrifuging the SGP-heterophile granulocyte mixture at 250g, the sediment was resuspended in 4 ml of Hank's Balanced Salt Suspension (HBSS) containing 100 μl / ml of gentamicin and incubated for 10 minutes.
3. Dopo centrifugazione, il surnatante è stato coltivato in terreno colturale per accertare l’assenza di SGP extracellulari vive non fagocitate. 3. After centrifugation, the supernatant was cultured in culture medium to ascertain the absence of non-phagocytosed live extracellular SGP.
4. Il sedimento del punto 3 è stato risospeso in HBSS contenente 100μl di Kanamicina ed è stato incubato per 10 minuti. 4. The sediment from step 3 was resuspended in HBSS containing 100μl of Kanamycin and incubated for 10 minutes.
5. Dopo centrifugazione, il sumatante è stato coltivato in un terreno colturale per accertare l’assenza di SGP extracellulari vive non fagocitate. 5. After centrifugation, the sumatant was grown in a culture medium to ascertain the absence of non-phagocytosed live extracellular SGP.
6. Il sedimento del punto 4 è stato risospeso in HBSS non contenente antibiotici ed è stato incubato per 10 minuti. 6. The sediment from step 4 was resuspended in HBSS containing no antibiotics and incubated for 10 minutes.
7. Dopo centrifugazione, il sumatante è stato coltivato in un terreno colturale per accertare l’assenza di SGP extracellulari vive non fagocitate. 7. After centrifugation, the sumatant was grown in a culture medium to ascertain the absence of non-phagocytosed live extracellular SGP.
8. Il sedimento cellulare del punto 6 è stato Usato incubandolo per 3 minuti in una soluzione all’ 1% di saponina ed acqua distillata. Il sedimento residuo è stato inoculato in TSA (tripticase soy agar) ed incubato 24 ore al fine di permettere la raccolta di colonie di SGP fagocitate. 8. The cellular sediment of step 6 was used by incubating it for 3 minutes in a 1% solution of saponin and distilled water. The residual sediment was inoculated in TSA (tripticase soy agar) and incubated 24 hours in order to allow the collection of phagocytosed SGP colonies.
9. 10 colonie distinte del punto 8 sono state somministrate a pulcini di 1 giorno per uso orale ad una concentrazione di 10<7 >CFU al fine di accertarne l’avirulenza. 9. 10 distinct colonies of point 8 were administered to 1-day-old chicks for oral use at a concentration of 10 <7> CFU in order to ascertain their avirulence.
Un clone (singola colonia) è definito avirulento se 5 pulcini di 1 giorno sopravvivono dimostrando un buono stato di salute per 10 giorni in seguito all’ inoculazione di SGP fagocitate. A clone (single colony) is defined avirulent if 5 1 day old chicks survive demonstrating a good state of health for 10 days following the inoculation of phagocytosed SGP.
10. Il clone avirulento, ottenuto con i passaggi 1-10 stato nuovamente sottoposto ai passaggi sopra descritti per riconfermarne l’avirulenza. 10. The avirulent clone, obtained with steps 1-10, was again subjected to the steps described above to reconfirm its avirulence.
Nel caso specifico: In the specific case:
Si è partiti da un ceppo di Salmonella gallinarum-pullorum virulento (SGP695V) che è stato sottoposto al trattamento costituito dai 10 passaggi prima descritti. Alla fine di ogni ciclo di fagocitosi è stato selezionato un clone sul quale è stata condotto l’ulteriore ciclo di fagocitosi. Il ceppo risultante SGP695AV è stato testato in 47 pulcini di un giorno. I test comparativi con il ceppo virulento di origine eseguiti su 25 polli di 1 giorno mostrano che il ceppo SGP695AV è avirulento e non è trasmissibile a soggetti suscettibili posti nello stesso ambiente come sentinelle. Al contrario, il ceppo di origine SGP695V, dopo 10 giorni dall’infezione, ha causato la morte del 64% dei soggetti, l’invasione degli organi interni nell’ 89% dei casi e la trasmissione dell’infezione a soggetti sentinella suscettibili. We started from a virulent strain of Salmonella gallinarum-pullorum (SGP695V) which was subjected to the treatment consisting of the 10 steps described above. At the end of each phagocytosis cycle, a clone was selected on which the further phagocytosis cycle was conducted. The resulting SGP695AV strain was tested in 47 day-old chicks. Comparative tests with the virulent parent strain performed on 25 1 day old chickens show that the SGP695AV strain is avirulent and is not transmissible to susceptible individuals placed in the same environment as sentinels. On the contrary, the SGP695V strain of origin, after 10 days of infection, caused the death of 64% of subjects, invasion of internal organs in 89% of cases and transmission of the infection to susceptible sentinel subjects.
I dati sono riportati nelle tabelle 1 e 2. The data are shown in Tables 1 and 2.
Tabella 1: esito dell'infezione di pulcini EOP (esenti da organismi patogeni) di 1 giorno con il ceppo virulento ed attenuato. Table 1: Infection outcome of 1 day old EOP (pathogen-free) chicks with the virulent and attenuated strain.
Tabella 2: Confronto della trasmissione orizzontale dei ceppi SGP695AV e SGP695V (virulento) dagli animali vaccinati ad animali sentinella. Table 2: Comparison of horizontal transmission of SGP695AV and SGP695V (virulent) strains from vaccinated to sentinel animals.
Claims (8)
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ITMI20010555 ITMI20010555A1 (en) | 2001-03-16 | 2001-03-16 | REDUCED STRAW OF SALMONELLA GALLINARUM-PULLORUM AND VACCINE THAT CONTAINS IT |
TR2002/00659A TR200200659A3 (en) | 2001-03-16 | 2002-03-13 | Salmonella gallinarum-pullorum weakened strain and rebel containing them. |
HU0200984A HU226257B1 (en) | 2001-03-16 | 2002-03-14 | Salmonella gallinarum-pullorum attenuated strain and vaccine containing it |
PL352815A PL202301B1 (en) | 2001-03-16 | 2002-03-15 | Attenuated strain of Salmonella gallinarium and vacine incorporating such strain |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ITMI20010555 ITMI20010555A1 (en) | 2001-03-16 | 2001-03-16 | REDUCED STRAW OF SALMONELLA GALLINARUM-PULLORUM AND VACCINE THAT CONTAINS IT |
Publications (1)
Publication Number | Publication Date |
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ITMI20010555A1 true ITMI20010555A1 (en) | 2002-09-16 |
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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ITMI20010555 ITMI20010555A1 (en) | 2001-03-16 | 2001-03-16 | REDUCED STRAW OF SALMONELLA GALLINARUM-PULLORUM AND VACCINE THAT CONTAINS IT |
Country Status (4)
Country | Link |
---|---|
HU (1) | HU226257B1 (en) |
IT (1) | ITMI20010555A1 (en) |
PL (1) | PL202301B1 (en) |
TR (1) | TR200200659A3 (en) |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ES2150419T3 (en) * | 1990-11-01 | 2000-12-01 | Univ Iowa State Res Found Inc | BACTERIAL AND VACCINE ATTENUATION PROCEDURE. |
JP2000502257A (en) * | 1995-12-19 | 2000-02-29 | アイオワ ステート ユニバーシティー リサーチ ファウンデーション,インコーポレーテッド | Heterofil-compatible poultry vaccine |
-
2001
- 2001-03-16 IT ITMI20010555 patent/ITMI20010555A1/en unknown
-
2002
- 2002-03-13 TR TR2002/00659A patent/TR200200659A3/en unknown
- 2002-03-14 HU HU0200984A patent/HU226257B1/en unknown
- 2002-03-15 PL PL352815A patent/PL202301B1/en unknown
Also Published As
Publication number | Publication date |
---|---|
HU226257B1 (en) | 2008-07-28 |
PL202301B1 (en) | 2009-06-30 |
HU0200984D0 (en) | 2002-05-29 |
PL352815A1 (en) | 2002-09-23 |
TR200200659A2 (en) | 2002-10-21 |
TR200200659A3 (en) | 2002-10-21 |
HUP0200984A2 (en) | 2006-12-28 |
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