CA1059915A - Growth promotor and processes - Google Patents
Growth promotor and processesInfo
- Publication number
- CA1059915A CA1059915A CA238,049A CA238049A CA1059915A CA 1059915 A CA1059915 A CA 1059915A CA 238049 A CA238049 A CA 238049A CA 1059915 A CA1059915 A CA 1059915A
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Abstract
ABSTRACT
A method for stimulating growth in animal or poultry which involves feeding a complete feed to the animal or poultry. The complete feed contains (i) gentian violet and (ii) the remainder inert ingredients. The inert ingredients include all of the Inert ingredients normally in a complete or basal animal or poultry feed. The gentian violet is a growth promoter for animal or poultry. The gentian violet is present in the feed in an amount between 0.00077 and 0.01 percent by weight, based on the weight of the feed. The growth of the animal or poultry is increased at a rate significantly faster than if the animal or poultry had not been fed the complete feed containing the gentian violet. A
method for treating or preventing Candida albicans infections in animals and/or poultry.
A method for stimulating growth in animal or poultry which involves feeding a complete feed to the animal or poultry. The complete feed contains (i) gentian violet and (ii) the remainder inert ingredients. The inert ingredients include all of the Inert ingredients normally in a complete or basal animal or poultry feed. The gentian violet is a growth promoter for animal or poultry. The gentian violet is present in the feed in an amount between 0.00077 and 0.01 percent by weight, based on the weight of the feed. The growth of the animal or poultry is increased at a rate significantly faster than if the animal or poultry had not been fed the complete feed containing the gentian violet. A
method for treating or preventing Candida albicans infections in animals and/or poultry.
Description
Field o f this Invention This invention relates to animal and poultry feeds, poultry drinking water and poultry water sanitizer which ~ontains a growth promoter that is a selective fungicidal mold-inhibitor of Candida ,a~bicans. This invention also relates to the use of such a growth promoter-containing feed'to stimulate the arowth of ani~als and poultry. This invention further relates to the use of such a feed ~or treating and, preventing Candida albicans infections in animals and/or poultry. -Broad Descripti.on of This'Invention An object of this invention is to provide a growth promotercomposition for non-humans animal~ and/or poultry. Another object. o. this invention is'to provide a growtl, pro~l~ot r COtpO-sition for non-human animals and/or poultry that sirnultaneously treats or prevents Candida aibicans infections in non-human . --~~ .
animals and/or poultry. A further object of this invention is to provide a proce.ss for using such growth promotion'compositiorls Ahother object of this invention is to provide animal and poultry ,feeds, poultry drinking water and poultry water sa~itizer that ,20 achieve the above process ob}ectives; Other objects and advan-tages are set out herein or are obvious'to one ordinarily skilled in the art herefrom.
Such objects and inventions are achieved by this i.nvent:ion.
~OS99~5 One embodiment of this invention is a method for stimulating growth in non-h~man animals and/or poultry.
The method includes feeding a complete feed to the non-human animal and/or poultry. The complete feed consists only of (i) gentian violet and (ii) the remainder inert ingredients. The inert ingredients include all of the inert ingredients normally found in a complete or basal non-human animal poultry feed. The gentian violet is present in the feed in an amount between 0.00077 and 0.01 percent by weight, based on the weight of the complete feed. The growth of the non-human animal or poultry is increased at a rate faster than if the non-human animal or poultry had not been fed the complete feed containing the gentian violet. The gentian violet is a growth promoter for said non-human animal or poultry.
Consumption by non-human animals and poultry of the gentian violet composition of this invention in the stated levels causes significant growth increase. The gentian violet is in effect a growth factor, i.e., is a substance which promotes growth in the poultry and non-human animals, particularly the young and immature ones.
For the best effects the feed composition of this invention is fed to poultry, hogs or cattle.
Preferably the preparation of the feed involves pre-paring a premix concentrate for addition to the feed which consists only of between 0.1 and 10 percent by weight of gentian violet and the remainder inert ingredients and thén adding enough of the premix concentrate to the other ingredients of feed to obtain the recited amount of gentian violet in the feed. Preferably inert ingredients used in the premix concentrate are diatomaceous silica, ` \
corn cob fractions, and vegetable oil. Most preferably, the feed contains 0.00156 percent by weight of gentian violet, based on the weight of the complete feed.
Other substituted benzophenone azoanilide dyes or rosaniline base dyes can be used in place of the gentian violet. Such are also known as triaminotritane dyes related to rosaniline. The -N(CH3)2 groups of gentian violet can be replaced by other auxochrome groups such as -OH, -NH2 -N(C2H5)2, etc. The other various ,, substituents, and the substituents on gentian violet~ can be located at any position on the three benzyl rings.
The other dyes must have at least one amino chloride group and/or substituted amino chloride group.
Examples of such other substituted benzophenone azoanilide dyes are: pentamethylpararosaniline chloride, tetramethylpararosaniline~chloride; methyl green or light green, which is the methyl chloridé add~ition product of gentian violet; ethyl green, which is the ethyl chloride addition product of gentian violet; sulfonic acids of the rosanilines, such as, water blue and patent blue, phenyl-ated rosanilines, such as, triphenyl fuch~sin, aniline blue, diphenylamine blue and spirit blue; pararosaniiine;
and mixtures of pararosaniline with its methyl homologs, such as fuchsins and magentas. More broadly, any tri-phenylmethane dye that contains an amino group or substituted amino group can be used in place of ~ 1059915 gentian violet. Examples o such tripbenylmethane dyes are the malachite green seri.es of dyes, such as, malachite green.
Most o~ such aniline dyes are rather toxic, but can be readily used if they are buffered by any generally non-toxic conventional bueren~ system to reduce their toxicity. So when such aniline dyes (other than genti.an violet) are used, the feed compositi.ons of this invention incl.ude a bufering a~ent in the inert ingredi-ents The term inert in~redients excludes therapeutically and pharmaceutically active ingredients ~e g., broad spectrum baci-~-riostatic or bacteriocidal ingredients), but does not exclude vitamins and minerals.
The gentian violet is preferably used in the form of a premix concerltrate, which can ~ dded t-o the con.plete feed as needed.
The most preferred prem.ix concentrate contai.ns corn cob ~ractions, 7r~-J~ ~t1a~1~
white mineral oil, diatomaceous silica (e.g., Micro-Cel ~j~ dis tilled water and 1.55 percent of gentian violet.
rrhe concent:ration of gentian violet in the prerni.x concentrate can easily be as high as 60 percent; it is preferably between 0 1 and 10 percent and is most preferably 1.55 percent.
The prelnix concentrate can contain any materlal that is used in a complete or basal non-human animal or poultry eed. The premix concentrate can be used in the form of a liquid or solid admixture~ The liqllid is best in the form of a ver~ viscous susrension or ~i,nilar ~emi-~luid. Preferclbly a solid premix concen~rate is used even thou~h it may contain a liquid material _5-. .
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such as white mineral oil which reduces the dust factor in the premix.
... The premix concentrate is used by addin~ it to the complete poultry or non-human feed as long as growth stimulation is wanted.
When the premix concentrate contains 1.55 percent of gentian ~iole~
preferably two pounds of the premix is used per ton of complete poultry feed for growth stimulation of non-human animals or poultry--this reflects a preferred concentrat~on of about 0.00155 percent of gentian violet in the complete poultry or non-human animal feed. Effective concentrations of gentian violet are within the range of 0.00077 to 0.005 percent of gentian violet per ton of complete poultry feed or non-human animal feed. The preferred concentrate ranqe of gentian violet is from 0.00085 to 0.01 per-: cent of gentian violet per ton of complete poultry or non-human animal feed. It has been found that most female poultry preferably are fed at a gentian violet level of 0.00155 perce.nt per ton of complete poultry.feed, and male poultry are fed at a gentian violet eve~ of 0.00385 percent per ton of complete poultry feed.
. The above concentrations in the complete poultry feed apply even when the gentian violet is directly admixed with the complete .
poultry.or non-human animal feed without going through the ilite~-mediate premix concentrate form. .
The premix is used for growth stimulation by adding i.t to t}-e complete feed on a continuou~ basis.
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Any complete poultry feed or basal poultry feed can be used. It can contain, for example, ground yellow corn, soybean oil meal, steamed bone meal, ground lime-stone, iodized salt, manganese sulfate, Vitamin A oil, dry Vitamin D-3, riboflavin, Vitamin B-12 and niacin. It can also contain, for example, fish meal and meat meal.
Any complete non-human animal feed/or basal non-human animal feed can be used; it can contain, for example, any of the following ingredients: mogul starch, iodized salt, dry Vitamin D-3, riboflavin, Vitamin B-12, niacin, meat meal, calcium pantothenate, cracke~ or milled grains such as corn, wheat, oats, barley and the like, dried molasses, dried sorghum, soybean meal, cottonseed meal, peanut meal, ; fish meal, essential amino acids such as lysine, peptides and polypeptides containing essential amino acids, casein, soya bean protein, vitamins such as Vitamins A, D, E and K, mineral nutrients such as~sodium chloride, ferrous salts, magnesium sulfate and calcium salts, proteins, buffers, dextrose, sucrose, lactose, maltose, corn syrup solids, hydrolized cereal solids, hay, etc.
The following is a typical or standard vitamin premix that is used in a complete poultry feed:
gm/100 lb.
Manganese sulfate (feed grade)11.4 Vitamin A oil (10,000 I.U./gm)22.7 Dry Vitamin D-3(1,6550~I.C.U./gm) 22.7 Riboflavin 0.15 Vitamin B-12 (3 mg. lb.) 45.4 Niacin 1.0 An exemplary cattle feed is 73 percent rolled shelled 105~91S
corn, 20 percent ground corn cobs, and a supplement containing soybean meal, alfalfa meal, cane molasses, urea, salt, dicalcium phosphate, Vitamin A concentrate and Vitamin D concentrate.
The greatest growth stimulation is achieved in young, maturing non-human animals or poultry, although excellent growth stimulation is achieved in grown mature non-human animals or poultry.
An important feature of this invention is that there is little or no residue of the gentian violet in the tissue of the non-human animals or poultry fed the com-positions of this invention--this is apparently so due, in part, to the levels of gentian violet used in the compositions of this invention. This means that there - is no toxicological danger to animals or humans from ingestion of the edible tissues of poultry or non-human animals which have been fed the compositions of this invention.
The concentrations ranges in the non-human animal and poultry feeds of this invention are important re efficacy, growth stimulation and toxicity in poultry or non-human animals. If amounts lower than ranges of~this invention are used, there is little or no efficacy in inhibiting Candida albicans in poultry or non-human animals and -little or no growth stimulation of non-human animals or poultry. If an amount of gentian violet is used which is higher than the range (in feed) of this invention then toxic effects in the poultry or non-human animals can be . encountered. The toxic effect can kill the poultry or non-human animals, or cause a decrease in the weight gain of the non-human animals and poultry, which represents a ~: .
serious economic loss to commercial poultry or non-human animal raisers (and loses the advantage of the gentian violet being a growth promoter). The residue build up is a potential health hazard when such higher levels are used.
The growth promoter compositions of this invention can be used to simultaneously treat non-humans and/or poultry infected with Candida albicans., This results in the curing of the candidiasis infection of the non-human animals and/or poultry and a simultaneous stimulation of their growth. The use of the growth promoter compositions of this invention also results in non-infected non-human animals and/or poultry from becoming infected with Candida albicans with a simultaneous stimulation of their growth.
Candida albicans infection in non-human animals or poultry has a growth suppressing nature, but feed con-taining 7 to 35 p.p.m. of gentian violet stimulates non-human animal or poultry growth even in the face of such active infections.
Gentian violet inhibits the growth of and destroys Candida albicans without materially reducing the growth of and activity o$ the normal coliform bacteria present in the intestinal tract, which are necessary in syn`thesizing important nutritional factors.
Another name for Candida albicans is Monila albicans.
Candida albicans, a yeast-like organism, is the usual cause of moniliasis pheumonia, meningitis and other forms of moniliasis. It is normally saprophytic but may become pathogenic after the administration of certain antibiotics.
Broad-spectrum antibiotics are often used in relatively large amounts in animal and poultry feeds. The use of the antibiotics allow molds and fungi to grow. The above processes of this invention are useful in eliminating and preventing internal infestation of Candida albicans, for example, in the intestines and infestation of Candida albicans in drinking water.
Another embodiment of this invention includes the complete or basal poultry or non-human animal feed which consists of 0.00077 to 0.01 percent by weight of gentian - violet and the remainder inert ingredients. This fe~d is decribed in detail herein. Another substituted benzo-phenone azoanilide dye or rosaniline base dye can be used in place of the gentian violet.
Another embodiment of this invention involves the use of gentian violet, a selective fungicidal mold inhibitor, as an additive to non-human animal or poultry feed to treat or prevent Candida albicans infections in poultry or non-human animals. The ~ c gentian violet specifica~ly is a selective ungicidal mold inhibitor of Candida albicans Another 3ubstituted benzophenone azoanilide dye or rosaniline dye can be used in place of the gentian violet. -The gentian violet is preferably used in the form of a pre-mix concentrate, which can be added to the complete feed as needed.
The disclosure elsewhere herein concerning the premix is applicable to this embodiment of this invention.
The premix concentrate is used for treatment of Candida ~0 albicans by adding it to the complete non-human animal or poultry . . ._ feed or a period of about seven days, although the time can be more or less as needed. When the premix concentrate contains 1.55 percent o~ gentian viol~t, preferabiy t~!o pounds of the premix is used per ton of complete poultry or non-human animal feed. That reflects a preferred concentration of about 0.00155 percent of gentian violet in the complete non-human animal or poultry feed ; Effective concentrations of gentian violet for treatment are from 0.00077 to 0.01 percent of gentian violet per ton of cornplete non-human animal or poultry feed.
ao The premix is used for prevention of Candi~ ~lbi~n~ by adding it to the complete non-human animal or poultry feed on a continuous basis. (After treatment with the premix, it should be used on a continuous basis to achieve prevention.) When the pre-mix sontains 1.55 percent of gentian vio]et, preferably one pound of the premix is used per ton of complete poultry or non-human ` 11 .
~c animal feed. That reflects a preferred concentration of about 0.00~77 percent of ~entian violet in the complete non-human animal or poultxy feed. Effective concentrati~ns o~ gentian violet for prevention are from 0.00077 to 0.005 percent of yentian violet per ton of complete non-human animal or poultry feed.
The above treatment and prevention times and concentrations in the complete poultry or non-human animal feed apply even when the gentian vio~et is directly admixed with the complete poultry or non-human animal feed without going through the intermediate ~0 premix concentrate form.
Any basal or complete non human animal or poultry feed or basal poultry feed can be~used. The disclosure elsewhere herein concerning the premi~ is applicable to 1this embodiment of this invention.
A further aspect o this invention involves the use of gentia violet, a selective fungicidal mold inhibitor, as an additive to poultry or non-human animal water.
The gentian violet is preferably used in the form of a liquid premix concen'rate, which can be added to the poultry or non-human animal water as needed or made into a sbDck solution and added as needed. The most preferred concentrate contains water and 1.55 percent of gentian violet, but the concentration usually ranges from O.l to 10 percent, although any concentration can be used.
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~The k2y 8 to get a suf~iciently high concentration in the non- ¦
human animal or poultry water itsel. The concentration of gen-tian violet in the poultry or non-human animal water itself should be between 0.001 and 0.1 percent. ' The treatment period is preferably seven days but cn be any length of time needed to achieve the desired effect.
' The gentian violet can be added directly to the'''nOn-hull:an animal or poultry water, in which case the above treatment times and concentrations also apply.
Another a~pect of this invention involves the use of gentian violet, a selective fungicidal mold inhibitor, as a non-human animal or poultry wate sanitizer. It aids in the reduction of Cc~ndida albicans contamination in water linos and poultry or non-human ani~al watering equip~ent.
The gentian violet is preferably used in the form of a liquid premix concentra~e, which can be added to the~ poultry or non'-human animal water on a regulax basis, for example, once a week. It can also be made up into a stock solution and added to the poultry or non-hu~an ani~al water The most preferred concentrate conta;ns 8Q water and 0,387~ percent of gentian violet, but the concentration usually ranges from 0.05 to S percent The concentration of gentlan violet in the poultry water itself should be between 0.001 and 0.0 percent, ' ' -13-.' ' ':, ' , .
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. ' ' 1~. 1 The treatment period is preferably one day every week but can be on any basis as needed to achieve the desired effect, The gentian violet can be added directly to the non-human animal or poultry water, in which case the above treatment times and concentrations also apply, .
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DETAILED DECRIPTION OF THIS INVENTION
As used herein, the term "poultry" means domestic fowls including chickens, ducks, turkeys, geese, etc.
As used herein, the term "non-human animal" includes domestic pigs, other swine, cattle, sheep, goats, rabbits, etc.
As used herein, the term "gentian violet" is hexa-- methylpararosaniline chloride or a mixture of at least 96 percent of hexamethylpararosaniline chloride with minor portions of pentamethylpararosaniline chloride and/or tetramethylpararosaniline chloride, and meeting all spec-ifications of quality for USP grade as listed in the United States Pharmacopeia XIV.
Gentian violet is a dark green powder or greenish, glistening pieces with a metallic luster. Gentian violet is soluble in water, chloroform and ethanol. Other names for gentian violet are methylrosaniline chloride and crystal violet.
Gentian violet, both as a contaminant in water and as a residue in poultry manure, is biodegradable in the presence of ultra-violet rays of sunlight.
The use of wide-spectrum bacteriostatic or bacterio-cidal agents, such as, sodium propionate, in con~unction with the gentian violet would have an adverse effect on -the growth stimulation of non-human animals or poultry.
There is a natural balance between mold entities and bacterial entities in the gut of poultry or non-human animals. When this balance is disturbed by the use ! ~ .
~05~915 of wide-spectrum antibacterial druys, the mold entities are allowed to pro.iferate tc the poultry's or non-human animal's detriment.
The wide-spectrum antibacterial druy has in effect reduced a natural (in v-vo) barrier in the poultry or non-human animal against the proliferation of molds, particularly Candida albicans.
Sodium propionate is a~ antibacterial drug, whose use would allow the growth o Candida alb;.cans by destroying the natural balance in the gut of poultr~ or non-human animals.
Experime~ts have shown that gentian violet is effective in .
treating a pre-existing Candida _lbicans infection when the gentian violet is used in poultry or non-human animal feed at certain levels. Sodium propionate, at the same levels, was not a good mold .
inhibitor, and calcium propionate, at the same levels, was even less effective than the sodium propionate, The gentian violet levels o~ ~his invention lowered the degree of mold growth ~Candid~.
albicans) in in~ected chicks to the.degree of mold growth in non-infected, non~medicated chicks, A~!so, the sodium and calcium propionate each caused a decrease in the weigh$ gain of the chicks, which represents a serious economic loss to commercial poultry raisers ~alons wi~h no Candids albicans ~nhibition).
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I 105~915 ! ~
I ~erein all parts, ratios, proportions and percentages are on a wo,ght basis, unless otherwise state~ or obvious to one ordinaril ll skilled in the art. .
¦ The following examples illustrate this invention.
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L~ 160 sexed, day-old broilers chickens (Hubbard~ x Hubbard) were randomly divided into eight groups (four male and four female) i o 20 chickens each. The chickens were hatch r~n and all from the same breeder flocks. All of the chickens were wing banded and ~'0 debeaked at one day of age, All of the chickens were ~D vaccinated on day 1 and were ND-IB vaccinated as'indicated below The feed was prepared by preparing 1,600 pounds of a 25 per-I cent prot2in broiler mash. The 1,600 pounds of feed were divided into four equal batches of 400 lbs. each and the ViGen premix added to three of the batches according to th'e following table:
Control level0 ppm400 lbs. no ViGen premlx added I 1 x use level- 7 ppm400 lbs. 9~.8 gm of ~7igca pre ' ' mix added ' 2 x use level14 ppm400 lbs. 181 6 gm of 7~ e~ 'pre-mix added ~0 5 x use level35 ppm400 lbs. 454 gm of ViGen premix . added 1,l ,' . . ' .
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-1059~115 , The Vigen premix contains l.SS percent of gentian violet, 1 per-cent of vegetable oil, 1.65 percent o~ Micro-Cel E and 91.~ percent ~ ~f corn cob fractions and 4% water and iæ manufactured by A.H.P., ¦ Inc., Gainesville, Ga., U.S.A. Each 400 lb. batch of feed was di-vided into ? lb, lots and placed in the designated houses. -A
¦ ,sample (1. lb) from each~feed lot was set aside for ~-uture testing purposes, Feed remaining in each house at termlnation of thë
trial (day 56) was weighed to determine the feed consumed by each qroup of birds. The treatment levels for the chickens in the ~0 houses were as follows:
House 1- 14 p.p.m. of ViGen premix - male ' House 2- 14 p.p.m. of ViGen premix - female House 3~ 35 p.p.m. of ViGen premix - male House 4- 35 p.p.m. of ViGen premix - female House 5- 7 p.p.m. of ViGen premix - male House 6- 7 p.p.m. of ViGen premix - female House 7~ 0 p.p.m. of ViGen premix - control - male ,House 8- 0 p.p.m. of Vigen premix - control - female This allows testing of male and female chickens at the following ~0 tre~tment levels:
O ppm - unmedicated controls 7 ppm - 1 x normal use level 14 ppm - 2 x normal use level 35 ppm - 5 x normal use level The test was conducted for 56 days. The parameters measured during the test were: (1) weight gains - all o the qroups vs. the ~ ' controls - by pen at the conclusion of the trial: and (2) eed ! conversion - all of the groups vs. the controls - by pen at the l conclu~ion o the trial.
Durina the ~est, ~ ntinued observation was made of mortalLty, feed consumption, diarrhea, etc, On the tenth and thirty second day after the start of the test, the chickens were vaccinated I ND-IB by treatment with Broilerbron spray. The chickens were weighed on the twenty eighth day. The chickens and the remaining feed were weighed on the fifty sixth day, The question of whether or not the gentian violet (ViGen premix) in the poultry feed was a growth promoter for the poultry was determined by examining the weight ~ain patterns of the groups ~0 of broiler chic~ens fed different amounts of ViGen premix during the growing period. The ~uestion of whether or not the amount; of gentian violet in the poultry feed was toxic for the poultry was dete mined ~y examining the weight gain patterns and death rates o~ groups of br~oiler chickens fed different amounts of ViGen during the growing period, The parameters dealt with to determine such questions were feed conversion ratios, weight (days 1, 28 and 56), and death rate. The feed conversion ratios wer~ determined by di*iding the kilograms of feed consumed by the total weight of broilers produced (gained) in each group. The mean, ~variance ~0 stat~dard error of the mean and 9S percent confidence interval were calculated from the data Additionally, a chi-square test was calculated to determine if any feed conversion ratio was signifi-cantly different. The weight data were examined utilizing an analysis of variance (one-way) for each group for each tirne period.
., ' , The death rate data was analyzed using a chi-square test (one-way analysis of variance).
The mean feed conversion ratio was 2.3110. A 95 percent confidence interval was calculated around this mean and was determined to have a lower limit of 2.2486 and an upper limit of 2.3734. Because two observations (males 14 ppm - 2.1960 and males 35 ppm - 2.2349) did not fit within the upper and lower limits, further calculation was done. The standard deviation was calculated to be 0.0787itherefor, two standard deviations are 0.1574.
Subtracting this ligure from the mean indicated that the feed conversion ratios of both of these groups were less than two standard deviations away from the mean and therefore are not significantly di,fferent from the other feed conversion ratios. A chi-square test confirmed this observation.
The death pattern of the groups of chickens were examined using a chi-square one-way analysis of variance.
The results demonstrated that no significant difference existed in the death rate of the groups under test.
The male group of chickens receiving 35 ppm ViGen premix feed was significantly heavier than any other group. The female groups receiving 7 and 14 ppm ViGen premix feed were significantly heavier than the female control group. This shows that the use of gentian violet (in the amounts of this invention) in poultry feed results in growth promotion or stimulation of poultry - gentian violet is a growth promoter for poultry.
To answer the question of whether gentian violet in poultry feed was a toxic agent to poultry three parameters were examined. These parameters were feed conversion ratios, weight gains, and deaths. In no case was there any indication that significant differences existed which were detrimental. In fact, the opposite was true. For the reasons expressed above, gentian violet in poultry feed in the amounts claimed in this invention does not have any effect which could be considered toxic to poultry.
Gentian violet, in the amounts of this invention, have a strong stimulatory or promotive effect on the weight ! growth or production of chickens. This example shows that such amounts of orally ingested gentian violet (e.g., carried in inert carriers) in a poultry feed is a good growth promotor for poultry, such as, chickens.
; At day 56, the average weight of the control male chickens was 2.000 kg. and the average weight of the 35 ppm male chickens was 2.132 kg. The average weight difference from the control is very significant. At day 56, the average weight of the control female chickens (hens) was 1.657 kg., the average weight of the 7 ppm ; female chickens was 1.740 kg and the average weight of the 14 ppm female chickens was 1.747 kg. The average weight differences from the control is very significant.
This example establishes that gentian violet, at the levels of this invention, in poultry feed is non-toxic to poultry and is a growth promoter for the poultry.
720 sexed, day-old broiler chickens (Hubbard x Hubbard) were assigned to pens according to the following chart. All chicks in all groups were MD vaccinated and - wing-banded for positive identification. Chicks were weight averaged by sex and the top and bottom weights (approximately 10%) discarded to provide maximum weight uniformly in the groups.
Pen 1 - 60 males Pen 2 - 60 females Pen 3 - 60 males 60 females Pen 4 - 60 males 60 females Pen 5 - 60 males 60 females Pen 6 - 60 males 60 females Pen 7 - 60 males Pen 8 - 60 females (The chicks were debeaked at one day of age, and were obtained from a single breeder flock. Sufficient chicks were obtained to allow for weight averaging of the groups.).
The toxicity study which is part of Example 2 may not have provided completely accurate toxicity data due to excessive mortality and undiagnosed illness in one group of the birds receiving 7 ppm ViGen premix feed. However, it was noted that the groups receiving 35 ppm (5 X treat-ment level) showed no evidence of a toxic effect. Weight gain was equal to and often greater than nonmedicated controls. Droppings remained normal in appearance and consistency as well as general condition. It was apparent that 35 ppm was not a toxic level. When repeated at 0, 7, 14 and 35 ppm (in Example 1), there was again no evidence of toxic symptoms or change in droppings at the higher levels. The average weights were comparable and feed consumption per bird was the same (35 ppm) or less (7 and 14 ppm) than the controls. Necropsy of 4 male and 4 female birds from each dosage level at 56 days showed no gross differences in intestines or abdominal organs.
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Example 1 conirmed that as much as 35 ppm ViGen premix ' in the feed was not a toxic level.
' ' EX~PLE 3 '- -. \ ~, Chickens infected with Ca,ndida a~bic~ were fed, for eight weeks, a complete-poultr'y feed containing two pounds of premix concentrate per ton o a 20 percent protein poultry starter feed. The premix concentrate contained 1 percent of white mineral oil, 1.65 percent of Micro-Cel E, 1.55 percent of gentian violet, 91,8 percent of corn cob fractions 'and 4 percent of water. The ~1~ complete poultry feed contained~
, ' , ' percent ' by weight, Ingredients weight pounds Gro~md yellow corn ' 62.50 1250 Soybean oil meal (44V/o protein) , 30.00 600 Meat scraP (50% protein-) 2.00 40 M~nhaden fish meal (60% protei'n) 2.00 40 Dica].cium phosphate , . ' 1.90 3~
Calcium carbonc~e 1.00 20 ~0 Standard vitamin premix 0.50 10 Gen~ian violet premix concentrate 0.01 2 ' Total......... 100.00 ~b~' . . ' ''''" . ' ,' . .
, The!~aL~lida ~lb~Q~ infection was successfully treated and the gro~th of the poultry was significantly stimulated.' , EXAMPLE 4 . . : ' , ' .
Chickens not infected with',~ ~h~c ~ were fed,for eight weeks, a complete poultry feed containing one pound of th~
. gcntian violet premix concentrate described in Example 3 per ton ,~, of complete poultry feed (described in Example 3, except that it ~0 con~ained 39 pounds of dicalciu~.n phosphate). There ~as no occ~ur~
c~lce of ~ ,]h~ infectioll cluring the treatment pcriod and the grow~ll of the poul~ry was significantly stilnulate~.
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-i EX~MPLE 5 A ~eries of trials were conducted to evaluate the efficiency of gentian violet on an artifically induced outbreak of candidiasis in broilers, This organism was obtained from a clinical case of crop mycosis; grown in liquld Sabaurads media and diluted to con-tain one million organisms per dose Crop swabs were taken when the birds were 7 days, 14 days, and 21 days~of age and grown on ~ Sabaurads media. Groups o the broiler were fed at levels of l i pound of gentian violet per ton of feed, 2 pounds of gentian violet per ton of feed, and 5 pounds of gentian violet per ton of feed Controls were run against other proaucts and against no treatment. (The other products were copper suifate, Mycostatin, Mold Curb, sodium propionate and calcium propionate.) The birds fed gentian violet had a marked decrease of Candida recovery when compared to the no treatment controls. The birds started on the gentian violet containing feed had the lowest incidence of Candida recovery compared to the controls using the other products and had weights on about the same level as some non-infected, non-treatea controls (although there was a slight weight dcprPssion at the ; five pound per ton level), ~. ~XAMPLE 6 ;~ Hogs infected with Candida albicans were fed, for seven days, a complete hog feed containing two pounds of premix concentrate per ton of completc hog feed The premix contained 1.0 percent of .. . . .. . . . . . . .. .. . . . . . . . . . .... ... . .. ..
1~5991S
vegetable oil, 4.45 percent of Micro-Cel E, 1.55 percent of gentian violet and 94 percent of corn cob fractions.
The complete hog feed was the same as the one used in Example 3. The Candida albicans infection was success-fully treated.
The hogs of Example 6 were fed thereafter a complete hog feed containing one pound oÇ the premix concentrate (described in Example 6) per ton of complete hog feed (described in Example 6). There was no occurrence of the Candida albicans infection during the treatment period.
240 ml. of a premix concentrate containing water and 1.55 percent of gentian violet were diluted to make one gallon of stock solution. The gallon of stock solution and a number of gallons of water were metered through a proportioner, at a ratio of one ounce of stock solution per gallon of water, to poultry drinking trays as needed over a seven-day period.
240 ml. of a premix concentrate containing water and : 0.3875 percent gentian violet were diluted to make one gallon of stock solution. The gallon of stock solution and a number of gallons of water were metered through a proportioner, at a ratio of one ounce of stock solution per gallon of water delivered. The gentian violet addition was one day every week.
Cattle infected with Candida albicans were fed, for seven days, a complete cattle feed containing two pounds of premix concentrate per ton of complete cattle feed.
The premix contained 1.0 percent of vegetable oil, 4.45 percent of Micro-Cel E, 1.55 percent of gentian violet and 93 percent of corn cob fractions. The Candida albicans infection was successfully treated.
The cattle of Example 10 were fed thereafter a complete cattle feed containing one pound of the premix concentrate (described in Example 10) per ton of complete cattle feed. There was no occurrence of the Candida albicans infection during the treatment period.
Turkeys infected with Candida albicans were fed, for seven days, a complete turkey feed containing two pounds of premix concentrate per ton of complete turkey feed.
The premix contained 1.0 percent of vegetable oil, 1.65 percent of microcel E, 1.55 percent of gentian violet, 91.8 percent of corn cob fractions and 4%~water. The Candida albicans infection was successfully t-reated.
The turkeys of Example 12 were fed thereafter a complete turkey feed containing one pound of the premix concentrate (described in Example 12) per ton of completè
turkey feed. There was no occurrence of the Candida albicans infection during the treatment period.
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A ViGen premix ormulation was prepared which had the follot~ing components: 1.55 percent of ViGen (gentian violet from A.H.P. Inc.); l percent of white mineral oil, NF-70 Food Grade, from Standard Oil Co., 1.65 percent of ~icro-Cel E, synthetic cal-cium silicate Code 21-E,'from the Celite Division of Johns-Manvill~
Products Co.; 91.8 percent of Grit-O-Cob, Grade 2040 corn cob ractions, from Anderson Cob Mills, Inc. and 4 percent of distille water. The corn cobs and calcium silicate are inert carriers;
white oil reduces the dust in the premi,x and distilled water is th solvent for the gentian violet ~or adsorption o' the gentian viole onto the inert carriers. The ViGen premix formulation was found to be stable or over one year. ' ' In al~ of the above pertinent examples, other ingredient such as cereal grains and grain by-products, can be substituted in part for the ground yellow corn i.n these rations as the market price of feed ingredients 1uctuates since all poultry'feeds are formulated on a least-cost-basis, ~ rK
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lOS99~5 EXAMPJ,E 15 Several experiments were conducted to determine the efficiency of gen$ian violet on candidia~is in broiler chickens. The experi-ments or study had a 56-day duraction. Broadly, the experiments involved reproducing candidiasis in broiler chickens by challenge per os with a dose of approximately one million Candida organisms ¦ per bird and then treatment o some of the broiler chickens to determine the efficiency of gentian violet against the artifically induced outbreak of c~ndidiasis.
'~l~ The gentian violet was used in the form of ViGen premix.
About 6000 lbs. of 25 percent protein broiler feed obtain~d rom Kreamer Co., Kraemer, Pa. (this feed was fed throughout the 56-day eeding pexiod) was ~elleted and crumbled. 2,998.5 lbs.
of the fecd containing 7 ppm (1.516 lb~.) of ViGen premix were admixed--this was termed V feed. 3,000 lbs. of the feed was used for the controls, and was termed C-feed.
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The following parameters were measured during the experiments:
1. Weights - reported in grams.
a. Day one - chicks were weight averaged with top and bottom weights (approximately 10%) discarded to give maximum sex weighted average by group. Weight of each chick in each group was recorded.
b. Day 28 - weighed all birds in all groups, and reported weiqht in grams.
c. Day 56 - weighed all birds in all groups, and reported weight in grams.
animals and/or poultry. A further object of this invention is to provide a proce.ss for using such growth promotion'compositiorls Ahother object of this invention is to provide animal and poultry ,feeds, poultry drinking water and poultry water sa~itizer that ,20 achieve the above process ob}ectives; Other objects and advan-tages are set out herein or are obvious'to one ordinarily skilled in the art herefrom.
Such objects and inventions are achieved by this i.nvent:ion.
~OS99~5 One embodiment of this invention is a method for stimulating growth in non-h~man animals and/or poultry.
The method includes feeding a complete feed to the non-human animal and/or poultry. The complete feed consists only of (i) gentian violet and (ii) the remainder inert ingredients. The inert ingredients include all of the inert ingredients normally found in a complete or basal non-human animal poultry feed. The gentian violet is present in the feed in an amount between 0.00077 and 0.01 percent by weight, based on the weight of the complete feed. The growth of the non-human animal or poultry is increased at a rate faster than if the non-human animal or poultry had not been fed the complete feed containing the gentian violet. The gentian violet is a growth promoter for said non-human animal or poultry.
Consumption by non-human animals and poultry of the gentian violet composition of this invention in the stated levels causes significant growth increase. The gentian violet is in effect a growth factor, i.e., is a substance which promotes growth in the poultry and non-human animals, particularly the young and immature ones.
For the best effects the feed composition of this invention is fed to poultry, hogs or cattle.
Preferably the preparation of the feed involves pre-paring a premix concentrate for addition to the feed which consists only of between 0.1 and 10 percent by weight of gentian violet and the remainder inert ingredients and thén adding enough of the premix concentrate to the other ingredients of feed to obtain the recited amount of gentian violet in the feed. Preferably inert ingredients used in the premix concentrate are diatomaceous silica, ` \
corn cob fractions, and vegetable oil. Most preferably, the feed contains 0.00156 percent by weight of gentian violet, based on the weight of the complete feed.
Other substituted benzophenone azoanilide dyes or rosaniline base dyes can be used in place of the gentian violet. Such are also known as triaminotritane dyes related to rosaniline. The -N(CH3)2 groups of gentian violet can be replaced by other auxochrome groups such as -OH, -NH2 -N(C2H5)2, etc. The other various ,, substituents, and the substituents on gentian violet~ can be located at any position on the three benzyl rings.
The other dyes must have at least one amino chloride group and/or substituted amino chloride group.
Examples of such other substituted benzophenone azoanilide dyes are: pentamethylpararosaniline chloride, tetramethylpararosaniline~chloride; methyl green or light green, which is the methyl chloridé add~ition product of gentian violet; ethyl green, which is the ethyl chloride addition product of gentian violet; sulfonic acids of the rosanilines, such as, water blue and patent blue, phenyl-ated rosanilines, such as, triphenyl fuch~sin, aniline blue, diphenylamine blue and spirit blue; pararosaniiine;
and mixtures of pararosaniline with its methyl homologs, such as fuchsins and magentas. More broadly, any tri-phenylmethane dye that contains an amino group or substituted amino group can be used in place of ~ 1059915 gentian violet. Examples o such tripbenylmethane dyes are the malachite green seri.es of dyes, such as, malachite green.
Most o~ such aniline dyes are rather toxic, but can be readily used if they are buffered by any generally non-toxic conventional bueren~ system to reduce their toxicity. So when such aniline dyes (other than genti.an violet) are used, the feed compositi.ons of this invention incl.ude a bufering a~ent in the inert ingredi-ents The term inert in~redients excludes therapeutically and pharmaceutically active ingredients ~e g., broad spectrum baci-~-riostatic or bacteriocidal ingredients), but does not exclude vitamins and minerals.
The gentian violet is preferably used in the form of a premix concerltrate, which can ~ dded t-o the con.plete feed as needed.
The most preferred prem.ix concentrate contai.ns corn cob ~ractions, 7r~-J~ ~t1a~1~
white mineral oil, diatomaceous silica (e.g., Micro-Cel ~j~ dis tilled water and 1.55 percent of gentian violet.
rrhe concent:ration of gentian violet in the prerni.x concentrate can easily be as high as 60 percent; it is preferably between 0 1 and 10 percent and is most preferably 1.55 percent.
The prelnix concentrate can contain any materlal that is used in a complete or basal non-human animal or poultry eed. The premix concentrate can be used in the form of a liquid or solid admixture~ The liqllid is best in the form of a ver~ viscous susrension or ~i,nilar ~emi-~luid. Preferclbly a solid premix concen~rate is used even thou~h it may contain a liquid material _5-. .
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such as white mineral oil which reduces the dust factor in the premix.
... The premix concentrate is used by addin~ it to the complete poultry or non-human feed as long as growth stimulation is wanted.
When the premix concentrate contains 1.55 percent of gentian ~iole~
preferably two pounds of the premix is used per ton of complete poultry feed for growth stimulation of non-human animals or poultry--this reflects a preferred concentrat~on of about 0.00155 percent of gentian violet in the complete poultry or non-human animal feed. Effective concentrations of gentian violet are within the range of 0.00077 to 0.005 percent of gentian violet per ton of complete poultry feed or non-human animal feed. The preferred concentrate ranqe of gentian violet is from 0.00085 to 0.01 per-: cent of gentian violet per ton of complete poultry or non-human animal feed. It has been found that most female poultry preferably are fed at a gentian violet level of 0.00155 perce.nt per ton of complete poultry.feed, and male poultry are fed at a gentian violet eve~ of 0.00385 percent per ton of complete poultry feed.
. The above concentrations in the complete poultry feed apply even when the gentian violet is directly admixed with the complete .
poultry.or non-human animal feed without going through the ilite~-mediate premix concentrate form. .
The premix is used for growth stimulation by adding i.t to t}-e complete feed on a continuou~ basis.
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Any complete poultry feed or basal poultry feed can be used. It can contain, for example, ground yellow corn, soybean oil meal, steamed bone meal, ground lime-stone, iodized salt, manganese sulfate, Vitamin A oil, dry Vitamin D-3, riboflavin, Vitamin B-12 and niacin. It can also contain, for example, fish meal and meat meal.
Any complete non-human animal feed/or basal non-human animal feed can be used; it can contain, for example, any of the following ingredients: mogul starch, iodized salt, dry Vitamin D-3, riboflavin, Vitamin B-12, niacin, meat meal, calcium pantothenate, cracke~ or milled grains such as corn, wheat, oats, barley and the like, dried molasses, dried sorghum, soybean meal, cottonseed meal, peanut meal, ; fish meal, essential amino acids such as lysine, peptides and polypeptides containing essential amino acids, casein, soya bean protein, vitamins such as Vitamins A, D, E and K, mineral nutrients such as~sodium chloride, ferrous salts, magnesium sulfate and calcium salts, proteins, buffers, dextrose, sucrose, lactose, maltose, corn syrup solids, hydrolized cereal solids, hay, etc.
The following is a typical or standard vitamin premix that is used in a complete poultry feed:
gm/100 lb.
Manganese sulfate (feed grade)11.4 Vitamin A oil (10,000 I.U./gm)22.7 Dry Vitamin D-3(1,6550~I.C.U./gm) 22.7 Riboflavin 0.15 Vitamin B-12 (3 mg. lb.) 45.4 Niacin 1.0 An exemplary cattle feed is 73 percent rolled shelled 105~91S
corn, 20 percent ground corn cobs, and a supplement containing soybean meal, alfalfa meal, cane molasses, urea, salt, dicalcium phosphate, Vitamin A concentrate and Vitamin D concentrate.
The greatest growth stimulation is achieved in young, maturing non-human animals or poultry, although excellent growth stimulation is achieved in grown mature non-human animals or poultry.
An important feature of this invention is that there is little or no residue of the gentian violet in the tissue of the non-human animals or poultry fed the com-positions of this invention--this is apparently so due, in part, to the levels of gentian violet used in the compositions of this invention. This means that there - is no toxicological danger to animals or humans from ingestion of the edible tissues of poultry or non-human animals which have been fed the compositions of this invention.
The concentrations ranges in the non-human animal and poultry feeds of this invention are important re efficacy, growth stimulation and toxicity in poultry or non-human animals. If amounts lower than ranges of~this invention are used, there is little or no efficacy in inhibiting Candida albicans in poultry or non-human animals and -little or no growth stimulation of non-human animals or poultry. If an amount of gentian violet is used which is higher than the range (in feed) of this invention then toxic effects in the poultry or non-human animals can be . encountered. The toxic effect can kill the poultry or non-human animals, or cause a decrease in the weight gain of the non-human animals and poultry, which represents a ~: .
serious economic loss to commercial poultry or non-human animal raisers (and loses the advantage of the gentian violet being a growth promoter). The residue build up is a potential health hazard when such higher levels are used.
The growth promoter compositions of this invention can be used to simultaneously treat non-humans and/or poultry infected with Candida albicans., This results in the curing of the candidiasis infection of the non-human animals and/or poultry and a simultaneous stimulation of their growth. The use of the growth promoter compositions of this invention also results in non-infected non-human animals and/or poultry from becoming infected with Candida albicans with a simultaneous stimulation of their growth.
Candida albicans infection in non-human animals or poultry has a growth suppressing nature, but feed con-taining 7 to 35 p.p.m. of gentian violet stimulates non-human animal or poultry growth even in the face of such active infections.
Gentian violet inhibits the growth of and destroys Candida albicans without materially reducing the growth of and activity o$ the normal coliform bacteria present in the intestinal tract, which are necessary in syn`thesizing important nutritional factors.
Another name for Candida albicans is Monila albicans.
Candida albicans, a yeast-like organism, is the usual cause of moniliasis pheumonia, meningitis and other forms of moniliasis. It is normally saprophytic but may become pathogenic after the administration of certain antibiotics.
Broad-spectrum antibiotics are often used in relatively large amounts in animal and poultry feeds. The use of the antibiotics allow molds and fungi to grow. The above processes of this invention are useful in eliminating and preventing internal infestation of Candida albicans, for example, in the intestines and infestation of Candida albicans in drinking water.
Another embodiment of this invention includes the complete or basal poultry or non-human animal feed which consists of 0.00077 to 0.01 percent by weight of gentian - violet and the remainder inert ingredients. This fe~d is decribed in detail herein. Another substituted benzo-phenone azoanilide dye or rosaniline base dye can be used in place of the gentian violet.
Another embodiment of this invention involves the use of gentian violet, a selective fungicidal mold inhibitor, as an additive to non-human animal or poultry feed to treat or prevent Candida albicans infections in poultry or non-human animals. The ~ c gentian violet specifica~ly is a selective ungicidal mold inhibitor of Candida albicans Another 3ubstituted benzophenone azoanilide dye or rosaniline dye can be used in place of the gentian violet. -The gentian violet is preferably used in the form of a pre-mix concentrate, which can be added to the complete feed as needed.
The disclosure elsewhere herein concerning the premix is applicable to this embodiment of this invention.
The premix concentrate is used for treatment of Candida ~0 albicans by adding it to the complete non-human animal or poultry . . ._ feed or a period of about seven days, although the time can be more or less as needed. When the premix concentrate contains 1.55 percent o~ gentian viol~t, preferabiy t~!o pounds of the premix is used per ton of complete poultry or non-human animal feed. That reflects a preferred concentration of about 0.00155 percent of gentian violet in the complete non-human animal or poultry feed ; Effective concentrations of gentian violet for treatment are from 0.00077 to 0.01 percent of gentian violet per ton of cornplete non-human animal or poultry feed.
ao The premix is used for prevention of Candi~ ~lbi~n~ by adding it to the complete non-human animal or poultry feed on a continuous basis. (After treatment with the premix, it should be used on a continuous basis to achieve prevention.) When the pre-mix sontains 1.55 percent of gentian vio]et, preferably one pound of the premix is used per ton of complete poultry or non-human ` 11 .
~c animal feed. That reflects a preferred concentration of about 0.00~77 percent of ~entian violet in the complete non-human animal or poultxy feed. Effective concentrati~ns o~ gentian violet for prevention are from 0.00077 to 0.005 percent of yentian violet per ton of complete non-human animal or poultry feed.
The above treatment and prevention times and concentrations in the complete poultry or non-human animal feed apply even when the gentian vio~et is directly admixed with the complete poultry or non-human animal feed without going through the intermediate ~0 premix concentrate form.
Any basal or complete non human animal or poultry feed or basal poultry feed can be~used. The disclosure elsewhere herein concerning the premi~ is applicable to 1this embodiment of this invention.
A further aspect o this invention involves the use of gentia violet, a selective fungicidal mold inhibitor, as an additive to poultry or non-human animal water.
The gentian violet is preferably used in the form of a liquid premix concen'rate, which can be added to the poultry or non-human animal water as needed or made into a sbDck solution and added as needed. The most preferred concentrate contains water and 1.55 percent of gentian violet, but the concentration usually ranges from O.l to 10 percent, although any concentration can be used.
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~The k2y 8 to get a suf~iciently high concentration in the non- ¦
human animal or poultry water itsel. The concentration of gen-tian violet in the poultry or non-human animal water itself should be between 0.001 and 0.1 percent. ' The treatment period is preferably seven days but cn be any length of time needed to achieve the desired effect.
' The gentian violet can be added directly to the'''nOn-hull:an animal or poultry water, in which case the above treatment times and concentrations also apply.
Another a~pect of this invention involves the use of gentian violet, a selective fungicidal mold inhibitor, as a non-human animal or poultry wate sanitizer. It aids in the reduction of Cc~ndida albicans contamination in water linos and poultry or non-human ani~al watering equip~ent.
The gentian violet is preferably used in the form of a liquid premix concentra~e, which can be added to the~ poultry or non'-human animal water on a regulax basis, for example, once a week. It can also be made up into a stock solution and added to the poultry or non-hu~an ani~al water The most preferred concentrate conta;ns 8Q water and 0,387~ percent of gentian violet, but the concentration usually ranges from 0.05 to S percent The concentration of gentlan violet in the poultry water itself should be between 0.001 and 0.0 percent, ' ' -13-.' ' ':, ' , .
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. ' ' 1~. 1 The treatment period is preferably one day every week but can be on any basis as needed to achieve the desired effect, The gentian violet can be added directly to the non-human animal or poultry water, in which case the above treatment times and concentrations also apply, .
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DETAILED DECRIPTION OF THIS INVENTION
As used herein, the term "poultry" means domestic fowls including chickens, ducks, turkeys, geese, etc.
As used herein, the term "non-human animal" includes domestic pigs, other swine, cattle, sheep, goats, rabbits, etc.
As used herein, the term "gentian violet" is hexa-- methylpararosaniline chloride or a mixture of at least 96 percent of hexamethylpararosaniline chloride with minor portions of pentamethylpararosaniline chloride and/or tetramethylpararosaniline chloride, and meeting all spec-ifications of quality for USP grade as listed in the United States Pharmacopeia XIV.
Gentian violet is a dark green powder or greenish, glistening pieces with a metallic luster. Gentian violet is soluble in water, chloroform and ethanol. Other names for gentian violet are methylrosaniline chloride and crystal violet.
Gentian violet, both as a contaminant in water and as a residue in poultry manure, is biodegradable in the presence of ultra-violet rays of sunlight.
The use of wide-spectrum bacteriostatic or bacterio-cidal agents, such as, sodium propionate, in con~unction with the gentian violet would have an adverse effect on -the growth stimulation of non-human animals or poultry.
There is a natural balance between mold entities and bacterial entities in the gut of poultry or non-human animals. When this balance is disturbed by the use ! ~ .
~05~915 of wide-spectrum antibacterial druys, the mold entities are allowed to pro.iferate tc the poultry's or non-human animal's detriment.
The wide-spectrum antibacterial druy has in effect reduced a natural (in v-vo) barrier in the poultry or non-human animal against the proliferation of molds, particularly Candida albicans.
Sodium propionate is a~ antibacterial drug, whose use would allow the growth o Candida alb;.cans by destroying the natural balance in the gut of poultr~ or non-human animals.
Experime~ts have shown that gentian violet is effective in .
treating a pre-existing Candida _lbicans infection when the gentian violet is used in poultry or non-human animal feed at certain levels. Sodium propionate, at the same levels, was not a good mold .
inhibitor, and calcium propionate, at the same levels, was even less effective than the sodium propionate, The gentian violet levels o~ ~his invention lowered the degree of mold growth ~Candid~.
albicans) in in~ected chicks to the.degree of mold growth in non-infected, non~medicated chicks, A~!so, the sodium and calcium propionate each caused a decrease in the weigh$ gain of the chicks, which represents a serious economic loss to commercial poultry raisers ~alons wi~h no Candids albicans ~nhibition).
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I ~erein all parts, ratios, proportions and percentages are on a wo,ght basis, unless otherwise state~ or obvious to one ordinaril ll skilled in the art. .
¦ The following examples illustrate this invention.
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L~ 160 sexed, day-old broilers chickens (Hubbard~ x Hubbard) were randomly divided into eight groups (four male and four female) i o 20 chickens each. The chickens were hatch r~n and all from the same breeder flocks. All of the chickens were wing banded and ~'0 debeaked at one day of age, All of the chickens were ~D vaccinated on day 1 and were ND-IB vaccinated as'indicated below The feed was prepared by preparing 1,600 pounds of a 25 per-I cent prot2in broiler mash. The 1,600 pounds of feed were divided into four equal batches of 400 lbs. each and the ViGen premix added to three of the batches according to th'e following table:
Control level0 ppm400 lbs. no ViGen premlx added I 1 x use level- 7 ppm400 lbs. 9~.8 gm of ~7igca pre ' ' mix added ' 2 x use level14 ppm400 lbs. 181 6 gm of 7~ e~ 'pre-mix added ~0 5 x use level35 ppm400 lbs. 454 gm of ViGen premix . added 1,l ,' . . ' .
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-1059~115 , The Vigen premix contains l.SS percent of gentian violet, 1 per-cent of vegetable oil, 1.65 percent o~ Micro-Cel E and 91.~ percent ~ ~f corn cob fractions and 4% water and iæ manufactured by A.H.P., ¦ Inc., Gainesville, Ga., U.S.A. Each 400 lb. batch of feed was di-vided into ? lb, lots and placed in the designated houses. -A
¦ ,sample (1. lb) from each~feed lot was set aside for ~-uture testing purposes, Feed remaining in each house at termlnation of thë
trial (day 56) was weighed to determine the feed consumed by each qroup of birds. The treatment levels for the chickens in the ~0 houses were as follows:
House 1- 14 p.p.m. of ViGen premix - male ' House 2- 14 p.p.m. of ViGen premix - female House 3~ 35 p.p.m. of ViGen premix - male House 4- 35 p.p.m. of ViGen premix - female House 5- 7 p.p.m. of ViGen premix - male House 6- 7 p.p.m. of ViGen premix - female House 7~ 0 p.p.m. of ViGen premix - control - male ,House 8- 0 p.p.m. of Vigen premix - control - female This allows testing of male and female chickens at the following ~0 tre~tment levels:
O ppm - unmedicated controls 7 ppm - 1 x normal use level 14 ppm - 2 x normal use level 35 ppm - 5 x normal use level The test was conducted for 56 days. The parameters measured during the test were: (1) weight gains - all o the qroups vs. the ~ ' controls - by pen at the conclusion of the trial: and (2) eed ! conversion - all of the groups vs. the controls - by pen at the l conclu~ion o the trial.
Durina the ~est, ~ ntinued observation was made of mortalLty, feed consumption, diarrhea, etc, On the tenth and thirty second day after the start of the test, the chickens were vaccinated I ND-IB by treatment with Broilerbron spray. The chickens were weighed on the twenty eighth day. The chickens and the remaining feed were weighed on the fifty sixth day, The question of whether or not the gentian violet (ViGen premix) in the poultry feed was a growth promoter for the poultry was determined by examining the weight ~ain patterns of the groups ~0 of broiler chic~ens fed different amounts of ViGen premix during the growing period. The ~uestion of whether or not the amount; of gentian violet in the poultry feed was toxic for the poultry was dete mined ~y examining the weight gain patterns and death rates o~ groups of br~oiler chickens fed different amounts of ViGen during the growing period, The parameters dealt with to determine such questions were feed conversion ratios, weight (days 1, 28 and 56), and death rate. The feed conversion ratios wer~ determined by di*iding the kilograms of feed consumed by the total weight of broilers produced (gained) in each group. The mean, ~variance ~0 stat~dard error of the mean and 9S percent confidence interval were calculated from the data Additionally, a chi-square test was calculated to determine if any feed conversion ratio was signifi-cantly different. The weight data were examined utilizing an analysis of variance (one-way) for each group for each tirne period.
., ' , The death rate data was analyzed using a chi-square test (one-way analysis of variance).
The mean feed conversion ratio was 2.3110. A 95 percent confidence interval was calculated around this mean and was determined to have a lower limit of 2.2486 and an upper limit of 2.3734. Because two observations (males 14 ppm - 2.1960 and males 35 ppm - 2.2349) did not fit within the upper and lower limits, further calculation was done. The standard deviation was calculated to be 0.0787itherefor, two standard deviations are 0.1574.
Subtracting this ligure from the mean indicated that the feed conversion ratios of both of these groups were less than two standard deviations away from the mean and therefore are not significantly di,fferent from the other feed conversion ratios. A chi-square test confirmed this observation.
The death pattern of the groups of chickens were examined using a chi-square one-way analysis of variance.
The results demonstrated that no significant difference existed in the death rate of the groups under test.
The male group of chickens receiving 35 ppm ViGen premix feed was significantly heavier than any other group. The female groups receiving 7 and 14 ppm ViGen premix feed were significantly heavier than the female control group. This shows that the use of gentian violet (in the amounts of this invention) in poultry feed results in growth promotion or stimulation of poultry - gentian violet is a growth promoter for poultry.
To answer the question of whether gentian violet in poultry feed was a toxic agent to poultry three parameters were examined. These parameters were feed conversion ratios, weight gains, and deaths. In no case was there any indication that significant differences existed which were detrimental. In fact, the opposite was true. For the reasons expressed above, gentian violet in poultry feed in the amounts claimed in this invention does not have any effect which could be considered toxic to poultry.
Gentian violet, in the amounts of this invention, have a strong stimulatory or promotive effect on the weight ! growth or production of chickens. This example shows that such amounts of orally ingested gentian violet (e.g., carried in inert carriers) in a poultry feed is a good growth promotor for poultry, such as, chickens.
; At day 56, the average weight of the control male chickens was 2.000 kg. and the average weight of the 35 ppm male chickens was 2.132 kg. The average weight difference from the control is very significant. At day 56, the average weight of the control female chickens (hens) was 1.657 kg., the average weight of the 7 ppm ; female chickens was 1.740 kg and the average weight of the 14 ppm female chickens was 1.747 kg. The average weight differences from the control is very significant.
This example establishes that gentian violet, at the levels of this invention, in poultry feed is non-toxic to poultry and is a growth promoter for the poultry.
720 sexed, day-old broiler chickens (Hubbard x Hubbard) were assigned to pens according to the following chart. All chicks in all groups were MD vaccinated and - wing-banded for positive identification. Chicks were weight averaged by sex and the top and bottom weights (approximately 10%) discarded to provide maximum weight uniformly in the groups.
Pen 1 - 60 males Pen 2 - 60 females Pen 3 - 60 males 60 females Pen 4 - 60 males 60 females Pen 5 - 60 males 60 females Pen 6 - 60 males 60 females Pen 7 - 60 males Pen 8 - 60 females (The chicks were debeaked at one day of age, and were obtained from a single breeder flock. Sufficient chicks were obtained to allow for weight averaging of the groups.).
The toxicity study which is part of Example 2 may not have provided completely accurate toxicity data due to excessive mortality and undiagnosed illness in one group of the birds receiving 7 ppm ViGen premix feed. However, it was noted that the groups receiving 35 ppm (5 X treat-ment level) showed no evidence of a toxic effect. Weight gain was equal to and often greater than nonmedicated controls. Droppings remained normal in appearance and consistency as well as general condition. It was apparent that 35 ppm was not a toxic level. When repeated at 0, 7, 14 and 35 ppm (in Example 1), there was again no evidence of toxic symptoms or change in droppings at the higher levels. The average weights were comparable and feed consumption per bird was the same (35 ppm) or less (7 and 14 ppm) than the controls. Necropsy of 4 male and 4 female birds from each dosage level at 56 days showed no gross differences in intestines or abdominal organs.
1~ . ' ' .
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Example 1 conirmed that as much as 35 ppm ViGen premix ' in the feed was not a toxic level.
' ' EX~PLE 3 '- -. \ ~, Chickens infected with Ca,ndida a~bic~ were fed, for eight weeks, a complete-poultr'y feed containing two pounds of premix concentrate per ton o a 20 percent protein poultry starter feed. The premix concentrate contained 1 percent of white mineral oil, 1.65 percent of Micro-Cel E, 1.55 percent of gentian violet, 91,8 percent of corn cob fractions 'and 4 percent of water. The ~1~ complete poultry feed contained~
, ' , ' percent ' by weight, Ingredients weight pounds Gro~md yellow corn ' 62.50 1250 Soybean oil meal (44V/o protein) , 30.00 600 Meat scraP (50% protein-) 2.00 40 M~nhaden fish meal (60% protei'n) 2.00 40 Dica].cium phosphate , . ' 1.90 3~
Calcium carbonc~e 1.00 20 ~0 Standard vitamin premix 0.50 10 Gen~ian violet premix concentrate 0.01 2 ' Total......... 100.00 ~b~' . . ' ''''" . ' ,' . .
, The!~aL~lida ~lb~Q~ infection was successfully treated and the gro~th of the poultry was significantly stimulated.' , EXAMPLE 4 . . : ' , ' .
Chickens not infected with',~ ~h~c ~ were fed,for eight weeks, a complete poultry feed containing one pound of th~
. gcntian violet premix concentrate described in Example 3 per ton ,~, of complete poultry feed (described in Example 3, except that it ~0 con~ained 39 pounds of dicalciu~.n phosphate). There ~as no occ~ur~
c~lce of ~ ,]h~ infectioll cluring the treatment pcriod and the grow~ll of the poul~ry was significantly stilnulate~.
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-i EX~MPLE 5 A ~eries of trials were conducted to evaluate the efficiency of gentian violet on an artifically induced outbreak of candidiasis in broilers, This organism was obtained from a clinical case of crop mycosis; grown in liquld Sabaurads media and diluted to con-tain one million organisms per dose Crop swabs were taken when the birds were 7 days, 14 days, and 21 days~of age and grown on ~ Sabaurads media. Groups o the broiler were fed at levels of l i pound of gentian violet per ton of feed, 2 pounds of gentian violet per ton of feed, and 5 pounds of gentian violet per ton of feed Controls were run against other proaucts and against no treatment. (The other products were copper suifate, Mycostatin, Mold Curb, sodium propionate and calcium propionate.) The birds fed gentian violet had a marked decrease of Candida recovery when compared to the no treatment controls. The birds started on the gentian violet containing feed had the lowest incidence of Candida recovery compared to the controls using the other products and had weights on about the same level as some non-infected, non-treatea controls (although there was a slight weight dcprPssion at the ; five pound per ton level), ~. ~XAMPLE 6 ;~ Hogs infected with Candida albicans were fed, for seven days, a complete hog feed containing two pounds of premix concentrate per ton of completc hog feed The premix contained 1.0 percent of .. . . .. . . . . . . .. .. . . . . . . . . . .... ... . .. ..
1~5991S
vegetable oil, 4.45 percent of Micro-Cel E, 1.55 percent of gentian violet and 94 percent of corn cob fractions.
The complete hog feed was the same as the one used in Example 3. The Candida albicans infection was success-fully treated.
The hogs of Example 6 were fed thereafter a complete hog feed containing one pound oÇ the premix concentrate (described in Example 6) per ton of complete hog feed (described in Example 6). There was no occurrence of the Candida albicans infection during the treatment period.
240 ml. of a premix concentrate containing water and 1.55 percent of gentian violet were diluted to make one gallon of stock solution. The gallon of stock solution and a number of gallons of water were metered through a proportioner, at a ratio of one ounce of stock solution per gallon of water, to poultry drinking trays as needed over a seven-day period.
240 ml. of a premix concentrate containing water and : 0.3875 percent gentian violet were diluted to make one gallon of stock solution. The gallon of stock solution and a number of gallons of water were metered through a proportioner, at a ratio of one ounce of stock solution per gallon of water delivered. The gentian violet addition was one day every week.
Cattle infected with Candida albicans were fed, for seven days, a complete cattle feed containing two pounds of premix concentrate per ton of complete cattle feed.
The premix contained 1.0 percent of vegetable oil, 4.45 percent of Micro-Cel E, 1.55 percent of gentian violet and 93 percent of corn cob fractions. The Candida albicans infection was successfully treated.
The cattle of Example 10 were fed thereafter a complete cattle feed containing one pound of the premix concentrate (described in Example 10) per ton of complete cattle feed. There was no occurrence of the Candida albicans infection during the treatment period.
Turkeys infected with Candida albicans were fed, for seven days, a complete turkey feed containing two pounds of premix concentrate per ton of complete turkey feed.
The premix contained 1.0 percent of vegetable oil, 1.65 percent of microcel E, 1.55 percent of gentian violet, 91.8 percent of corn cob fractions and 4%~water. The Candida albicans infection was successfully t-reated.
The turkeys of Example 12 were fed thereafter a complete turkey feed containing one pound of the premix concentrate (described in Example 12) per ton of completè
turkey feed. There was no occurrence of the Candida albicans infection during the treatment period.
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~ 1~59915 .
! , F,X~MPLr. 14 ' ' . . .
A ViGen premix ormulation was prepared which had the follot~ing components: 1.55 percent of ViGen (gentian violet from A.H.P. Inc.); l percent of white mineral oil, NF-70 Food Grade, from Standard Oil Co., 1.65 percent of ~icro-Cel E, synthetic cal-cium silicate Code 21-E,'from the Celite Division of Johns-Manvill~
Products Co.; 91.8 percent of Grit-O-Cob, Grade 2040 corn cob ractions, from Anderson Cob Mills, Inc. and 4 percent of distille water. The corn cobs and calcium silicate are inert carriers;
white oil reduces the dust in the premi,x and distilled water is th solvent for the gentian violet ~or adsorption o' the gentian viole onto the inert carriers. The ViGen premix formulation was found to be stable or over one year. ' ' In al~ of the above pertinent examples, other ingredient such as cereal grains and grain by-products, can be substituted in part for the ground yellow corn i.n these rations as the market price of feed ingredients 1uctuates since all poultry'feeds are formulated on a least-cost-basis, ~ rK
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lOS99~5 EXAMPJ,E 15 Several experiments were conducted to determine the efficiency of gen$ian violet on candidia~is in broiler chickens. The experi-ments or study had a 56-day duraction. Broadly, the experiments involved reproducing candidiasis in broiler chickens by challenge per os with a dose of approximately one million Candida organisms ¦ per bird and then treatment o some of the broiler chickens to determine the efficiency of gentian violet against the artifically induced outbreak of c~ndidiasis.
'~l~ The gentian violet was used in the form of ViGen premix.
About 6000 lbs. of 25 percent protein broiler feed obtain~d rom Kreamer Co., Kraemer, Pa. (this feed was fed throughout the 56-day eeding pexiod) was ~elleted and crumbled. 2,998.5 lbs.
of the fecd containing 7 ppm (1.516 lb~.) of ViGen premix were admixed--this was termed V feed. 3,000 lbs. of the feed was used for the controls, and was termed C-feed.
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lOS991S
The following parameters were measured during the experiments:
1. Weights - reported in grams.
a. Day one - chicks were weight averaged with top and bottom weights (approximately 10%) discarded to give maximum sex weighted average by group. Weight of each chick in each group was recorded.
b. Day 28 - weighed all birds in all groups, and reported weiqht in grams.
c. Day 56 - weighed all birds in all groups, and reported weight in grams.
2. Feed conversions.
a. By pen at termination of the trial.
a. By pen at termination of the trial.
3. Crop lesions.
a. Severity - as outlined below.
a. Severity - as outlined below.
4. Recovery of organism.
a. 10 percent of the birds in each group were sampled.
a. 10 percent of the birds in each group were sampled.
5. Histopathology.
a. As outlined below.
a. As outlined below.
6. Mortality.
a. First 7 days - recorded date of death and determining cause whenever possible.
b. Day 8 to 56 - birds dying after day 7 were necropsied and cause of death determined whenever possible.
The Candida organism was obtained from a clinical case of candidiasis. The organism was grown in liquid , Sabouraud's media and each bird in each designated group - was challenged with approximately one million Candida organisms per bird.
The challenge dose was administered per os when the birds are 7 days of age.
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~11 '. . l ~ " lOS99~S
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The high inection groups were further challenged by a daily dose of one million Candida organi~ms administered via water contamination from day 8 to 42.
. The birds whose crops were swabbed with S~ h~ culture were marked on the head with dye.
Trial ~o. 1 involved four pens consisting of 60 males and G0 females, broiler chicks were housed 1~0 per pen. The chicks were fed and challenged as listed below:
Pen 3 - Preventative qro~p.
.
P0 The chic~s were placed on V-feed at one day of age and maintalned on the ViGen feed until termination o the trail at day 56. This group was challenged per os on day 7 with one million ~andida albicans or~anism~ per bird (singl~ challenge).
Pen 4 - Treatn~ uP.
The chicks were placed on unmedicated feed ~C-feed) at day one and continued on unmedicated feed until day 14 (seven days post-challenge), at which time this group was placed on feed con-taining the premix ViGen at 7 ppm and maint2ined on the medicated feed from day 14 ~o day 56. ~nis group was chailenged per os on .
2n ; day 7 with one million C?ndi~da albicans organisms per hird (single ch3 llenge), .
Pen 5 - Infected non-medicated grou~
The chicks were placed on unmedicated feed (C-feed) on day one and continued on unmedicated feed to day 56. This group was challenged per os on day 7 with one million Candida albicans organisms per bird (single challenge).
Pen 6 - Non-infected, non-medi~ted group.
The chicks were placed on unmedicated feed (C-feed) on day one and continued on unmedicated feed to day 56. This group was not challengaed during the 56-day feeding period (negative control).
Trial No. 2 involved foor pens containing 60 chicks each, segregated by sex. The chicks were fed and challenged.
Pen 1 - High infection preventive group.
Sixty male broiler chicks were placed on V-feed (i.e., feed containing the ViGen premix at 7 p.p.m.) on day one and continued on the V-feed (medicated feed) until day 56. At day 7 this group received per os a challenge with one million Candida organisms per chick. On day 8 this .
group received a further challenge with one million Candida orgaisms per chick via water contamination. This daily challenge continued until day 42, at which time the challenge ceased and the chicks were allowed to stabilize for 14 days prior to sacrifice on day 56.
Pen 2 - High infection preventive group.
` Sixty female broiler chickens were feed and challenged exactly as Pen 1 above.
1~ l lOS9915 Pen 7 ~ h in~ection unnedicated control ~rou~
Sixty male broiler chicks were placed on unmedicated feed on day one and continued on unmedicated feed to day 5~, This group was challenged exactly as Pen 1 above.
Pen 8 - Hiqh ~infection unmedicated control qrouP, Sixty female broiler chicks were fed and challenged exactly as Pen 7.
Trial 2 was done to study the effects of continuous high level challenge of broiler chickens over a prolonged portion of `10 their growing life. This trial Eurther tested the ability of gentian violet, in the ViGen premix, to protect against high level challengecontinuously by ~ albicans.
After ten days, the birds were re-vaccinated with ND-IB
treatment with Broilerbron spray; and again, after thirty days, the birds were re-vaccinated with ND-IB by treatment with Polybron spray.
During tHe experiments the birds were checked twice weekly for diarrhea, cannibalism, feather dropping, leg weakness, feed consumption and general appearance.
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.~ lOS9915 . - The following is a summary of the total feed consumed in each pen during the experlments (trials): , Pen No, Total Feea Consumed 1 465 lbs, 2 468 lbs. ' 3 939 lbs.
' 4 -804 lbs.
925 lbs, 6 857 lb~.
~0 7 , 4~5 lbs.
8 , 425 lbs. ' The Candida organisms were recovered frpm the appropriate birds.' Ten percent of the birds in each group were sampled, Swa~s were taken from the crops of the same birds in each group at day
a. First 7 days - recorded date of death and determining cause whenever possible.
b. Day 8 to 56 - birds dying after day 7 were necropsied and cause of death determined whenever possible.
The Candida organism was obtained from a clinical case of candidiasis. The organism was grown in liquid , Sabouraud's media and each bird in each designated group - was challenged with approximately one million Candida organisms per bird.
The challenge dose was administered per os when the birds are 7 days of age.
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~11 '. . l ~ " lOS99~S
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The high inection groups were further challenged by a daily dose of one million Candida organi~ms administered via water contamination from day 8 to 42.
. The birds whose crops were swabbed with S~ h~ culture were marked on the head with dye.
Trial ~o. 1 involved four pens consisting of 60 males and G0 females, broiler chicks were housed 1~0 per pen. The chicks were fed and challenged as listed below:
Pen 3 - Preventative qro~p.
.
P0 The chic~s were placed on V-feed at one day of age and maintalned on the ViGen feed until termination o the trail at day 56. This group was challenged per os on day 7 with one million ~andida albicans or~anism~ per bird (singl~ challenge).
Pen 4 - Treatn~ uP.
The chicks were placed on unmedicated feed ~C-feed) at day one and continued on unmedicated feed until day 14 (seven days post-challenge), at which time this group was placed on feed con-taining the premix ViGen at 7 ppm and maint2ined on the medicated feed from day 14 ~o day 56. ~nis group was chailenged per os on .
2n ; day 7 with one million C?ndi~da albicans organisms per hird (single ch3 llenge), .
Pen 5 - Infected non-medicated grou~
The chicks were placed on unmedicated feed (C-feed) on day one and continued on unmedicated feed to day 56. This group was challenged per os on day 7 with one million Candida albicans organisms per bird (single challenge).
Pen 6 - Non-infected, non-medi~ted group.
The chicks were placed on unmedicated feed (C-feed) on day one and continued on unmedicated feed to day 56. This group was not challengaed during the 56-day feeding period (negative control).
Trial No. 2 involved foor pens containing 60 chicks each, segregated by sex. The chicks were fed and challenged.
Pen 1 - High infection preventive group.
Sixty male broiler chicks were placed on V-feed (i.e., feed containing the ViGen premix at 7 p.p.m.) on day one and continued on the V-feed (medicated feed) until day 56. At day 7 this group received per os a challenge with one million Candida organisms per chick. On day 8 this .
group received a further challenge with one million Candida orgaisms per chick via water contamination. This daily challenge continued until day 42, at which time the challenge ceased and the chicks were allowed to stabilize for 14 days prior to sacrifice on day 56.
Pen 2 - High infection preventive group.
` Sixty female broiler chickens were feed and challenged exactly as Pen 1 above.
1~ l lOS9915 Pen 7 ~ h in~ection unnedicated control ~rou~
Sixty male broiler chicks were placed on unmedicated feed on day one and continued on unmedicated feed to day 5~, This group was challenged exactly as Pen 1 above.
Pen 8 - Hiqh ~infection unmedicated control qrouP, Sixty female broiler chicks were fed and challenged exactly as Pen 7.
Trial 2 was done to study the effects of continuous high level challenge of broiler chickens over a prolonged portion of `10 their growing life. This trial Eurther tested the ability of gentian violet, in the ViGen premix, to protect against high level challengecontinuously by ~ albicans.
After ten days, the birds were re-vaccinated with ND-IB
treatment with Broilerbron spray; and again, after thirty days, the birds were re-vaccinated with ND-IB by treatment with Polybron spray.
During tHe experiments the birds were checked twice weekly for diarrhea, cannibalism, feather dropping, leg weakness, feed consumption and general appearance.
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.~ lOS9915 . - The following is a summary of the total feed consumed in each pen during the experlments (trials): , Pen No, Total Feea Consumed 1 465 lbs, 2 468 lbs. ' 3 939 lbs.
' 4 -804 lbs.
925 lbs, 6 857 lb~.
~0 7 , 4~5 lbs.
8 , 425 lbs. ' The Candida organisms were recovered frpm the appropriate birds.' Ten percent of the birds in each group were sampled, Swa~s were taken from the crops of the same birds in each group at day
7, day 14 and day 56. Swabs were placed'on Pagano-Levin media plates and grown for 7 days at room tempe'rature and colony counts made on the 7 day growths. '' , The day 7 swabs were taken immediately pre-challenge of Candida occurrence from natural exposure to feed, water and environment. The day 14 swabs were taken 7 days post-cl~allenge to determine the degree of candidiasis occurring due to-the challen~e dose of Candida. The day'56 swabs were taken at te~mination of the 56-day trial to determine the degree of control of candidiasis occurring from the incorporation of the premix ViGen into the feed of the preventive and treatment,group.
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~ ~ OS9915 l . . , . ~
Pos morten examination of the birds in each group ~rom which crop swabs were taken were made by a poultry pathologist to r determine the crop lesions and the intestinal (gut) lesion scores.
The parameters dealt with ln this example were feed conversio~ t ratios, weight (day 1, 28, and 56-day), lesions (crop and gut) scored durin~ necropsy.
¦ Prior to examination of the raw data it was decided that if a bird was found crippled during the course of these trials, it .
was to be dropped from consideration after the first day that a trial commenced. Only three birds were found to be crippled at the end of Trial l; in Trial 2, only one bird from the preventive group was ound to be crippled. In a practical sense, because of the s~all nun~er of c~ippled birds and the size of the groups involved, this decision or exclusion had little to no beariny on . : , .
the analy~is.
Trial l the mean feed conversion ratio was 2.1663. A 95%
confidence interval was calculated around this mean. Since each of the observatlons fell~within these limits, no significance could be attached to this finding. In trial 2 the mean feed con- i~
~0 version ratio was ?.1383. Again, each of the observations fell within the 95% confidence interval limits and no significance coul~
: be a ch-d to th finding.
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The ana~ysi~ of variance of the weight data for Trial 1 ¦demonstrates significant differerlces in the weights of each group ¦to exist at day 1, 28 and 56 of the trial. Further comparison ¦indicates that at day 1 the treated group of ~roiler chicks had a ¦mean weight significantly larger than the mean weights of the other ¦three groups. No difference existed in the mean weights of the ¦preventive, non-infectea control or the infected control group.
¦At day 28, the situation changed. The preventative group had a ¦mean weight (641 gms) which was significantly larger ( P~ 0.01) ¦than the other three groups (treated--601,38 ~ms, non-infected ¦control--598.92 gms, infected control--59~.88 gms). In turn, the ¦mean weights of the other three groups were not si~nificantly ¦different from each other.
This situation again changed at day 56. At ~his time inter-val, the mean weiyhts of the preventative~(1686.6 gms) and the treated group~(l638.~) were not different from each other.
However, the mean weight of the preventative group was signifi-cantly larger than the non-infected controls (1586.8 gms), while the mean weight of the treated group was not. Both the treated 2~ and non-infected control groups had mean;~eights significatnly larger than the infected controL (1496.5 gms) Concerning the weight data for Trial 2, with the exception of day 2~, the calculated F-ratios was again highly significant.
¦Asain, further co~.parisons of th- ~ean weiaht demonstrated signi ~icant dif~erences to cxist between the groups o~ chicks in . question. The preventative group (males) ha~ a mean weight of ~ : . ' , ' .
~: ~ lOS9915 .- .
37.2 gms which was larger than the preventative group females (34.8 gms), infected control males (34 7 gms) and the infected control females (34.4 gms). In turn, the preventative group females had a mean weight no different from the infected control ~- males but a weight significantly larger than the infected control _ females. Finally, the infected control males had a mean weight significantly larger than the infected control females.
By day 28 this situation hsd changed drastically. As fore-. told by the F-ratio, only one significant difference existed in the '-~ 10 ~ean weights of the group in question, that of the infected control ; ¦males (660.4 gms) as compared to the infected control females (636,4 gmsl. ~
`~ On day 56 the mean weight of preventative group males (1764.3 ~ qms) was significantly larger thsn any other group. The.mean i- weight of the preventative group females (1676.3 gms) while not significantly larger than the infected control males (1666 9 gms) ; was significantly larger than the infected control females (1407.
gms). Lastly, the mean weights of the infected control males were ~ .
significantly larger than the infected control females.
~20 ¦ The analysis of ~ariances concerned with the crop and gut _ lesions was made. In all cases the F-ratios were significant.
_ ~ - Re Trial 1 with regard to crop lesions, the infected control i had a mean lesion score (2.5) which was significantly larger than any other mean lesion scroe. There was no significant difference ; between the mean lesion scores of the treated group (1.7S) and .,,..
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(the preventative group (1.33). However, the treated group had a mean lesion score significantly larger than the non-infected control (1.17). No significant differ-ence existed between the mean lesion scores of the preventative and the non-infected control groups.
Re Trial 2, with regard to crop lesions, the mean lesion score of the infected control females (3.38) was not different from that of its male counterpart (2.75).
However, this mean lesion score of 3.38 was significantly larger than both of the preventative groups (males, 1.38;
~ females, 1.0). In turn, the mean lesion score of the infected control males (2.7~ was significantly larger than both of the preventative groups. Lastly, no sig-' nificant difference existed between the mean lesion scores of the male and female preventative groups.
The ~significant" weight differences manifest in the groups of broiler chicks in both Trials 1 and 2 at one day of age are consistent with past experience and should not be considered an impediment to the results of these efficacy trials. It would be unlikely that the division of a finite number of chicks into groups of any size on a randomized basis would result in these groups being equal in any respect.
In Trial 1, the efficacy of ViGen premix becomes manifest when weights are considered at days 28 and 56 and when lesion scores are examined.
ViGen was efficacious both as a preventative and a treatment. Neither the ViGen-treated or preventative group developed any gut lesions.
In Trial 2, the mean lesion scores, either crop or gut, were significantly larger in the infected groups 10599~5 than in the preventative regimen groups.
The results of Teial 2 reaffirmed the findlng in Trial 1 that ViGen was efficacious. A direct comparison of the mean weights on basis of sex shows that males and females on a preventative regimen of ViGen were significantly heavier than their counterparts of the infected control.
Secondly, sex appeared to be a factor in candidiasis when weight gains were considered. In both the infected control and the preventative regimen groups the males weighed significantly more (P 0.01) than their female counterparts. One expects a group of males to be heavier ' than their female counterparts. However, the group of females on the preventative regimen of ViGen did not weigh significantly less than the males of the infected control group. This lack of difference appeared to be caused by the prophylactic action of YiGen.
The mean lesion scores (crop or gut) between the sexes of the same group was not significantly different. It would appear than that sex was not a factor with regard to the resulting lesions of a Candida infection.
The feed conversion ratios of both trials were dis-tributed evenly in a biostatistical sense. In Trial 1 the feed conversion ratio of the treated group was 13 percent better than the standard, the preventation group was 28 -percent better, the infected control was 11 percent worse. These figures have real meaning to a poultry ` producer.
In summation, when the parameters of weight and lesion - . scores are examined, ViGen (gentian violet) was demon-strably efficacious in the prevention and treatment ofcandidiasis.
" 105991S
Gross observation throughout the trial showed no detectable difference between groups with respect to general growth and condition; and no Glinical signs could be detected in infected birds as compared to controls.
But this does not consider the weights, feed consumptions, culturings for Candida and necropsies which are also part of the test data and results.
The challenge with 106 Cand_da at 8 days established infection which persisted to 8 weeks. Although swab counts were low, there were definite lesions in both the crops and intestines. Slight infection (low swab counts) occurred in nonchallenged birds but there was no evidence of lesions. Medication with 7 ppm from one day of age prevented establishment of infection as evidenced by low swab counts at 14 days of age. Culture indicated a mild infection had developed by 56 days but only slight crop lesions could be detected. Delay of treatment until 14 days allowed establishment of infection following challenge at 8 days. However, swab counts and lesions at 56 days were almost identical to those in the group medicated from one day of age. This would indicate treatment as well as prophylacitc activity. The infected, nonmedicated birds had a persistent infection from the single exposure at 8 days but swab counts at 56 days were much lower than expected considering the lesions observed in most birds. Three-plus crop score with a count of only 30 and then a negative crop with a count of 70 are examples of this variability. It appeared that older birds with "chronic" infection do not have as much mucus and active organisms in the crop and thus lower counts from the relatively dry swab.
Daily challenges with 106 Candida per bird resulted in high counts and severe crop lesions in most birds.
When medicated, very few birds showed slight lesions and in general relatively low counts. The efficacy of ViGen premix at 7 ppm to control infection even with repeated exposure is obvious.
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~ ~ OS9915 l . . , . ~
Pos morten examination of the birds in each group ~rom which crop swabs were taken were made by a poultry pathologist to r determine the crop lesions and the intestinal (gut) lesion scores.
The parameters dealt with ln this example were feed conversio~ t ratios, weight (day 1, 28, and 56-day), lesions (crop and gut) scored durin~ necropsy.
¦ Prior to examination of the raw data it was decided that if a bird was found crippled during the course of these trials, it .
was to be dropped from consideration after the first day that a trial commenced. Only three birds were found to be crippled at the end of Trial l; in Trial 2, only one bird from the preventive group was ound to be crippled. In a practical sense, because of the s~all nun~er of c~ippled birds and the size of the groups involved, this decision or exclusion had little to no beariny on . : , .
the analy~is.
Trial l the mean feed conversion ratio was 2.1663. A 95%
confidence interval was calculated around this mean. Since each of the observatlons fell~within these limits, no significance could be attached to this finding. In trial 2 the mean feed con- i~
~0 version ratio was ?.1383. Again, each of the observations fell within the 95% confidence interval limits and no significance coul~
: be a ch-d to th finding.
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The ana~ysi~ of variance of the weight data for Trial 1 ¦demonstrates significant differerlces in the weights of each group ¦to exist at day 1, 28 and 56 of the trial. Further comparison ¦indicates that at day 1 the treated group of ~roiler chicks had a ¦mean weight significantly larger than the mean weights of the other ¦three groups. No difference existed in the mean weights of the ¦preventive, non-infectea control or the infected control group.
¦At day 28, the situation changed. The preventative group had a ¦mean weight (641 gms) which was significantly larger ( P~ 0.01) ¦than the other three groups (treated--601,38 ~ms, non-infected ¦control--598.92 gms, infected control--59~.88 gms). In turn, the ¦mean weights of the other three groups were not si~nificantly ¦different from each other.
This situation again changed at day 56. At ~his time inter-val, the mean weiyhts of the preventative~(1686.6 gms) and the treated group~(l638.~) were not different from each other.
However, the mean weight of the preventative group was signifi-cantly larger than the non-infected controls (1586.8 gms), while the mean weight of the treated group was not. Both the treated 2~ and non-infected control groups had mean;~eights significatnly larger than the infected controL (1496.5 gms) Concerning the weight data for Trial 2, with the exception of day 2~, the calculated F-ratios was again highly significant.
¦Asain, further co~.parisons of th- ~ean weiaht demonstrated signi ~icant dif~erences to cxist between the groups o~ chicks in . question. The preventative group (males) ha~ a mean weight of ~ : . ' , ' .
~: ~ lOS9915 .- .
37.2 gms which was larger than the preventative group females (34.8 gms), infected control males (34 7 gms) and the infected control females (34.4 gms). In turn, the preventative group females had a mean weight no different from the infected control ~- males but a weight significantly larger than the infected control _ females. Finally, the infected control males had a mean weight significantly larger than the infected control females.
By day 28 this situation hsd changed drastically. As fore-. told by the F-ratio, only one significant difference existed in the '-~ 10 ~ean weights of the group in question, that of the infected control ; ¦males (660.4 gms) as compared to the infected control females (636,4 gmsl. ~
`~ On day 56 the mean weight of preventative group males (1764.3 ~ qms) was significantly larger thsn any other group. The.mean i- weight of the preventative group females (1676.3 gms) while not significantly larger than the infected control males (1666 9 gms) ; was significantly larger than the infected control females (1407.
gms). Lastly, the mean weights of the infected control males were ~ .
significantly larger than the infected control females.
~20 ¦ The analysis of ~ariances concerned with the crop and gut _ lesions was made. In all cases the F-ratios were significant.
_ ~ - Re Trial 1 with regard to crop lesions, the infected control i had a mean lesion score (2.5) which was significantly larger than any other mean lesion scroe. There was no significant difference ; between the mean lesion scores of the treated group (1.7S) and .,,..
' ,~""~
.. ~ ' ' . " ~ '- ~ ~- . -.
(the preventative group (1.33). However, the treated group had a mean lesion score significantly larger than the non-infected control (1.17). No significant differ-ence existed between the mean lesion scores of the preventative and the non-infected control groups.
Re Trial 2, with regard to crop lesions, the mean lesion score of the infected control females (3.38) was not different from that of its male counterpart (2.75).
However, this mean lesion score of 3.38 was significantly larger than both of the preventative groups (males, 1.38;
~ females, 1.0). In turn, the mean lesion score of the infected control males (2.7~ was significantly larger than both of the preventative groups. Lastly, no sig-' nificant difference existed between the mean lesion scores of the male and female preventative groups.
The ~significant" weight differences manifest in the groups of broiler chicks in both Trials 1 and 2 at one day of age are consistent with past experience and should not be considered an impediment to the results of these efficacy trials. It would be unlikely that the division of a finite number of chicks into groups of any size on a randomized basis would result in these groups being equal in any respect.
In Trial 1, the efficacy of ViGen premix becomes manifest when weights are considered at days 28 and 56 and when lesion scores are examined.
ViGen was efficacious both as a preventative and a treatment. Neither the ViGen-treated or preventative group developed any gut lesions.
In Trial 2, the mean lesion scores, either crop or gut, were significantly larger in the infected groups 10599~5 than in the preventative regimen groups.
The results of Teial 2 reaffirmed the findlng in Trial 1 that ViGen was efficacious. A direct comparison of the mean weights on basis of sex shows that males and females on a preventative regimen of ViGen were significantly heavier than their counterparts of the infected control.
Secondly, sex appeared to be a factor in candidiasis when weight gains were considered. In both the infected control and the preventative regimen groups the males weighed significantly more (P 0.01) than their female counterparts. One expects a group of males to be heavier ' than their female counterparts. However, the group of females on the preventative regimen of ViGen did not weigh significantly less than the males of the infected control group. This lack of difference appeared to be caused by the prophylactic action of YiGen.
The mean lesion scores (crop or gut) between the sexes of the same group was not significantly different. It would appear than that sex was not a factor with regard to the resulting lesions of a Candida infection.
The feed conversion ratios of both trials were dis-tributed evenly in a biostatistical sense. In Trial 1 the feed conversion ratio of the treated group was 13 percent better than the standard, the preventation group was 28 -percent better, the infected control was 11 percent worse. These figures have real meaning to a poultry ` producer.
In summation, when the parameters of weight and lesion - . scores are examined, ViGen (gentian violet) was demon-strably efficacious in the prevention and treatment ofcandidiasis.
" 105991S
Gross observation throughout the trial showed no detectable difference between groups with respect to general growth and condition; and no Glinical signs could be detected in infected birds as compared to controls.
But this does not consider the weights, feed consumptions, culturings for Candida and necropsies which are also part of the test data and results.
The challenge with 106 Cand_da at 8 days established infection which persisted to 8 weeks. Although swab counts were low, there were definite lesions in both the crops and intestines. Slight infection (low swab counts) occurred in nonchallenged birds but there was no evidence of lesions. Medication with 7 ppm from one day of age prevented establishment of infection as evidenced by low swab counts at 14 days of age. Culture indicated a mild infection had developed by 56 days but only slight crop lesions could be detected. Delay of treatment until 14 days allowed establishment of infection following challenge at 8 days. However, swab counts and lesions at 56 days were almost identical to those in the group medicated from one day of age. This would indicate treatment as well as prophylacitc activity. The infected, nonmedicated birds had a persistent infection from the single exposure at 8 days but swab counts at 56 days were much lower than expected considering the lesions observed in most birds. Three-plus crop score with a count of only 30 and then a negative crop with a count of 70 are examples of this variability. It appeared that older birds with "chronic" infection do not have as much mucus and active organisms in the crop and thus lower counts from the relatively dry swab.
Daily challenges with 106 Candida per bird resulted in high counts and severe crop lesions in most birds.
When medicated, very few birds showed slight lesions and in general relatively low counts. The efficacy of ViGen premix at 7 ppm to control infection even with repeated exposure is obvious.
. ~
.
Claims (8)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A supplemental feed for animals or poultry which contains a complete animal or poultry feed or a basal animal or poultry feed and between 0.00077 and 0.005 percent by weight, based on the weight of the complete or basal animal or poultry feed, of gentian violet.
2. A supplemental feed according to claim 1 which further contains inert ingredients selected from diatomaceous silica, corn cob fractions and vegetable oil.
3. A supplemental feed according to claim 1 wherein said feed contains 0.00156 percent by weight of gentian violet.
4. A method of preparing a supplemental feed for animals or poultry, which comprises preparing a premix concentrate for addition to complete or basal animal or poultry feed which premix consists only of between 0.1 and 10 percent by weight of gentian violet and the remainder inert ingredients, and adding enough of said premix concentrate to the other ingredients of the feed to obtain between 0.00077 and 0.005 percent by weight of gentian violet in said feed.
5. A method as described in claim 4 wherein said inert ingredients are selected from diatomaceous silica, corn cob fractions and vegetable oil.
6. A premix concentrate for addition to animal and poultry feed which consists essentially of gentian violet, which is a selective fungicidal mold inhibitor of Candida albicans, and the remainder inert ingredients selected from diatomaceous silica, corn cob fractions and vegetable oil.
7. A premix concentrate as described in claim 6 wherein about 1.55 percent of gentian violet is present.
8. A premix concentrate as described in claim 6 wherein between 0.1 and 10 percent by weight of gentian violet is present.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US59587675A | 1975-07-14 | 1975-07-14 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1059915A true CA1059915A (en) | 1979-08-07 |
Family
ID=24385055
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA238,049A Expired CA1059915A (en) | 1975-07-14 | 1975-10-21 | Growth promotor and processes |
Country Status (2)
| Country | Link |
|---|---|
| AU (1) | AU8595575A (en) |
| CA (1) | CA1059915A (en) |
-
1975
- 1975-10-21 CA CA238,049A patent/CA1059915A/en not_active Expired
- 1975-10-23 AU AU85955/75A patent/AU8595575A/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| AU8595575A (en) | 1977-04-28 |
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