Studies on Control of Fowl Cholera T.A

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Agricultural Experiment Station Technical Bulletins SDSU Agricultural Experiment Station

1959 Studies on Control of Fowl Cholera T.A. Dorsey

G.S. Harshfield

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Recommended Citation Dorsey, T.A. and Harshfield, G.S., "Studies on Control of Fowl Cholera" (1959). Agricultural Experiment Station Technical Bulletins. 34. http://openprairie.sdstate.edu/agexperimentsta_tb/34

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Studies on Control of Fowl Cholera

Veterinary Department CONTENTS

Introduction ------1 Nature of Disease______1

Occurrence ------1 Symptoms and Lesions______2

Epizootiology ------2

Experimental Studies ------4

Studies of Pasteurella Multocida Strains______4

Detection of Carriers by Agglutination Test______6

Immunization ------8

Treatment ------12 Summary and Conclusions______16

Bibliography ------16 lM-5-59-6661 Siudies on Conlrol of Fowl Cholera 1

T. A. DORSEY and G. S. HARSHFIELD 2

INTRODUCTION NATURE OF THE DISEASE Fowl cholera is a contagious dis Occurrence ease of domestic fowls and many Fowl cholera ranked with fowl wild birds caused by a bacterium, leuko.sis and coccidiosis to consti Pasteurella multocida. It is one of tute the three most frequently oc the oldest known infectious diseases curring diseases in poultry exam of poultry. Many reports on different ined at the South Dakota State phases of research on the disease College veterinary diagnostic labo have been published, dating to a ratory. It occurred at all seasons of period before Louis Pasteur, whose the year but was most prevalent name is associated with early during late summer, fall, and early studies of fowl cholera . Although winter months from August through the disease has received much at January ( figure 1 ). tention for over 100 years, its oc Most of the outbreaks of the dis currence is still widespread and ease in South Dakota poultry flocks control measures have generally were acute. In outbreaks of this na lacked effectiveness. ture, a heavy death loss occurred In South Dakota, fowl cholera early in the outbreak; then less has been a poultry disease of major acute, and sometimes chronic, cases importance for many years. Experi developed after the disease had mental work was undertaken at this been in progress for several days. station in 1944 to study fowl chol A few outbreaks were more chronic era as related to this area and to 1This study was financed in part by funds investigate the development or im from the North Central Regional Experi- provement of control measures to ment Station Cooperative Research Proj more effectively prevent losses from ect IC-6. 2Associate Veterinarian and Veterinarian, this disease. This bulletin is a re respectively, South D akota Agricultural port of that work. Experiment Station. 1 2 South Dakota Experiment Station Technical Bulletin 23 10

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0 J F M A M J J A s 0 N D Figure 1. Average number of diagnoses of fowl cholera by month, 1949-58. from the onset. With these, the lameness resulting from swollen death rate was not so rapid, al joints ( figure 3). though duration of the outbreak Birds that died suddenly, showed was more prolonged. It was not un few lesions on necropsy. A few usual in flocks affected with chronic petechiae were found on the heart cholera for the disease to suddenly and serous membranes, particularly become acute with a heavy death over the gizzard fat. Small necrotic loss. foci also were found in the liver in Symptoms a nd Lesions less acute cases ( figure 4). Lesions Symptoms were lacking in most associated with the chronic disease acute cases. Birds would be found were characterized by accumula dead with no earlier sign of dis tions of a yellowish, caseous exu ease. In the chronic form, birds date in affected parts such as the air were affected in different ways. sacs, ears, ear lobes, wattles, or Respiratory involvement was com joints. mon and torticollis ("wry-neck") Epizootiology was observed occasionally ( figure Histories which were obtained 2). Some birds had swollen eyes, concerning individual outbreaks wattles, or ear lobes. Some showed of fowl cholera often suggested that Figure 2. Chronic infection in the ear causes torticollis.

Figure 3. Localized chronic infection in the foot.

Figure 4. Acute fowl cholera showing petechiae on the heart and small, ne the source of infection could have c11otic foci on the liver. been birds which were apparently normal, but were carriers of the spe cific organism. Some outbreaks oc curred after birds of a healthy flock had been exposed to another flock in which there may have. been birds carrying infection acquired during a previous outbreak. Others fol- 4 South Dakota Experiment Station Technical Builetill 23 lowed the introduction of cockerels Rosenbusch and Merchant ( 24 ) or pullets from another flock. Many made a study of a number of P. mul of the outbreaks occurred in late tocida strains of mammalian and summer or fall months after the as. avian origin and divided them into sociation of susceptible pull~ts three groups according to their raised on the farm with the older ability to ferment xylose, arabinose, flock, on range or in the poultry and dulcitol. This grouping was house. In some instances, a history correlated by tube agglutination of an earlier outbreak in the older tests. birds was provided but this was not A similar study was made at this always the case. laboratory with 364 P. multocida A source of infection could not strains of fowl origin. Fermentation always be determined, nor could it of xylose, arabinose, and dulcitol be explained why a latent infection was determined by color change in in a flock would suddenly develop Difeo purple broth base to which into an explosive outbreak with a 0.5% of each sugar that was filter heavy death loss. It was not deter sterilized had been added. Three mined whether such occurrences tubes, each containing 2.5 milliliters resulted from a sudden increase in of purple broth and the respective the virulence of P. multocida in the sugar, were inoculated with each P. carri r birds, from lowered resist multocida strain and incubated at ance of the birds, or perhaps a com 37°C. The final reaction was re bination of these and other fatcors. corded on the seventh day. Fmther EXPERIMENTAL STUDIES incubation was not carri d out be cause of evaporation. Studies of Pasteurella Multocida Table 1 shows the basis for Strains grouping these strains according to Most of the strains of P. multo fermentation reactions, and the cicla isolated from fowls submitted number and percent of strains for for diagnosis during the past 10- each group. year period were retained. Isola A few more xylose - negative tions were made from blood, liver, strains could have been included in or suggestive lesions on blood agar group I but were omitted because and identification was made bv their biochemical characteristic{s Table 1. D istribution of Avian P. multo and morphology. Subcultures from cida Strains According to Fermentation each new isolation were made on Reaction Difeo stock culture agar slants and Fermentation incubated 24 hours at 37°C. The Reaction Number tubes were sealed with waxed corks Xy- Ara- Dul- of Per- and then stored at room tempera Group lose binose c:tol Strains cent ture. Many strains were held in this I + + 306 84.06 manner for 3 years or longer with II + + 53 14.57 out transfer. Only a few strains be or - came nonviable by this procedure. III + + + 5 1.37 Studies on Control of Fowl Cholera 5 of their inability to ferment arabi lated intravenously with two 1 milli nose and/ or dulcitol. liter doses of each culture given 5 Reciprocal agglutination tests days apart. These birds were nega were made to determine whether tive to agglutination tests with the agglutination reactions might be different antigens prior to inocula correlated with biochemical reac tion. About 1 week after the last in tions in classifying these P. multo oculation the birds were bled from cida strains. Rapid, somatic anti the heart. The .serum was separated gens were prepared from group I, from the blood and preserved with II, and III P. multocida strains. The merthiolate. Agglutination tests method for preparing these anti were conducted by mixing one gens is described under the section, drop of antigen with .02 milliliter "D etection of Carriers by Aggluti of serum. Antisera of strains of each nation Test." group were tested with each anti The same strains of P. multocida gen. The results of these tests are that were used to prepare the rapid shown in table 2. antigens were used to inoculate These tests showed that the so chickens to . produce agglutinins. matic antigens of strains of group I Difeo tryptose phosphate broth was and group III were very similar. inoculated with the different strains They were not closely related to and incubated 24 hours at 37°C. those of group II. Group II strains, Incubation was then continued for 33 and 338, fermented xylose but 96 hours at 56°C. to remove the not arabinose and dulcitol. The capsular substance and kill the cul other group II strain, 63, fermented tures. Chickens were then inocu- xylose and arabinose. However, on

Table 2. Results of Reciprocal Agglutination Tests Between P. multocida Groups

Antigen Group I Group II Group III Antiserum 5 99 33 338 63 97 283 Group I 1 + + + + I* 5 + + + + + 99 + + + I + Group II 33 + + + 338 + + + 63 I + + Group III

I 97 i + + + + 283 + + + + + *Tn co mplete. 6 South Dakota Experiment Station Technical Bulletin 23 the basis of agglutination tests, growths of the autogenous strain their somatic antigens could not be suspended in physiological saline distinguished. A slight relationship solution containing 1% formalin. of strains of different groups for one The suspension was adjusted to a another was shown, .g. strain 33 to turbidity of 75xl by the McFarland 99 and strain 338 to 283. This would ne helometer scale and stained indicate that some avian P. multo with 0.1% of crystal violet. The au cida strains of different groups thors reported control of the fowl might have related somatic anti cholera problem in this flock by re g ns. moval of the reactors. A comparison of death losses, Antigens prepared h re by the from different fowl cholera out Shook and Bunyea procedure some breaks in South Dakota, showed no times proved unstable because of appreciable difference in the viru autoagglutination. A different pro lence of the causative strains of P. cedure of preparation was finally multocida, whether they belonged evolved which resulted in an anti to group I or II. There were not gen that was stable and maintained enough outbreaks caused by group its sensitiveness for several months III strains to be considered in mak when stored at 45°F. This proce ing this comparison. Greater varia dure was as follows: Three serial tion in virulence could be found passages of the antigen culture of among the strains within one group. P. multocicla were made in 10-day old chicken embryos. One liter of Detection of Carriers by Agglutina- tryptose phosphate broth was in tion Test oculated with 1 milliliter of allantoic Several investigators ( 5, 11, 15, fluid from a third passage embryo 18, 21, 22, 23, 26, 28) working on and incubated 24 hours at 37°C. fowl cholera found that apparently Then, 10 milliliters of 0.1% blue tet healthy birds may cany P. multo razolium, an intravital dye, was add cida. Most commonly the organism ed to the antigen culture. This was was harbored in the upper respira incubated at 37°C. for an additional tory tract. Because the source of in 72 hours. The bacteria were staineq fection in many fowl cholera out dark blue shortly after adding the breaks can be traced to exposu~e blue tetrazolium. Further incuba to such carriers, the detection and tion for 24 hours at 56 °C. was used removal of carriers would be of to insure that the organisms were value as a control measure. killed and that the capsular sub Shook and Bunyea ( 26) pre stance had been removed. The bac pared a P. multocida antigen for a teria were sedimented in an angle rapid, whole-blood agglutination centrifuge and the broth was dis test and used it to test a flock in carded. The organisms were resus which a carrier problem had ex pended and washed in phenolized isted over several years. The anti ( 0.25%) saline solution, after which gen consisted of 24-hour agar they were centrifuged again and the Studies on Control of Fowl Cholera J saline wash was discarded. The bac and P. multocida was isolated from teria were resuspended in 20 milli seven. No attempt was made to cul liters of phenolized, buffered saline ture the negative birds. After re which was prepared from Soren moval of reactor birds, a second son's standard solutions as follows: course of oxytetracycline was given.

M/ 15 KH2P04-90 milliliter, M/ 15 When the antibiotic was discon

Na:., HP04-10milliliters, 0.85%NaCl, tinued, losses recurred in one flock. and 0.25% phenol. The pH of the Poor management and insanitation antigen was 5.7. Tests conducted in were probably responsible for con developing this method of prepara tinued loss. tion showed that as higher pH A more critical evaluation of the values were used the antigen be antigen to detect carriers was made came less sensitive and inagglutin on an adult flock of nearly 100 hens. able with weak P. multocida anti Fowl cholera had been a problem serum. The density of the antigen for several years on the farm where was about the same as th stained, this flock was raised. Losses from rapid pullorum antigen ( figure 5 ). cholera occurred in pull ts which natural outbreak of fowl chol had been in association with the era of low virulenc occurred in a older birds prior to the acquisition small group of hens in a breeding of the mature birds for this study. experiment. Most of the cases were They were removed from the farm chronic but occasionally a bird to the laboratory where they were would die of acute cholera. The 29 housed as a group and given aver surviving b i r d s composing this age care. group were tested with antigen, fol The hens were tested with two lowed by necropsy and bacteriolog antigens, one prepared from a ical examinations. Fifteen of the group I strain and the oth r from a birds reacted, with 11 ( 73%) yield group II strain. Earlier studies ( 9, ing cultures of P. multocida. Isola 10) had shown that most cases of tions were made from lesions which were generally characterized by ac Figure 5. Negative and positive tests cumulations of a yellowish, caseous with a rapid, whole-blood P. multocida exudate. None of the 14 negative antigen. birds were positive on culture. Two farm flocks were blood test ed when fowl cholera recurred after oxytetracycline (Terramycin ) treat ment. Facilities on the farms did not permit detailed study of the effectiveness of removing reactors in the control of the disease. Thirtv one reactors were found in the 421 birds comprising the two flock s. Eight of the reactors were cultured 8 South D akota Experiment Station ; echnical B ulletin 23 fowl cholera in South Dakota were cultural examinations of the indi caused by strains of these two vidual birds, excluding those birds groups of P. multocida. There were which were negative both to the 13 reactors, seven reacting only to agglutination test and bacteriologi the group I, five only to the group cal examination. II, and one to both antigens. These In all, 30 cultures of P. multocicla reactors were left in the flock. were recovered from swabs and During the next 6 months, swabs organ cultures from this flock. On were made from the palatine cleft the basis of their ability to ferment of each living bird. These were cul xylose, arabinose, and dulcitol, all tured in broth and the culture inoc belonged to group I. One of the cul ulated into mice. The mice that tlues, inoculated into a normal bird. died were subcultured to attempt resulted in an antiserum which ag the isolation of P. multocida. By glutinated both the group I and this procedure isolations were made group II antigens us ed in tests of from two of the reactors and from the flock. five of the negative birds. Although the correlation be Seventeen hens negative to the tween reactions with P. multocicla test died during the interval. P. mul antigen and recovery of the organ tocida was isolated on culture from ism was good in the earlier trials , seven of these, four having died· of results were not encouraging with acute cholera. Positive isolation this flock. Not only were isolations was made from one of three reactors from reactor birds at a low level, that died. but positive isolations were made from the palatine clefts of five liv A second test of the remaining ing birds which did not react. In birds was made 4}~ months after the ability of the test to detect all first at which time there were seven carriers would reduce the value of reactors. Three reacted only with the test as a control measure. From group I antigen, two only with the experience in the small group group II antigen, and two showed of 29 birds undergoing the outbreak reactions with both. P. multocida of chronic cholera, the antigen isolations were not made from any would appear to have some value of these birds. Three birds that as a diagnostic test to detect birds were negative on the first test re with chronic lesions. acted on the second. Six birds showing reactions on the first test Immunization were negative on the second. An Although Pasteur ( 19 ) in 1880 isolation of P. multocida had been demonstrated immunity in fowls made from the nasal cleft of one of inoculated with attenuated cultures these earlier, but it was negative on of P. multocida, workers since that cultures of the nasal cleft and or time have had irregular results with gans at necropsy following the last various vaccines and bacterins. test ( No. 44). Table 3 shows the Generally, no protection was pro results of agglutination tests and vided in the vaccinated fowls, or Studies on Control of Fowl Cholera 9 the resulting immunity was of a low for immunizing White Pekin ducks level and of short duration. Immuni against cholera. zation has never been accepted as a A chicken embryo (CE ) vaccine dependable control measure for similar to that used by Carter was fow 1 cholera. prepared to compare with a formal More recently, Carter ( 4 ) found a ized broth culture bacterin as an chicken-embryo vaccine effective immunizing agent in chickens. A against experimental pasteurellosis group I strain ( No. 21 ) was carried in mice, and D augherty ( 6 ) com through three passages in 10-day pared a duck-embryo vaccine with chicken embryos to enhance the four commercial bacterins, finding virulence of the organism. The en the duck-embryo vaccine superior tire contents of two last-passage

Table 3. Test and Cultural Record of Individuals from Adult Flock, Excluding the Negative Birds

1st Test, 11 -5-57 2nd Test, 3-19-58 P. multocida Isolation Bird No. Group I Group II Group I Group II Nasal Swab* Necropsyt 10 + 15 + Died 17 + Died + 26 + 28 + 32 Died + 34 Died + 37 + 40 + Died + 42 + + + 44 + + 47 Died + + 51 Died - + 53 + 57 + 67 + 75 + + + 80 + 81 Died + 82 + + 84 + + 86 + 89 + + 91 Died + + 93 + 96 Died + *Cultu res made when birds w ere alive. -!- Cultures made from affected o rgans a nd/or palatine clefts of birds that died and surviving reactors which w ere kill ed at th e terminati o n o( the experiment. 10 South D akota Experiment Station Technical Bulletin 23

ggs were added to 100 milliliters of 100 sterile physiological saline solution 9Q 1----+~--+-~-+-~-L-~l----l--~~ and mixed in a Waring blendor. Formalin was added to a 0.25%con e ntration. The broth culture (BC) bacterin was prepar d from the same strain by 24-hour incubation 'cl- of tryptose phosphate broth inocu ~ 601---+--+-- lated with allantoic fluid from the >- ~ 50 third passage embryo. This also .=! was formalized. Both preparations ~40 were held for 72 hours in the refrig ~ erator and sterility tested in mice. w 30 > Three pens of 5-week-old chick ~20 :..i ens were used in this trial. Pens 1 ::::, , \ cEN 2 and 2, each with 35 birds, were ~ 10 / .f t :fJlERIN) given two intramuscular injections <.) ~ 0 ------' in 1.0 milliliter amounts a week apart 0 I 2 3 4 5 6 7 of GE vaccine and BC bacterin, re . DAYS POST INOCU LATION spectively. The third pen of 36 Figure 6. Immunity from broth culture birds was not vaccinated and bacterin and chicken embryo vaccines. s rved as a control. Two weeks fol lowing the second injection, all the chickens were used. Pens 1 and 2 birds were challenged with P. mul received two weekly 1.0 milliliter tocicla of the same strain. The or intramuscular injections of the BC ganism for challenge was grown in and commercial bacterins, respec chicken embryos. Dilutions of em tively, and pen 3 served as the con bryo suspension in peptone water trol. A 1: 10,000 dilution of a 24- of 1: 1,000 or 1: 10,000 were inocu hour broth culture of the organism lated intramuscularly in 1.0 milli in a 1.0 milliliter dose served as the liter amounts. challenge 16 days after the second As shown in fi gure 6, the mortal immunizing dose. After 9 days, the ity was 46% in pen 1, 6% in pen 2, mortality was 27%in pen 1, and 64% and 83%in pen 3, the control group. in both pens 2 and 3. No benefit was Although mortality was reduced by shown with the commercial bac both preparations, the BC bacterin terin and the prepared BC bacterin was superior to the CE vaccine in did not provide immunity of suffi this trial. cient level for practical purposes. In a second immunization trial, a The third immunization trial was formalized BC bacterin prepare9- designed not only to compare CE as above from strain No. 1 was com vaccines and BC bacterins but also pared with a commercial P. multo to determine whether vaccination cicla bacterin. Three pens, each with group II organisms would pro with 26 four-week-old Whit Rock vide cross protection for the more Studies on Control of Fowl Cholera 11 prevalent group I P. multocida in group II CE vaccine, respectively. fections. Group I ( Strain No. 4 ) and The birds were challenged 16 group II ( Strain No. 10 ) CE vac days after the second vaccination cines and BC bacterins were pre with a 1: 10,000 dilution of a 24- pared separately and vaccination hour broth culture of group I P. was carried out by two weekly in multocida ( strain T ) . As shown in jections as before. Five pens of 4- figure 7, the challenge dose resulted week-old New Hampshire chickens in 100% mortality in the control were used, pen 5 serving as the con group and in pen 3, vaccinated with trol group. Pens 1, 2, 3, and 4, each group II BC bacterin. The death with 28 birds, were vaccinated with loss was complete in those two pens group I BC bacterin, group I CE by the fourth day. A high mortality vaccine, group II BC bacterin, and also occurred in the other three

Figure 7. Immunity from broth culture bacterins and chicken embryo vaccines prepared from group I and II strains.

100 PEN 5 (CONTROL)' ,~ /y \ 90 V / PEN 3 ~ y/ (BROTH BACTERIN - GROUP II) I 80 -, I I ~------== I /~Ett4---r-- I 70 -/ ;;---:=j;::;/-~ (CHICKEN EMBRYO I / VACCINE - GROUP Il) - 60 - - - I-~/- --- _..., ~l-l PEN 2 ---~-----,,~- I (CHICKEN EMBRYO ,,------~1 VACCINE- ,\ t, / jl GROUP I) / .,- ···-·--·---- 1,// /y ... .. ? ... '/ ----·;... --:l4 ------... -······<-c__pEN I __L, I (BROTH I , BACTERIN- - I GROUP I) 1/ I I ,------"' µ I I ,' J ;, /, / { ,// / I_____ ... ---·-1/ .. 0 0 2 3 4 5 6 7 8 9 10 DAYS POST INOCULATION 12 South D akota Experiment Station Technical B ulletin 23 pens over a 10-day period, but the Since these trials were com greater protection was provided in pleted, Heddleston and Hall ( 13 ) p ns 1 and 2 which received the reported more favorable results in group I preparations. The survival limited trials on fowl cholera im again was slightly better in p n 1, munization with a bacterin con which received the group I BC bac taining an adjuvant. Not only was terin. A similar trial using a group the immunity stronger but also of II P. multocida challenge was not longer duration than that provided made. Yaw, Briefman, and Kakavas by a chicken embryo vaccine. ( 29) have sugg sted that the im Treatment munogenic specificity of types of Pasteurella should be considered Effective treatment of poultry when immunizing agents are pre flocks for fowl cholera was not pared. The results in thjs trial tend possible until sulfonamide drugs to support that belief. became available. A number of workers ( 1, 2, 7, 8, 16, 25 ) found In each of the three trials, there that different sulfonamides wer was a high incidence of crippled effective in reducing mortality. and sick birds among the survivors They were of particular value in at the termination of the experi acute outbreaks if treatment was ments, which further reflects the started early. However, consider low level of protection provided by able death loss usually occurred be these immunizing agents. fore the drug became effective. The fourth trial dealing with im Prolonged medication produced munization involved a group of 72 toxic reactions and it was found 5-month old pullets. Thirty-eight of necessary to stop treatment every them received two weekly injec few days to avoid this. During the tions of BC bacterin. Seven weeks period when the flock was not following the second injection, a treated, the disease often recurred, simulated fowl cholera outbreak making it necessary to sta1t treat was initiated in the pull ets by plac ment again. Some flocks do not re ing with them six birds which were spond well to sulfonamide treat swabbed in the palatine cleft with ment, particularly after one or two a virulent culture of P. multocida courses fail to check the disease. ( Strain S). All six of the swabbed Outbreaks of a chronic nature sel birds had died of fowl cholera dom respond. Other workers ( 3, within 48 hours. Losses began to 14) reported that certain sulfona occur in the exposed birds 24 hours mides cause a marked drop in egg later and continued through the fol production. Despite these disad lowing 30 days. Thirty of the 34 vantages, the sulfonamides have ( 88%) nonvaccinated and 24 of the been the drugs chiefly used against 38 ( 63%) vaccinated birds died. cholera. Again, th slight protection which Antibiotics have not been used app ared to have been provided extensively to treat flocks affect d was not of practical value. with the disease because of their Studies on Control of Fowl Cholera 13 relatively high cost. Antibiotics are compared penicillin, streptomycin, used widely in poultry production oxytetracycline, sulfamerazine, and as growth stimulants and for a sulfaquinoxaline. The two sulfona favorable effect on egg production. mid s, shown to be effective against The cost is not high for this pur cholera, were included as a basis for pose, as only small amounts of anti comparison. The antibiotics were biotics are needed and generally mixed in the mash at a level of 1.0 the more economical residues are gram per kilogram ( 1: 1,000 ). Sul used. Several tiials using penicillin, famerazine was used at a 0.5% and streptomycin, and chlortetracycline sulfaquinoxaline at 0.05% level in (Aureomycin) at growth promot the mash. ing levels were run at this station to Treatment was started about 48 determine whether they would pro hours before the birds were chal tect against experimental fowl chol lenged. Each treatment was used era. Results were irregular when on approximately 65 seven-week trials were compared but it was old chickens. A pen wit.h a similar concluded that such low levels of number of untreated birds was the these antibiotics were of no value in control. The birds were challenged preventing or controlling outbreaks by intramuscular inoculation with of the disease. 1.0 milliliter of a 1: 10,000 dilution McNeil and Hinshaw ( 17 ) re of a 24-hour tryptose phosphate ported that streptomycin complete broth culture of P. multocida ( strain ly inhibited the growth of P. multo 18 ). An LD50 test, usjng brooder cida in vitro. The antibiotic also mates, indicated that each bird re prevented mortality in experimen ceived approximately 100 LD50 tal fowl cholera in turkeys, but the doses. protection was not complete. Prier As shown in figure 8, oxytet ( 20) found that chlortetracycline racycline and both sulfonamides was more effective against P. multo afforded c o m p 1 e t e protection cida in vitro than penicillin or against the challenge. Penicillin and streptomycin. He also found that streptomycin were not effective; chlortetracycline, given at the rate the birds receiving these antibiotics of 250 and 500 milligrams per kilo djed about the same rate as the gram of mash, materially reduced controls. the mortality rate in chickens to ex Similar trials have been run with perimental infection. Gualandi ( 12 ) chlortetracycline, tetracycline, and used oxytetracycline in the mash a furazolidone concentrate ( NF- successfully against experimental 180 ). Chlortetracycline was com Pasteurella infection in birds. pared to oxytetracycline at 1 gram Trials were conducted at this sta and 0.5 gram per kilogram of mash. tion to determine the efficacy and The 1: 2,000 level of oxytetracycline the level at which some common provided practically as much pro antibiotics would be effective tection as the 1: 1,000 level of chlor against fowl cholera. The first trial tetracycline against xperimental 14 South D akr:ta E xp eriment Station Technical Bulletin 23 100 I . 1 = 90 _:.---- I I ~- - '\EN , (CONTROL) 1~--+--cf J 80 - ~ ~-' I ,,., ... ,------I I I 'cl PEN 6 (STREPTOMYCfN ~ ::>' - ~70 / -- TREATMENT)

~ t: 60 - ...J r/!,,, / ~ / 1· ~50 - ~ r"~ PJN 2 (PENICILLIN TREATMENT) ''/ ~40 -

~ J. t I 530 - ~ => 820 - <3: - i . rEN 3 (OXYTETRACYCLiNE TREATMENT) ii/ - 10 PEN 4 (SULFAMERAZINE TREATMENT) \ fEN ~ (SU~FAQU!NOX~LINE TRE~TMENT) 11 I 1 .. 0 0 2 3 4 5 6 7 8 9 IO II 12-20 DAYS POST INOCULATION Figure 8. The efficacy of differen't treatments against fowl cholera.

infection ( figure 9 ). One-half gram Oxytetracycline checked losses in of pure tetracycline p r kilogram of a simulated natural outbreak of mash was ineffective against experi cholera in a pullet flo ck. The out mental fowl cholera. This may have break was started in the 103-bird been due to faulty mixing as a very flock by swabbing seven birds in small amount was used, compared the palatine cleft with a virulent to the relatively large amount of P. multocida culture ( Strain G ). residues of oxytetracycline and When 20 birds had died the re chlortetracycline that was needed mainder of the flock was divided to prepare a similar concentration. into two nearly equal groups by A soluble oxytetracycline prepa partitioning the pen. The birds on ration used in the drinking water at one side of the pen were treated 0.5 gram per liter was more effec with oxytetracycline in the mash at tive against the experimental dis the 1: 1,000 level for 9 days and at ease than, 0.033%furazolidone in the 1:2,000 for 11 succeeding days. mash. The challenge in this tria] Losses stopped in the treated was so severe that 100% of the con group the first day after treatment trol and furazolidone-treated birds was begun and over a period of 50 died, compared to 22% of those days, five ( 12%) of the treated birds treated with oxytetracycline. had died, compared to 33 ( 80%) of Studies on Control of Fowl Cholera 15

100

I I I - 90 ~ EN I (CONTROL)-- I I _ _!_ ___ ------

/ / ---- // ~70 / - / >- / ~ / :J 6 0 / - ~ / Q: / / ~50 / - / / / / - ~40 / .=:; / / <[ / ~ (CH LORTETRACYCLINE TREATMENT- .I_ 5 3 0 / ~ I => I I I j I -~~->. _ J_ j_ = ..... I --, ------:- I 820 I I I I <[ I JT PEN2 (CH LORTETRACYCLINE TREATMENT-I: 1000) I I I I I I I I I I I 10 - - I / ,,V-} PE-~~~;-~~:1TETRf CYC11N~--~~-~~~~ ~-~~~:~=~~J~~ = / / 1 -- 1 --- - ' I ' I I I I I l~i _..y:_ ___ )--- •• ,-PEN 4 (OXYJETRACYCLINE TREATMENT- I: 1000) 0 0 2 3 4 5 6 7 8 9 10 11 12-20. DAYS POST INOCULATION Figure 9. The efficacy of two levels of chlortetracycline and oxytetracycline against fowl cholera. the untreated group. No toxic about one per day for 5 days and ffects were observed during the 20 stopped. The disease recurred about days of treatment and egg produc 2~f months after treatment was tion was not depressed. stopped. Losses stopp d in the Oxytetracycline was used to treat third flock in 2 days and did not six farm flocks affected with chol recur. The fourth flock was blood era. Four of these flocks had been tested with a P. multocida antigen treated unsuccessfully with sulfona when the disease recurr d after a mides. Most of the flocks were course of treatment. The reactors treated with oxytetracycline in the were removed and the flo ck was mash at the 1:1,000 level for 3 days, given a second course of treatment. then at the 1:2,000 level for 3 days . Losses stopped without r currence. Losses stopped in the first flock Losses in the fifth flock were r - the first day after treatment was duced but continued at about one started and did not recur. In the per day during a cours of treat second flock losses dropped from 18 ment. A check revealed that the birds the first day of treatment to birds were getting grain in addi three the second day and continued tion to the treated mash and that 16 South Dakota Experiment Statiou Tech1tical Bulletin 23 so e old hens had been kept from isolat d in South Dakota belong to th previous flock. These were group I. The strains of both groups culled and losses stopped after t~e ar qually virulent. first day when they received only A whole-blood agglutination test treated mash. The outbreak did was developed and used to detect not recur. The sixth flock was han chronic carriers of fow 1 cholera. dled in about the same manner as Although studies showed that some the fourth flock. Although treat of the reactors were carriers, it also m nt stopped losses, the disease was found that some nonreactors re urred even after a blood test was carried the infection in the upper conducted and the reactors were respiratory tract. It is doubtful that removed. These results will have to removing reactor carriers would b considered with reservation as halt cholera outbreaks as long as the farm facilities did not provide noni-eactor carriers remain sources for establishing control groups. of infection. Smith ( 27 ) reported that one in A satisfactory immunizing agent tramuscular injection of 25 milli for fowl cholera was not developed. grams of oxytetracycline per kilo Results tended to support the pro gram of body weight was usually posal that consideration should be effective against experimental fowl given to immunogenic specificity of cholera in chicks and chickens. A th two main groups of P. multocicla limited trial was conducted at this when immunizing agents are pre laboratory with an experimental pared. injectable form of oxytetracyclim: Oxytetracycline (Terr am y c in) for poultry. None of 20 birds was the most effective antibiotic treated with this preparation was tested against experimental fowl affected by a challenge inoculation cholera. It was as effective as the of P. multocicla that killed 70% of a sulfonamides experimentally and similar control group. This type of also was effective in fi eld trials treatment might prove of value for where sulfonamides had failed. Al checking early losses in acute fowl though oxytetracycline offers addi chol ra outbreaks. tional advantages, it i not used ex tensively against fowl cholera be SUMMARY AND CONCLUSIONS cause of the cost. Most outbreaks of fowl cholera in BIBLIOGRAPHY South Dakota poultry flocks are acute and cause heavy death loss. l. ALBERTS, J. 0. "The prophylac Chronic outbreaks are less common. tic and therapeutic properties of The disease is caused by strains· sulfamerazine in fowl cholera." American Journal of Veterinary of P. multocida that belong to at least t w o different serological Research, XI:414-420. 1950. groups. These groups can be identi 2. ARMSTRO G, W. H. and R. A. fied roughly by biochemical reac BANKOWSKI. "Sulfaquinoxaline tions. Over 80% of the avian strains treatment in an acute outbreak of fowl cholera." Veterinary South Dakota Farm and Home Medicine, 44:306-307. 1949. Research, 111:23-24. 1951. 3. BANKOWSKI, R. A. "A decrease 10. ----- "Fowl cholera." South in egg production following sul Dakota Farm and Home Re famerazine and sulfamethazine search, IX: 10-13. 1958. medication." Journal of the 11. GoGOASA, V. "Frecventa purta American V ete1'inary Medical torilor de Pasteurella intr-o en Association, 113:49-50. 1948. zootie de holera aviara la rate 4. CARTER, G. R. "Studies on a in plina evolutie." Probl. Epiz. Pasteurella multocida chicken Microbial., Inst. Pat. Igien. embryo vaccine I. The compara Anim., Bucresti, 5: 13-18. 1956. tive immunizing value of broth bacterins and a chicken embryo 12. GuALANDI, G. "La terramicina vaccine in mice." American nello lotta contro il colera avi Journal of Veterinary Research, are." Arch. Vet. Ital., 4:539-543. XI:252-255. 1950. 1953. 5. CERNAIANU, C. "Ueber die Gel 13. HEDDLESTON, K. L. and W. J. Hugel cholera, sine durch Hal HALL. "Studies on pasteurellosis tung, Emahrung a n d Aus II. Comparative efficiency of bentung bedingte Krankheit." killed vaccines against fowl Zeitschr. Infektionskrankh. par cholera in chickens." Avian Dis asitaire Krank. u. Hyg. Haus eases, 2:322-335. 1958. tiere, 58:142-159. 1942. 14. HINSHAW, W. R. and E. Mc 6. DAUGHERTY, E. 3rd. "The effica NEIL. "Experiments with sulfa cy of several immunizing agents nilamide for turkeys." Poultry for the control of fowl cholera in Science, 22:291-294. 1943. the White Pekin duck." Cornell 15. HUGHES, THOMAS P. and IDA W. Veterinarian, XLID : 421 - 426. PRITCHETT. "The epideiniology 1953. of fowl cholera. III. Portal of en 7. DELAPLANE, JoHN P. "Sulfaqujn try of P. avicida; reaction of oxaline in preventing upper host." I ournal of Experimental respiratory infection of chickens Medicine, 51:239. 1930. inoculated with infective field 16. KlsER, J. s., J. PmER, C. A. BOT material containing Pasteurella TORFF, and L. M. GREENE. "Treat avicida." American Journal of ment of experimental and natu Veterinary Research, VI: 207- rally occurring fowl cholera with 208. 1945. sulfamethazine." Poultry Sci 8. DELAPLANE, J. P. and T. C. HIG ence, 27:257-262. 1948. GINS. "Sulfaquinoxaline in the 17. McNEIL, ETHEL and W. R. HIN prevention and control of chron SHAW. "The effect of streptomy ic fowl cholera." Cornell Veter .cin on Pasteurella multocida in inarian, 38: 267-272. 1948. vitro and on fowl cholera in tur 9. DoRSEY, T. A. "Vaccination and keys." Cornell Veterinarian, 38: blood tests for fowl cholera." 239-246. 1948. 18. NOBREGA, R. and R. C. BUENO. hemorrhagic septicemia Pas ( Inst. Biol., Sao Paulo.) "Sohre teurellae." Journal of Bacterio.Z o reconhecimento de porto ogy, 37:69-89. 1939. dores na colera aviaria." Arq. 25. SAURAT, P. "De utilisation des Inrt. Biol. ( Sao Paulo.) 15:339- sulfamides dans la pasteurellose 342. 1944. aviaire. Leu association aves la 19. PASTEUR, L. "De I' attenuation vaccination." Rev. Med. Vet., du virus du cholera des poules." Lyon et Toulouse, 95: 106-115. Compt. Rend. Acad. Sci., 91: 1944. 673. 1880. 26. SHOOK, WARREN H. and HUBERT BUNYEA. "The detection of car 20. PRIER, J. E. "The in vivo and in riers of fowl cholera and its con vitro effect of aureomycin upon trol by means of a stained-anti Pasteurella multocida." Veteri gen, rapid, whole-blood test." nary Medicine, 45 : 243 - 245. Poultry Science, 18:146. 1939. 1950. 27. SMITH, H. WILLIAMS. "The 21. PRITCHETI, IDA w., F. R. BEAU chemotherapy of experimental DETTE, and THOMAS R. HUGHES. fowl cholera in fowls (Gallus 'The epidemiology of fowl chol Domesticus)." Journal of Com era. IV. Field observations of parative Pathology and Thera the spontaneous disease." Jour peutics, 65:309-316. 1955. nal of Experimental Medicine, 28. VAN Es, L. and F. OLNEY. 51:249. 1930. J. "An inquiry into the influence of 22. ----- 'The Epidemiology of environment on the incidence of fowl cholera. V. Further field poultry diseases." University of observations of the spontaneous Nebraska Agricultural Experi disease." Journal of Experimen ment Station. Research Bulle tal Medicine, 51:259. 1930. tin, 118: 17-21. 1940. 23. REIS, J. and T. C. BUENO. ( Inst. 29. YAw, K., L. BRIEFMAN, and J.C. Biol., Sao Paulo. ) "Vacinacao e KAKA VAS. "A comparison of vir mortalidade na colera aviaria." ulence for mice and chickens of Arq. Inst. Biol. ( Sao Paulo.) different colonial variants of the 4:81-86. 1943. three serological types of Pas teurella multocida." American 24. RosENBUSCH, CARLos T. and I. Journal of Veterinary Research, A. MERCHANT. "A study of the XVII:157-159. 1956.

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