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Diaz, R., and N. Bosseray (1974) Effect of infection with B. abortus on hypersensitivity and hyperreactivity of the mouse to endotoxin of Brucella melitensis, Yersinja enterocolitica and Escherichia coli. Annales de Recherches Veterinaires 5:41-46. Frasch, C.E., ReM. McNelis, and E.C. Gotschlich (1976) Strain-specific variation in the protein and lipopolysaccharide composition of the group B meningococcal outer membrane. Journal of Bacteriology 127:3:973-981. Frasch, C.E., and E.C. Gotschlich (1974) An outer memkrrane protein of Neisseria menjngitidis group B responsible for serotype specificity. Journal of Experimental Medicine 140:87-104. Freeman, BeA., JeR. McGhee, and R.E. Baughn (1970) Some : physical, chemical, and taxonomic features of the soluble antigens of the Brucellae. Journal of Infectious Diseases 121: 522-527. Hase, S., and E. Th. Rietschel (1976) Isolation and analysis of the lipid A backbone. European Journal of Biochemistry 63: 101-107. Hinsdill, R.D., and D.T. Berman (1967) Antigens of Brucella abortus I. Chemical and immunoelectrophoretic characterization. Journal of Bacteriology 93:544-549, Hinsdill, R.D., and D.T. Berman (1968) Antigens of Brucella abortus. II. Toxicity for macrophages in culture. Journal of Infectious Diseases 118:307-316. Hurvell, B., P. Ahvonen, and E. Thal (1971) Serological cross-reactions Lketween different Brucella species and Yersinia enterocolitica. Agglutination and complement fixation. Acta Veterinaria Scandinavia 12:86-94. Hurvell, B., and A.A. Lindberg (1973) Serological cross- reactions between different Brucella species and Yersinia enterocolitica. Immunochemical studies on phenol-water extracted lipopolysaccharides from Brucella abortus and Yersinia enterocolitica type IX. Acta Pathologia et Microbiologia Scandinavia Section B. 81:113-119. Jones, L.M., D.T. Berman, and R. Diaz (1973a) Factors influencing biometric potency assays of Brucella allergens in guinea pigs. British Journal of Experimental Pathology 54: 368-379. | 56
Jones, L.M., R. Diaz, and A.G. Tayler (1973b) Characterizations of allergens prepared from smooth and rough strains of Brucella melitensis. British Journal of Experimental Pathology 54:492-508. Jones, L-M. (1974) Specificity of Brucella protein antigens and role of lipopolysaccharide antigens in eliciting delayed hypersensitivity reactions in sensitized guinea pigs. Annales de Recherches Vétérinaires 5: 189-199. Jones, L.M., and D.T. Berman (1975) Antibody-mediated and delayed-type hypersensitivity reactions to Brucella skin test antigens in guinea pigs. Infection and Immunology 11:3360-364. Jones, L.M.e, and D.T. Berman (1976) Studies of Brucella lipopolysaccharide. International Symposium on Brucellosis (II), Developments in Biological Standardization 31:62-67. Jones, L.M., R-e Diaz, and D.T. Berman (1976) Endotoxic activity of rough organisms of Brucella species. Infection and Immunology 13:1638-1641. Jones, L.M. (1976) A potency assay for Brucella allergens in guinea pigs. International Symposium on Brucellosis (II), Developments in Biological Standardization 31: 165- 170. Kiss, Z. (1971) Experimental study of the Duphavac N.A. (BR45/20) brucellosis vaccine. I. Brucellosis responses of animals in brucellosis-free herds. Acta Veterinaria (Budapest) 21:171-174. Lehrer, S., and A. Nowotny (1972) Isolation and purification of endotoxin by hydrolytic enzymes. Infection and Immunology 6: 928-933. McCullough, N.B. (1970) Microbial and host factors in the pathogenesis of brucellosis. JIn Mudd, S. Infectious agents and host reactions. W.B. Saunders Co., Philadelphia. Miles, A.A., and N.W. Pirie (1939a) The properties of antigenic preparations from Brucella melitensis. I. Chemical and physical properties of bacterial fractions. British Journal of Experimental Pathology 20:83-98. Miles, A.A., and N.W. Pirie (1939b) The properties of antigenic preparations from Brucella melitensjs. II. Serological properties of the antigens. British Journal of Experimental Pathology 20: 109-121. 57
Miller, J.K., J.I.e Kelly and J.H.G. Roerink (1976) A complement fixation method for quantitative differentiation of reactions to 45/20 vaccine and Brucella infection. Veterinary Record 98:210-214. Myers, D.M., LeM. Jones, and V.M. Varela-Diaz (1972) Studies of antigens for complement fixation and gel diffusion tests in the diagnosis of infections caused by Brucella Ovis and other Brucella. Applied Microbiology 23:894- 902. Nowotny, A. (1971) Chemical and biological heterogeneity of endotoxins in microbial toxins, Vol. 4 Academic Press, New York, N. Y. Pe 309-329. Olitzki, A. (1970) Immunological methods in brucellosis research. Part 1. JIn vitro procedures. S. Karger, New York, N.Y. Pe 1-59. Raff, R.A., and R.W. Wheat (1968) Carbohydrate composition of the phenol soluble lipopolysaccharides of Cjtrobacter freundji. Journal of Bacteriology 95: 2035-2043. Reid, M.A., and P.R. Harvey (1972) The use of Brucella abortus 45/20 adjuvant vaccine as a diagnostic aid in the brucellosis eradication campaign in Papua, New Guinea. Australian Veterinary Journal 48:495-499. Renoux, Ge, Me Renoux, and R. Tinelli (1973) Phenol-water fractions from smooth Brucella abortus and Brucella melitensis: immunochemical analysis and biologic behavior. Journal of Infectious Diseases 127: 139-148. Roerink, J.H.G. (1967) Investigation into usefulness of the non-agglutinogenic Brucella abortus adjuvant vaccine Duphavac N.A. in the control of bovine brucellosis. Veterinary Record 80;727-733. Saunders, G.C., and E.H. Clinard (1976) Rapid micromethod of screening for antibodies to disease agents using the indirect enzyme-labeled antibody test. Journal of Clinical Microbiology 3(6) :604-608. Schnaitman, C.A. (1974) Outer membrane proteins of Escherichia coli IV. Differences in outer membrane proteins due to strain and culture differences. Journal of Bacteriology 118:454-464. Spink, W.W. (1956) The Nature of Brucellosis. University of Minnesota Press. 58
Sterne, M., G. Trim, and E.S. Broughton (1971) Immunization of laboratory animals with non-agglutinogenic extracts of Brucella abortus Strain 45/20. Journal of Medical Microbiology 4:185-194. Swanson, J., G. King, and B. Zeligs (1975) Studies on gonococcus infection. VIII. 12S5Iodine labeling of gonococci and studies on their in vitro interactions with eukaryotic cells. Infection and Immunity 11:453- 459. Volk, We (1966) Cell wall lipopolysaccharides from Xanthomonas species. Journal of Bacteriology 91:39-42. 59
CHAPTER 6 PREVENTION STRAIN 19 VACCINE In 1919 the United States Bureau of Animal Industry (BAI) permitted the licensing of commercial biological concerns to produce and distribute Brucella vaccine. Vaccination on a large scale was carried on in the U.S. and in other countries with virulent cultures of B. abortus. Several institutions in various parts of the world, but especially in the U.S., conducted studies on field and experimental herd vaccination. Generally, results indicated the subcutaneous injection of the virulent cultures of B. abortus in non-pregnant cows would protect the majority of vaccinated animals so that they would not abort in subsequent pregnancies. Investigations by Cotton (1932) and Haring and Traum (1937) pointed out that in vaccinating lactating cows, B. abortus frequently became established in the udder and the vaccinated animals became carriers, shedders, and spreaders of virulent B. abortus. Furthermore, during the period when live virulent cultures were being used, several reports indicated that B. sujs had been isolated from cow's milk, and BAI found vaccines on the market which contained B. suis (Mohler et al. 1941). It was mainly for this reason that the use of virulent B. abortus cultures was discouraged, and in 1932 the BAI (Mohler et al. 1941) no longer permitted the use of live B. abortus cultures in the production of vaccine, excepting those made from BAI Strain 19, or some other strain of low virulence. Buck (1930) administered 20 cc of a three-day arowth (Gates nephelometer reading of 1 cm) of B. abortus, Strain 19, subcutaneously to each of three calves 5-8 months of age. Each of the three vaccinates and five controls were bred and at the beginning of the fourth month of pregnancy, they were given approximately 150 cc of fetal stomach and intestinal contents orally. The fetus was from an infected herd and B. abortus was isolated from the exposure material. The three Strain 19 vaccinates calved normally, while three of five non-vaccinated controls became infected. Buck 61
concluded that the immunity afforded by early vaccination continued through the second gestation, although it may have been somewhat reinforced by ingestion exposures. In follow-up experiments, Cotton et al. (1933), and Buck et al. (1934) concluded that a strain of B. abortus of low virulence (Strain 19) conferred as much, or better, immunity to calves against conjunctival exposure during their first pregnancy as more virulent strains. In subsequent experiments, Buck et al. (1938), Haring and Traum (1937, 1943) and Haring (1938) verified earlier results and concluded that Strain 19 vaccine engendered distinct immunity when administered to calves and this immunity persisted when the animals matured. It was also reported that certain objectionable features which accompanied the vaccination of unbred cows and heifers near breeding age were eliminated by the administration of the vaccine in animals 4-8 months of age. Following Buck's discovery of Strain 19 as a promising immunizing agent, Cotton et al. (1934a, b) made a number of experiments to further determine its efficacy. A total of 70 calves between four and eight months of age were vaccinated, of which only eight (11.4 percent) became infected following severe exposure during their first pregnancy. Only one abortion occurred that could be traced to B. abortus. Of 73 control animals, 57 (78 percent) became infected, of which 55 (75 percent) aborted. The BAI in 1936 carried out a large-scale field trial with vaccine prepared from Strain 19 (Mohler et al. 1941). The study involved 260 infected herds in 24 states. Of the 8,182 calves vaccinated, 7,872 or 96.2 percent calved normally. On the basis of the blood agglutination test, only 1.6 percent of the abortions involved in three pregnancies could be attributed to brucellosis. These results were so encouraging that calfhood vaccination was adopted as an adjunct to the official "Test and Slaughter" policy. Characterization of Strain 19 Characterization studies by Brown et al. (1972) indicated that Strain 19 cultures differed from Strain 2308 (biotype 1) and most field strains, in that they did not grow on media containing thionin blue (1:500,000) or penicillin (5y/ml). The âoriginalâ Strain 19 cultures dispensed prior to 1956 varied in their ability to grow on medium containing erythritol (1 mg/ml) and in their ability to oxidize erythritol. Seed cultures dispensed after 1956 did not grow on medium containing erythritol and were unable 62
to oxidize erythritol, whereas B. abortus Strain 2308 grew very well on medium containing erythritol and its rate of Oxidation (QO2N) of erythritol was consistently over 300. There were consistent quantitative differences between Strain 19 and other strains of B. abortus in the oxidative rates on certain substrates. The oxidation rate of Strain 19 on L-alanine was approximately double the rate on D- alanine; the rate (Q0OsN) on L-glutamic acid was invariably over 500. The rates of d¢+)-galactose and d-ribose were approximately equal. Collectively, the pattern of utilization of these substrates could be used as an additional criterion for differentiation of Strain 19 and field strains of B. abortus. Strain 19, like various organisms, is capable of existing in several variant forms having differing immunizing values (Henry 1933; Mingle and Manthei 1941) and this presents difficult problems incident to the manufacture of the vaccine. The smooth, highly antigenic type is considered the most desirable for vaccine production. Irregularities in technique, in the culture media used, and the dissociative potentialities of various substrains are all factors influencing the possible dissociation of vaccine serials. Strain 19 vaccine is a suspension of live organisms in 0.85 percent sodium chloride solution. The fact that the Organisms are alive and maintained in a medium supplying no nutrients makes the vaccine a highly perishable product subject to deterioration if handled under adverse conditions (Love and Mingle 1941). The USDA supplies the biological companies with cultures of Strain 19 at various intervals and the vaccine is prepared from these cultures. Every lot of the vaccine is tested by USDA before being released for distribution. It should contain at least 10 billion viable organisms per cubic centimeter on initial test and not less than 5 billion at the expiration date. It should contain no more than 5 percent dissociates. Strain 19 vaccine is available in liquid or lyophilized forms. The advantage of lyophilized over liquid vaccines is that the dried form is more stable under varying temperature and storage conditions which adversely affect liquid vaccine. Unfortunately, in itsS preparation a 50 percent loss occurs in the original concentration of cells. This has led the manufacturers to compensate by the addition of Many extra cells thereby yielding a final dose with at least the required number of living cells but also containing many dead cells that contribute endotoxins to the dose. This 63
results in endotoxin~-like disease and death that has been observed in four month old calves. Pathogenicity of Strain 19 Strain 19 differs from more virulent strains in that even in large doses it produces in guinea pigs no more than a Slight enlargement of the spleen and a blood serum agglutinating titer seldom over 1:50, and organisms are rarely found in the spleen six weeks after injection. In cattle, Strain 19 did not become localized in the udder in lactating cattle excerpt in rare cases where high doses were injected into pregnant cows (Haring and Traum | 1937, Haring 1938, Traum and Maderious 1947). Mingle et al. (1941) failed to increase virulence of Strain 19 in six serial passages through normal rfregnant animals by intravenous injections of very large doses. These injections did produce abortions in which Strain 19 was isolated. Controlâ pregnant cattle in close contact with vaginal discharges, rlacentas and fetuses showed no more than a transient blood agglutination titer of 1:25. These investigators also passed Strain 19 through a series of guinea pigs without changing its virulence. Birch et al. (1943) produced brucellosis syndrome in only one of eight pregnant cows and transient agglutination titers of 1:100 in fcur other cows by feeding large doses of Strain 19 for 3-4 months. Seven of eight cows calved normally with no evidence of infection of Strain 19 in the genital tracts or milk. The seven cows were in close contact with the only cow which aborted and was shedding Strain 19 organisms. Jones et al. (1965) studied over 100 cultures of Brucella sp. in order to determine the characteristic of carbon dioxide-independent cultures isolated from cattle vaccinated with Strain 19. They reported that Strain 19 and 19 of 21 cultures isolated from the milk of vaccinated cows were inhibited by erythritol, thionin-blue, penicillin and Saffrinin O. Only virulent Strain 2308 grew in the presence of all four substances, suggesting that all four substances be used to check batches of vaccines and identify carbkon dioxide-independent cultures suspected of originating in the vaccine. Meyer and Nelson (1969) reported that residual localization of Strain 19 organisms occurred following immunization of female cattle, but was infrequent in occurrence. The residual infection involved the mammary glands primarily and did not spread or involve the reproductive tract. Residual localization of the organisms 64
following immunization was not found to be entirely attributable to overage vaccination. Heifers that maintained a post-immunization infection into sexual maturity remained chronically infected and excreted the Organisms via the mammary gland for long periods of time. Danks (1943) reported the occurrence of orchitis and subsequent infertility in a bull previously vaccinated at Six months of age with Strain 19 vaccine. Approximately one year post-vaccination, an organism indistinguishable from Strain 19 was isolated from the epididymis of the right testicle. Lambert et al. (1964) reported the development of orchitis and isolation of Strain 19 organisms from the testicles of two bulls vaccinated at five and six months of age. More recently, Lambert et al. (1965) administered Strain. 19 vaccine to 15 bulls when 4-10 months of age. A mild transient post-vaccinal orchitis was detected in eight bulls. Post-vaccinal blood agglutination titers persisted at diagnostic levels in bulls longer than in heifers vaccinated at the same age in a previous experiment. The bulls were sacrificed at 18 months of age and there was no evidence of gross or microscopic pathologic alterations; Strain 19 was not isolated from any of the tissues. Age of Vaccination The very early work of the USDA investigators suggested that vaccination with Strain 19 be done on cows and heifers before breeding (Cotton et al. 1933 and Cotton 1934). Buck et al. (1938) suggested that use of vaccination be limited to calves 4-8 months of age, mainly to avoid retention of blood agglutination titers caused by the administration of the vaccine in older animals. However, Haring (1938) and Haring et al. (1947) produced evidence that indicated effectiveness of the vaccine was enhanced when administered to animals nearing sexual maturity. The conclusions drawn by Haring (1938) and Haring et al. (1947) were questioned when Manthei et al. (1951) reported that no difference could be shown in the degree of resistance between animals vaccinated as calves and two-year-old animals when challenged to brucellosis during the third gestation. Later Gilman and Wagner (1959) determined there was no significant difference in the immunity induced in calves vaccinated at four and eight months of age. These findings were corroborated by King and Frank (1961), and Lambert et al. (1961), when it was reported that resistance produced in calves vaccinated at three months of age compared favorably with calves vaccinated at nine months of age. In addition, results confirmed earlier work that vaccination at three and four months of age materially reduced the problem of persistent post-vaccinal titers. Redman et al. (1967) reported that resistance to infection of two and three- 65
month-old vaccinated cattle compared favorably with resistance found in cattle vaccinated when 4-8 months of age. The vaccination of adults and yearlings has generally fallen in disfavor because: (1) the blood agglutination test is generally used as the final criterion of infection, (2) carefully controlled research results have indicated that resistance was as effective in cattle vaccinated at three and four months of age as those vaccinated at eight and nine months of age and (3) vaccination at three and four months of age materially reduced the problem of persistent post-vaccinal blood titers. Resistance Conferred by Strain 19 Vaccine A compilation of all data from research at the National Animal Disease Laboratory indicated that 65-75 percent of all vaccinated animals were completely protected against most kinds of exposure (Manthei 1959). Although the remaining 25-35 percent of the vaccinated animals became infected, many did not show clinical signs, such as abortion. This is evidence that there was partial protection in a relatively large percentage of animals other than those that were completely protected. This is comparable to the field results obtained in Montana (Safford 1959) and California (Stuart et al. 1959). For an excellent summary of the Wisconsin experience, retrospectively examined, the reader is referred to the role of living vaccines in prophylaxis by Berman and Jones (1976). The relationshir of infection to various amounts of exposure with highly virulent B. abortus 2308 is shown in Table 1. Curation of Resistance The length of time that resistance induced in cattle with Strain 19 vaccine will protect against virulent B. abortus infection has received considerable attention from livestock owners and disease control officials, since vaccination was included in the federal-state brucellosis control program in 1941. Early studies were conducted under semi-controlled conditions and the results were variable and contradictory. Because of these inconclusive results, it was impossible for disease control officials to give good advice on the length of time calfhood vaccinated cattle would be protected against virulent B. abortus infection and the necessity or desirability of revaccination. 66
TABLE 1 Relationship of Infection to Conjunctival Exposure with Brucella abortus, Strain 2308 ----254 Vaccinates----- --210 Non-vaccinates---- Exposure Dose Infection Abortions Infection Abortions Percent Percent Percent Percent 350,000 0 0 78 56 700,000 26 20 87 54 15,000,000 61 47 100 100 25,000,000 57 44 97 91 75,000,000 61 45 100 92 100,000,000 71 52 100 97 Natural 20 20 56 22 Natural = Contact exposure to infected cattle within a herd where the incidence of brucellosis was maintained at 50 percent or greater. Source: Proceedings, 63d Annual Meeting (1959), U.S. Livestock Sanitary Association, p. 93. 67
Manthei et al. (1951) carried out an extensive project involving 93 calves vaccinated subcutaneously with 5 ml Strain 19 and 59 unvaccinated or control animals. Resistance of various numbers of the animals was challenged during first, second, third, fourth and fifth pregnancies. They found that resistance induced with Strain 19 did not decrease with time. In vaccinated animals that received artificial exposure, the resistance was greatest in the Older animals and the least in younger ones. These findings were further supported by the results of controlled experiments on duration of immunity reported by Goode et al. (1956). This study indicated that there was no decrease in immunity with an increase in age of cattle vaccinated subcutaneously as calves with 5 ml of Strain 19. McDiarmid (1957) concluded from results of a seven-year experiment involving 500 cattle, that one dose of Strain 19 administered at about six months of age conferred adequate immunity for at least five pregnancies, and probably for the normal milking life of the cow. koutes of Administration The use of intracutaneous or intradermal administration of B. aboxytus vaccines as an irmunizing agent for brucellosis was first reported by Cotton et al. (1933). A small number of cattle were vaccinated with 0.5 ml of Strains 11, 19, and 484. Since Strain 19 and Strain 484 became localized in the udder in the intracutaneously- vaccinated animals, Eut not in those vaccinated subcutaneously, it was concluded that the danger of infecting milk causing a public health hazard was so great, the continuance of investigations on intracutaneous adult vaccination was not warranted. Rabstein and Cotton (1942) reported good results in field experiments by vaccinating calves intracutaneously using 0.2 ml of Strain 19. Since the report of that work, several investigators have used that technique in their studies. Campbell and Rodwell (1945) reported that the intracutaneous vaccination produced high agglutination titers in cattle similar to those obtained when 25 times the dose was administered subcutaneously. McDiarmid (1948) found that intracutaneous vaccination of guinea pigs produced as satisfactory immunity as did 25 times the dose given subcutaneously. These observations were confirmed by McDiarmid (1950) in heifers vaccinated at 15-18 months of age and challenged with 150 million organisms during the fifth month of pregnancy. Cotton (1953) reported that the production of opsonins and agglutinins in intracutaneously vaccinated calves was at least equal to, and in many cases greater than, those produced in subcutaneously vaccinated animals where 25 times the dose was used. Agglutinins in both groups were measurable on the fourth day after vaccination, and by the 68
tenth and eleventh days, titers had reached their peaks. At the end of six months, 97.3 percent of the intracutaneously vaccinated heifers were negative, but only 64.8 percent of those vaccinated subcutaneously had no titers. Berman and Beach (1954) found no apparent differences in the titer of agglutinins or the rate of disappearance of agglutinins when sexually mature cattle were vaccinated subcutaneously and intracaudally. There were no apparent differences in resistance between the two groups when challenged by the conjunctival route with 12 x 10° B. abortus Strain 2308 organisms. Results of studies on calves vaccinated 7-10 months of age by injection of 1.0 ml Strain 19 intracaudally or 5.0 ml subcutaneously indicated that the intracaudal method was as effective as the standard subcutaneous method in preventing brucellosis infection (Gregory 1953). In a recent report in which small numbers of cattle were used, Plommet and Fensterbank (1976) indicated that Strain 19 vaccine administered by the conjunctival route was at least as effective as the standard method of vaccination with Strain 19 vaccine. They also indicated that using this method, the vaccine could be administered at any age without risk of producing serological response that would interfere with diagnosis. Adult Vaccination The Subcommittee has examined a "Proposed Protocol for Adult Vaccination with Strain 19 in Brucellosis-Infected Problem Herds" (see text attached) approved by the U.S. Animal Health Association in November, 1976. The proposal has been sent to APHIS for adoption. The Association's action was based upon the paper delivered at the meetings by Dr. Paul Nicoletti ("A Preliminary Report on the Efficiency of Adult Cattle Vaccination Using Strain 19 in Selected Dairy Herds in Florida") and upon earlier controlled and field trials (Berman, Beach, and Irwin, Amer. J. Vet. Res. ,~-15:406, 1954; Moore and Mitchell, Canad. J. Comp. Med., (a) 12:278, 1948; (b) 14:209, 1950; Haring and Traum, Proc. U.S. Livestock San. A., (1043):42.3; Worthington, Mulders, McFarland and Becker, Onderstepoort J. Vet. Res., 40:7, It will be noted that the draft proposal is intended for use in chronically infected and heavily infected herds in high incidence areas and under strict guidelines. (Quarantined herds, branding of vaccinated animals so that 69
they will only move to slaughter as identified animals). It will also be noted that such qualifying dairy herds in Florida and Texas and beef herds in the South, will go to adult vaccination under strict control. The action described above reflects to some degree the conclusions of the NAS Subcommittee on Brucellosis Research where in this report comments were made about adjusting to. certain regional specifications of the brucellosis problem. It will be recalled that earlier studies in this country and abroad noted the effectiveness of Strain 19 vaccine in adult animals and at the same time noted also the persistence of serum titers that would, if the procedure were wide-spread, wreck the existing system whereby blood tests form the very backbone of a control program. Several of the references above spoke to the need, in certain well- specified herd infection situations, to use only for an emergency period the procedure of adult vaccination and further to specify what could be the allowable movement of such animals. It would seem that these precautions have been fully attended to in the draft proposal for what is essentially a special regional dispensation from the national control program. Field trials are inherently difficult to carry out and these difficulties are often compounded by problems of control groups. While criticisms have been voiced of the trial carried out by Dr. Nicoletti in Florida relative to controls (except for herd 5) and too few bacterial cultures drawn from the card test-positive -- CF and Rivanol tests negative animals, it is nevertheless our view that the major conclusions would not have been modified had these faults not been present in the Florida trials. Furthermore, these studies present the first data on titer recession in adults vaccinated with reduced doses and using the supplemented tests. The conclusion drawn by the Subcommittee is that a policy that makes good sense in some states should not be endangered on the grounds of certain weaknesses in Dr. Nicoletti's data since many earlier studies led to the same observations and similar conclusions as the Animal Health Association has recently drawn. 70
PROPOSED _ PROTOCOL FOR _ ADULT VACCINATION WITH STRAIN _19_IN_BRUCELLOSIS_INFECTED PROBL M HER S Herds to be adult vaccinated shall be selected jointly by the State Veterinarian and the Federal Veterinarian responsible for the brucellosis program in the state where the herd is located. Factors to be considered in choosing a herd for adult vaccination shall include the following: 1. Incidence of infection in the herd. 2- Problems in eliminating infection from the herd using test and slaughter procedures prescribed by the UMR. 3. Herd abortion rate. Vaccination, testing and supervision of herds selected shall be the responsibility of a veterinarian delegated by the State Veterinarian and Federal Veterinarian in Charge of the state's brucellosis progran. PROCEDURES: 1. 2e 3e Test all eligible cattle in the herd using UMR approved test(s). Remove reactors for slaughter. Vaccinate negative females with Strain 19 within 10 days of test. Identify all adult vaccinates by placing an âAv" (each letter at least 2 X 2 inches) hot brand on the right jaw and an officially coded eartag in the right ear. Add only cattle from non-quarantined herds which heave passed a negative brucellosis test (UMP approved) within the preceding 10 days. Vaccinate females with Strain 19 vaccine before being placed with the herd. Add only officially vaccinated calves to the herd when the supply becomes adequate, such adequacy to be determined jointly by the State Veterinarian and VIC. Quarantined, adult- vaccinated herds may be combined if jointly approved by the State Veterinarian and VIC. Vaccinate at UMR approved age all heifer calves raised on the farm. 71
Maintain herd in quarantine under UMR approved restrictions. "AV" branded cattle shall move only on permit to slaughter or quarantined feedlot, just as "S" branded cattle, except as provided under 3 above. Test all eligible cattle in herd at intervals not to exceed six (6) months (time may be extended or shortened by joint approval of State Veterinarian and VIC) using UMR approved test(s) and remove reactors. Testing should be continued until herd is eligible for release under the UMR. While the herd is under quarantine, calves under six (6) months of age may move in accordance with the UMR. Ddms found reacting to the UMR approved test shall be moved to slaughter with indemnity. Subject to future consideration by the USAHA Brucellosis Committee, and availability of official calf vaccinates as replacements, an adult-vaccinated herd shall at the end of 24 months following the first vaccination of adults be subject to all UMR requirements which apply to a non~adult vaccinated herd, and no further adult vaccination shall be permitted. However, previously adult-vaccinated animals will remain under restricted movement and special test requirements until sold to slaughter. 72
REFERENCES Berman, D.T., and B.A. Beach (1954) A comparison of the effects of subcutan2ous and intracaudal vaccination of | sexually mature cattle with Brucella abortus Strain 19. American Journal of Veterinary Research 25:406-411. Berman, D.T., and L. Jones (1976) The role of living vaccines in prophylaxis. International Symposium on Brucellosis 2:332-333. S. Kargar & Co., Munich. Birch, R.R., H.-L. Gilman, and W.S. Stone (1943) The pathogenicity of Brucella abortus Strain 19 for sexually mature cows. Cornell Veterinarian 33: 198-208. Brown, G.M., E.L. Love, D.E. Pietz, and C.R. Ranger (1972) Characterization of Brucella abortus Strain 19. American Journal of Veterinary Research 33 (4) 3759-763. Buck, J.M. (1930) Studies of vaccination during calfhood to prevent bovine infectious abortion. Journal of Agricultural Research 41(9) :667-689. Buck, JeM., WE. Cotton, and H.E. Smith (1938) Vaccination of calves and yearlings against Bang's Disease. USDA Technical Bulletin No. 658:1-6. Campbell, A.D. and A.W. Rodwell (1945) The relationship of dosage and site of inoculation to the agglutinin response to Brucella abortus Strain 19 vaccines a comparison of the subcutaneous, intracutaneous and intracaudal: routes. Journal of Comparative Pathology 55:277-289. Cotton, C.M. (1953) An intensive study of post-vaccination responses in groups of calves vaccinated intracutaneously and subcutaneously with Strain 19 Brucella abortus vaccine. American Journal of Veterinary Research 14:337-342. Cotton, W.E. (1932) Efficacy of different strains of Brucella abortus as immunizing agents against infectious abortion. Journal of Agricultural Research 45:12. Cotton, WE. (1934) New researches and developments on contagious abortion (Bang's disease). Proceedings 12th International Veterinary Congress 283-297. 73
Cotton, WeE., JM. Buck, and H.E. Smith (1933) Efficacy and safety of abortion vaccines prepared from Brucella abortus strains of different degrees of virulence. Journal of Agricultural Research 46:4. Cotton, W.E., JM. Buck and HE. Smith (1934a) Studies of five Brucella abortus strains as immunizing agents against Bang's disease (infectious abortion). Journal of the American Veterinary Medical Association 38 (2) $232-247. Cotton, W.E., JeM. Buck, and H.Ew Smith (1934b) Further studies of vaccination during calfhood to prevent Bang's disease. Journal of the American Veterinary Medical Association 38(2):. Danks, A.G. (1943) Byucella abortus Strain 19 and orchitis in a bull. Cornell Veterinarian 33:381-382. Gilman, H.L., and W.C. Wagner (1959) The evaluation of brucellosis vaccination at 4 and 8 months of age. Cornell Veterinarian 49: 399-408. Goode, E.R. Jre, CeA. Manthei, and T.E. Amerault (1956) Further studies on duration of immunity to brucellosis induced in calf-vaccinated cattle with Strain 19 vaccine. Proceedings, 60th Annual Meeting, U.S. Livestock Sanitary Association 89-96. Gregory, T.-S. (1953) A comparison of the effects of intracaudal and subcutaneous vaccination of calves with Brucella abortus Strain 19. 4. A general assessment of results three years after vaccination. Australian Veterinary Journal 29:117-121. Haring, C.M. (1938) Vaccination against Bang's disease in an infected dairy herd with United States Bureau of Animal Industry Brucella abortus Strain 19. Journal of the American Veterinary Medical Association 92:52-60. Haring, C.M. and J. Traum (1937) Observations of pathogenic and antigenic effects of Brucella abortus, United States Bureau of Animal Industry Strain 19. Journal of Agricultural Research IV:117-128. Haring, C.M., and J. Traum (1943) The effect of Brucella abortus Strain 19 on cattle of various ages and its bearing on adult cattle vaccination. Proceedings, United States Livestock Sanitary Association 47:42-46. Haring, C.M., J. Traum, and W.E. Maderious (1947) Vaccination against brucellosis. Journal of the American Veterinary Medical Association 90:103-107. 74
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