Bovine Fetal Inoculations with Calf Rotavirus

The serological and histopathological responses of bovine fetuses to in utero inoculation with virulent and attenuated strains of the calf rotavirus (reovirus-like agent of neonatal calf diarrhea) are described. Thirteen bovine fetuses, 63 to 190 days of gestation, were inoculated in utero with attenuated (three fetuses) or field strain virus (nine fetuses) or both (one fetus).

Serum-neutralizing antibody titers ranging from 1:16 to > 1:256 were detected in six of eight fetuses tested, demonstrating the ability of the bovine fetus to respond immunologically to this agent. The youngest fetus in the series was inoculated at 63 days of gestation and developed a titer of 128 in 64 days. This represents the earliest stage of gestation at which a bovine fetus has been inoculated with a bovine virus and found to produce antibody to it. Serum neutralizing titers in six of the eight dams tested increased significantly following the inoculations of their fetuses in utero.

Histological changes associated with viral replication and antigenic stimulation of the lymphoreticular system were observed. Pneumonic lesions consisting of both local and diffuse lymphoreticular proliferation were present in five of the nine fetuses that were alive at slaughter. Gliosis and perivascular cuffing were noted in the brains of two of these fetuses and meningitis was seen in one. No evidence of teratogenic change was found.

     

Drug Resistance Among Pathogenic Bacteria from Animals in Ontario

Prevalence of antimicrobial drug resistance among over 3000 clinical isolates of animal pathogens in Ontario during 1971-72 has been studied. A high number of multiple resistance patterns is prevalent among members of Enterobacteriaceae, especially Escherichia coli and Salmonella typhimurium. The most common resistance pattern among bovine strains was against not less than six drugs in common use. Among different animal species the bovine population was found to be the source of a high percentage of chloramphenicol resistant E. coli and S. typhimurium organisms. All the isolates resistant to this drug were in addition resistant to three or more other antibiotics of established therapeutic value. All the multiple resistant isolates of S. typhimurium tested had R factors and they transferred most of the resistance determinants including that for chloramphenicol to a recipient E. coli in vitro.

Penicillin resistance among Staphylococcus aureus of bovine mammary origin did not appear to be high. Antimicrobial drug susceptibility patterns of staphylococci, streptococci and Corynebacterium sp indicate effective in vitro activity with many antimicrobial agents.

     

Evaluation of a Macroscopic Plate Test and an Indirect Immunofluorescence Test to Detect Leptospiral Antibodies in Bovine Serum

A macroscopic plate test was found to be reliable and convenient for detection of Leptospira serotype pomona antibodies in bovine sera. However, the procedure was unreliable for detecting L. serotype canicola antibodies because of false positive reactions. An indirect immunofluorescence test and the microscopic agglutination test provided comparable results and they effectively detected serotype pomona antibodies in bovine sera.     

Escherichia coli Agglutinins in Cow Serum, Colostrum and the Nursing Calf

Immunochemical properties of Escherichia coli O antibodies present in bovine serum and colostrum were investigated. Dam and calf serum samples plus colostral whey samples were fractionated by gel filtration, and the 7S and 19S fractions isolated. Antibody activity against the O antigens of four recognized E. coli bovine pathogens was determined by the indirect hemagglutination test on the whole serum and colostral whey samples and the 7S and 19S fractions thereof. Mercaptoethanol reduction was used to chemically study the immunochemistry of the E. coli O antibodies.

The E. coli O antibodies in dam serum were entirely 19S macroglobulins and appeared to be IgM immunoglobulins. The antibodies in colostrum and calf serum were both 7S and 19S globulins. Reasons for believing these 7S antibodies may be IgG, and the 19S antibodies IgA, immunoglobulins are presented.

     

A Comparison of Various Haemophilus somnus Strains

Sixty-eight Haemophilus somnus strains isolated from the bovine in Canada and the U.S.A. were compared. In media enriched with 5% ovine serum, 5% bovine serum and 10% yeast extract, H. somnus fermented glucose, levulose, maltose, mannitol, mannose, sorbitol, trehalose and xylose, but failed to ferment arabinose, dulcitol, galactose, inositol, lactose, raffinose, rhamnose, salicin and sucrose. The organisms acidified litmus milk, produced cytochrome oxidase, indole and hydrogen sulfide (H₂S) and reduced nitrates to nitrites. The motility, methyl-red, acetylmethyl-carbinol urease catalase, citrate, malonate, lysine, ornithine and arginine tests were negative. Haemophilus somnus was resistant to lincomycin, neomycin and triple sulfa, but susceptible to ampicillin, chloramphenicol, streptomycin, penicillin and tetracycline. No antigenic differences were noted between strains when tested against rabbit antisera of eight strains using agglutination, complement-fixation, immunodiffusion and counterimmunoelectrophoresis tests. Low titre cross-reactions were found in the agglutination tests with some of the anti-H. somnus rabbit sera with Actinobacillus lignieresi and Moraxella bovis. No distinct antigenic similarities to nine other species of pathogenic bacteria of animal origin were found. No difference was observed between H. somnus isolates from Ontario and those from western Canada and the U.S.A.     

Epizootiologic Studies of Shipping Fever of Cattle: I. The Microbial Agents Isolated

Nineteen strains of Pasteurella spp., but no viruses cytopathogenic for bovine embryonic kidney cells were isolated from pneumonic lesions present in “normal” veal calves at slaughter.

In studies on two herds of native cattle and six lots of western feeder calves, Pasteurella spp. were isolated from nasal swabs from healthy cattle and those with shipping fever. Viruses of the psittacosis-lymphogranuloma group were isolated from nasal swabs from animals in five groups. Viruses provisionally identified as bovine enteroviruses were isolated from nasal swabs of calves in two lots.

There was serologic evidence of a temporal association of myxovirus para-influenza 3 (PI3) with shipping fever in three lots of calves. From two of these three lots, strains of PI3 were isolated from ten animals, four of which had clinical shipping fever at the time of virus isolation.

     

A Three Year Comparison of Acute Respiratory Disease, Shrink and Weight Gain in Preconditioned and Non-preconditioned Illinois Beef Calves Sold at the Same Auction and Mixed in a Feedlot

During 1969 to 1971, 78 preconditioned (PC) and 79 non-preconditioned (NPC) beef calves were purchased at the same auction and mixed in a feedlot. Preconditioned calves were weaned 30 days before the sale, used to drinking from a tank, and vaccinated against blackleg, malignant edema, infectious bovine rhinotracheitis (IBR), parainfluenza-3 (PI3) and bovine virus diarrhea (BVD) in 1970 and 1971, and Pasteurella hemolytica and multocida in 1971. All vaccinations were completed two to three weeks before the sale. PC calves were given thiabenzadole. PC calves had significantly less shrink after shipment and in 1971 significantly more rapid daily gain during the first weeks of the feeding period. In 1969 more PC calves were treated for acute respiratory disease than NPC calves during an outbreak of PI3 and BVD infection. In 1970 and 1971 fewer PC than NPC calves were treated for acute respiratory tract disease during outbreaks of PI3 infection. The differences in clinical respiratory disease were significant in 1971. Inclusion of two doses of P. hemolytica and P. multocida bacterin before the sale in 1971 and use of an intranasal PI3 vaccine was considered to improve the PC program. Fecal egg counts for gastrointestinal nematodes were much lower in PC calves treated with thiabenzadole than untreated NPC calves.     

A Mycoplasma Isolated from Cattle with Infectious Bovine Keratoconjunctivitis

A mycoplasma has been recovered from the eyes of calves in two naturally-occurring outbreaks of infectious bovine keratoconjunctivitis; also from a third group of calves accidentally exposed to an animal which had ocular exudates from one of the outbreaks instilled into its eyes.

The severity of the ocular lesions in infectious bovine keratoconjunctivis outbreaks may be related to a mixed infection with the mycoplasma and Moraxella bovis.

Preliminary typing studies indicate the mycoplasma is not serologically related to any known bovine mycoplasma.

     

Enzootic Pneumonia of Pigs: Identification of a Causative Mycoplasma in Infected Pigs and in Cultures by Immunofluorescent Staining

Immunofluorescent staining has been used to identify Mycoplasma hyopneumoniae in smears of broth cultures, in infected pig testicle cell cultures, and in frozen cut sections of pneumonic lungs from field and experimentally produced cases of enzootic pneumonia. In the pneumonic pig lung, fluorescent staining was limited to the surface of the bronchial and bronchiolar epithelium and to the contained exudate. In a series of trials using experimentally infected pigs fluorescence was not detected until 25 days post-infection and was regularly seen in pigs killed thereafter. Porcine immune globulin precipitated from the serum of experimentally infected pigs and conjugated with fluorescein isothiocyanate was reactive and specific for the detection of M. hyopneumoniae. Immune globulin conjugates prepared from the serum of hyperimmunized rabbits were reactive but in some cases produced a faint non-specific staining of frozen tissue sections. No such non-specific reactions were noted on stained culture smears or cell cultures.

Fluorescence was not seen in known positive preparations stained with non-immune pig globulin conjugates or in preparations from uninoculated cell cultures or pigs, stained with non-immune or immune globulin conjugates.

Mycoplasma hyorhinis was detected by immunofluorescent staining with homologous conjugates, in smears of broth cultures and in tissue sections from pigs with polyserositis.

Immunofluorescent staining was found to be species specific and useful for the early species identification of mycoplasma isolated from pigs.

     

Wide Distribution of Pasteurella haemolytica Type 1 Over the Nasal Mucosa of Cattle

Studies on the site of proliferation of Pasteurella haemolytica in the bovine nasal cavity have been carried out.

P.haemolytica were isolated from 15 selected major anatomical areas of the nasal cavity in calves with high numbers of P.haemolytica following shipment from Western Canada. When the organisms were present in the nasal cavity of live animals in low numbers, they were isolated from many, but not all, areas. P.haemolytica was isolated post mortem from one or more selected areas of several nasal cavities in spite of negative antemortem cultures.

By the direct fluorescent antibody technique, P.haemolytica was demonstrated at the surface of nasal epithelial cells. Organisms were not seen in or between epithelial cells nor in the ducts nor alveoli of glands. The findings were similar when high and low numbers of P.haemolytica were present in the nasal cavity.

     

Preliminary Observations on the Isolation of Escherichia coli Bacteriophages from the Intestinal Contents of Chickens

Eschericia coli bacteriophages were isolated from the intestines of chickens. These phages had different lytic patterns, and were propagated in nutrient broth containing 0.4 gm calcium chloride/litre. The agar layer technique was used to determine the routine test dilution (RTD) and plaque morphology. The phages differed in their 1) morphology, 2) RTD values, and their ability to lyse E. coli strains from various animals. All phages isolated lysed human K12 E. coli strains, whereas only two phages lysed the chicken E. coli strain. Phages isolated lysed E. coli from chicken, bovine, ovine, equine, and human, but not from porcine, canine and other avian species.     

Occurrence of Blood Cells and Serum Proteins in Bovine Fetuses and Calves

Hematological values of peripheral blood were determined for bovine fetuses and calves of various ages. Erythrocyte values increased through gestation. Fetuses 100 days or older had total values within the ranges of those reported for normal adult cattle. Mature erythrocytes were not observed in embryos and only a few were observed in fetuses 40 days of age. Fetuses 250 days or older had only a few rubricytes (<10/100 WBC). Leukocytes were first identified in the peripheral blood of a 45-day old fetus. Absolute leukocyte values increased through gestation and reached maximum values shortly before parturition. Granulocytes were first observed at 130 days of gestation and reached maximum values near parturition.

Total serum protein and gamma-globulin concentrations of colostrum-deprived calves were similar to serum protein and gammaglobulin concentrations of fetuses older than 265 days and were lower than values for the colostrum-fed calves. The immunoelectrophoretic pattern of 59-day old fetuses, the earliest age at which serum samples were obtained, demonstrated albumin, an α1 globulin and a β globulin, possibly transferrin. Additional α and β globulins appeared in the older fetuses and by 175 days of gestation serum electophoretic patterns of the fetuses were similar to patterns normally found with adult bovine serum except for the absence of the gammaglobulins in fetal serum. Immunoglobulin M was detected in 39 of 95 fetal serum samples by radial diffusion and in 13 of 95 samples by immunoelectrophoresis. Immunoglobulin G was detected in ten of 95 fetal serum samples by radial diffusion and in six of 95 samples by immunoelectrophoresis.

     

Pasteurella haemolytica of Cattle: Serotype, Production of Bete-galactosidase and Antibacterial Sensitivity

Two hundred and three isolates of Pasteurella haemolytica from cattle were studied. They originated from the nasal cavity of cattle in housed herds; the nasal cavity and pneumonic lungs of experimental feedlot calves and from pneumonic bovine lungs submitted for bacteriological diagnosis.

To determine whether a single characteristic or combination of characteristics might be a feature of isolates collected from animals with pneumonic pasteurellosis (Shipping Fever), the following tests were made. Cultures were serotyped by indirect haemagglutination; the ability to produce beta-galactosidase was examined in the ortho-nitrophenyl-beta-D-galactopyranoside (ONPG) test and antibacterial sensitivity tests were done. None of these factors could be directly related to the role of P. haemolytica in “Shipping Fever”.

     

Studies of Escherichia Coli in Gnotobiotic Pigs: V. Evaluation of the Effects of Orål and Parenteral Administration of Immune Serum

The effects of orally and/or parenterally administered immune serums were evaluated in 36 gnotobiotic pigs infected with an enteropathogenic strain of Escherichia coli. Pigs were euthanatized at predetermined time intervals between 6 and 48 hours prostinfection. The results were evaluated on the basis of: 1) clinical observations, 2) necropsy observations, 3) counts of viable E. coli in segments of the small intestine, 4) attempts to isolate E. coli from the heart, liver, and bile, 5) microscopic examination of fixed intestinal sections to determine the location of E. coli and morphologic evidence of the host response, and 6) determination of the pH of the various portions portions of the gastrointestinal tract.     

Necrotic Enteritis in Broiler Chickens II. Pathology and Proposed Pathogenesis

The intestines from 124 dead, sick and normal broiler chickens from 24 cases of necrotic enteritis were subjected to histological examination. Tissue sections from the duodenum, jejunum, ileum and ceca from each broiler were examined histologically for lesions of necrotic enteritis and the presence of coccidia. Lesions of necrotic enteritis were present in one or more areas of the intestine in all but six of 94 dead or sick birds and they were most common and severe in the jejunum. Coccidia were found in only small numbers in both diseased and normal birds.

Brown and Brenn stained sections showed Gram-positive bacilli intimately associated with early necrotic lesions on the tips of villi. Tissue sections from the intestines of sick birds permitted a proposed pathogenesis for this disease with the lesion starting at the tips of villi.

The similarity in pathogenesis and pathological lesions in this disease of broilers and Clostridium perfringens type C enteritis in baby pigs is discussed.

     

A Suggested Mechanism for the Pathogenesis of Infectious Atrophic Rhinitis

Boivin extracts of Bordetella bronchiseptica inhibited or uncoupled the energized processes of bovine heart and pig heart mitochondria. Energy-dependent accumulation of calcium phosphate by both types of mitochondria was markedly inhibited by the extracts. The thesis is advanced that Boivin extracts of B. bronchiseptica may contain the membrane-damaging component responsible for infectious atrophic rhinitis. The action of the extracts was decreased by heat but seemed rather insensitive to digestion by trypsin or extraction by lipid solvents. Attempts to resolve the active component from the extracts were unsuccessful.     

Influence of Immunization Procedures on Upper Respiratory Tract Immunity in Cattle

Aspects of respiratory tract immunity have been investigated in the bovine species. Using Past. hemolytica type I as the antigen for this model the relationship of nasal and serum antibody production to the route of vaccination and type of vaccine was investigated in a series of 15 dairy calves from two to four months of age. Experimental results indicated that an aerosol vaccination with live Past. hemolytica resulted in a significant nasal antibody response while parenterally vaccinated gave calves with equivalent serum titers had no significant nasal antibody response.     

Bovine Infectious Keratoconjunctivitis: Serological Aspects of Moraxella bovis Infection

A modification of a gel diffusion precipitin test (GDPT) was used to detect antibodies for Moraxella bovis (M. bovis) in the sera of cattle affected with bovine infectious keratoconjunctivitis (BIK). The test was also used for the detection of sequential antibody development in cattle vaccinated with cultures of M. bovis. Also, strains of M. bovis isolated from cattle herds affected with BIK were characterized serologically as a part of an identification scheme using the test.

A comparison of the antigenic properties of various strains of M. bovis and M. bovis-like organisms was conducted using the test. The results indicated that there might be antigenic relationships between M. bovisand M. bovis-like organisms such as Moraxella liquefaciens, Moraxella nonliquefaciens, an unidentified hemolytic diplococcus, Mima polymorpha, Mima polymorpha var. oxidans and Herellea vaginicola

The authors suggest that the GDPT can be used for serological studies of BIK, and the identification and antigenic analysis of M. bovis. They indicate, however, that a more definitive study is needed to evaluate the reliability of the test for quantitative work.

     

Acute Atypical Bovine Pneumonia Caused by Ascaris Lumbricoides

A case of acute atypical pneumonia (bovine asthma, pulmonary emphysema, or pulmonary adenomatosis) occurred in a group of cross-bred beef type yearlings in late autumn. Clinical signs included a forced expiratory grunt, excessive salivation, ruminal stasis and, on auscultation over the lungs, pulmonary emphysema and oedema. The cattle had been brought in from a poor summer pasture and housed in a pig pen heavily contaminated with Ascaris lumbricoides eggs as it had contained unwormed feeder pigs all summer.

Fifteen out of seventeen head were affected ten days following housing and all within twenty-four hours. One steer of the group died and at necropsy fourth stage A. lumbricoides larvae were isolated from lungs showing profuse oedema and some emphysema. Histopathological examinations of the lungs showed a diffuse interstitial pneumonia. All remaining animals appeared clinically normal six days following the outbreak.

     

Clinical, serologic, and cross-challenge response and virus isolation in cattle infected with three bovine dermatotropic herpesviruses.

The relationship of 3 dermatotropic bovine herpes-viruses (bovine mamillitis herpesvirus, dermatotropic bovine herpesvirus, and Allerton virus) was investigated; special emphasis was on the clinical response and the development of complement-fixation and precipitating antibodies in cattle. The immune respnse in steers was determined throughout the disease and after challenge exposure of the steers with 1 of the other 2 herpesviruses. Blood samples and sections of skin were collected intermittently from infected animals and examined for presence of virus.