A Survey of the Alimentary Tract of Cattle for Clostridium Perfringens

Contents of the rumen, abomasum, ileum, and colon of 100 fattened cattle were examined for the presence of Cl. Perfringens. Liquid medium iniculated with each sample of gut content was tested for the presence of toxins of Cl. Perfringens.

Identification of Cl. Perfringens was based on atmospheric requirements for growth, colonial morphology, and stormy fermentation in litmus milk. Identification of toxins was based on neutralization tests in guinea pigs and mice.

Cl. Perfringens was isolated from 202 of 399 samples. In 105 additional cultures, colonies characteristic of Cl. Perfringens were present but could not be isolated in pure culture.

Cl. Perfringens type D toxin was identified in only one culture, which was inoculated with ileum contents. Type A toxin was identified in eight of the 24 samples from the one lot of samples in which no type A antitoxin was used. There were no identifications of toxigenic types B, C, or E.

The results indicate that an isolation from necropsy specimens of untyped Cl. Perfringens or type A Cl. Perfringens is in itself of little significance. The infrequency of occurrence of the other toxigenic types in this survey of healthy cattle indicates that recovery of these types from necropsy specimens may be of more significance in determining the cause of death.

     

Colony Formation in Culture by Bovine Granulopoietic Progenitor Cells

Calf bone marrow cells cultured in a semi-solid medium of 0.8% methyl cellulose produced colonies of granulocytic cells and macrophages by seven days. A prerequisite for colony growth was the presence of serum obtained from a calf three hours after intravenous injection of endotoxin. Three morphological types of colonies were seen but cell types within these types of colonies did not differ. Cultured cells were identified by morphological and cytochemical characteristics.

Optimum growth occurred when serum from endotoxin stimulated calves and fetal calf serum were present in a volumetric ratio of 7:3. Inhibition of colony growth occurred when endotoxin-stimulated serum was present at greater than optimum concentration. Normal calf serum, fetal calf serum, mouse L-cell conditioned medium and bovine urine did not stimulate significant colony growth when 8.0 x 10⁴ marrow cells were cultured.

There was a linear relationhip between the number of marrow cells in the cultures and the number of colonies produced. Colony forming efficiency ranged from 13 to 59 colonies per 10⁵ cells plated.

The behaviour of calf colony forming units in suspension culture was similar to that reported for mouse colony forming units.

     

Aerobic bacterial isolates in horses in a university hospital, 1986-1988

Bacterial isolations were reviewed from equine trachea, guttural pouch, uterus, wounds, abscesses, blood, synovial fluid, and abdominal fluid submitted to the Clinical Bacteriology Laboratory of the School of Veterinary Medicine at the University of Montreal for aerobic bacterial culture from 1986 to 1988. Of the 733 samples submitted, 324 (44%) were positive for bacterial growth, and 233 antimicrobial sensitivity tests were performed. Seventy-six percent of all positive samples yielded one bacterial species and two were isolated from 22% of positive samples. Streptococcus zooepidemicus, Escherichia coli, and Actinobacillus spp. were isolated from 39%, 18%, and 15% of the samples, respectively.

Bacterial growth was most common from guttural pouches, wounds and abscesses, and transtracheal washes (TTW), but was less common from uterus, blood, abdominal fluid, and synovial fluids. Streptococcus zooepidemicus was the most common bacterium recovered from guttural pouches, TTW, uterus, and wounds and abscesses. Escherichia coli predominated in abdominal fluids, blood, and synovia. Bacterial sensitivities to common antimicrobials are presented.

     

The Isolation of Trypanosomes from Cattle in Ontario

A method for the recovery by blood culture of a T. theilieri-like flagellate from cattle is described.

In Ontario, 54% of 156 cows in 15 herds were found to have parasitemia.

     

The Effect of Edema, Hydrocortisone Acetate, Concurrent Viral Infection and Immunization on the Clearance of Pasteurella hemolytica from the Bovine Lung

The influence of pulmonary edema, hydrocortisone, immunization against Pasteurella hemolytica and concurrent infection with parainfluenza-3 virus upon pulmonary clearance of aerosolized P. hemolytica was studied in 31 calves. Following the various treatments calves were challenged with an aerosol of P. hemolytica. One control calf was killed immediately after the aerosol and the numbers of bacteria in the lung taken as 100%. Two calves were killed four hours after challenge and the numbers of bacteria in the lungs were compared to the 100% of the control calf. The result was the percentage clearance of bacteria at four hours.

Pulmonary edema was induced by three different methods: by an aerosol of histamine, by intravenous injection of endotoxin and by intravenous injection of croton oil emulsion. The edema impaired the clearance of P. hemolytica, which was reflected in high numbers of P. hemolytica present in the lungs at four hours after challenge: 260% after histamine, 300% and 400% after endotoxin and 92% after croton oil.

Six days of treatment of four calves with high doses of hydrocortisone acetate produced inconsistent results: two calves treated with a higher daily dose (36 mg/kg) had normal clearance whereas two calves treated with a lower dose had pulmonary edema and displayed lowered clearance with 111% and 31% respectively of P. hemolytica retained in the lungs four hours after challenge.

Immunization of calves by three different methods, a subcutaneously injected bacterin of P. hemolytica (2 calves), single aerosol (2 calves) and four aerosols (4 calves) of live P. hemolytica was reflected in an accelerated pulmonary clearance of P. hemolytica (with a mean of 1.55% of bacteria retained at four hours).

Concurrent infection with parainfluenza-3 virus did not lower the clearance of P. hemolytica in the lungs of 12 calves over 15 days except on the first day following the exposure to parainfluenza-3 virus. These calves had hemagglutinating antibodies against P. hemolytica before exposure.

     

Haematology and blood‐chemistry for predicting abscesses and other abnormalities in slaughtered pigs

Summary

Blood values of slaughtered pigs were determined in an attempt to relate the blood profile with the prevalence of abscesses. To this end 319 pigs were selected and grouped into classes on the basis of pathological ante‐ and post‐mortem findings. The classes thus distinguished were (i) no pathology, (ii) pleurisy or pneumonia and (iii) abscesses occurring singly, metastasised or in combination with other abnormalities.

By stepwise discriminant analysis it was established that the blood variables ln(fibrinogen) and albumin were particularly suitable for the detection of abscesses in slaughtered pigs. In this way a correct classification of 89.3% of affected pigs is possible. The numbers of false‐positives and false‐negatives were 19.3% and 10.7%, respectively.

For meat‐inspection purposes application of blood analyses is promising. For application in meat‐inspection practice rapid on‐line methods need to be devised.     

Swine Interferon I. Induction in Porcine Cell Cultures with Viral and Synthetic Inducers

The production of interferon by porcine kidney (PK₁₅) cell culture in response to viral and synthetic inducers was studied. The inducers used included a synthetic double-stranded polyribonucleotide, polyriboinosinic-polyribocytidylic acid (Poly I:C), swine influenza virus and three strains of pseudorabies virus. Following exposure to these inducers cell culture fluids were examined for interferon by the plaque-reduction method.

The Poly I:C and the swine influenza virus induced production of interferon by PK₁₅ cell cultures, whereas, all three strains of pseudorabies virus at the two concentrations tested failed to induce production of interferon in vitro.

The antiviral substance produced in PK₁₅ cells was identified as an interferon because it was pH stable, non-dialyzable, sensitive to trypsin, non-sedimentable, relatively heat stable, host-species specific and it possessed broad-spectrum antiviral activity. The latter was demonstrated by inhibition of vesicular stomatitis, vaccinia and pseudorabies viruses. Differences in interferon activity against the different viruses were observed.

     

Studies of Escherichia coli in Gnotobiotic Pigs VI. Effects of Feeding Bacteria-Free Filtrates of Broth Cultures

Nine gnotobiotic pigs derived from one gilt were fed bacteria-free filtrates prepared from: 1) cultures of an enteropathogenic strain of Escherichia coli 09:K·:NM (Strain 340), 2) cultures of a nonenteropathogenic strain of E. coli 08.K·.H16 (Strain CDC-1466-56), and 3) uninoculated culture medium.

Diarrhea was observed initially two to four hours after feeding the filtrate prepared from the enteropathogenic E. coli. The duration of diarrhea was five to ten hours. No diarrhea was observed after feeding filtrate prepared from uninoculated medium or cultures of nonenteropathogenic E. coli.

The pH values of the feces increased with the onset of diarrhea and decreased to normal after diarrhea stopped.

No histopathological lesions were found.

     

Experimental Colibacillosis in Gnotobiotic Baby Pigs II. Pathology

Sixty-two neonatal gnotobiotic pigs were used in three experiments to determine the lesions produced by two closely related strains of Escherichia coli O138:K81:NM (of Michigan origin) and O138:K81 (of Minnesota origin). Exposure was by subcutaneous injection of bacterial culture into the umbilical stump or by oral inoculation.

Gross signs common to monocontaminated pigs included distention of the flaccid small and large intestines with fluid contents. Edema was prominent in various tissues of most pigs exposed via the umbilical stump but not in those exposed orally.

Histological lesions were predominantly in the gastrointestinal tract and were variable. At one extreme acute hemorrhagic enteritis was present in two pigs, while at the other extreme in a few pigs it was difficult to distinguish tissues of infected pigs from those of noninfected germfree pigs. Significant histological lesions common to monocontaminates included mild inflammatory reaction, hydropic degeneration of the intestinal epithelium, evidence of interference with normal function of the villus-draining mechanisms, and vascular changes generally indicated by edema.

The findings suggest that interference with normal absorption of nutrients plays at least some role in the pathogenesis of colibacillosis in young gnotobiotic pigs.

     

Pasteurella haemolytica in the Tracheal Air of Calves

Pasteurella haemolytica was shown to be present in the tracheal air of calves and was likely transported in droplet nuclei formed in the nasal passages. The number of colonies of P. haemolytica found in the tracheal air of the calves ranged from 1.9 to 12.5 colonies per cu ft of air. As long as P. haemolytica colonized the nasal passage in numbers detectable in nasal swabs it could be found in the tracheal air but there was no direct correlation between the numbers in the nasal flora and the numbers found in the tracheal air. Of the P. haemolytica which travel via the tracheal air 47.8% were in droplets of the aerodynamic size of from less than one to five microns, the size range which is considered hazardous for lung penetration in man.

The technique used demonstrated the presence of P. haemolytica in the tracheal air of calves and provides a useful tool for monitoring and determining the phase in the colonization of the respiratory tract in which the majority of the potential pathogen P. haemolytica pass from the nose to the tracheal air and presumably to the lung.

     

Autolytic Changes in the Digestive System of Germfree, Escherichia coli Monocontaminated, and Conventional Baby Pigs

Tissues from three groups of pigs (germfree, Escherichia coli monocontaminated, and healthy conventional pigs) were collected at intervals between 24 minutes and 7 hours 12 minutes after death. Histological differences between the three groups were present in the alimentary mucosa and were most striking in the ileum and colon.

Autolytic change was detected only in the digestive system and the sequence of autolytic events was similar in all groups. The time of onset and rate of progress differed markedly between groups and between segments of the alimentary canal. In the conventional group, autolysis started quickly and progressed rapidly. In the germfree group it was later in onset and slower in progress. The E. coli infected group was intermediate. The initial visible change occurred in the small intestine.

The morphological differences and autolytic changes were described.

     

The Incidence and Significance of Clostridia Isolated from Ready-to-eat Meats and Rendered Products

Approximately 26% of rendered and 10% of ready-to-eat products contained species of the genus Clostridium.

Eleven species of clostridia were isolated from a total of 524 products tested. Some products harboured more than one species.

C. sporogenes, one of the most heat resistant organisms, was the most common type isolated.

C. sordellii produced a transient illness in guinea pigs while all other species were innocuous.

     

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.

     

The Detection of Specific Complement-Fixing Antibodies in Serum of White-Tailed Deer (Odocqileus Virginianus) with Theileria Infection

A complement-fixation (CF) antigen which has been prepared from Theileria infected erythrocytes is capable of reacting to specific serum antibodies of deer acutely infected with Theileria.

No sera from 17 deer known to be free of Theileria infection reacted positively to the CF test. Of 35 tests on sera from 12 infected deer having a parasitemia of 2% or less and no accompanying anemia, only 10 (29%) were positive, 2 (6%) were suspicious, and 23 (66%) were negative. Of 65 tests on 8 acutely infected deer, 49 (75%) were positive, 4 (6%) were suspicious and 12 (18%) were negative. Of the 8 deer in which acute theileriasis occurred all reacted to Theileria antigen at one time or another.

A significant correlation was found between CF titers and the degree of parasitemia in acute infections.

Rabbits were hyperimmunized using erythrocytes from either normal or Theileria infected deer. Reciprocal absorption of the hyperimmune sera with Theileria and normal erythrocytic antigens demonstrated the presence of antibodies specific for Theileria.

     

The Anthelmintic Activity of Cambendazole in Calves and Lambs

Tests were carried out on the efficacy of Cambendazole (5-isopropoxy carbonylamino-2-(4-thiazolyl) benzimidazole) against lungworm and gastrointestinal parasites of calves and lambs. Against Dictyocaulus viviparus the compound was highly effective (80%) against mature worms at levels of 35 mg/kg and over and immature “arrested” worms at 60 mg/kg (71% efective).

In the Calves the compound was very effective (90 to 99%) against adult Haemonchus placei, Ostertagia spp., Trichostrongylus axei and Cooperia oncophora at all levels tested (15, 20, 25, 30, 40 and 60 mg/kg). Against adult Nematodirus spp. dosages of 30, 40 and 60 mg/kg were 81%, 94% and 99% effective respectively

Against arrested Ostertagia spp. larvae, a dosage of 60 mg/kg was 90% effective. Immature stages of Cooperia spp. were susceptible at all levels used but those of Nematodirus spp. required levels of 60 mg/kg for 99% removal.

In the lambs the compound was highly effective against immature and mature nematodes when given orally at all levels tested from 15 mg/kg to 30 mg/kg. Significant reductions (81%) in Moniezia expansa scolices were also observed at dosages from 15 mg/kg to 30 mg/kg.

     

Immunization of Mice and Chinchillas Against Pseudomonas aeruginosa

The possibility of production of an effective vaccine against Pseudomonas aeruginosa infections in fur-bearing animals was investigated. Twenty-three strains of Pseudomonas aeruginosa isolated from diseased chinchillas and mink were tested in mice for their immunogenic properties. Nineteen of these strains produced good immunity against homologous strains, and three of these produced also good immunity against heterologous strains. Of the remaining four strains two produced moderate immunity and two no immunity.

It was found that 0.05% or 0.5% formalin added to suspensions of Pseudomonas aeruginosa or ultrasonification of the suspension produced better results than 0.5% phenol, 0.3% alcohol or heat at 100°C for half an hour.

Chinchillas vaccinated with two doses of formolized Pseudomonas aeruginosa bacterins were immune for 36 weeks after the second dose, while all controls died within 48 hours after being challenged.

It was found that the protection afforded by the polyvalent bacterin extended beyond the strains included in the vaccine.

A field survey on 34 ranches which included over 7,700 chinchillas showed very promising and encouraging results.

     

Studies with Radioactive Endotoxin III. Localization of 3H-Labelled Endotoxin in the Formed Elements of the Blood and Detection of Endotoxin in Calf Blood with the Limulus Amebocyte Lysate

³H-labelled Pseudomonas endotoxin was incubated in vitro with blood from nontolerant and endotoxin tolerant calves. Formed elements were separated from serial samples of the incubated mixtures.

The labelled endotoxin became associated with neutrophils, monocytes, lymphocytes, platelets and erythrocytes. Association of ³H-endotoxin with formed elements of the blood occurred during the first five minutes of incubation and did not significantly change over the course of a three hour incubation period. Tolerance did not result in increased uptake of ³H-endotoxin by formed elements of the blood. Tolerance of calves to endotoxin is apparently not due to increased uptake of endotoxin by formed elements of the blood.

The Limulus amebocyte lysate assay was unreliable for the detection of endotoxin which was present in calf blood in vitro and requires further modification.

     

Entérite nécrotique chez le poulet de gril II. Caractères des souches de Clostridium perfringens isolées

A Gram positive bacillus, strictly anaerobic, was isolated from the viscera of all diseased birds showing lesions of necrotic enteritis. Its morphology and biochemical reactions, the presence of alpha and thêta hemolysins and the production of a lecithinase-C in vitro, all these characteristics indicated a similarity to those belonging to the group of Clostridium perfringens.

The two hemolysins were neutralized in vitro only by the antitoxin A. Broiler chickens injected I.V. with a Viande-Foie (VF) broth culture of Clostridium perfringens together with the antitoxin A survived, whereas those receiving antitoxin C died. These results seem to indicate that this organism belongs to the type A. This bacillus was sensitive to a great variety of antibiotics, except neomycin.

     

The Anthelmintic Activity of 0,0-dimethyl-0-1,2-dibromo-2,2-dichloroethyl phosphate for Canine Hookworms

In a statistical study involving 18 adult dogs an enterically coated, tableted formulation of 0,0-dimethyl-0-1,2-dibromo-2,2-dichloroethyl phosphate was found to clear 86.5 per cent of the dogs of hookworm infections when given orally at a level of 15 mg./kg. for 2 consecutive days and 50 per cent when it was given at the same level once. The per-cent efficacy for hookworm removal was 70 and 78 per cent respectively. The dogs were infected with both A. caninum and U. stenocephala. Other concurrent helminth infections were also present.

No vomitions, symptoms, or lesions of drug toxicity were observed in the medicated dogs.

     

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.