Contribution à l'étude quantitative de la flore bactérienne du gros intestin des porcs dysentériques

The bacterial flora and the pH of the large intestine of dysenteric swine during acute subacute and chronic phases have been submitted to quantitative and qualitative studies. The methods used are based on primary isolation and differentiation of the bacteria by the use of selective media and the subsequent differentiation using the replica plating technique. The most characteristic changes are the following:

1. A significant increase of the pH of the chyme in the large intestine during acute dysentery

2. A significant increase of Vibrio, Escherichia coli and Staphylococcus in the colon and cecum during acute dysentery.

3. A significant increase of Shigella in the colon and cecum during subacute dysentery.

4. The almost total disappearance of Aeromonas and of the yeasts in the large intestine during acute, subacute and chronic dysentery.

5. A significant decrease of Klebsiella, in the cecum, during acute dysentery and of the fungi during subacute dysentery.

6. Decrease of Streptococcus in the colon during acute dysentery.

7. The total quantitative flora of the large intestine do not change very much.

     

Parasites of Dogs from Indian Settlements in Northwestern Canada: A Survey with Public Health Implications

A total of 959 faecal samples were obtained from dogs in 12 native communities in Northern Saskatchewan, Central and Northern Alberta and the Northwest Territories. All samples were examined using a flotation technique. Samples from an area of endemic human amoebic infections were also examined by a formol-ether sedimentation method. Eighteen necropsies were performed.

Entamoeba histolytica cysts were recovered from dog faeces at Loon Lake, Saskatchewan.

Toxocara canis had low incidence in Saskatchewan and Central Alberta, and appeared to be almost non-existent further North. Toxascaris leonina was found in all areas surveyed. Canine hookworm infections were plentiful in all areas, the highest incidence being recorded from Northern Alberta and Northwest Territories. Many Taenia (or Echinococcus) infections were found consistently in all areas. Only one infection with Dipylidium caninum was discovered.

Metorchis conjunctus infections were found to be common in the Saskatchewan reserves. Infections with Diphyllobothrium sp. were found in all communities with access to good fishing. One specimen of Dioctophyma renale was recovered at necropsy.

Infections with parasites of no known zoonotic importance such as Trichuris, Alaria and Isospora species were also recorded.

     

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.

     

Screening Test of Animal Sera for the Cultivation of Leptospires

The nutritive value of 8 kinds of animal's sera for the growth of Leptospires was studied by a screening method and compared with that of rabbit serum.

The sera of dairy cattle and goats were higher than rabbit serum in average growth number, and thus gave great value for the cultivation of Leptospires.

In eight kinds of animals other than dairy cattle, the growth promoting action of the pooled sera for Leptospires were lower than that of the individual sera of the same kind of animals.

The serum which is to be added to the medium for the cultivation of Leptospires should be tested by a screening test for the absence of growth inhibitory action against Leptospires, no matter from what kind of animal the serum originated.

     

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.

     

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 Effect of Medicated Feed on the Nasal Microflora and Weight Gain of Pigs

Antimicrobial agents were added to the feed of swine for three weeks to determine the interrelationships of potentially pathogenic agents in the nasal tract, turbinate atrophy and weight gains.

Bordetella bronchiseptica was not isolated from the groups fed the combination of chlortetracycline, penicillin and sulfamethazine. B. bronchiseptica was found in some pigs after the feeding trail, but this organism was not significantly associated with turbinate atrophy at the time of slaughter.

Mycoplasma hyorhinis was not found in the nasal passages of the pigs that received feed containing high concentration chlortetracycline but was found in pigs that received other diets. Hemophilus suis was not significantly reduced by any of the treatments used.

The organisms studied in the pigs were not isolated from the personnel handling the pigs.

     

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.

     

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.

     

Differentiation Among Strains of Goat Mycoplasma by Incident Light Immunofluorescence

Antigenic differentiation between strains of goat mycoplasma was studied by direct fluorescent antibody reactions employing incident (vertical) ultraviolet light. Agar colonies of the mycoplasma grown in petri dishes were fixed by alcohol in situ, and stained with conjugated globulin before examination with ultraviolet light.

The fluorescent antibody (FA) conjugate against Vom strain of Mycoplasma mycoides var. capri was Vom strain-specific, no cross reaction with Mexico, Connecticut, or Maryland strains. Similarly, the Mexico strain conjugate was specific for colonies of Mexico, and did not cross with the Vom, strain. Additionally, the conjugate of the PG-2 strain of Mycoplasma agalactiae, which was specific for the colonies of PG-2 was refractory for the strain #99 of M. agalactiae.

It was therefore possible to utilize an immunofluorescent technique (incident ultraviolet light) to demonstrate differences among strains of M. mycoides var. capri and M. agalactiae.

     

Mycoplasma and Associated Bacteria Isolated from Ovine Pink-eye

A mycoplasma was recovered from the untreated conjunctival membranes of nine sheep affected by Pink-eye. It was neither isolated from the conjunctiva of treated animals which were affected nor from the conjunctiva of normal animals either in contact or not in contact with affected animals.

Bacteria found on normal conjunctival membranes were Neisseria ovis, Escherichia coli, Staphylococcus epidermididis, Streptococcus and Bacillus spp.

Bacteria found in clinical cases of Pink-eye were N. ovis, E. coli, a Streptococcus and Pseudomonas spp.

     

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.

     

Incident Light Immunofluorescence of Alcohol-fixed Colonies of Ruminant Mycoplasma

Two species of ruminant mycoplasma colonies had to be fixed in ethyl alcohol so that incident immunofluorescence method could be applied. In addition, the stain reaction had to be kept for 90 minutes at 37°C.

This fluorescent antibody (FA) method was developed to identify colonies of Vom strain of Mycoplasma mycoides var. capri, V-5 strain of M. mycoides var. mycoides, and PG-2 strain of M. agalactiaeon agar, using fluorescent ultraviolet light. Fluorescence was not demonstrated when heterologous conjugates or normal rabbit serum conjugate were applied but the reaction appeared to be specific for each strain of mycoplasma.

The FA method was able to differentiate specific mycloplasma colonies in mixed cultures.

     

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.

     

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.

     

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 Influence of CO2 Supply on Colonial Formation of Leptospires

The colonial formation of three serotypes of Leptospires on Cox's solid medium was promoted by microaerophilic incubation of one to three per cent of CO₂ supplied by carbon dioxide cylinder, sodium carbonate oxalic acid, and candle method. In anaerobic incubation Leptospira pomona grew the same as with CO₂ incubation.

The pH of the medium was an important influence on the rate of colonial formation of Leptospires.

Addition of hemoglobin and inactivation of rabbit serum was not an essential condition for rapid colonial formation. It was found that variation in the morphology of leptospiral colonies occurred with hemoglobin from different species and individuals.

     

Une étude expérimentale sur la variation du taux d'anticorps sériques suivant le cycle évolutif des oestres du cheval

The changes of antibody titers in the sera of colts infested naturally or artificially with Gasterophilus have been determined in relation to the life cycle of this arthropod using passive hemagglutination, complement fixation, double diffusion techniques and saline extracts of antigens from the third larval stages of Gasterophilus intestinalis and G. nasalis.

In the sera of the infected animals the hemagglutinating antibodies were present at low titers at the third week post-infestation by using somatic extract of G. intestinalis and at the seventh week in case of G. nasalis. At eight weeks post-infestation the antibody titers reached their maximum 1:8192 (G. intestinalis) and 1:4096 (G. nasalis), then dropped at 12 weeks post-infestation.

The complement fixing antibodies were present occasionally between the seventh and 11th weeks after infestation. Precipitating antibodies were absent in all sera.

     

Minimal Inhibitory Concentrations of Twenty Antimicrobial Agents to Animal Pathogens

Minimal inhibitory concentration (M.I.C.) values as determined by an agar-plate-dilution method for 60 bacterial isolates, consisting of Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus of animal origin against 20 antimicrobial drugs are presented. Of all the drugs, gentamicin had the best in vitro antibacterial activity in terms of M.I.C. when considering all the species of organisms together, while spectinomycin had the least activity.

An inoculum replicator was a convenient tool in carrying out the agar-plate-dilution method.

A comparison of the M.I.C. values of 42 isolates of S. typhimurium with the results obtained by low level method and the Bauer-Kir-by method showed that with few exceptions, there is a general agreement.