Prevalence and Species Distribution of Yeast in Mammary Glands of Dairy Cows in Minnesota

A study of the prevalence of yeast-like fungi in the mammary glands of dairy cattle was conducted in Minnesota. Quarter samples from 6,020 cows were cultured for yeast. Growth of organisms was obtained from 3.2% of the quarter milk samples. The rate of yeast infection for Minnesota dairy cattle in this study was 2.0%.

The majority of the yeast isolated belonged to one of four species of the Candida genus. Candida krusei, Candida parakrusei, Candida guilliermundi, and Candida tropicalis, comprised 89% of the yeasts isolated. All of these species have been reported to cause clinical mastitis (1, 7, 9, 10, 12, 13, 15, 16).

It would appear that yeast-like fungi are of sufficient prevalence in mammary glands that yeast infection would be considered in the differential diagnosis in cases of clinical mastitis.

     

Serological Identification of Escherichia coli Isolated From Cats and Dogs

In a group of 112 cats, examined during an outbreak of diarrhea, 104 (92.8%) yielded Escherichia coli with identifiable O groups as compared to 12 (21.8%) of 55 control cats apparently free of intestinal ailments. E. coli group O6 occurred in 49.1% of the cats with diarrhea, and persisted in this group of cats at about the same rate when examined for a period of ten months. E. coli group O6 was not isolated from the control group.

Specimens were also collected from dogs with various clinical signs. E. coli O4 and O6 strains appear to be important potential pathogens for canine species and were found to spread from one dog to another when in close contact. E. coli group O4 was identified in ten and O6 in three of 14 dogs examined. All isolates in which the O group could be identified were hemolytic.

     

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.

     

Isolation of Western Equine Encephalomyelitis (Wee) Virus From Mosquitoes In Saskatchewan, 1962

Saskatchewan, in the summer of 1962, was the scene of an extensive outbreak of western equine encephalomyelitis (WEE) in horses. The results of mosquito survey work showed Culiseta inornata and Culex tarsalis respectively to be the two most abundant mosquito species during midsummer. These species are those reported to be most commonly associated with outbreaks of WEE. Five hundred and sixty-four pools of mosquitoes were examined for the presence of WEE virus. Six pools, three of C. tarsalis and one each of C. inornata, Aedes flavescens and Aedes dorsalis, yielded WEE virus. Positive mosquitoes were from St. Walburg (C. inornata), Saskatoon (C. tarsalis — two, A. dorsalis — one), Outlook (C. tarsalis) and Kisbey (A. flavescens).     

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.

     

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.

     

Bovine Viral Diarrhea Virus and Escherichia Coli in Neonatal Calf Enteritis

This study was initiated to determine the etiologic and pathogenic significance of an American strain of bovine viral diarrhea (BVD) virus (strain NADL-MD) in enteritis of neonatal calves (calf scours).

Three colostrum-fed calves from dams exposed intravenously to BVD virus at 6, 16 and 25 days prepartum, respectively, had moderate diarrhea persisting until the eighth day of life. The BVD virus was isolated from all 3 calves and persisted up to 93 days in 1 calf, indicating either that BVD was transmitted in utero or via the dam's milk.

Three specific pathogen free (SPF) calves permitted dams' colostrum for the first 4 feedings and then given milk replacer were exposed orally on the day of birth to BVD virus. One calf died of neonatal enteritis 28 hours post-exposure and at necropsy the BVD virus was isolated from several of its organs. The remaining 2 calves had a mild diarrhea persisting to the eighth day of age.

Two calves permitted dams' colostrum ad lib. for 72 hours, and then weaned, were exposed orally to BVD virus. Both calves had a mild persistent diarrhea and BVD virus was isolated from their blood for 56 days post-exposure.

Of 13 SPF, colostrum-deprived calves exposed orally or intranasally at birth to the BVD virus, 4 had severe diarrhea and died of neonatal enteritis from 38 hours to 13 days postexposure. Isolations of BVD virus were made from several of the organs of the calves at necropsy. All of the 9 surviving calves had a moderate to severe diarrhea frequently persisting for 7 to 10 days, and BVD virus was isolated from the survivors up to 103 days postexposure.

Several strains of Escherichia coli were isolated from calves after the second day of life, but were neither pathogenic for mice, nor serologically related to strains of E. coli usually associated with outbreaks of calf scours. Four colostrum-deprived SPF calves were exposed orally at birth to a strain of E. coli isolated from the intestine of the calf with the most acute symptoms and fatal neonatal enteritis. None of the four calves receiving the E. coli had diarrhea. One calf, however, had respiratory distress and died on day 5.

Two SPF colostrum-deprived control calves had neither diarrhea nor respiratory distress.

The above findings support the conclusion that BVD virus should not be overlooked as a primary cause of the neonatal calf enteritis complex.

     

Experimental Studies on the Pathogenesis of Corynebacterium equi Infection in Foals

Four month-old foals were infected orally with 75 mL of a suspension of 5.0 × 10⁸ Corynebacterium equi per mL. Two foals were killed after ten days and had scanty number of C. equi in the caeco-colic lymph nodes. No C. equi were recovered from the other two foals, killed 20 days after infection. No gross pathological change was detected in these four foals, although mild microscopic lesions were seen in the ileum of one foal. Results of lymphocyte blastogenesis using peripheral blood lymphocytes and C. equi antigens showed, however, that lymphocytes became sensitized to C. equi following this challenge.

In a second experiment four month-old foals were given orally the same dose of organisms but on five consecutive days. Two foals were killed ten days after infection and showed mild histological changes in the large bowel mucosa and C. equi could be recovered from all intestinal lymph nodes cultured. In one of these foals moderate numbers of C. equi were present in the bronchial lymph node. Of the other two foals, one died after 22 days with severe ulcerative enterocolitis and intestinal lymphadenitis. Only one small pulmonary abscess was detected despite large numbers of C. equi in the lungs. The other foal developed similar intestinal changes and was euthanized 25 days after infection. No C. equi were detected in the lungs or bronchial lymph node. Lymphocyte blastogenesis in these animals showed a rapid rise in response to C. equi antigens.

These studies suggest that C. equi pneumonia in foals does not always arise from an intestinal infection, that minor intestinal infection causes a cellular immune response and that massive exposure of the bowel over a sustained period is necessary to induce intestinal lesions.

     

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.

     

Vibriosis: Demonstration of Vibrio fetus and Vibrio bubulus Organisms in Preputial Fluid by Immunofluorescence and Cultural Techniques

Fluorescent conjugates were prepared from the sera of calves immunized with four Vibrio fetus strains and one Vibrio bubulus strain. The fluorescent antibody technique (FAT) was then used to detect vibrio organisms in preputial fluid collected from 67 bulls belonging to a Canadian artificial insemination (AI) unit. The V. fetus conjugates reacted with both V. fetus var venerealis and V. fetus var intestinalis. V. fetus was found in 20 animals (29.9%), 13 of which also harboured V. bubulus. In two cases, the FAT failed to detect V. fetus which was isolated by concurrent bacteriological examinations.

It was concluded that the FAT can be a rapid method of detecting some carrier bulls but more reliable results are obtained when a combination of FAT and bacteriological methods is employed. It was found that a single sample giving negative results is inconclusive and additional tests are required before making a final diagnosis. The FAT can also be used to differentiate V. fetus isolates from V. bubulus.

     

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.

     

Polyerositis and Arthritis Due to Escherichia coli in Gnotobiotic Pigs

Forty gnotobiotic pigs from six litters were exposed orally to Escherichia coli 083:K·:NM at 69 to 148 hours of age, while 17 pigs from the same litters served as unexposed controls. Clinical signs of infection included fever, anorexia, diarrhea, lameness, and reluctance to move.

Eighty-four percent of the exposed pigs in four litters died, while only 13% in two litters died. Gross and microscopic lesions included serofibrinous to fibrinopurulent polyserositis in 96% of the exposed pigs in four litters and 33% of the exposed pigs in two litters. A few pigs had gross and/or microscopic lesions of arthritis. Escherichia coli was routinely isolated from the serous and synovial cavities of infected pigs.

Anti-hog cholera serum administered orally as a colostrum substitute gave partial protection against E. coli infection.

     

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.

     

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.

     

Présence de facteurs R chez des souches d'entérobactéries pathogènes isolées d'animaux domestiques, et plus particulièrement du chien

Strains of enterobacteria (nine Escherichia coli and two Salmonella) isolated from primary or secondary infections in the dog, cat, pig, calf and kangaroo were studied for the presence of extrachromosomal drug resistance factors (R factors). Seven strains of E. coli and two strains of Salmonella transferred resistance involving the following antibiotics: streptomycin, ampicillin, chloramphenicol, neomycin and tetracycline. All strains harboring R factors transferred streptomycin resistance and the identified resistance patterns were as follows: Sm Am, Sm Te, Sm Neo, Sm Am Te, Sm CI Neo and Sm Am CI Te. The levels of resistance observed were comparable for all donor strains and their converted recipients.

Strains of E. coli harboring R factors were isolated from three dogs that had died of either otitis (followed by a generalized infection), enteritis or bronchopneumonia — secondary to distemper. The bacteria isolated from cats were recovered at the necropsy of animals that had died of purulent pleuresy and feline panleukopenia. The other strains (two Salmonella and one E. coli were isolated from fatal enteric diseases in the pig, calf and kangaroo.

     

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 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.

     

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.

     

Studies on the Susceptibility of Water Buffaloes to Leptospira

Eight mature farming type, Taiwan, water buffaloes were inoculate with L. australis A while six received L. canicola. Before inoculation all animals were negative to the microscopic-agglutination test (agglutinationlysis test) using the above species as antigen.

No sign of clinical leptospirosis was observed although four animals developed temperatures.

Cultures made from buffalo blood, kidneys and urine and from blood of guinea pigs inoculated with kidney emulsion and urine from the inoculated buffalo were all negative for leptospiral organisms.

Blood samples drawn from the water buffalo 2, 3 and 4 weeks post inoculation were negative to the microscopic-agglutination test except for one animal. Blood from the animal taken two weeks post-inoculation was positive at 1:100 dilution with L. australis A antigen but that taken at 3 and 4 weeks was negative.

     

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.