Correlation of Bedrock Type with the Geography of Leptospirosis

Leptospirosis occurs enzootically over most of Southern Ontario. Leptospira pomona is the serotype most commonly found in outbreaks. Antibodies to L. pomona occur frequently in the sera of deer in wilderness areas. The geographic location of leptospirosis presents a pattern which closely parallels the distribution of Paleozoic bedrock. By contrast, L. pomona infection is absent from areas underlain by Precambrian bedrock. Comparisons of water chemistry, soil type, habitat, and host and pathogen availability in these two geologically distinct environments have not defined the mechanisms involved in the disease pattern. Leptospires resembling saphophytic strains occur widely, regardless of bedrock type. High titers to L. biflexa, a saprophytic serotype, were found frequently in deer sera from a Precambrian area which was surveyed intensively. Antibodies to L. hardjo and L. sejroe occur in many bovine sera from a predominantly Precambrian area where Paleozoic outliers are numerous. Colloidal clay is common to leptospiral habitats. A microenvironment structured by the surface activity of clay is likely to be a key ecological factor in the landscape epizootiology of leptospirosis. In Ontario, bedrock composed of limestone and dolomite formed in the Paleozoic era appears to be a reliable ecological marker for Leptospira pomona infection.     

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.

     

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.     

Response of Gnotobiotic Pigs to Escherichia coli

In a study of the response of gnotobiotic pigs to coliform infections, 45 one-week-old germfree pigs were divided into five groups and each group was inoculated orally with a different strain of Escherichia coli. Three of these were enteropathogenic swine strains, P307[08:K87(B), K88 a,b (L):H19]; P570 [0138:K81]; P568[0141:K85a,b(B), K88a,b(L):H4], one was a virulent human strain, H224, [026:K60(B6)], and one was a non-enteropathogenic swine strain, P581[OX13:K68]. It was attempted to protect a portion of the pigs with orally administered specific antisera and sera from non-immunized specific pathogenfree (SPF) pigs. Observations were made on the clinical response, bacterial counts of feces and intestinal contents, gross pathological changes, distribution of the organisms in organs and serum hemagglutinin titers.

Infection with E. coli P307 resulted in diarrhea, dehydration and death, unless the pig was protected with specific antiserum. The pigs infected with E. coli P570 had a transient diarrhea but retained their appetites and recovered. Those infected with the other three strains remained healthy throughout. No circulating hemagglutinating antibody against the test strains of E. coli could be detected in any of the pigs seven days or earlier post-inoculation.

Relationship could not be established between the numbers of viable E. coli in the feces and the presence of clinical colibacillosis. Orally administered specific antiserum afforded protection against strain P307, but did not reduce the number of E. coli in the gut or alter their distribution in the internal organs. This suggested that the protective effect of specific antibody in the intestine was due to its action on a metabolite (enterotoxin) produced by E. coli P307 rather than the organism itself.

     

Experimental Atrophic Rhinitis in Gnotobiotic Pigs

Twenty-nine caesarian derived colostrum deprived germfree pigs were reared in isolators in groups of three to four per isolator. At seven days of age each group was inoculated intranasally with one of four strains of Bordetella bronchiseptica (designated B, J, L and 55B), or Pseudomonas aeruginosa or a mucoid strain of Escherichia coli, all previously isolated from nasal mucus of pigs affected with clinical atrophic rhinitis. Another group was inoculated simultaneously with B. bronchiseptica B and Pasteurella multocida. The animals were observed for clinical signs of atrophic rhinitis and monitored bacteriologically at weekly intervals for seven weeks. Then they were bled for serology and killed and their respiratory organs examined for gross and histopathological lesions.

All of the pigs inoculated with the Bordetellae had inflammation of the nasal mucosa and developed positive serum antibody titers against all four of the Bordetella strains used in this study. Strain J caused sneezing and turbinate atrophy in three of four pigs. One of the three pigs inoculated with strain L died in ten days from bronchopneumonia and pericarditis and had turbinate atrophy. Strains B and B55 caused no turbinate atrophy, but two out of three pigs inoculated with both B. bronchiseptica B and P. multocida had turbinate atrophy. No nasal lesions were observed in the pigs inoculated with E. coli or P. aeruginosa or in the noninoculated germfree controls.

The results indicate a variation in the ability of different strains of B. bronchiseptica to cause turbinate atrophy in pigs and demonstrate that nasal infections by these organisms stimulate serum antibody response. Presence of P. multocida appears to increase the severity of the lesions. As the E. coli and Pseudomonas failed to produce atrophic rhinitis, they are probably of no significance as primary etiological agents in the atrophic rhinitis syndrome in swine.

     

Anomalous Origin of the Coronary Arteries in Two Calves

Anomalities of the coronary arteries may include one or more of the following variations: abnormal number, abnormal course, or abnormal origin. Many of these anomalies have been described in man.⁽²⁸⁾ Anomalous origin of the left coronary artery from the pulmonary artery (A.O.L.C.A.) has been estimated ⁽¹⁹⁾ to occur in one of 300,000 children and to represent approximately 0.5% of all types of congenital heart disease. In a recent review⁽²⁸⁾ of these conditions in man, anomalous origin of the right coronary artery from the pulmonary artery (A.O.R.C.A.) had been reported 16 times, A.O.L.C.A. 39 times, and anomalous origin of both coronary arteries from the pulmonary artery (A.O.B.C.A.) 5 times.     

Some Observations on the Diagnosis and Epidemiology of Leptospirosis in Swine

The results of combined epidemiological, clinical, serological, bacteriological and histopathological studies following an outbreak of disease caused by L. pomona on a farm stocked with cattle, sheep, pigs, goats and horses maintained for experimental purposes, are reported.

The incidence of infection was high in horses, cattle and pigs. A few low titres were seen in sheep. The goats were not infected. Apart from a single bovine abortion all the clinical symptoms observed occurred in pregnant sows. Seven of these aborted or gave birth to stillborn pigs within a six week period.

Fifteen species of wildlife were trapped or shot on the farm during the year following the outbreak. L. pomona was isolated from four skunks and a porcupine. Epidemiological studies indicated that wildlife reservoir hosts were the primary source of infection for the domestic livestock.

Leptospiruria and the serological response were studied in a group of eight infected sows. Microscopic agglutination titres of 10² or less could not be associated with leptospiruria and the duration of leptospiruria was found to range from a few weeks to over two years in individual sows. Direct dark-field examination of urine proved superior to guinea-pig inoculation as a method of detecting leptospiruria and it is suggested that the former technique could be adopted with advantage as a routine aid to diagnosis.

     

Humoral Antibody Responses of Swine Infected Experimentally with Group E Streptococcus I. Whole Cell Agglutinin Response

Reference streptococcal antisera and sera collected from swine infected experimentally (by intranasal inoculation or contact exposure) with group E Streptococcus (GES) were studied in a tube agglutination system using whole GES cells.

Specificity studies revealed common group specific antigen among GES serotypes I and III, GES strains devoid of type specific antigen (untypable by ring precipitin testing) and group P and group U Streptococcus. The group specific antigens were not agglutinated by GES type specific antisera or by group specific antisera against Streptococcus groups A, B, C, D, F, G, H, K, L, M, N, or O. Results of the study suggested that GES serotypes I and III are invalid; i.e., they are devoid of type specific antigen.

Groug E Streptococcus type specific antigens II, IV, and V were agglutinated significantly only by their homologous antisera.

Experimentally infected swine developed significant titers against both the group and type specific antigen of GES. Antibodies appeared from three to eight weeks postexposure and persisted for the duration of the experiment (six months). The potential utilization of the whole cell agglutination (WCA) test for detection of GES carrier swine is discussed.

     

Effect of Ampicillin on E. Coli of Swine Origin

The in vitro susceptibility of 103 cultures of E. coli isolated from scouring and nonscouring pigs, and four cultures of Salmonella isolated from a case of necrotic enteritis was tested against Ampicillin contained in nutrient broth at concentrations of 0, 0.1, 1.0 and 5.0 uG per ml. of the medium. All but three cultures of E. coli were found to be susceptible to 5.0 uG/ml., all Salmonella isolates were also susceptible to this concentration of the antibiotic. Susceptibility of E. coli was also tested by plating dilutions of fecal samples obtained from either a scouring or a nonscouring pig, with E.M.B. agar containing 0, 0.1, 1.0, 2.5, 5.0 and 10.0 uG Ampicillin per ml. of the medium. No difference in the growth of E. coli was observed at 0, 0.1 and 1.0 uG concentrations. The three higher concentrations of the antibiotic inhibited the growth of E. coli proportional to the amount of Ampicillin in each concentration.

Ampicillin proved very effective in alleviating the symptoms of hemorrhagic enteritis in a 11-week old pig. The disappearance of scours was associated with the replacement of the previously existing sero-biotypes of fecal E. coliwith another aberrant type of E.coli which produced H₂S. No Ampicillin resistant strains of E. coli emerged following treatment of the animal with this antibiotic.

     

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.

     

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.

     

Prediction of Future Carcass Traits in Stocker Cattle at the Conclusion of Grazing

Prediction of eventual carcass traits in stocker cattle at the conclusion of grazing could be useful for culling, co-mingling of animals, feedlot pen assignments, and making management decisions in the feedyard. Ultrasound measures of 12th to 13th rib longissimus area (ULA) and fat thickness (UFAT), and off-pasture BW (OPBW) were collected from yearling cattle (n = 261) at the conclusion of grazing in two experiments that evaluated stocking rate and grazing management effects on rye (Secale cereale L.) annual ryegrass (Lolium multiflorum Lam.) pastures. Carcasss data were subsequently recorded at harvest following feedyard finishing to a visual 1-cm backfat. Correlations were analyzed to determine relationships between carcass traits and ULMA, UFAT, body condition measure (BCM), and OPBW. Ultrasound measures, breed type (BRDT; n = 4), gender (steers and heifers), and feedlot days on feed (DOF) were evaluated in multiple regression models to determine whether these variables influence eventual carcass percentage retail product, kilograms retail product (KRP), hot carcass weight (HCW), and marbling score. Ultrasound FAT and BCM were negatively correlated with percentage retail product, KRP, and HCW and were positively correlated with marbling score. All reduced regression models had R² values of between 0.15 and 0.63, and models with inputs of UFAT and OPBW consistently had the numerically greatest R² values and least RMSE. Multiple regression analyses indicated that prediction of carcass traits from stocker cattle ultrasonic measurements at the conclusion of grazing were possible, but improvement in the models will be necessary to reduce error and improve reliability.     

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.

     

Chronological Development of Mycoplasma hyorhinis and Mycoplasma hyosynoviae Infections in Cultures of a Swine Synovial Cell Strain

The sequential development of Mycroplasma hyorhinis and Mycoplasma hyosynoviae was observed in cultures of a swine synovial fluid cell strain. An early transitory filamentous phase was observed with M. hyorhinis infection followed by the development of cell-associated, relatively large, round structures and some ring forms. Infection with M. hyorhinis was characterized by a generalized distribution of the organism and a severe cytopathic effect.

Infection with M. hyosynoviae was represented by the development of circumscribed foci of small pleomorphic structures and a milder effect on the cells. At a high multiplicity of infection, this organism became associated with the cytoplasmic membranes of the cells.

     

Comparison of the Virulence of Various Strains of Moraxella bovis

The relative virulence of various strains of Moraxella bovis (M. bovis) was studied using the eyes of mice and cattle. The investigation consisted of three separate experiments. Experiments I and II involved a study on the effects of (1) different methods of growth and (2) serial blood agar passaging on the virulence of M. bovis. Experiment III involved a study on the relative virulence of different strains of M. bovis and M. bovis-like organisms.

Strains of M. bovis and M. bovis-like organisms varied in their pathogenicity for mice. However, different methods for preparation of exposure cultures of M. bovis did not influence the disease produced.

     

A Study of Complement-fixation and Serum Agglutination Tests on Vibrio fetus Infected and Immunized Cattle

Complement-fixation (CF) and tube agglutination (TA) tests for demonstration of Vibrio fetus antibodies were conducted on the sera of three groups of ten heifers. One group was vaccinated subcutaneously with a commercial V. fetus var venerealis bacterin and challenged intra-utero, at the external os cervicus one month later; the second was infected only and the third vaccinated only.

The vaccinated cattle developed high CF serum titers, but no such increase was observed in animals infected only. A moderate increase in serum antibody titers was demonstrated by the TA test following either infection or vaccination; although titers observed were not higher than those observed in the sera of some apparently normal uninfected animals. The group receiving both vaccine and challenge was the only one in which significant serum antibody titers were demonstrable by the TA test. The sera of these animals also had significant titers in the CF tests.

The CF and TA tests detected serum antibodies produced by the parenteral inoculation of V. fetus antigen. These two tests were of limited value in detecting serum antibodies from animals with genital V. fetus var venerealis infection, although the formation of local antibodies was demonstrable by the vaginal mucus agglutination test.

     

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.

     

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.

     

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.

     

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.