Replication of the bovine immunodeficiency-like virus in diploid and aneuploid cells: permanent, latent and virus-productive infections in vitro.

Bovine immunodeficiency-like virus (BIV) is an infectious and leukotropic retrovirus, the sole lentivirus candidate which has been isolated from cattle. Although BIV has recently been shown to be related to the human immunodeficiency virus, there is very limited information on the replication and the pathogenesis of BIV. It is reported here that BIV can permanently infect diploid and aneuploid cells from four different species: bovine, canine, ferret and ovine.

Characterization of an atypical biotype of Brucella abortus.

Brucella abortus strains were isolated from bovine tissue and milk samples from seven Ontario herds. The isolates were characterized by colonial morphology, requirement of CO2 for growth, lysis by Tbilisi phage, biochemical tests and agglutination in monospecific sera. They resembled B.

A comparison of the bovine leukemia and bovine syncytial virus status in utero-tubal cells recovered from fluids used to flush the uterus and oviducts of BLV-infected, superovulated cattle.

Twenty-four cell lines were established from uterine-oviductal flush fluid (UOFF) cells from 20 bovine leukosis virus (BLV)-infected embryo-donor cows and 4 BLV-free control cows harvested by the Ficoll-gradient technique. Similar epithelial-like and fibroblast-like cells were observed in the primary cultures of UOFF from both groups. BLV-antigens were not detected with direct immunofluorescence test in any of the cell-lines from the 20 positive BLV-cows but a positive reaction was observed with the competitive radioimmunoassay in one cell line only.

A comparison of five serological tests for bovine brucellosis.

Five serological assays: the buffered plate antigen test, the standard tube agglutination test, the complement fixation test, the hemolysis-in-gel test and the indirect enzyme immunoassay were diagnostically evaluated. Test data consisted of results from 1208 cattle in brucellosis-free herds, 1578 cattle in reactor herds of unknown infection status and 174 cattle from which Brucella abortus had been cultured. The complement fixation test had the highest specificity in both nonvaccinated and vaccinated cattle.

The persistent infection of a canine thymus cell line by equine infectious anaemia virus and preliminary data on the production of viral antigens.

Equine infectious anaemia virus (EIAV) was adapted to the Cf2Th cell line, a heterologous malignant line from canine thymus. A persistent infection was monitored for 100 serial passages by demonstrating the presence of virus and viral antigens at each 10th passage by electron-microscopy, immunodiffusion and immunofluorescence. Chromosome analysis of EIAV-infected cells indicated they had a karyotype resembling the control cells of similar passage history.

Embryo transfer in relation to bovine leukemia virus control and eradication.

Two hundred and seven, zona pellucida-intact bovine embryos were collected from bovine leukemia virus-infected donors, washed, and transferred to uninfected recipients: 111 of these embryos were sired by bovine leukemia virus-infected bulls. Fifty live calves were obtained from the 57 pregnancies resulting from the transfers. None of the recipients or calves developed antibodies to bovine leukemia virus.

Campylobacter fetus in artificial insemination unit and slaughterhouse bulls in Ontario.

Preputial fluid samples were collected from 90 bulls in two Ontario artificial insemination units using a penial glove swab technique previously developed by one of us for use in donor bulls. No Campylobacter fetus organisms were identified from the prepuce or from samples of semen collected at the same time from these bulls. The distal genitalia of 200 bulls were collected at a slaughter house.

Isolation, characterization, and serotyping of Campylobacter jejuni and Campylobacter coli from slaughter cattle.

A total of 525 specimens from 100 slaughter beef cattle were examined for the presence of Campylobacter jejuni and Campylobacter coli by direct plating and enrichment techniques. Isolates were identified by cultural, biochemical, antibiotic sensitivity, and immunofluorescence tests and further characterized with the aid of recently developed biotyping and serotyping methods. Fifty animals were positive for C.

Quantitative evaluation of a transport-enrichment medium for Campylobacter fetus.

The enrichment feature of a selective serum-based transport medium for Campylobacter fetus was quantitatively examined. Preputial samples from artificial insemination bulls were spiked with known numbers of C fetus strains and inoculated into transport-enrichment medium (TEM). The survival and multiplication of these strains in TEM under different incubation periods and temperatures were assessed by plate counts.

Isolation of bovine syncytial virus from lymphocytes recovered from fluids used to flush uterus and oviducts of superovulated cattle.

A Ficoll-gradient method was applied to isolate lymphocytes from fluids used to flush the uterus and oviducts of superovulated cows. Bovine syncytial virus antigens were demonstrated in 15 of 19 cows by cocultivation of lymphocytes with fetal lamb spleen cells and examining them with direct immunofluorescence. Viral serum antibodies were found in the same 15 of 19 cows as above by the modified direct complement-fixation test.

An hemolysis-in-gel test for bovine brucellosis.

An hemolysis-in-gel test (HIGT) for bovine antibody against Brucella abortus was developed and evaluated. Sera to be tested were placed in wells in an agarose gel containing guinea pig complement and J-negative bovine erythrocytes coated with lipopolysaccharide prepared from B. abortus biotype 1.

Detection of Campylobacter fetus in artificial insemination bulls with a transport enrichment medium.

One hundred and five bulls from an artificial insemination unit were tested for the presence of Campylobacter fetus subspecies venerealis. The method involved the inoculation of preputial samples into a new transport enrichment medium prior to culture and immunofluorescence tests. Seventeen bulls (16%) were found to be either positive or suspected carriers of C.

Attempts to isolate bovine leukemia and bovine syncytial viruses from blood, uterine flush fluid, unfertilized ova and embryos from infected donor cattle.

Blood leucocytes, sediments of uterine flush fluid (UFF), eggs and embryos from 25 BLV-positive donor cows were tested for bovine leukemia (BLV) and bovine syncytial (BSV) viruses by cocultivation with fetal lamb spleen cells and by applying syncytium induction and immunofluorescence tests. BLV was diagnosed in 11/15 (73.3%) leucocyte and 4/25 (16.

Demonstration of parvovirus in Canadian swine and antigenic relationships with isolates from other countries.

A Canadian isolate of porcine parvovirus, isolated from cultured pig thyroid cells, was shown to be antigenically indistinguishable from a British (59e/63) and a German (G10/1) strain when treated by the modified direct complement-fixation, the hemagglutination-inhibition and the fluorescent antibody tests. These tests also revealed that antibodies to parvoviruses were detectable in a large proportion of the conventionally raised pigs in the provinces of Quebec and Ontario. Cell cultures, prepared from tissues collected in a slaughterhouse, were often found to be infected with parvovirus.

Transmissible gastroenteritis: demonstration of the virus from field specimens by means of cell culture and pig inoculation.

Isolation of transmissible gastroenteritis virus was attempted from segments of jejunum collected from piglets submitted for diagnosis of transmissible gastroenteritis. The virus was isolated more frequently in susceptible piglets than in pig kidney or pig thyroid cells. Practically, both cell systems were equally capable of demonstrating the virus when the tissue suspensions were sonicated.

A comparison of some serological tests for bluetongue virus infection.

The plaque neutralization, complement fixation, and agar gel precipitin tests were compared by measuring bluetongue virus antibody in 137 serums from experimental animals (cattle and sheep) and suspected field reactors (cattle and deer). In general, the tests agreed well with each other. Plaque neutralization titers began earlier than the other two and went much higher than the complement fixation titers.

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.

Anaplasmosis: control of the first outbreak in Canada by serological identification and slaughter.

On August 13, 1968 Canada experienced its first outbreak of anaplasmosis. The initial diagnosis based on hematological and clinical evidence was made by the Provincial Veterinary Laboratory, Winnipeg, Manitoba, and later confirmed in our laboratory by use of the complement-fixation test, hematology, and animal transmission studies. Sixteen herds (1,717 cattle) were examined but the outbreak was found to be localized mainly in one herd of 830 cattle.

Studies on bovine virus diarrhea: serum neutralization, complement-fixation and immunofluorescence.

The complement-fixation, the serum neutralization tests and the fluorescent-antibody technique were the serological methods applied in this laboratory for the detection of antigens for bovine virus diarrhea (BVD). As observed previously, the modified direct complement-fixation (MDCF) test was required to demonstrate antibodies against virus infections of cattle. At a certain stage of infection, the MDCF test was found to be as accurate and less time-consuming than the serum neutralization test for the detection of antibodies in bovine sera.

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.

Equine infectious anemia: preliminary investigation of the complement-fixation test for the demonstration of antibodies and antigen.

Clinical field cases of equine infectious anemia were studied and the disease was reproduced experimentally in horses. Attempts were made to adapt the complement-fixation test to the detection of antibodies in the serum of infected animals and to the demonstration of antigens in tissue extracts.A moderate complement-fixing antibody response was demonstrated in the serum of horses shortly after primary exposure to the infectious agent.

Studies on bluetongue. VI. Animal transmission studies.

The Cyprus strain of bluetongue virus was successfully transmitted through six passages and the Station strain through one passage in calves. Although the animals developed no visible evidence of infection, viremia as shown by both passage and fluorescent antibody examination of infected foetal bovine kidney culture, and by serological conversion was nevertheless demonstrated. No enhancement of virulence for calves or sheep was shown following bovine passage.