A total of five data sets obtained from four separate sheep flocks were used to evaluate the effects of subclinical maedi-visna virus infection (as determined by serological procedures) on productivity in ewes. In general, infection with maedi-visna virus had a detrimental effect on productivity. Infection resulted in a reduction in conception rates, with the odds of an infected ewe conceiving being only approximately 0.67 times those of a non-infected ewe. For ewes which did conceive, there was no detrimental effect of infection on the number of lambs born. The birth weights of lambs born to middle-aged (3–4 years) infected ewes were 3–6% lower than the weights of lambs born to non-infected ewes of the same age. There was very little apparent effect of infection in younger (1–2 years) and older (5 + years) ewes: some factors which may have biased these results are discussed. Lamb weights at 25–50 days of age appeared to be reduced by maedi-visna infection in the ewe but the results were not statistically significant.
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. After incubation, zones of hemolysis were produced by positive sera. The activity of some positive sera was heat labile, but was restored to heated sera by addition of a crude preparation of the first component of bovine complement. The HIGT was compared with the complement fixation test (CFT), the buffered antigen plate test (BPAT), the standard tube agglutination test (STAT) and the standard plate agglutination test (SPAT), using sera from 1041 brucellosis-free cattle, from 51 cattle infected with B. abortus biotype 1 or biotype 4, and from six heifers vaccinated with strain 19. Three of 1041 sera (0.29%) from brucellosis-free cattle were HIGT-positive, ten (0.96%) were BPAT-positive, and none were positive in the CFT, STAT, or SPAT. The HIGT was more sensitive, and detected infection earlier than the other tests in the case of B. abortus biotype 1 infection, but was less sensitive for biotype 4 infection. In vaccinated heifers, HIGT reactions appeared later than reactions to the other tests, and persisted as long as 565 days. Further studies are needed to standardize the HIGT test conditions and to improve its sensitivity for B. abortus biotype 4 infections. Attempts to determine the effects of LPS antigen prepared from different biotypes of B. abortus are in progress.
Since the identification of approximately 1 400 bovine serological reactors to bluetongue virus in the Okanagan Valley of British Columbia in 1976, there has been no evidence of virus establishment in Canada. No clinical signs suggestive of bluetongue were observed. It was not possible to demonstrate viral activity at the time the seropositive animals were detected and subsequent serological testing supports the hypothesis that the virus has not become endemic or indeed survived in Canadian cattle populations. This combined with the dramatic reduction in prevalence of serological reactors in the years following the initial slaughter suggests that viral activity occurred in the Okanagan Valley prior to 1976 and disappeared. There has been no evidence for transmissions different from that expected of a classical arbovirus; that is, no evidence of "vertical" transmission.
Live virus vaccines against bovine and porcine diseases were examined for the presence of adventitious pestivirus RNA or pestiviruses by reverse transcription-polymerase chain reaction (PCR). Pestivirus RNA was detected in the live virus vaccines against Akabane disease, Ibaraki disease, infectious bovine rhinotracheitis, porcine parvovirus infection, transmissible gastroenteritis and Japanese encephalitis. Pestivirus RNA or pestivirus in the fetal bovine serum used to grow the host cells used to prepare the bovine and swine viral vaccines is a likely source of the contamination. Nucleotide sequence analysis of the PCR products suggests that modified live virus vaccines being used for immunization of cattle against bovine viral diarrhoea was not responsible for the contamination of the vaccines examined.
One hundred and fifty-one, day 6 or 7, embryos collected from cattle infected with bovine leukaemia virus (BLV) were transferred to uninfected recipients. Thirty-two pregnancies resulted. Two animals aborted at seven months. Three sets of twins and one single calf were still-born. The remaining 26 pregnancies produced 27 live calves which were raised to six months of age. All of the recipients, pregnant and non-pregnant, and all of the calves remained serologically negative for antibodies to BLV-glycoprotein antigen.
Ten strains of Yersinia enterocolitica belonging to ten various serogroups isolated from raw milk were inoculated into groups of five guinea pigs and five calves. Y. enterocolitica serotype 0:16 was the only serotype tested that induced an antibody response to Brucella abortus in calves. No anti-Brucella response could be demonstrated serologically in guinea pigs. Activity of the anti-Y. enterocolitica 0:16 calf sera against B. abortus antigen was shown by the tube agglutination test, and by the complement fixation test. The early agglutinating antibody response was partly sensitive to reduction by 2-mercaptoethanol. This sensitivity decreased later in the response. This is the first report of anti-Brucella responses induced by a serotype of Y. enterocolitica other than 0:9; sera from a group of five calves inoculated with 0:9 were tested by the same serological techniques for comparison.
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 (16.0%) UFF-sediment specimens as compared to BSV in 14/15 (93.3%) and 21/25 (84.0%) of the similar specimens and neither BLV or BSV were found in 26 eggs and 60 embryos collected from 20 of the 25 cows. Detection of BLV antigens by immunofluorescence was hampered by the competitive replication of both BLV and BSV and competitive growth in indicator cells and uterine cells. As BLV has not been observed in cells of UFF sediments, it was probably isolated from leucocytes present in the lumen of uterus or from blood seeping out from inapparent vessel damage during flushing. Isolation of BLV in UFF sediments gives additional evidence to the concept of a transplacental transmission by a not yet elucidated mechanism. The high rate of BSV recovery from cells of UFF sediments indicated that this virus is more wide-spread than previously shown and that it may play a role in causing disorders of the reproductive tract.
Infection of normal adult cattle and sporadic serological Brucella abortus reactors, which no longer had diagnostic titers to B. abortus, with a culture of Escherichia coli isolated from a cow (or its environment) resulted in the production of a primary IgM serum antibody response to both B. abortus and E. coli. Cattle injected with heat killed E. coli and guinea pigs or young calves lacking natural agglutinins to B. abortus injected with live E. coli, produced serum antibody only to E. coli. The subsequent reinjections of live E. coli into the former two groups of cattle resulted in all but one animal in each group producing IgM ‘secondary’ responses to B. abortus of decreasing magnitude, while the anti-E. coli responses increased and eventually switched to synthesis of IgG class antibody. The remaining two animals produced a series of ‘secondary’ responses of IgM antibody to B. abortus similar in amplitude and duration to the primary immune response. The anti-E. coli agglutinins of these two animals increased in titer and IgG class antibody to E. coli was evident after several injections of E. coli. These results indicate that infection of cattle by E. coli can cause a problem in the serological fiagnosis of B. abortus infection. Speculations on the cause of this cross-reaction and ways of minimizing misdiagnosis are discussed.
A bluetongue virus plaque neutralization test, done in small tissue culture wells (2.0 cm2) using minimal antibody-virus contact time, was found to be of comparable sensitivity to more classical methods involving preincubation. By using the cell culture plates as serum dilution wells and adding cells directly to the virus-antibody mixture, a considerable saving in time is realized. Because of the small size of the wells, reagent cost and incubator space requirements are reduced making the test much more attractive for wide scale use, either as a screening test or one that yields dose-response lines.
Six hundred and fifty-three alkaligenic and psychrophilic organisms were isolated from 136 samples of milk and milk products, and 182 belonged to the genus Pseudomonas. The Pseudomonas, with 16 known species, were further classified into four groups for 97 features by Adansonian's classification. By S values Group I comprised 98 cultures identified as P. fluoresceins and “psychrophilic P. aeru-ginosa-like” organisms. Group II consisted of 80 cultures, with fluorescents characterized as P. putida species, with an intermediate group between P. putida and P. fluorescens. The nonfluorescents were related to P. fragi and P. putrefaciens. Group III had only 15 cultures which could not be associated with any group or species. Group IV comprised 5 cultures which were related to P. alkaligenes. The known type species, members of Groups III and IV, by S and M comparisons were markedly dissimilar.
1.1. Chromatographically purified preparations of diphtheria toxin and chicken antibody to mouse lactate dehydrogenase (L-lactate:NAD+ oxidoreductase, EC 1.1.1.27) isoenzyme 5, were conjugated by p,p′-difluoro-m,m′-dinitrodiphenyl-sulphone and used to study the value of antilactate dehydrogenase as a carrier of toxic compounds into malignant cells.2.2. The conjugate, following chromatographic separation from nonconjugated molecules, was shown by double immunodiffusion to contain both antienzyme and toxin, and to give significant inhibitions of lactate dehydrogenase activity and protein synthesis in cell-free systems.3.3. Antilactate dehydrogenase and conjugate bearing this antienzyme were each shown, by means of fluorescein-conjugated rabbit antichicken γ-globulin, to have been taken up and accumulated within Ehrlich ascites cells in cell culture but not by mouse kidney cells. Conjugate bearing toxin, but not toxin alone, was similarly shown, by means of rhodamine-conjugated antitoxin, to have been taken up and accumulated in these cells.4.4. The cytotoxicity of the toxin, as measured by inhibition of both protein synthesis and glycolysis, was decreased for kidney cells and increased for Ehrlich cells as a result of conjugation with antilactate dehydrogenase.