Natural mode of transmission of the bovine leukemia virus: role of bloodsucking insects.

The development of bovine leukemia virus (BLV) infection was studied in 14 noninfected young adult cattle exposed to 25 to 30 BLV-infected cows in an area of approximately 0.5 ha. Of 7 cattle (group 1) exposed beginning in July and August (midsummer) of 1976, 4 were infected by October, and all 7 by November (4 months' exposure). Of 7 cattle (group 2) exposed from February 1977 (midwinter), all remained negative for 3 months, and only 1 was positive after 6 months. By October 1977, however, 4 cattle in this group were infected, indicating that contact transmission of BLV is prevalent during the summer months. This, and the fact that BLV-infected lymphocytes were recovered from tabanids allowed to feed on a BLV-positive cow, supports the idea that bloodsucking insects play a major role in the spread of BLV.     

Fluorescence, browning index, and color in infant formulas during storage

Free and total fluorescent compounds, browning index, and color formation were measured in milk-based powdered infant formulas (IF) during 2 years of storage at 20 and 37 degrees C. The excitation spectra from 415 nm emission show three peaks (ex lambda1 = 270 nm, lambda2 = 325/315 nm, lambda3 = 350 nm) and from 347 nm excitation two emission peaks (415 and 520 nm), and no wavelength shifts were observed. Temperature and time of storage exert in general no significant effect on the development of fluorescence emission intensity and browning index. However, an important increase in pentodilysine was recorded-probably because of the iron and ascorbic acid contents of the samples-as well as in browning index in adapted IF. In both IF a color increase (deltaE) throughout storage was observed, this increase being greater in samples stored at 37 degrees C than in those stored at 20 degrees C. The increase in color with time fitted a linear regression model. Color appeared to be an indicator of sufficient sensitivity to measure the effect of temperature or storage time.     

Jaagsiekte sheep retrovirus found in milk macrophages but not in milk lymphocytes or mammary gland epithelia of naturally infected sheep

Jaagsiekte sheep retrovirus (JSRV) causes ovine pulmonary adenocarcinoma. JSRV can be transmitted via infected colostrum or milk, which contain somatic cells (SCs) harboring JSRV provirus. Nevertheless, the cell types involved in this form of transmission and the involvement of the mammary gland remain unknown. We separated adherent cells (macrophages and monocytes) by plastic adherence, and lymphocytes (CD4+ and CD8+ T cells, and B cells) by flow cytometry, from SCs in milk samples from 12 naturally infected, PCR blood test JSRV–positive, subclinical ewes. These cell populations were tested by PCR to detect JSRV provirus. The ewes were euthanized, and mammary gland samples were analyzed immunohistochemically to detect JSRV surface protein. We did not detect JSRV provirus in any milk lymphocyte population, but milk adherent cells were positive in 3 of 12 sheep, suggesting a potential major role of this population in the lactogenic transmission of JSRV. Immunohistochemistry did not reveal positive results in mammary epithelial cells, pointing to a lack of participation of the mammary gland in the biological cycle of JSRV and reducing the probability of excretion of free viral particles in colostrum or milk.     

Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Development and evaluation of a highly sensitive and specific blocking enzyme-linked immunosorbent assay and polymerase chain reaction assay for diagnosis of bovine leukemia virus infection in cattle

OBJECTIVE: To develop a blocking ELISA for detection of bovine leukemia virus (BLV) antibodies that is comparable to a radioimmunoprecipitation (RIP) assay, to evaluate use of this ELISA for identification of BLV-infected herds, and to develop a polymerase chain reaction (PCR) assay for direct diagnosis of infection with BLV. SAMPLE POPULATION: Serum samples and pooled bulk-tank milk samples from cattle. PROCEDURE: The blocking ELISA was developed, using BLV gp51 as antigen, captured by a selected bovine polyclonal serum. A nested PCR was conducted with primers specific for a segment of the pol region of the BLV genome. RESULTS: Sensitivity and specificity of the ELISA were comparable to those of the RIP assay. Use of the ELISA on pooled milk samples allowed identification of herds in which prevalence of BLV infection among lactating cows was as low as 2.5%. Pooled milk samples from BLV-free herds did not react in the ELISA. All cattle that had positive results for the nested PCR had BLV antibodies, but cattle with consistantly low antibody titers required examination of sequential DNA samples to detect viral sequences. None of the 63 antibody-negative cattle had positive results for the PCR. CONCLUSIONS AND CLINICAL RELEVANCE: This ELISA is a highly specific and sensitive assay for the detection of BLV antibodies in serum and milk samples of cattle. Examination of pooled milk samples with the ELISA provides a reliable, practical, and economic procedure for identification of BLV-infected herds. The nested PCR also constitutes a specific procedure for direct diagnosis of infection with BLV.     

Evaluation of the efficacy of two leishmanins in asymptomatic dogs.

There are few studies in dogs concerning leishmanin skin test. We evaluated and compared the efficacy of two leishmanin preparations for the detection of dog Leishmania cellular-mediated immunity. Clinically healthy dogs living in an endemic area were studied. A leishmanin preparation 1 (3 x 10(8) promastigotes/ml) was superior to a leishmanin preparation 2 (5 x 10(6) promastigotes/ml), measured as the percentage of positive reactions and the diameter of the induced induration. The leishmanin skin test is a valuable tool, although the results show that the degree of response, as it has been shown in human beings, depends on the preparation used.     

Taenia saginata derived synthetic peptides with potential for the diagnosis of bovine cysticercosis

Immunity in Taeniids is predominantly antibody mediated and thus many serological immuno-determinants will have potential in both protection and diagnosis. The antigenicity of six peptides derived from four potentially protective molecules cloned from a Taenia saginata oncospheres cDNA library have been evaluated as targets for the specific diagnosis of bovine cysticercosis. The six peptides consist of: two peptides (HP6-2 and HP6-3) derived from the sequence of the 18 kDa surface/secreted oncospheral adhesion antigen identified by McAb-HP6, two peptides (Ts45W-1 and Ts45W-5) derived from the sequence of the T. saginata homologue of the T. ovis 45W protective gene family, one peptide (TS45S-10) derived from a T. saginata sequence with significant similarity to the T. ovis 45S protective antigen, and one peptide (TEG-1) derived from the sequence of the T. saginata homologue of Echinococcus spp. main surface protein. Longitudinal studies indicate that T. saginata infected cattle respond to all six peptides by 3-4 weeks post-infection and that the antibody levels remain high for at least 12 weeks post-infection. As protection against Taeniid parasites is predominantly antibody mediated, some of these six peptides may be of value as immuno-prophylactic tools and hence also in assays to determine resistance to infection with the parasite. For diagnosis, on the other hand, only three peptides (HP6-2, TEG-1 and Ts45S-10) performed with the necessary sensitivity and specificity to determine exposure to infection with T. saginata, and now merit an exhaustive evaluation prior to employment as routine diagnostic tools.     

Quantitative study of 'flame follicles' in skin sections of Shar-pei dogs

This study determined the frequency of occurrence and the mean number of 'flame follicles' per skin section and assessed their diagnostic significance in cutaneous biopsies of Shar-pei dogs. The number of 'flame follicles' per section was recorded in skin sections from 42 Shar-pei dogs, of which 40 had non-neoplastic skin disease and non-atrophic dermatoses and 2 had healthy skin. Forty-two skin sections from dogs of different breeds served as control specimens, 28 of which were examples of non-neoplastic and non-atrophic dermatoses and 14 were from dogs with healthy skin. Differences among groups were analysed by means of the unpaired Student's t-test. It was concluded that 'flame follicles' were more frequent and found in significantly higher numbers in the Shar-pei group when compared with the control group suggesting that 'flame follicles' in skin sections from Shar-pei dogs do not have the same diagnostic significance as in other breeds.