Vaccination of the brushtail possum (Trichosurus vulpecula) against Mycobacterium bovis infection with bacille Calmette-Guérin: the response to multiple doses.

In New Zealand, the brushtail possum (Trichosurus vulpecula) is the principal wildlife vector of bovine tuberculosis. Control of infected possum populations contributes to the control of tuberculosis in domestic livestock. Vaccination is potentially a complementary strategy to population control, but to be cost-effective, administration of the vaccine to possums would need to be from an appropriately designed automatic vaccinator. Possums themselves would activate the vaccinator so that it would deliver an aerosol spray of vaccine. There would be no direct way to prevent possums receiving multiple doses of vaccine. This study examined the effect on protective immunity of repeated vaccination. Captive possums were vaccinated with BCG strain pasteur 1173P2 either 12 times at weekly intervals, twice at 6-weekly intervals, or once. Vaccination was by a combination of intranasal aerosol and conjunctival instillation. Eight weeks after the last dose of vaccine, all possums were challenged intratracheally with Mycobacterium bovis strain 83/6235. Vaccination induced a significant immune response as measured by the lymphocyte proliferation assay (LPA). A significant level of protection, as measured by the response to challenge, developed in all the vaccinated possum groups, but protection was greatest in the group vaccinated 12 times. It was concluded that protection would be enhanced if vaccinations were repeated at short intervals (weekly), but no benefit or detriment resulted from revaccination after longer intervals (1-2 months).     

Bovine sire effects on daughters' in vitro blood neutrophil functions, lymphocyte blastogenesis, serum complement and conglutinin levels

Blood neutrophil functions, lymphocyte blastogenic responses, serum complement, and serum conglutinin activity of 98 lactating Holstein cows from two genetic lines were evaluated. The genetic lines were produced in a selection experiment that created and perpetuated genetic differences in milk production for up to seven generations. No significant differences between the two genetic lines of cows were found for neutrophil function, lymphocyte blastogenic responses, serum complement levels, or serum conglutinin levels. Significant differences between sire progeny groups within lines were found for unstimulated and mitogen-stimulated lymphocyte blastogenesis (P less than 0.0001), and almost all neutrophil functions (antibody independent neutrophil cytotoxicity, antibody dependent neutrophil cytotoxicity, ingestion of bacteria, iodination, chemiluminescence, chemokinesis, and chemotaxis (P less than or equal to 0.05)). Sire progeny group differences (P less than or equal to 0.0001) within lines for serum complement and conglutinin activity were also found. Neutrophil chemiluminescence activity (positive relationship; P less than or equal to 0.001), concanavalin A-stimulated lymphocyte blastogenesis (positive relationship; P less than or equal to 0.004), and serum conglutinin activity levels (negative relationship; P less than or equal to 0.01) each had small but significant associations with the total milk somatic cell count. Cows seropositive for bovine leukosis virus had increased resting and mitogen-stimulated lymphocyte blastogenic activity and were associated with increased in vitro neutrophil random migration and production of superoxide anion. Estimates of genetic parameters of various immune cell functions, of serum complement and of conglutinin levels for daughters of 11 sires with 4-6 daughters in the data set were determined. In this report, genetic variation was demonstrated for nonspecific humoral and cellular immunity.     

Culture of bovine bone marrow progenitor cells in vitro.

In vitro methylcellulose cultures of bovine bone marrow progenitor cells were developed. An existing technique described for bovine species was compared to a method for human tissue and further adapted during subsequent experiments. Bovine bone marrow samples were collected at the slaughterhouse, and mononuclear cells were separated by gradient centrifugation (1.077 g/ml specific density and 400 g). The use of 3% bovine leucocyte-conditioned medium, produced by stimulation of blood lymphocytes with 4 microg/ml concanavalin A and harvested on day 4 of culture, gave better results than the use of supernatant of the human bladder carcinoma 5637, which is widely used in human bone marrow cultures. However, bovine leucocyte-conditioned medium was not added to erythroid cultures because inhibitory effects were observed. Erythroid colonies were stimulated with erythropoietin, and hemin was added to enable microscopic identification. Reduced oxygen tension was necessary to induce growth of erythroid colonies. This was not necessary for myeloid cultures. In conclusion, the results of this study show that the growth of myeloid and erythroid colonies in methylcellulose-based medium requires different culture conditions, which are different from the culture conditions for human cells.     

The Application of GIS in Small Watershed Classification in Loess Plateau

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Disappearance of acetic acid from the bovine reticulorumen at basal and elevated concentrations of acetic acid.

Disappearance of acetic acid was quantified to determine whether removal of this acid from the reticulorumen is altered when ruminal acetic acid concentrations are elevated. Ruminally fistulated beef steers (n = 3 per experiment; BW = 320 +/- 9 kg) were fed eight times daily a 46% corn-based concentrate:54% mixed hay diet to meet maintenance energy requirements (3.5 kg of DM/d). In situ production of acetic acid, determined by pulsed-continuous infusion of [1-14C]acetic acid, was 530 mmol/h (CV = 12%). Disappearance from the reticulorumen (i.e., presumed absorption) was 460 mmol/h (CV = 2%) or 87% (CV = 2%) of ruminal production. Variation is described within an operational steady state. Acetic acid concentrations were increased by continuous infusion of unlabeled acetic acid into the reticulorumen. Ruminal disappearance (mmol/h) increased when this simulated production was elevated up to and exceeding in situ rates reported previously (530 to 2,700 mmol/h). These data suggest that two-thirds to three-fourths of ruminal acetate production disappeared across the reticuloruminal wall when concentration was elevated; the complement exited from the rumen with the liquid phase. The reticulorumen has an additional capacity to remove acetic acid; however, it does so with an apparent reduced efficiency at higher production rates. Liquid out-flow may affect the efficiency of reticuloruminal disappearance. Sites distal to the rumen become quantitatively important when ruminal acetic acid concentrations are increased.     

Toxoplasmosis in Pallas' cats (Otocolobus felis manul) at the Denver Zoological Gardens.

In May 1996 the Denver Zoological Gardens obtained two male and two female Pallas' cats (Otocolobus felis manul) that were wild-caught in the Ukraine. These animals were part of a group of 16 wild-caught adults (eight male and eight female) imported to the United States and Canada between 1995 and 1996. The Denver Zoological Gardens cats were quarantined at the zoo hospital for approximately I mo. During the quarantine period they were immobilized for physical examination, and sera were obtained from them to evaluate for exposure to Toxoplasma gondii. All cats were positive for T. gondii antibodies by latex agglutination (titers from 1:512 to 1:1,024). After being paired for breeding, one pair produced two litters, and another pair produced four litters, a total of 17 kittens between 1997 and 2001. Four kittens and two young adults died from a disseminated granulomatous and necrotizing inflammation consistent with toxoplasmosis. Toxoplasma gondii infection was confirmed in all six deceased cats by polymerase chain reaction performed on formalin-fixed tissues. An additional five kittens disappeared and were not available for necropsy. The fatality rate from toxoplasmosis was 35.3% (6/17) for cats that were available for necropsy and could have been as high as 64.7% (11/17) if it were assumed that the disappeared kittens were also affected. The Pallas' kitten survival rate at the Denver Zoological Gardens was 35.3%. This article describes the clinical and pathologic features of toxoplasmosis in a group of Pallas' cats at the Denver Zoological Gardens.     

Relating Kirtland's warbler population to changing landscape composition and structure

The population of male Kirtland's warbler (Dendroica kirtlandii) in the breeding season has averaged 206 from 1971 to 1987. The Kirtland's warbler occupies dense jack pine (Pinus banksiana) barrens from 5 to 23 years old and from 1.4 to 5.0 m high, formerly of wildfire origin. In 1984, 73% of the males censused were found in habitat naturally regenerated from wildfire or prescribed burning. The rest were in plantations (11%) or in harvested, unburned jack pine stands stocked by natural regeneration (16%). Twenty-two percent (630 of 2,886) of the Kirtland's warbler males counted in the annual censuses from 1971 through 1984 were found in 26 stands that were unburned and naturally regenerated following harvest. From 1982 to 1987, suitable regenerating areas were barely sufficient to replace currently occupied maturing stands, so population growth was impeded. Ecosystems of suitable size and regeneration characteristics (wildfire and plantation) doubled in area by 1989. In response, the population of Kirtland's warblers increased from 167 to 398 males between 1987 and 1992, but they withdrew almost entirely from the unburned, unplanted barrens by 1989 when the area of more suitable regeneration types increased. Minimum (368 males) and maximum (542 males) population estimates for 1996 were calculated based on 1984 average density (1.9 males per 40 ha) and peak population in burns (2.8 males per 40 ha).     

Bovine spongiform encephalopathy in Northern Ireland: epidemiological observations 1988-1990.

This study describes the epidemiological features of bovine spongiform encephalopathy (BSE) in Northern Ireland where the first case occurred in November 1988. They were very similar to those observed in Great Britain except that the annual incidence of BSE in 1990 in Northern Ireland, 2.3 confirmed cases per 10,000 adult cows, was approximately one 10th of that in Great Britain. The findings were also consistent with the current hypothesis that affected cattle had been exposed to a scrapie-like agent via cattle feedstuffs containing ruminant-derived protein. However, a preliminary investigation of the potential sources of infection for cattle in Northern Ireland did not provide any conclusive evidence.