Flow cytometric analysis of intracellular complexity and CD45 expression for use in rapid differentiation of leukocytes in bovine blood samples.

OBJECTIVE: To develop an efficient and reliable method that accurately differentiates bovine lymphocytes from monocytes in leukograms. SAMPLE POPULATION: Blood samples from 30 healthy cows and 1 calf with bovine leukocyte adhesion deficiency. PROCEDURE: Flow cytometric analysis of intracellular complexity and CD45 expression on bovine leukocytes was compared with results for conventional light microscopy methods. Verification of leukocyte subpopulations determined by intracellular complexity and CD45 expression was conducted, using 2-color phenotypic analysis with selected monoclonal antibodies. RESULTS: The CD45 and side-scatter properties of bovine leukocytes clearly differentiated cell types, including neutrophils, eosinophils, monocytes, and lymphocytes. CONCLUSIONS AND CLINICAL RELEVANCE: This is a rapid assay that is simple to use. More importantly, it is more accurate than the conventional method that involves the use of blood slides and light microscopy, because of the ability of the assay to readily distinguish bovine monocytes and lymphocytes. Rapid preparation of samples and short analysis times allow for efficient and reliable examination of a large number of samples, and the task of viewing slides by light microscopy is eliminated. The labor-savings benefit of this procedure is most apparent in research environments that require frequent processing of batches of blood samples.     

Feeding Frequency Affects Growth, Not Fillet Composition, of Juvenile Sunshine Bass Morone chrysops×M. saxatilis Grown in Cages

Abstract.— In an effort to feed sunshine bass Morone chrysops × M. saxatilis efficiently, promote optimal growth, and reduce labor costs associated with feeding, sunshine bass were grown in cages and fed one of four feeding frequencies: once/d, twice/d, once every other day (I X/EOD), and twice every other day (2X/EOD) for 21 wk. Juvenile sunshine bass were fed a commercial floating diet containing 40% protein and 11.5% lipid. One hundred fish were hand-counted and stocked into each of 12 3.5-m³ cages with three replications per treatment. At the conclusion of the study, percentage weight gain of sunshine bass fed twice/d was significantly (P < 0.05) higher (1,850%) compared to fish fed all other feeding frequencies. Specific growth rate (SGR) of fish fed twice/d was significantly higher (2.1%/d) compared to fish fed all other feeding regimes, while fish fed once/d had a higher SGR (2.0%/d) compared to fish fed I X/EOD (1.6%/d) and 2X/EOD (1.8%/d). Percentage survival was not significantly different (P > 0.05) among all treatments and averaged 70.4%r. Feed conversion ratio (FCR) of fish fed twice/d was significantly higher (2.40) compared to fish fed all other feeding regimes. Percentage fillet weight of fish fed twice/d was significantly higher (27.8%) compared to all other treatments. Percentage moisture, protein, lipid, and ash in fillet were not significantly different among all treatments and averaged 75.7%, 19.4%, 3.5%, and 1.2%n, respectively (wet-weight basis). Based upon data from the present study, it appears that producers growing juvenile sunshine bass in cages may want to feed fish twice daily. This feeding regimen allows for higher growth rates, without adverse effects on body or fillet compositions. However, economic analysis needs to be conducted to determine if feeding twice/d is profitable.     

Fate of Escherichia coli and Escherichia coli O157 in soils and drainage water following cattle slurry application at 3 sites in southern Scotland

Abstract. Slurry from farm animals may contaminate water supplies, rivers and bathing waters with faecal coliforms, such as Escherichia coli . Where animals harbour the O157 strain the hazard to human health is particularly high, but both the hazard level, and the low incidence and sporadic nature of the excretion of E. coli O157 make it difficult to study this strain under field conditions. The survival of total E. coli and of E. coli O157 were compared in the laboratory for two soils under controlled temperature and moisture. E. coli O157 die-off rate was the same as or quicker than for total E. coli . This result meant that field experiments studying the fate of total E. coli should give a satisfactory evaluation of the risk of water contamination by the O157 strain. In four field experiments at three sites, slurry containing total E. coli numbers of 2.2 × 10⁴ to 5.7 × 10⁵ colony forming units per mL (c.f.u. mL^–1) was applied to drained field plots. Field die-off was faster than expected from laboratory experiments, especially in one experiment where two weeks dry weather followed application. In all but this experiment, the first drain flow events after slurry application led to very high E. coli concentrations in the drains (10³ to 10⁴ c.f.u. mL^–1). E. coli O157 was present in the slurry used for two of the experiments (33 c.f.u. per 100 mL in each case). However the proportion of E.coli O157 was very low (about 1 in 10⁵) and it was not detected in the drainage water. After the first week E. coli drainage water numbers decreased rapidly but they were 1–10 c.f.u. mL^–1 for much of the sampling period after slurry application (1–3 months).     

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.     

An unusual tumor of the nictitating membrane gland of dogs-- a case report]

A tumor of the third eyelid gland was diagnosed in a 13 year old female Chihuahua. Histopathology of the excised tissue revealed an apocrine adnexoma with squamous metaplasia. The literature regarding prolapse and tumors of the third eyelid gland was reviewed. No case with similar morphology to this tumor was described to our knowledge.     

A comparison between the nutritive value of short-cutting cycle, high temperature-dried alfalfa and timothy hay for horses.

The objective was to evaluate the nutritive value of short-cutting cycle, high temperature-dried (SCCHTD) alfalfa compared to timothy hay. This was achieved by carrying out 4 x 4 Latin Square digestibility trial using 4 Thoroughbred (one three-quarter Thoroughbred) horses (mean liveweight, 531 kg). The four dietary treatments were 0AA (timothy hay only), 33AA (0.33 alfalfa: 0.67 timothy hay), 67AA (0.67 alfalfa: 0.33 timothy hay) and 100AA (alfalfa only). Digestibility data were obtained by using acid-insoluble ash to estimate apparent digestibility coefficients of nutrients. Rate of passage of the feedstuff was determined using chromium-mordanted hay. Plasma triglyceride and cholesterol concentrations were estimated. The digestibilities of organic matter (0.63), energy (0.57) and crude protein (0.74) of the alfalfa were significantly (P less than 0.001) higher than those for the hay (0.45, 0.43 and 0.36 respectively). The fibre components of alfalfa and hay were digested to the same extent but the ether extract of alfalfa was less well digested. Alfalfa saponins had no consistently significant effects on plasma cholesterol and triglyceride values but may contribute to the negative digestibility of alfalfa ether extract. We conclude that SCCHTD alfalfa is of much higher nutritive value than timothy hay when fed to Thoroughbred horses.     

Antibody responses to avian encephalomyelitis virus vaccines when administered by different routes

Antibody responses to a commercial avian encephalomyelitis virus (AEV) vaccine administered by different routes were measured by an enzyme-linked immunosorbent assay (ELISA). Responses to single doses of vaccine administered by the ocular route to 10% of a flock were comparable with those obtained when all birds received a single dose in the drinking water. However, ocular vaccination of 5% of the flock resulted in significantly lower responses than those obtained when 10% were vaccinated. Maternal antibody was shown by the ELISA to persist in chickens from vaccinated flocks for up to 21 days after hatching. Day-old chickens with serum absorbances of < 0.3 at 492 nm, as determined by the ELISA, were shown to be susceptible to intracerebral challenge with the neurotropic Van Roekel strain of AEV.