<Emphasis Type="Italic">Streptococcus uberis</Emphasis>-specific T cells are present in mammary gland secretions of cows and can be activated to kill <Emphasis Type="Italic">S. uberis</Emphasis>

The presence, phenotype and function of Streptococcus uberis-specific T cells in the mammary gland secretion (MGS) and blood of cows exposed to S. uberis were assessed. MGS T cells in the udder were purified and incubated with autologous blood monocytes as antigen-presenting cells (APC). Most cows, irrespective of prior S. uberis infection status and lactation status, were shown to have S. uberis-specific T cells both in MGS and in the blood. When cells from a subgroup of cows were studied, it was found that the S. uberis-specific T cells produced high levels of interferon-gamma (IFN-γ), but low levels of interleukin-10 (IL-10). A high percentage of responding T cells were of the CD8 ⁺ memory (CD45RO) subset. T cells from the MGS specific for S. uberis were propagated from animals during the drying off period and expanded in vitro using interleukin-2 (IL-2) and S. uberis antigens. This led to the accumulation of T cells of the CD8 ⁺ subset bearing memory cell markers (CD45A ⁻ , CD45RO ⁺ ), which released high levels of IFN-γ. Four of the five T cell lines derived from the MGS of three animals had substantial direct killing activity towards S. uberis in vitro. It is concluded that there is an emergence of S. uberis-specific bactericidal T cells in the MGS of cows after infection or environmental exposure to S. uberis. Vaccines aimed at activating and expanding this T cell population in the mammary glands of cattle may offer an avenue for the prevention of mastitis caused by S. uberis.     

Effect of Season on Travel Patterns and Hoof Growth of Domestic Horses

Four Morgan mares and five Morgan geldings ranging in age from 5- to 12-years-old were fitted with Global Positioning System units to determine if season has an influence on travel pattern. Body and hoof growth measurements were obtained so that the influence of season on body condition and hoof growth could be determined. Waist and heart circumference, cresty neck score, and body condition score were determined in each season. The ambient temperature and precipitation was recorded for each season. Waist circumference was the greatest (P < .05) in the spring and summer and the least in the fall and winter. Hoof growth was the greatest (P < .05) in the fall and the least in the winter. The front and rear hooves grew at similar rates in all horses. Hoof growth in geldings and mares were also similar. The average distances traveled were similar across seasons; however, the horse did numerically travel more in the spring and summer compared with the fall and winter. The range of the travel pattern was influenced by season with the horses traveling significantly less in the winter, although the average travel distances were similar. In conclusion, season in temperate zones will influence body condition, hoof growth, and pattern of travel, but the total distance traveled will be similar. Further research needs to be conducted to determine the influence of season on hoof growth and travel patterns.     

Conceptus elongation in ruminants: roles of progesterone,prostaglandin, interferon tau and cortisol

The majority of pregnancy loss in ruminants occurs during the first three weeks after conception, particularly during the period of conceptus elongation that occurs prior to pregnancy recognition and implantation. This review integrates established and new information on the biological role of ovarian progesterone(P4), prostaglandins(PGs),interferon tau(IFNT) and cortisol in endometrial function and conceptus elongation. Progesterone is secreted by the ovarian corpus luteum(CL) and is the unequivocal hormone of pregnancy. Prostaglandins(PGs) and cortisol are produced by both the epithelial cells of the endometrium and the trophectoderm of the elongating conceptus. In contrast, IFNT is produced solely by the conceptus trophectoderm and is the maternal recognition of pregnancy signal that inhibits production of luteolytic pulses of PGF2αby the endometrium to maintain the CL and thus production of P4. Available results in sheep support the idea that the individual, interactive, and coordinated actions of P4, PGs, IFNT and cortisol regulate conceptus elongation and implantation by controlling expression of genes in the endometrium and/or trophectoderm. An increased knowledge of conceptus-endometrial interactions during early pregnancy in ruminants is necessary to understand and elucidate the causes of infertility and recurrent early pregnancy loss and provide new strategies to improve fertility and thus reproductive efficiency.     

Optimization and validation of a flow cytometric method for immunophenotyping peripheral blood lymphocytes from cynomolgus monkeys (Macaca fascicularis)

BACKGROUND: Guidelines published by the Food and Drug Administration and Center for Human Medicinal Products describe the need to assess immunotoxic effects in nonclinical studies that evaluate drug toxicity, including the use of immunophenotyping to measure immunotoxicity. We are not aware of previous studies, however, that have validated methods for immunophenotyping peripheral blood lymphocyte subsets in whole blood samples from cynomolgus monkeys. OBJECTIVE: The purpose of this study was to optimize and validate a flow cytometric assay for immunophenotyping lymphocytes in the peripheral blood of cynomolgus monkeys. METHODS: A series of prevalidation experiments were done to determine optimal reagents, volumes, timing, and other procedural details of the flow cytometric assay. Using the optimized method, we then determined precision, interindividual variation, laboratory-to-laboratory variability, and sample stability. Stabilized human blood was used as a positive control for staining, processing, and analysis. The percentage and number of pan-T cells (CD3+), T-helper cells (CD3+4+), T cytotoxic/suppressor cells (CD3+8+), natural killer cells (CD3-16+), and B-cells (CD3-20+) were determined in 146 male and 140 female, clinically healthy monkeys and reference intervals were calculated. RESULTS: By doing 4-color staining with a lyse-wash method, intra- and interassay precision were <5% for all lymphocyte subsets. Variability between technicians and laboratories was minimal (CVs<3%). Samples were stable for up to 24 hours after staining and fixing. CONCLUSIONS: The validated method is extremely robust and can be performed under good laboratory practice conditions to support nonclinical studies. Reference intervals for lymphocyte subsets were similar to those previously reported.     

Cryptosporidiosis in buffalo calves (<Emphasis Type="Italic">Bubalus bubalis</Emphasis>): Prevalence and potential risk factors

The objective of the present study was to describe the prevalence and risk factors associated with cryptosporidiosis in buffalo calves in Middle Egypt. During one year, 458 fecal samples were collected from buffalo calves less than 3 month age in 55 small scale herds and examined for the presence of Cryptosporidium oocysts. Data describing age, gender, season, and herd management practices were gathered to assess potential risk factors. Fecal examination showed that 14.19% of the examined calves were positive for Cryptosporidium spp. Calves at 1-15 days were at the highest risk (P < 0.001), and a significant relationship between season and infection (P < 0.05) was recorded. A significant association between infection and hygiene (P < 0.001), type of floor (P < 0.01) and source of water (P < 0.01) was also recorded. Statistical analysis concerning the clinical signs and fecal characteristics revealed a significant association with fecal consistency (P < 0.001), presence of blood (P < 0.01) and mucous (P < 0.01). Moreover, a significant association was found between infection and the desire for suckling (P < 0.05) and tenesmus (P < 0.05). The results of the present study demonstrated the strong relation between infections by Cryptosporidium spp. and diarrhea in buffalo calves.     

Circulating oxidative stress status in dromedary camels infested with sarcoptic mange

Oxidative stress is an imbalance between radical-generating and radical-scavenging activity, resulting in oxidation products and tissue damage. This study was aimed to evaluate the status of oxidative stress indices in blood of camels naturally infested with S. scabiei. Forty-seven male camels (Camelus dromedaries) were divided according to the extent of the infested area with Sarcoptes scabiei into four groups, mild (MID, n = 12), moderate (MOD, n = 10), severely infested (SEV, n = 10) and healthy control group (n = 15). Blood was used for determination of red cell count (RBC), hemoglobin (Hb), packed cell volume (PCV), serum nitric oxide (NO^•, a free radical), ascorbate and albumin concentrations, and erythrocytic values of malondialdehyde (MDA, a marker of lipid peroxidation), protein carbonyls (PC, an indicator of protein oxidation), glutathione (GSH) superoxide dismutase (SOD) and catalase (CAT). Decreased levels (P < 0.05) of RBC, Hb, PCV, albumin and ascorbate were noticed in MOD and SEV compared to controls with the lowest values (P < 0.05) in SEV except for ascorbate, where MOD did not differ from SEV. Compared to controls, NO^• gradually increased (P < 0.05) in MID followed by MOD and SEV, whereas MDA and PC were higher (P < 0.05) in MOD and SEV. PC was higher (P < 0.05) in MOD than SEV. In addition, the antioxidants GSH, SOD and CAT were higher (P < 0.05) in MID and lower (P < 0.05) in MOD and SEV compared to controls. GSH was lower (P < 0.05) in SEV compared to MOD. Besides, Hb was negatively correlated with NO^• (r = −0.68, P < 0.001), MDA (r = −0.53, P < 0.001) and PC (r = −0.73, P < 0.001). In conclusion, dromedary sarcoptosis is accompanied by a state of oxidative stress process, which increased by increasing the area of infestation, and may contribute to the pathogenesis of the disease.     

Adherence to various host cell lines of Mycoplasma bovis strains differing in pathogenic and cultural features

Mycoplasma bovis is known to be responsible for pneumonia and arthritis in calves, as well as mastitis in dairy cows. Despite clear evidence of its pathogenic potential, little is known about mechanisms of cytadherence and the molecular factors involved. The purpose of this work was to compare adherence rates of M. bovis field strains to different host cell lines and study the effects of cloning and sub-culturing M. bovis strains on their adherence properties. Eighteen metabolically labeled M. bovis strains isolated from different pathological backgrounds were examined in adherence trials using four different host cell lines, i.e. embryonic bovine lung (EBL), embryonic bovine trachea (EBTr), Madin Darby bovine kidney (MDBK) and rabbit kidney (RK) cells. Although large interstrain variations in adherence rates (3.4-19.1%) were measured they could not be correlated to the pathological background (pneumonia, arthritis or mastitis). Adherence rates to the fibroblast cell line (EBTr) were significantly lower than those to the three epithelial cell lines (EBL, MDBK and RK). The only non-pathogenic strain (221/89) exhibited lower adherence rates than three isolates from clinical mastitis. Interestingly, adherence rates were significantly reduced after in vitro passaging. In contrast, no effect of single cloning of strains on adherence was observed. There was no general correlation between expression of variable surface proteins (Vsps) as monitored by immunoblotting and adherence rates, although alterations in Vsp expression profiles were seen as a consequence of passaging. As there is probably a large number of adhesins, variable and non-variable, on the surface of M. bovis cells the issue is very complex, and the most active components have yet to be identified.