The aim of the study was to assess the influence of the oestrous cycle phase on neutrophil secretory activity and to extrapolate it to susceptibility to uterine infections. The obtained results indicate that the highest enzyme release seen in the late follicular phase (elastase release was 42.18 ±3.11% of maximal release, myeloperoxidase was 45.0 ±5.12%, and alkaline phosphatase was 44.75 ±9.0%) was related to the level of 17β-oestradiol in plasma. Similarly, a free radical generation was also the most enhanced during this phase. Significantly lower values were obtained from sows during the luteal phase in regard to both enzyme release (36.62 ±3.58% for elastase, 27.87 ±8.7% for myeloperoxidase, and 22.12 ±2.4% for alkaline phosphatase), and that of free radicals (2.28 1.6 μM/106 cells for nitric oxide and 2.47 0.6 nM/106 cells for superoxide).

A total of 142 chicken blood samples were collected and a specific primer set was used to amplify a fragment of growth hormone locus using PCR. PCR products were digested with SacI and MspI restriction endonucleases. The amplified fragment digested with SacI enzyme revealed two “+” (wild type) and “-” (normal type) alleles with the frequency of 0.898 and 0.102, respectively. The amplified fragment digested with MspI enzyme revealed three A, B and C alleles with the frequency of 0.599, 0.102, and 0.299, respectively. Frequencies of +/+, +/- and -/- were 0.817, 0.162, and 0.021, respectively, and those of AA, AB, AC, BB, BC, and CC were 0.338, 0.113, 0.409, 0.007, 0.070, and 0.063, respectively, in the studied population. The results of 2 and likelihood ratio tests showed that this population was at Hardy-Weinberg equilibrium with respect to the marker locus. Marker-trait association analysis revealed statistically significant differences between “SacI-RFLP” genotypes for egg production and rate of laying eggs. The relationship between the molecular marker and these traits can be useful to improve the chicken breeding programmes.

The purpose of the experiment was to compare apoptosis induced by equine influenza virus (EIV A1 and EIV A2) infection in MDCK, RK13, and NEURO-2A cell lines. Flow cytometry was used to observe two symptoms of apoptosis: phosphatidylserine translocation in plasmalemma (annexin V assay) and the fragmentation of DNA generated by endonuclease activity (TUNEL assayterminal deoxynucleotidyl transferase biotin-dUTP nick end labelling). The differences in the onset of apoptosis in the studied cells was observed. In MDCK cells infected with EIV A1 and A2, a weak signal of the phosphatidylserine translocation was observed but more cells showed the DNA fragmentation. An opposite effect was observed in case of RK 13 cells. NEURO-2A cells displayed a similar number of annexin V and TUNEL positive cells after the infection with EIV A2, while in case of EIV A1 infection, only the early symptoms of apoptosis were noted. Differences between both viral serotypes could originate from functioning of viral proteins responsible for induction or inhibition of apoptosis. The differences between cell types may result from the activation of cellular pro or anti-apoptotic mechanisms.

The aim of the study was to determine serum free amino acid concentration in male dogs suffering from benign and malignant perianal tumours. Serum concentrations of cysteic acid, glutamic acid, glutamine, aspartic acid, glycine, γ-aminobutyric acid, tryptophan, methionine, arginine, taurine, threonine, tyrosine, alanine, valine, isoleucine, and leucine were significantly changed in dogs suffering from benign and/or malignant tumours when compared to the control group (P<0.05). Serum concentration of serine, phenylalanine, lysine and histidine was not influenced in dogs with neoplastic disease (P>0.05). The evaluation of serum free amino acid concentration, has shown that threonine, glutamic acid, alanine, valine, isoleucine, leucine, γ-aminobutyric acid, and glycine have the highest diagnostic and prognostic value in dogs suffering from benign and malignant perianal tumours. Highly diagnostic and prognostic value in relation to benign tumour growth was also found while evaluating branched chain and acidic groups of amino acids. Serine, phenylalanine, lysine, and histidine, as well as alkaline amino acids were proved not to have diagnostic and prognostic value in dogs with the tumours. Thus, the evaluation of free amino acid concentration may serve diagnostic purposes and help in tumour malignancy differentiation. The elaborated experimental model may be used in further studies on neoplastic disease development and monitoring of applied treatment efficiency.

The study was conducted on 20 clinically healthy stallions, aged 3-4 years. The evaluation of the degree of adaptation of stallions to exercise loads in the course of a 100-d training programme was conducted three times: (I) - on the 1st d of training, (II) - on the 46th d of training, and (III) - on the 97th d of training. Rectal temperature (T) and number of heart and respiratory rates were measured. Blood was sampled before exercise (1), after exercise (2), and 30 min after rest (3). The following indices were determined in the blood: concentration of haemoglobin, haematocrit level, number of red blood cells, white blood cells, lymphocytes, neutrophils, eosinophils, basophils, monocytes, and platelets. Post-exertion changes in the evaluated indicators showed moderate intensity to the applied loads with respect to functional potential of tested horses. The effect of the standard 100-d training was to stimulate the body's adaptation to functional tasks, which was evidenced, among others, by lower variability of post-exertion T and the number of red blood cells. The highest T increase was found at the beginning of the programme (about 2.1%), whereas during the subsequent studies (II and III) it was significantly (P<0.05) lower (0.98% and 0.84%, respectively). The variability in red blood cell count in the course of training showed a downward trend (16.4%, 13.7%, and 11.0%, respectively). The results confirm the relatively low exercise loads and good health of young stallions during the 100-d performance test.

Efficacies of hCG and GnRH with respect to follicular diameter in Cosynch protocol were investigated. Lactating cows (n=119) received injections of GnRH and PGF2 7 d apart. Norgestomet ear implant was inserted at GnRH administration and removed prior to PGF2 injections. Presence of the corpus luteum (CL) at GnRH and PGF2 injections, and diameter of preovulatory follicle [small (SPF) <8 mm or large (LPF) ≥8 mm] at PGF2 were determined with transrectal ultrasonography (USG). The animals were randomly treated with GnRH (0.01 mg; GPG, n=62) or hCG (1,500 IU; GPH, n=57) at timed artificial insemination (TAI) 56 h after injection of PGF2α. Pregnancies were diagnosed with USG 32-35 d after TAI. Pregnancies per TAI (P/TAI) were significantly (P<0.05) lower in GPH cows (28.1%; 16/57) compared to those in GPG animals (46.8%; 29/62). P/TAI with respect to follicular size did not differ between GPG and GPH cows, although P/TAI was numerically higher in LPF (30.0%; 15/50) than in SPF (14.3%; 1/7) in GPH cows. P/TAI in cows without CL at PGF2 was 2.5 (1.0-6.7) times higher (P<0.01) in GPG (20.0%; 3/15) compared to GPH (10.5%; 2/19). In conclusion, use of hCG in milieu of the second GnRH in Cosynch+progesterone insert protocol did not have any advantage in lactating cows.

Changes in selected blood lymphocyte subpopulations in dogs administered with a soluble parasitic antigen (SPA) derived from a supernatant of 18S RNA-A and 18S RNA-B Babesia canis cell culture were investigated. The studies included 20 dogs divided into three groups: group I (n=8) - comprised of dogs receiving SPA twice, at 3 week intervals; group II (n=5) - nonvaccinated control dogs, and group III (n=7) - dogs vaccinated twice with a commercial B. canis vaccine. Cytometric analysis revealed that vaccination with SPA derived from B. canis culture had similar effects to the vaccination with a commercial vaccine. The vaccination lowered the percentage of T lymphocytes (CD3+), T helper cells (CD4+), cytotoxic/supressor T cells (CD8+), B lymphocytes (CD21+), and MHC II lymphocytes in the blood in comparison to non-vaccinated dogs. Statistical analysis of the results demonstrated that mean values of the tested parameters at each stage of the study were similar in groups I and III and significantly higher in group II. The lowered level of the lymphocyte subpopulations in groups I and III persisted during the whole period of the study. The results presented that SPA has immunosuppressive effect in the first period after being administrated.

Objective-To determine the efficiency of a novel point-of-care gravitational marrow separator and bone marrow aspiration needle for concentrated bone marrow production and bone marrow-derived mesenchymal stem cell (MSC) separation and assess the effect of repeated bone marrow collections in horses. Animals-8 healthy adult horses. Procedures-Bone marrow aspiration was performed twice (1 month apart) from sternebral bodies with a standard or prototype multidirectional needle. Concentrated bone marrow was obtained by gravitational marrow separation and evaluated for WBC and platelet counts, automated and cytomorphologic cell differential counts, MSCs, and cell viability. Results-Concentrated bone marrow samples obtained with the marrow separator had 5- to 19-fold bone marrow-derived MSC, WBC, and platelet counts, compared with original bone marrow samples. Use of a multidirectional needle increased the frequency of obtaining MSC-richer concentrated bone marrow. Repeating bone marrow aspiration at 1 month yielded greater MSC numbers but slightly lower cell viability after processing. Conclusions and Clinical Relevance-The gravitational bone marrow separator and multidirectional needle were used to effectively harvest bone marrow and improve the quality of concentrated bone marrow. Comparable, or even greater, numbers of bone marrow-derived MSCs were collected by repeated bone marrow aspiration after a 1-month interval from the same aspiration sites. Use of the marrow separator and multidirectional bone marrow aspiration needle can facilitate a 1-step, point-of-care, nonlaboratory method to obtain concentrated bone marrow as a mixture of bone marrow-derived MSCs and growth factors from platelets and plasma.

Objective: To evaluate fecal calprotectin concentrations in healthy dogs and dogs with chronic diarrhea, to identify cutoff values for fecal calprotectin concentrations for use in differentiating dogs with chronic diarrhea and a canine chronic enteropathy clinical activity index (CCECAI) < 12 from dogs with chronic diarrhea and a CCECAI ≥ 12, and to evaluate the association between histologic evidence of intestinal mucosal changes and fecal calprotectin concentrations in dogs with chronic diarrhea. Sample: Fecal samples from 96 adult dogs (27 dogs with chronic diarrhea and 69 healthy control dogs). Procedures: Severity of clinical signs was evaluated on the basis of the CCECAI scoring system. Endoscopy was performed in all dogs with chronic diarrhea, and mucosal biopsy specimens were evaluated histologically. Fecal calprotectin concentration was quantified via radioimmunoassay. Results: Fecal calprotectin concentrations were significantly higher in dogs with chronic diarrhea than in healthy control dogs. Fecal calprotectin concentrations were also significantly higher in dogs with a CCECAI ≥ 12, compared with concentrations for dogs with a CCECAI between 4 and 11. Fecal calprotectin concentrations were significantly higher in dogs with chronic diarrhea associated with histologic lesions, compared with concentrations in control dogs, and were significantly correlated with the severity of histologic intestinal lesions. Among dogs with chronic diarrhea, the best cutoff fecal calprotectin concentration for predicting a CCECAI ≥ 12 was 48.9 μg/g (sensitivity, 53.3%; specificity, 91.7%). Conclusions and Clinical Relevance: Fecal calprotectin may be a useful biomarker in dogs with chronic diarrhea, especially dogs with histologic lesions.

Objective-To evaluate the antinociceptive and sedative effects and duration of action of hydromorphone hydrochloride after IM administration to American kestrels (Falco sparverius). Animals-11 healthy 2-year-old American kestrels. Procedures-Hydromorphone (0.1, 0.3, and 0.6 mg/kg) and an equivalent volume of saline (0.9% NaCl) solution (control treatment) were administered IM to kestrels in a masked randomized complete crossover study design. Foot withdrawal response to a thermal stimulus was determined 30 to 60 minutes before (baseline) and 0.5, 1.5, 3, and 6 hours after treatment administration. Agitation-sedation scores were determined 3 to 5 minutes before each thermal test. Results-Hydromorphone at 0.6 mg/kg, IM, significantly increased the thermal foot withdrawal threshold, compared with the response after administration of saline solution, for up to 3 hours, and hydromorphone at 0.1, 0.3, and 0.6 mg/kg, IM, significantly increased withdrawal responses for up to 6 hours, compared with baseline values. No significant differences in mean sedation-agitation scores were detected between hydromorphone and saline solution treatments; however, appreciable sedation was detected in 4 birds when administered 0.6 mg of hydromorphone/kg. Conclusions and Clinical Relevance-Hydromorphone at the doses evaluated significantly increased the thermal nociception threshold for American kestrels for 3 to 6 hours. Additional studies with other types of stimulation, formulations, dosages, routes of administration, and testing times are needed to fully evaluate the analgesic and adverse effects of hydromorphone in kestrels and other avian species and the use of hydromorphone in clinical settings.