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 study included 40 strains of Malassezia pachydermatis obtained in pure culture from external ear canal of dogs and the reference strain. Protein fractions were received by mechanical disruption of the fungal cells. Denaturing protein electrophoresis was performed according to Laemmli method. More than 90% of the all tested strains were characterised by the presence of the following protein fractions: 83.0; 77.0; 59.0; 55.0; 48.0; 38.0; 35.0; 28.0, and 27.0 kDa. In other regions of electrophoregrams, a relatively high differentiation was observed. The strains isolated from animals with otitis externa were characterised by the presence of the following protein fractions: 75.0; 61.0; 52.0; 36.0; 19.5; 16.0; 13.8, and 13.0 kDa. These fractions were absent in the commensal strains. The use of statistical analysis confirmed the obtained results and allowed to generate dendrogram grouping potentially pathogenic and commensal strains on two different branches. Such results may indicate significant differences between protein profiles of strains of M. pachydermatis isolated from healthy and diseased animals.

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 estimate prevalence of tricuspid dysplasia (TD) in dogs with respect to breed, age, sex, clinical signs, and echocardiographic findings and to compare this data with literature. TD was found in 15 dogs (6.5% of congenital cardiac disease) of 215 dogs with congenital heart defects. All dogs had right heart enlargement on thoracic radiographs, echocardiography, and electrocardiography. Doppler echocardiography revealed tricuspid valve regurgitation. Seven dogs presented no clinical symptoms to date. TD took the form of Ebstein anomaly in all Labrador Retrievers, one Boxer, and one German Shepherd dog. TD predominated in males (11 males vs. three females). The body weight of the affected dogs, with the exception of the Miniature Schnauzer, exceeded 20 kg. Two dogs (Boxer and Bull Terrier) had additional congenital cardiac lesions in the form of mitral valve dysplasia. The most affected breeds in the study were the Labrador Retriever and Boxer.

Sixteen white Wistar female rats were divided into two equal groups. Experimental group received per os 40 mg/kg b.w. of L-arginine, every other day for 2 weeks and were decapitated after 3 weeks of the experiment. Control rats received in the same manner 2 ml of distilled water and were decapitated after 3 weeks of the experiment. The renal lesions observed under electron microscope were of focal character and concerned only the experimental group. The tubules with necrotic cells were observed among normal tubules or single normal epithelial cells of the tubular wall. The boundaries between epithelial cells of the tubule wall were blurred. The mitochondria indicated abnormal structure. Numerous lysosomes and peroxysomes with dark, homogenous content were observed. The rough endoplasmic reticulum had widened channels and was focally completely destroyed. The nucleus of damaged cells was most commonly located in one of the cell poles; its shape was changed and visibly smaller than the nuclei of normal cells. Condensation and peripherally located chromatin were noticed. The lesions observed were characteristic for apoptotic cells.

The study included a sheep flock comprising five genetic groups. The ELISA was applied to perform constant monitoring (every six months) for the infection of ewes with small ruminant lentivirus (SRLV). The research results demonstrated a negative effect of SRLVs infection on lamb rearing that, depending on the genetic group, proved to be lower 1.3%-1.4% compared to the seronegative mothers. At relatively equal fertility (94%-100%) and more differentiated prolificacy (179%-198%) in all the examined groups (except the Suffolk breed), a rearing index was higher in the seronegative animals 6.8%-24.1% compared to the seropositive mothers. The Suffolk breed proved to be the genetic group most susceptible to SRLV infection. A prolificacy of infected ewes was 10% lower, a lamb rearing rate was 13% lower , and a general reproductive performance was 18% lower in comparison to healthy ewes.

Objective—To determine an efficient method for the collection of semen samples by means of electroejaculation, characterize spermatozoa quality and quantity, and determine the effect of refrigerated storage on motility of spermatozoa obtained from green iguanas (Iguana iguana). Animals—18 adult green iguanas. Procedures—Green iguanas were anesthetized, and semen samples were obtained by means of electroejaculation. Up to 3 series of electrostimulations were performed; the procedure was stopped after a semen sample was obtained. Various semen sample variables were evaluated. Results—Semen samples were obtained from 16 iguanas; most (n = 10) iguanas produced a semen sample after the second series of electrostimulations. Median semen sample volume was 0.05 mL. Mean spermatozoa concentration was 2 69.0 × 10(6) spermatozoa/mL. Median percentage of motile spermatozoa was 78%. The only morphological abnormality of spermatozoa was bent tails (mean percentage in a semen sample, 5.7%). Spermatozoa motility decreased significantly during refrigeration (4°C); median percentage motility after 24, 48, and 72 hours of refrigeration was 60%, 33%, and 0%, respectively. Conclusions and Clinical Relevance—Results of this study suggested electroejaculation can be performed to collect semen samples from green iguanas, characteristics of iguana semen samples are similar to those for semen samples obtained from other reptiles, and motility of iguana spermatozoa decreases during refrigeration within 48 to 72 hours.

Abundant lymphocyte infiltration is frequently found in canine malignant mammary tumors, but the pathological features and immunophenotypes associated with the infiltration remain to be elucidated. The aim of the present study was to evaluate the relationship between lymphocyte infiltration, histopathological features, and molecular phenotype in canine mammary carcinoma (MC). The study was done with archived formalin-fixed, paraffin-embedded samples (n = 47) by histologic and immunohistochemical methods. The degree of lymphocyte infiltration was evaluated by morphologic analysis, and the T- and B-cell populations as well as the T/B-cell ratio were evaluated by morphometric analysis; results were compared with the histologic features and molecular phenotypes. The degree of lymphocyte infiltration was significantly higher in MCs with lymphatic invasion than in those without lymphatic invasion (P < 0.0001) and in tumors of high histologic grade compared with those of lower histologic grade (P = 0.045). Morphometric analysis showed a larger amount of T-cells and B-cells in MCs with a higher histologic grade and lymphatic invasion, but the T/B ratio did not change. Lymphocyte infiltration was not associated with histologic type or molecular phenotype, as assessed from the immunohistochemical expression of epidermal growth factor receptor 2, estrogen receptor, cytokeratin 14, and p63. Since intense lymphocyte infiltration was associated with aggressive histologic features, lymphocytes may be important for tumor aggressiveness and greater malignant behavior in the tumor microenvironment.

Objective—To determine the effects of advanced age on whole-body protein synthesis and activation of the mechanistic target of rapamycin (mTOR) signaling pathway in skeletal muscle of horses Animals—Six 22- to 26-year-old (aged) and six 7- to 14-year-old (mature) horses. Procedures—Whole-body protein synthesis was measured with a 2-hour primed constant infusion of 13C sodium bicarbonate, followed by a 4-hour primed constant infusion of 1-13C phenylalanine. After the infusions, a biopsy specimen was obtained from a gluteus medius muscle and activation of protein kinase B (Akt), p70 riboprotein S6 kinase (S6K1), riboprotein S6 (rpS6), and eukaryotic initiation factor 4E binding protein 1 (4EBP1) was determined with western immunoblot analysis. For all horses, inflammatory cytokine expression in muscle and blood samples was measured with quantitative real-time PCR analysis. Results—Advanced age had no effect on whole-body protein synthesis or the phosphorylation of Akt, rpS6, and 4EBP1; however, muscle specimens of aged horses had 42% lower phosphorylation of S6K1 than did those of mature horses. Aged and mature horses had similar inflammatory cytokine expression in muscle and blood samples. Conclusions and Clinical Relevance—The lower S6K1 activation for aged horses, compared with that for mature horses, could be indicative of low rates of muscle protein synthesis in aged horses. However, advanced age had no effect on any other indicators of whole-body or muscle protein synthesis or on measures of systemic or muscle inflammation, which suggested that protein metabolism and subsequently requirements may not differ between healthy mature and aged horses.

Objective: To evaluate first-intention healing of CO2 laser, 4.0-MHz radiowave radiosurgery (RWRS), and scalpel incisions in ball pythons (Python regius). Animals: 6 healthy adult ball pythons. Procedures: A skin biopsy sample was collected, and 2-cm skin incisions (4/modality) were made in each snake under anesthesia and closed with surgical staples on day 0. Incision sites were grossly evaluated and scored daily. One skin biopsy sample per incision type per snake was obtained on days 2, 7, 14, and 30. Necrotic and fibroplastic tissue was measured in histologic sections; samples were assessed and scored for total inflammation, histologic response (based on the measurement of necrotic and fibroplastic tissues and total inflammation score), and other variables. Frequency distributions of gross and histologic variables associated with wound healing were calculated. Results: Gross wound scores were significantly greater (indicating greater separation of wound edges) for laser incisions than for RWRS and scalpel incisions at all evaluated time points. Necrosis was significantly greater in laser and RWRS incisions than in scalpel incision sites on days 2 and 14 and days 2 and 7, respectively; fibroplasia was significantly greater in laser than in scalpel incision sites on day 30. Histologic response scores were significantly lower for scalpel than for other incision modalities on days 2, 14, and 30. Conclusions and Clinical Relevance: In snakes, skin incisions made with a scalpel generally had less necrotic tissue than did CO2 laser and RWRS incisions. Comparison of the 3 modalities on the basis of histologic response scores indicated that use of a scalpel was preferable, followed by RWRS and then laser.