Abstract The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-old’s lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V–FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days’ culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs.

ABSTRACT. In total, 240 one-day-old Japanese quails (Coturnix Coturnix Japonica) allocated at random to 6 treatments with 4 replicates and 10 birds in each. Treatments used were: 1) Negative control (without any additives or AFB1); 2) Positive control (basal diet + 2.5 ppm AFB1; 2); 3) TA008 (positive control + 108 cfu/ml Bacillus. megaterium TA008); 4) TA049 (positive control + 108 cfu mL-1 Bacillus. subtilis TA049); 5) TA010 (positive control+ 108 cfu mL-1 Brevibacillus brevis TA010) and 6) P (positive control + 2.5 g kg-1 Polysorb® in feed). Hatchability and embryonic mortality were significantly influenced by additives and AFB1 (p < 0.05). Birds fed TA008 improved 12 % hatchability and reduced 10 % embryonic mortality in compared to positive control (p < 0.05). Weight gain and feed conversion ratio did not affected by treatments (p > 0.05). Feed intake was significantly improved in birds feeding by TA008 at 0-21 days (p < 0.05). There were significant differences on relative weights of carcass, gizzard and proventriculus among treatments (p < 0.05). Serum total protein, albumin, cholesterol, glucose, HDL, globulin and uric acid were significantly affected by treatments (p < 0.05). These results showed that the inclusion of bacillus megaterium as potential probiotic into contaminated diets could improve the adverse effects of AFB1 in Japanese quails.

ABSTRACT. Industrial effluents of alcohol production are problematic for the environment. One of the proposed solutions to the reduction of harmful effects of the wastewater is to convert it into vinasse. The purpose of the present study is to investigate effects of rations with different levels of vinasse as a source of protein in the diet of 28 male Holstein calves, previously health checked, with an average initial weight of 300 ± 22.56 kg and the maximum similarity of weight and age. The calves were divided randomly into four treatments varied in vinasse (substituted for cottonseed meal): 0 (control), 5, 10 and 15 % with seven replications and each group was given its own specific diet for 110 days. Growth performance, nutrient digestibility and feed intake were examined. Final body weight, average daily weight gain and dry matter intake were significantly higher in calves fed diet with 10% vinasse compared to other treatments (p < 0.05). Rumen pH significantly increased in treatment four (15% vinasse) (p < 0.05). Crude fat, NDF, non-fiber carbohydrate (NFC) and dry matter digestibility didn’t show any significant difference between treatments (p > 0.05), but treatments with 10 and 15% vinasse significantly increased organic matter andcrude protein digestibility compared with control (p < 0.05). Totally substituting cottonseed meal with 10% vainasse in calves diet improved body weight gain and organic matter digestibility without any significant effects on rumen pH.