BACKGROUND: Granuloca cells have a key role during estroeidogenesis. OBJECTIVES: The purpose of this study was to determine the effect of number of culture medium granulosa cells on estradiol concentrations and mRNA codding estrogenic and progestagenic enzyme. METHODS: Briefly, follicles between 2 and 5 mm diameter were dissected from the ovaries of adult cows and were collected by rinsing the follicle walls with Dulbecco Modified Eagle medium/F12 (DMEM/F12). The number of cells was counted with a haemocytometer and the viable cells were assessed by the dye exclusion method using 0.4% Trypan Blue. Treatments were 1) 500,000 cell/500 ml, 2) 250,000 cell/500 ml, 3) 500,000 cell/200 ml 4) 250,000 cell/ 200 ml.  All data were analyzed by JMP (SAS). RESULTS: Low plating density increased E2 secretion and mRNA encoding LHR, FSHR and estrogenic enzymes (17βHSD, CYP19), whereas decreased mRNA encoding GADD45β. There were no differences among treatments for RNA and protein concentration. Low plating density also decreased protein amount but there was no difference among treatments for RNA amount. In conclusion, decreased cell density cause increase in mRNA encoding codding estrogenic enzyme gene expression and decrease in mRNA encoding progestagenic enzyme gene expression. CONCLUSIONS: Protein concentrations did not changed with decreased cell density therefore we can save cells against harmful effect of increasing cell density.

BACKGROUND: Brucellosis is one of the most common zoonosis in Middle East and Iran. OBJECTIVES: The purpose of this study was genomic detection of Brucella spp. in sero-positive dairy cattle. METHODS: We have collected 28,519 blood samples from cows during 2012-2013. Samples were screened by Slide and tube agglutination and 2-Mercaptoethanol tests. Samples with anti-Brucella antibodies titer ≥ 1:80 and ≥1:40 in tube agglutination and 2-ME tests were considered as positive respectively. Tissue samples include: lymph nodes, liver, testicle and kidney from 122 samples of slaughtered cows were collected. The Sero-positive samples were examined by a collection of specific primers for Brucella abortus, Brucella melitensis, vaccinal strains included RB51 and Rev1 using PCR tests. RESULTS: Results showed that 450 samples were positive in slide agglutination test and 447 samples had anti-brucella antibodies titer equal to or more than 1:80. So they were positive by tube agglutination test. Three hundred eighty nine samples were positive by 2- mercaptoethanol test. PCR test results showed that 46 samples (37.7%) out of 122 samples had a specific sequence of Brucella or otherwise they have an active infection with Brucella species, whereas 62.3% of samples were negative. The PCR results showed that 2 samples (4.35%) were infected by B. melitensis, 2 samples (4.35%) infected by Rev1 strain and 42 samples (91.3%) were infected by B. abortus. CONCLUSIONS: The results showed that, as we had expected, the majority of cows were infected by B. abortus. Animals who infected by B. melitensis and Rev1 strain may be a result of contact with sheep or goats. We couldn’t find Brucella genome in 76 samples (62.3%) of sero-positive cows. It may be caused by cross reaction of sera with Brucella species in tests or activation of immune system response and elimination of organism from internal organs.

Pasteurella multocida type A is the etiologic agent of fowl cholera, a highly contagious and fatal disease of chickens. The present research work was performed for the isolation, identification and molecular detection of P. multocida Type A from chickens. Liver, heart and spleen of suspected dead chicken (n=35) were collected from Gazipur and Pabna districts in Bangladesh. The targeted bacteria from the samples were isolated, identified and characterized based on their morphology, staining, cultural, biochemical characters, pathogenicity test, histopathological study and Polymerase Chain Reaction (PCR). The P. multocida organism was isolated from 11.42% (n=4/35) samples. The organisms were gram negative, non-spore forming rod, non-motile, occurring singly or pairs in Gram staining, whereas in Leishman's stain, bipolar shaped organisms were observed. All the isolates were found positive for oxidase and catalase tests, produced indole, and fermented glucose, mannitol and sucrose. Necrotic foci in liver and congestion with hemorrhages in heart were found on necropsy. After pathogenicity test, the pathological changes were reconfirmed by histopathology depicting congestion, hemorrhage and lymphocyte infiltration in heart, liver and spleen tissues. In type specific PCR reaction, the organisms were confirmed as P. multocida Type A. In conclusion, P. multocida type A is prevalent among poultry in the studied regions; thus, care must be taken to control of the disease. [J Adv Vet Anim Res 2015; 2(3.000): 338-345]

Duck plague (DP) is the most feared duck disease in the world. For isolation, identification, molecular detection and characterization of DP virus (DPV), a total of 94 samples were collected from commercial farms (n=6) and households (n=13) from Rajshahi (n=37), Netrokona (n=35) and Mymensingh (n=22) districts of Bangladesh. The samples were processed and inoculated into 11-13 days old embryonated duck eggs for virus propagation. Virus was identified using agar gel immunodiffusion test (AGIT) and passive hemagglutination (PHA) test, and was confirmed by polymerase chain reaction (PCR) targeting DNA polymerase and gC genes, followed by sequencing. Pathogenicity tests were performed using duck embryos, ducklings and ducks. Among the 94 samples, 17 isolates were confirmed as DPV by PCR amplification of partial DNA polymerase (446-bp) and gC genes (78-bp), respectively. One of the isolates (Anatid herpes 1 BAU DMH) was sequenced and found to be closely related with a Chinese variant of DPV (GenBank: JQ647509.1). Thus, we assume that both Bangladeshi and Chinese isolates of DPV may have a common ancestor. [J Adv Vet Anim Res 2015; 2(3.000): 296-303]

The Kedah-Kelantan cattle (KK) is an indigenous cattle breed and is mainly kept for meat production in Malaysia. Due to lack of information about polymorphism of growth traits in these cattle, Insulin-like Growth Factor 1 (IGF-1) was chosen to be the candidate gene in this preliminary study. The aim of this study is to investigate the polymorphism of IGF-1 gene in KK and to show that the PCR-RFLP technique can be used as a basis for use as molecular markers in cattle. A total of 46 KK blood samples were collected for DNA extraction performed using a commercial kit. The exon 1 of IGF-1 gene was amplified to produce a 249 bp fragment. The amplified fragments were digested with Eco105I restriction endonuclease and then subjected to electrophoretic separation in Fluorosafe stained 2.5 % agarose gel. The result revealed two alleles, A and B. Threegenotypes were observed: AA, AB and BB. Frequencies were 0.07, 0.13 and 0.80 for AA, AB and BB, respectively. This gives frequencies of 0.13 and 0.87 for A and B alleles. It is concluded that the population is in Hardy-Weinberg disequilibrium (p<0.05). It is possible that this gene has been exposed to selection.