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: Essential oils as growth stimulant play an important role in improving intestinal microflora and morphological properties and immune system in broiler chickens. OBJECTIVES: The aim of this study was to investigate the effects of Artemisia sieberi oil on intestinal morphology, secum microflora and immune system in broiler chickens. METHODS: Two hundred day-old broiler chickens (Ross 308) were allocated to 5 treatments, 4 replications with a completely randomized design during 42 days of age. Experimental treatments consisted of control (basal diet) or basal diets containing different levels of Artemisia sieberi oil (100, 200 and 300 mg/kg) and 600 mg/kg flavophospholipol antibiotic. On day 42 days of age, intestinal morphology and microflora population and immune system were evaluated by measuring the weight of bursa of fabricious and spleen and antibody production. Results: The results indicated that the highest lactobacillus count and the lowest Escherichia coli count of the caecum was found by inclusion of 300 mg/kg Artemisia sieberi oil in the diet (p<0.0001). The antibiotic and Artemisia sieberi oil treatments showed higher villus height in the duodenum compared with control group (p<0.01). Artemisia sieberi oil and antibiotic supplementations significantly decreased epithelial thickness and goblet cell number of the small intestinal compared with control group (p<0.05). The dietary supplementation did not significantly affect the crypt depth and villus height to crypt depth ratio in small intestine. The level of 300 mg/kg Artemisia sieberi oil significantly increased antibody titration against Newcastle disease virus (p<0.01). Conclusions: Adding Artemisia sieberi oil at levels of 300 mg/kg to broiler chicken diets can improve gut microflora (as measured by changes in populations of Escherichia coli and lactobacillus).

BACKGROUND: Maturity of silver carp (Hypophthalmichthys molitrix) ovary takes place at 2-8 years old and it is depends on photoperiod and environmental water temperature. Objectives: The aim of this study was to identify  macroscopical and microscopical structure of silver carp  ovary in  two categories of  body weight, total body length and  at  2 and 4 years old  in Khuzestan province climatic conditions. Methods: 20 silver carp ovaries were studied in two groups: Group 1(G1): 10 fish with total body length 46 ±1.33 Cm and weight 1050±0.09 grand approximately 2 years old. Group 2(G1): 10 fish with total body length 86±3.64 Cm and weight 5460±800 gr and approximately 4 years old. For the histometrical studies, 5-6 µ sections were made by routine paraffin embedding method and stained by H&amp;E and PAS. Results: Microscopic results showed that ovaries of G1 covered by a thin loose connective capsule which numerous of egg-bearingtrabecula extended from capsule into ovaries. These trabecula contained primary germ cells, oogonium, chromatin nocleolus, perinocleolus and cortical alveolus follicles. Ovaries of G1 do not have yolk and mature follicles. But in ovaries of G2, growth and mature follicles were observed and mature follicles were most follicles. Histometrical results showed that there are not a significant differences in diameter of follicles, oocytes and oocytes nucleolus in G1 and 2. The maximum and minimum diameter of follicles were seen in mature and chromatin nucleolus follicles respectively. The Zona plucida thickness was increased from corticalalveolus (1.53±1.11 mm) to mature follicles (6.88±0.21 mm) significantly (p<0.05). ConclusionS: The most interesting finding of this study is that in Khuzestan Province climate, the ovary of silver carp fish in G1, with average weight 1050 ±0.09 gr and total length 46±1.33 Cm and approximately of 2 years old is immature but ovary of G2 with average weight of 5460± 800 gr and total length 86±3.64 Cm and approximately 4 years old is mature.

Neuroendocrine cells are of the peptide and amine secreting cells and because of endodermal origin, exist in respiratory and gastrointestinal system. The report confirms the presence of a tumor using immunohistochemical methods. A hepatic mass was detected in a 10-year-old male mixed–breed dog which was referred with clinical history of frequent vomiting, anorexia, lethargy and syncope. Biochemical and hematological abnormalities included elevation of ALT and AST levels, leukocytosis and anemia. Radiographically the large hepatic mass and multifocal metastatic nodular areas in the other organs were observed. At necropsy the liver had a multilobulated firm mass involving whole caudate lobe of the liver. On the cut section, the tumor was grey sanguineous with hemorrhagic surface and numerous necrotic foci. Microscopically, there were unencapsulated, highly cellular neoplastic proliferations with few hemorrhagic foci in the liver that had a trabecular pattern to Rosette formation which were separated by a fibrovascularstroma. The neoplastic cells were round to oval, with granular eosinophilic cytoplasm and hyperchromatic nuclei. Metastases were observed in lungs, kidneys, mesenteric lymph nodes and even in the arteries of the tunica albuginae of the testes. Immunohistochemically, neoplastic cells were immunoreactive for NSE and S100 protein. They were negative for CEA, Chromogranine A, CD34, AE1/AE3, CK20 and Hepatic Antigen (HA). Negative CEA, CK20, CD34 and HA disproved the probability for collangiocarcinoma, metastatic carcinoma, hematopoietic and vascular origin and hepatocellular carcinoma of this tumor respectively. Immunohistochemical demonstration of NSE and S100 protein supported the diagnosis of the Carcinoid like. However, negative reaction for Chromogranine A does not disprove neuroendocrine carcinoma diagnosis.

BACKGROUND: Newcastle disease is one of the most serious viral diseases in the poultry worldwide. OBJECTIVES: Since the traditional strategies have been hardly effective in controlling the disease, the purpose of this study was to introduce new methods for early and rapid diagnosis of Newcastle. The present study helps to reduce further damage to the poultry industry. METHODS: RNA extraction was performed, using RNease mini kit, according to the manufacturer’s instructions. Extracted RNA with 68.23×109 copy numbers was prepared as serial dilutions of 100 μL for RT-PCR and RRT-PCR reactions. RRT-PCR and RT-PCR were performed, using commercial kit and RNease mini kit, respectively.  RESULTS: Results showed that amplification was done according to prepared dilution equal 10-34 for RRT-PCR reaction and a visible band observed on 1.5% Agarose gel up to 10-20 for RT-PCR reaction. Based on the results observed, RRT-PCR and RT-PCR reactions are able to detect 10-34 and 10-20 copy numbers of primary sample, respectively. CONCLUSIONS: The sensitivity of RRT-PCR reaction is almost twice compared with RT-PCR reaction, also RRT-PCR reaction is able to diagnose Newcastle disease virus in infected samples with 10,000 copy numbers of the RNA virus less than RT-PCR.

BACKGROUND: Causing site direct mutation can be one of the efficient methods to evaluate the characteristics and properties of various genes. Brucellosis is the most common zoonotic infectious disease that would cause great economic losses. Thus, recognition of pathogenic and immunogenic factors in the genus Brucella can lead to control this health problem. Objectives: Considering the importance of site direct mutation in identification of genome structure and numerous ways to achieve this goal, Overlap Extension PCR is introduced as an improved technique for the removal and replacement of the gene target. Methods: For this study, with two-step PCR using specific primers, upstream and downstream fragments from target gene and antibiotic resistance cassette from plasmid pET28a (+), were reproduced and were connected to each other. The resulting fragment was cloned in specific position of pBluescriptIISK(-) plasmid by the restriction enzymes. Then, the construction was transferred into the genome of Brucella abortus by electroporation method. Results: Fusion PCR product was obtained without any change in the nucleotide sequence and then it was cloned into pBluescriptIISK (-) plasmid, finally the construction was replaced and the target gene was deleted. Conclusions: The results of this study show that the Overlap Extension PCR is an optimized and modified technique to create mutations in the bacterial genome structure and can easily be used in the family Brucella.

BACKGROUND: One of the most important heart diseases in dogs is valvular insufficiency, which can be evaluated by diagnosis ways such as phonocardiography, echocardiography, etc. OBJECTIVE: The purpose of the current study was to evaluate of valvular insufficiencies with phonocardiography and echocardiography and using phonocardiography technique in detection of cardiac valvular disease in practice. METHODS: This survey was done on 180 five-year-old dogs which 30 of them had valvular insufficiency. They have been referred to radiology section and echocardiography technique was used after listening to heart sounds and recording heart murmur and surveying by phonocardiography. The type and location of valvular insufficiency was diagnosed by phonocardiography and then echocardiography was used, the results from both techniques was compared afterwards. RESULTS: In all of these 30 dogs, murmur was systolic and mitral insufficiency and mitral regurgitation were diagnosed by phonocardiography. using echocardiography, the mitral insufficiency was confirmed in 28 dogs, one of them has been diagnosed to have tricuspid inssufiency and pulmonary stenosis in addition to mitral insufficiency. In two cases no abnormality sign has been detected. CONCLUSIONS: According to this study, it is recommended to use phonocardiography technique in order to pre-diagnose the valvular insufficiency, it's type and location and use echocardiography to determine the process of disease and control this progress.

BACKGROUND: Brucellosis is one of the most important zoonosis that is prevalent among human and animal. Today, a large percentage of animal and human population suffer from its side effects. OBJECTIVES: The purpose of the present study was to estimate the prevalence of anti-Brucella antibodies in flock- and animal-level in districts of south of Kerman province. METHODS: In this cross-sectional study, 300 herds of 7 districts in the area were selected randomly; 10 samples of sheep and goats in each flock were randomly selected. Out of 3000 samples, 2952 samples were examined using Rose-Bengal test; Wright and 2-ME tests were done on positive samples. Descriptive statistics and logistic regression in Stata 11.2 were used to analyze the data. RESULTS: The seroprevalence of anti-Brucella antibodies in animal- and flock-level was 6.4 and 25.3 percent, respectively. The chance of being infected in sheep was 2.12 times of goats. CONCLUSIONS: The prevalence of Brucella was considerably high in animal- and herd-level in this area. It is necessary to empower Iran Veterinary Organization in financial aspects to control this infection.

BACKGROUND: Adiponectin is one of the most important adipocytokines that regulate male and female fertility via AdipoR1 and AdipoR2 receptors. Recently expression of adiponectin system and its negative regulatory role on hypothalamus-pituitary axis have been confirmed. Objectives: No information is available about the expression pattern of adiponectin and its receptors in hypothalamus-pituitary axis in domestic animals. Here for the first time, we studied hypothalamus-pituitary adiponectin system gene expression in different stages of bovine estrous cycle. Methods: Anterior pituitary and hypothalamus were collected from Holstein cow at the local abattoir. The estrous cycle was classified to four phases (proestrous, metstrous, early luteal and late luteal) based on macroscopic examination of ovaries and uteri. Gene expression analysis of adiponectin and its receptors was done using quantitative real time PCR (qPCR Probe MasteKit) and according to the comparative 2-ΔΔCt method. E2 and P4 levels were measured using ELISA method. Results: Our results demonstrated that adiponectin and its two receptors were expressed in pituitary and hypothalamus of cyclic cow. Maximal expression of adiponectin was observed in early luteal phase, while it was expressed at minimal level during the proestrous stage. We observed no significant changes in the expression of AdipoR1 in both tissues at different stages of estrous cycle. The highest expression of AdipoRII in both tissues was detected during the proestrous stage, while it expressed at minimal level during the late luteal phase. E2 and P4 had respectively negative and positive correlations with adiponectin expression levels in hypothalamus and pituitary. Conclusions: Based on our results that demonstrated adiponectin was minimally expressed at proestrous stage and other data about the negative action of adiponectin on LH secretion from pituitary, we concluded that adiponectin may has role in the hormonal function of this axis during the estrous cycle.

BACKGROUND: Reptiles, especially turtles that inhabit both on land and water, have made some special adaptations. Many people keep turtles as pets. Therefore, the anatomical knowledge of turtles should be more carefully evaluated and used for therapeutic purposes. One of these turtles is European pond turtle (Emys orbicularis). Most of vital systems are enclosed by the carapace and the plastron so it cannot be examined customarily by clinicians. The noninvasive diagnostic imaging techniques provide detailed information concerning these organs. OBJECTIVES: This study was conducted to give complete topographic information and knowledge about the position of the non respiratory organs of the coelomic cavity in the European pond turtle using Computed Tomography (CT) and usual anatomic methods. METHODS: 10 adult turtles (5 female, 5 male) were selected. All scans were obtained on a two detector scanner. In anatomical study three female and three male turtles were dissected. Two other female and male turtles were sectioned transversely. RESULTS: The results showed some differences in the position of the organs including stomach, gall bladder, liver and heart with those of other species. Moreover, the topography of the organs is described in retracted and protruded neck in this article. Retraction of the neck had an influence on the position of the organs such as oesophagus, stomach, liver and heart. CONCLUSIONS: The general morphological features of the non respiratory organs of the coelonic cavity of European pond turtle were examined by CT images and macroscopically in this study. Significant differences were found compared with other species.