BACKGROUND: H9N2 avian influenza viruses (AIV) A have become panzootic in Eurasia over the last decade and have caused several human infections in Iran since 1998 and inactivated vaccine has been used in chickens to control the disease. OBJECTIVES: The purpose of this study was to analyze H9N2 viruses that have infected broiler in Tehran Province, Iran between 1998 and 2008 based on Matrix gene. METHODS:  The complete coding region of Matrix (M) gene from 8 of H9N2 subtype isolated from chicken flocks in Tehran Province during 1998-2007 was amplified and sequenced. RESULTS: Sequence analysis and phylogenetic studies of H9N2 viruses on the basis of data of viruses in this study and other selected strains available in the GenBank were conducted and determined variations among these sequences at different levels. Sequence analysis revealed a large number of similar substitution mutations and close evolutionary relation among sequences of M gene. Phylogenetic analysis showed that all our isolates belonged to the G1-like sublineage. In this study, it was determined that Iran’s isolates have been in two separate branches and have the most similarity with Pakistan, United Arab Emirate and occupied Palestine’s isolates. CONCLUSIONS: The available evidence indicates that M genes of H9N2 circulating in Iran during the past years were not well conserved. Our finding emphasizes the importance of reinforcing AIV surveillance.

BACKGROUND: Minerals are essential materials in animal reproduction. The determination of these elements is valuable in relation to different conditions in Arabian horses. OBJECTIVES: The purpose of this study was to evaluate the effects of pregnancy on certain macro and micro minerals in Arabian horses. METHODS: Blood samples were collected from one hundred apparently healthy mares from different regions of Ahvaz including 50 pregnant mares (light, medium, heavy) and 50 non-pregnant mares. Macro-minerals (calcium, phosphorus, magnesium, sodium and potassium) and micro-minerals (copper, zinc and iron) were measured, using standard common laboratory methods. Methods of O-cresol phetalein, Molybdate reduction, Xylidil blue were used for measurement of calcium, phosphorus and magnesium respectively. Flame photometry was used for measurement of sodium and potassium. Amounts of copper, zinc and iron were measured by atomic absorption spectroscopy.  RESULTS: In order to analyze the results, all the data was statistically compared by SPSS-17.0 software. The study of serum calcium levels between the pregnant and non-pregnant mares showed that the differences were not significant. In this study there was significant differences between serum phosphorus levels between pregnant and non-pregnant mares (p<0.05). The serum phosphorus levels during different stages of pregnancy was significantly different (p<0.05), and the level of serum phosphorus during pregnancy declined in total compared to non-pregnant mares. Serum magnesium did not show significant differences between two groups. Serum sodium levels, compared to the other two groups, did not show significant changes. Changes in serum potassium levels showed significant difference between two groups of pregnant and non-pregnant mares (p<0.05). The level of potassium in pregnant mare’s serum was less than non-pregnant, and the least level of this element was seen in medium pregnancy. CONCLUSIONS: Except phosphorus, potassium and zinc, other minerals did not affected different stages of pregnancy. Although the relationship between these changes and pregnancy can be the result of physiological processes, the other factors such as parity, age, nutrition, season, weather conditions and other factors should not be ignored in the evaluation.

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: Identifying the potential of livestock and forage plants of each region is a way to compensate the lack of forage, and efficient use of edible resources of the country. Milk Thistle is a medicinal plant which may be used as fodder for native livestock of Khuzestan Province. OBJECTIVES: The purpose of this research was to study  the effects of different levels of Milk Thistle on rumen fungi and whole rumen microorganisms (WRM) of Khuzestan buffalo, with diets containing grains with different degradation rate (barley and maize). METHODS: The impact of diets containing different levels of Milk Thistle (0, 100 and 200 g per kg DM) on rumen fungi and WRM of buffalo was measured by different techniques. RESULTS: Potential and rate of gas production from experimental diets by WRM were not significantly different. This parameters by buffalo rumen fungi were significantly differed and increased in diets containing Milk Thistle (p<0.05). Adding Milk Thistle in the barley-based diet increased dry matter and NDF digestibility numerically while in the corn-based diet dry matter and NDF digestibility was slightly reduced. In the SRFCM, NDF digestibility, in both basal diets in 9 days and during total period was affected by diets containing Milk Thistle and decreased NDF digestibility with increasing levels of Milk Thistle (p<0.05). Regardless of the type of basal diet, digestibility of NDF (p<0.05) and dry matter (p>0.05) for day 6, were increased with increasing the amount of Milk Thistle. CONCLUSIONS: In general, the use of Milk Thistle did not have negative effect on microorganisms and digestion of nutrients by them. Therefore, results suggest that Milk Thistle could be used up to 20 percentage in buffalo diet without any negative effect on digestion and fermentation characteristics by total microorganisms and fungi.

BACKGROUND: Inclusion of antioxidant sources in feed or water reduces the detrimental effects of free radicals and increases the antioxidative potential and stability of lipids in heat stressed broiler chicken. OBJECTIVES: This study was conducted to examine the effect of Satureja khuzistanica essential oils (SkEO) on early postmortem and ultimate pH as well as antioxidative potential of breast muscle in broiler chicken. METHODS: A total number of 720,1-d-old Arian broiler chicks were used in a 42-d trial to examine the effects of six experimental treatments consisting inclusion of 0 (control-), 200, 300, 400, 500 mg/L SkEO and 500 mg/L Polysorbate-80 (control+) in drinking water in six replicates of 20 birds each in a completely randomized blocks design. Catalase enzyme activity (by kinetic method) and glutathione peroxidase as well as superoxide dismutase activities (via Randox® kits) were measured spectrophotometerically. Lipid peroxidation also was measured in breast muscle by a chemical method with thiobarbituric acid. RESULTS: The breast percentage and early as well as ultimate pH of breast muscle was not differ for the birds received SkEO-treated water compared to the control birds (p>0.05). Addition of SkEO in drinking water at doses greater than 200 mg/L significantly decreased the thiobarbituric acid reactive substances values in breast muscle of the birds in comparison with the control birds (p<0.05). Catalase activity was significantly higher in breast muscle of the SkEO-treated groups than the control groups (p<0.05). Activity of glutathione peroxidase was significantly greater in the birds given 400 and 500 mg/L SkEO. Addition of SkEO into water up to 400 mg/L increased superoxide dismutase activity in breast muscle of the treated birds compared to the control birds. CONCLUSIONS: It was concluded that supplementation of drinking water with SkEO elevates the antioxidative potential and increases the lipid stability of breast muscle in heat stressed male broiler chicken.

BACKGROUND: Theileriosis is a blood protozoan disease with high mortality in cattle in tropical and subtropical regions of the world. Several studies were conducted to identify Theileria species in infected cattles and vector ticks by microscopic examination. However, microscopic technique has lower sensitivity compared to molecular method.                  OBJECTIVES: This study was carried out to identify Theileria species and its carriers in cattles of Yazd city, sing semi nested PCR. METHODS: Between June to September of 2012, 100 EDTA blood samples and 249 ticks were collected from Holstein breed with no history of vaccination against the ileriosis in Yazd area. The collected samples were transported to the laboratory, then prepared the blood smears and stained with Giemsa method. Also, the collected ticks were separated into 50 tick pools, according to their species. Then their salivary glands were removed using stereomicroscope in 0.85% saline. DNA of blood and salivary glands was extracted using a commercial kit and analyzed by Semi-nested PCR. PCR-RFLP was also used to differentiate Theileria lestoquardi from Theileria annulata in positive samples of ticks. RESULTS: Ring forms of Theileria spp. were found in 4 (4%) of blood smears. All ticks were Hyalomma a.anatolicum. Results of PCR were indicated that 11 (11%) of blood samples and three pools of tick’s salivary glands were infected with Theileria. Theileria annulata were only detected in all positive samples by Semi-nested PCR. The results of PCR-RFLP using MspI enzyme showed that the tick’s salivary glands were negative to Theileria lestoquardi infection. CONCLUSIONS: Based on the results, it is concluded that Theileria annulata and Hyalomma a.anatolicum are important agent and vector tick of tropical theileriosis in dairy cattles of Yazd area.

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: Outbreak and spread of bacterial infections resulted in increasing use of antibiotics which subsequently leaded to bacterial resistance to conventional antibiotics. Objectives: To evaluate the effects of organic acids (Selko-pH®), Probiotic (PrimaLac®) and Echinacea purpurea (EP) alcoholic extract on the immune system of the broiler chickens, a completely randomized design experiment was conducted over 120 one-day-old male broiler chicks (Ross 308) with 4 treatment groups that each treatment consisted of 3 replications with 10 birds in each pen. Methods: Groups were designed as follow: group one received organic acid constantly in drinking water (1:1000 v/v) until 14th day and then received 8 hours in a day consecutively (OA); group two received Probiotic (PrimaLac®) water soluble (by 14th day) and mix in the feed (until the end of the rearing period) (PM); group three received EP alcoholic extract in drinking water (1:1000 v/v) like organic acid group and group four were considered as control (received no feed additives). Results: All groups except the control increased the relative weight of the spleen and Bursa of Fabricius as lymphoid tissues; however, this increase was statistically significant only in the case of EP group (p≤0.001). Data of antibodies test against sheep red blood cells and heterophil to lymphocyte ratio also showed that all groups except the treatment of organic acid had significant differences in terms of specific and cellular immune responses (p<0.01). The data of total count of bacteria and also Lactobacilli bacteria in cecal’s content obviously showed that the number of Lactobacilli bacteria significantly increased in PM and OA treatments (p≤0.05). ConclusionS: Feed additives used in these experiments have the ability to improve the immune system.

BACKGROUND: The main part of ovary is consisted of follicles which certain drugs may cause change in them. OBJECTIVES: The purpose of this study was to investigate the effects of Methylphenidate on ovarian follicle of mice, treated by Ritalin before puberty. METHODS: 40 immature female mice at 3 weeks of age were divided into 4 groups, consisting of one control and 3 experimental groups. The experimental groups were gavaged by 2, 5 and 10 mg/kg methylphenidate respectively and the control group received only distilled water with the same method for 60 days. At the end of the experiment, the mice were weighed and then the serum levels of FSH and LH were assessed and structural changes of ovarian follicles and corpora lutea were studied. RESULTS: The mean difference of body weight in experimental groups compared with the control group which showed a significant reduction (p<0.05). In experimental groups compared with the control group, a significant reduction in pre enteral, enteral follicles, corpora lutea and a significant increase in atretic follicles were observed (p<0.05). CONCLUSIONS: Ritalin intake for a long period may increase the number of atretic follicles and decrease corpora lutea, so subsequently results in reduction of the growth of follicles and oocytes as well as inducing the atypical appearance of the cells in the luteinized cells.

BACKGROUND: K99 pilus antigen is one of the major adherence factors found on enterotoxigenic Escherichia coli (ETEC) of neonatal calves. It causes severe diarrhea in newborn calves via the production of heat-stable enterotoxin (STa).With increasing concern over the spread of antimicrobial resistance, the development of alternative to conventional antibiotics such as colicin is urgently needed. Colicin is an antimicrobial peptide produced by one strain of E. coli to suppress the growth of other strains of E.coli. Objectives: The purpose of this study was to investigate the control of E.coli k99 and the efficacy of colicinogenic E.coli (CEC) in adult mice. Methods: The mice, used antibiotic were divided into four groups. The first group did not receive any inoculation. The second group was fed just with 0.5 ml colicin solution. The third group was fed just with 0.5 ml E.coli k99 suspension. The fourth group was first fed by 0.5 ml E.coli k99 suspension immediately after oral administration of CEC suspension. Fecal samples of mice in four groups were taken 1, 2, 3, 4, 5, 6 and 7 days after inoculation and colony forming units (CFUs) were monitored per gram feces. ResultS: The results showed that CEC has inhibitory effect against E.coli k99. There were observed significant differences between the amounts of E.coli k99 recovered from the feces of mice in fourth group with the amount of E.coli k99 recovered from the feces of mice in third group. Conclusions: The data presented here support this claim that CEC plays a significant role against E.coli k99. Furthermore, the study suggested colicin warrants further evaluation as a potential alternative to conventional antibiotics for use to control of E.coli k99.