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: 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: Considering the difference between the rate of multiple ovulation and ovarian activities of different breeds of ewes in different regions, especially out of seasonal breeding condition, finding efficient and effective protocols to induce multiple ovulation out of seasonal breeding would be much appreciated. OBJECTIVES: This study was conducted to determine fertility and multiple ovulation rates during summer and investigate the effect of supplementary feeding, and eCG injection prior to mating in the reproductive performance of Zel ewes. METHODES: At the beginning of summer, 135 ewes in Savadkooh region (north of Iran) were randomly selected and divided into six groups. The estrus cycles of ewes, except in control groups, were synchronized using a vaginal sponge for 14 days. In groups 1 (n=22) and 2 (n=22) ewes were fed with supplementary feeding for 21 and 8 days, respectively, prior to mating. In Group 3 (n=26) ewes were fed with the same ration as Group 2 and were injected (IM) with 400 IU eCG at the time of sponge withdrawal. Ewes in Group 4 (n=23) just grazed on the pasture and injected with eCG as Group 3. In Group 5 (n=20) the estrus of ewes was synchronized as other groups, but they were not received any other treatment. Ewes in Group 6 (n=22, Control) received no treatment, and as the same Group 1 and 2 were mated with fertile rams for eight weeks. Experimental ewes were mated with fertile rams (1:8) 48 hrs after sponge removal for 96 hrs. RESULTS: The findings showed that the fertility in group 3 was significantly more than group 2 and 5, but there was no significant difference between the others (p>0.05). The prolificacy and fecundity were significantly higher in groups treated with eCG than the others (p<0.01), however there was no significant difference between groups 3 (with flashing) and 4 (without flashing). CONCLUSIONS: It could be concluded that vaginal sponge and eCG treatment may improve the multiple lambing and reproductive performance of Zel ewes during summer, and supplementary feeding prior to mating, as described in this experiment, cannot make any significant improvement in multiple lambing.

BACKGROUND: AmpC and ESBLs as mediated-plasmid extended spectrum β-lactabases are the main factors of resistance to extended-spectrum cephalosporins in enterobacteriacea especially E. coli and will follow treatment failure, high costs of treatment in human and economic losses in the poultry industry. OBJECTIVES: The purpose of this study was to screen and study the faecal E. coli isolates producing extended spectrum β-lactamases (ESBLs) and AmpC enzymes and related workers. METHODS: A total of 500 cloacal swab samples from broiler chickens and 25 rectal swab samples from workers were collected from five poultry houses around Tehran. All samples were seeded on MacConkey agar and identification of E. coli isolates were performed via biochemical tests. Antibiotic susceptibility was determined against 12 antibiotics using the disk diffusion method as recommended by the clinical and laboratory standard institute (CLSI2012). Ceftazidim / ceftazidim-clavolanic acid and cefoxitin / cefoxitin-EDTA disks were used for the detection of ESBL and AmpC phenotypes, respectively. phonetic analysis of the drug resistances was performed via SPSS software and Chi-square test. ESBL- producing E. coli screened by PCR for the presence of genes encoding beta-lactamases of TEM, CTX-M and SHV.       RESULTS: A total 467 E. coli isolates were isolated from 88.9% of the samples as 92% and 72.7% of isolates presenting MDR phenotype among chickens and workers respectively. ESBL phenotype detected in 5.5% (26) of poultry isolates while, none of the workers isolates have this phenotype. Six isolates carried both of TEM and CTX-M whereas, five and one isolates were detected only for TEM and CTX-M, respectively. Eighty-eight and nine-tenths percent of ESBL E. coli displayed AmpC phenotype. CONCLUSIONS: Since cephalosporins are not used in broilers in Iran, isolation of faecal E. coli isolates producing extended spectrum β-lactamases in broilerchickens can indicate transfer of the resistance genes via plasmids and other mobile genetic elements among Enterobacteriaceae.

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.

BACKGROUND: SDFT, DDFT and suspensory ligament are the most important tendons and ligament of the palmar aspect of the metacarpus that contribute to stability mechanism. OBJECTIVES: The purpose of this study was to describe the tendons and ligaments of the palmar surface of metacarpus in Anatoly donkey and compare them with those in horse. METHODS: 14 healthy Anatoly donkeys without lameness were selected to detect the tendons, ligaments and their accessories on the palmar surface of metacarpus in both left and right forelimbs after euthanasia. 4 horses were also selected and their tendons and ligaments in palmar surface of metacarpus were compared with those in Anatoly donkeys.  RESULTS: DDFT and suspensory ligament in this region were similar in Anatoly donkeys and horses but SDFT in Anatoly donkeys had an accessory ligament in the palmar surface of the metacarpus that was originated from the deep fascia of carp after the carpal joint and was joined to the SDFT. CONCLUSIONS: This second accessory ligament of SDFT has not been observed in the studied horses and has never been reported in the related references. The results of this study can be used in to diagnose and treat lameness in Anatoly donkeys by radiologists and surgeons.

BACKGROUND: Freshwater pulmonate family Lymnaeidae are well-known for their role in transmission of diginean trematodes worldwide. Objectives: The study was aimed to investigate the ecology and effects of physical and chemical components of the environment on their distribution and populaion density. Methods: The lymnaeid snails were randomly collected from 16 freshwater habitats in West Azarbaijan Province and water samples were also provided from the habitats for chemical analysis. Results: The distribution patterns of the lymnaeid snails in all the examined sites were almost identical throughout the year except in winter. The snails were mostly found in lentic waters or slow-moving streams with muddy beds. The population densities of Lymnaea auricularia, L. gedrosiana and L. stagnalis significantly differed among the investigated waters during the course of study. The concentration of nitrate had significant positive correlations with the snails’ density while there was no significant correlation between nitrite or phosphate concentration with the population density and body size. Conclusions: The results indicated that distribution and density of the snails were affected by season and physicochemical characteristics of environments. These results can be useful for launching the control programs against parasitic trematodes in the region.

BACKGROUND: Sarcocystis infection is one of the most common zoonotic protozoon diseases caused by different Sarcocystis spp. Objectives: Due to the importance of this infection in public health, the infection rate of macroscopic and microscopic cysts in sheep and cattle of abattoir of Shahriar, was investigated. Methods: 138 slaughtered sheep and cattle were selected randomly and their esophagus, diaphragm, heart, tongue, masseter and intercostal muscles were separated. In order to find cysts, the samples were examined by two methods: direct observation for macroscopic cysts and finding microscopics cysts by smear dab, Giemsa staining and microscopic investigation for bradyzoites of parasite. Results: In slaughtered samples, there was no macroscopic cyst but microscopic cysts were positive in 93.48% of cattle and 86.95% of sheep by impression smear method. The results showed the significant difference between different muscles and microscopic cysts (p<0.05) .Heart and esophagus were the most infected and tongue was the least infected part. Infections in males were more than females in both sheep and cattle. There was no significant different in various ages of cattle, however, infection in sheep less than one year old, were higher than the other ages. ConclusionS: Due to the heavy Sarcocystis infection in meat of cattle and sheep and the importance of this parasite in public health, it is suggested to avoid eating raw and undercooked meat and conduct preventive measures such as closer inspection of carcasses and local or total removal of slaughtered in abattoir.

BACKGROUND: Enhancement of the immune system seems to be the most promising method of preventing fish diseases and increasing growth rate. Objectives: The purpose of the present study was to investigate the influence of dietary administration of Zataria multiflora Boiss and Menthapulegium L extracts on phagocytosis, lysozyme, respiratory burst and total white and red blood cells (WBC/RBC) of rainbow trout (Oncorhynchus mykiss). Methods: Two hundred and ten fish (100±10 g) were used in a completely randomized design with 7 treatment and 3 replicates in a 2 weeks period (from 0 to 14 d). The basal diet supplemented with 0 (control), 20, 50 and 100 mg/kg food Z. multiflora and M.pulegium extracts. At the end of the experiment (after 14 days), samples from kidney and blood of the fish were collected in order to determine WBC/RBC (by neubauer chamber), serum lysozyme activity (by turbid metric assay, phagocytosic (by number of yeast cells phagocytosed method) and respiratory burst activities (by reduction of nitroblue tetrazolium method) of head kidney tissue. Results: The results indicated that the highest ratio of phagocytosis and respiratory burst activity was observed in 50 mg/kg extract concentration of Z. multiflora (p<0.05). The highest WBC lysozyme activities were seen   in 100 mg/ kg extract concentration of Z. multiflora. No significant difference was shown between RBC in treatment groups and control group (p>0.05). The highest ratio of phagocytosis activity was observed in 100 mg/kg extract concentration of M. pulegium (p<0.05). No significant difference was observed between WBC /RBC, lysozyme, respiratory burst means in treatment groups and control group (p>0.05). Conclusions: It can be concluded that 50 and 100 mg/ kg of the methanol Z. multiflora and 100 mg/ kg M. pulegium have positive effects on stimulating of innate immune system in O.mykiss, but the influence of Z. multiflora extract with100 mg/ kg  concentration is  better than  M. pulegium extract.