نشریه میکروبیولوژی دامپزشکی
سال سیزدهم شماره 1 (پیاپی 34، بهار و تابستان 1396)

Genetic diversity of rumen bacterial population in goats was identified using a modern molecular culture independent method. It is probable that structure of bacterial colonies of goats show special characters and some differences, in comparison to the other ruminants because of their distinct diet and habitat. So that obtaining enough knowledge about bacterial diversity in various breeds of goats and their variations in response to diverse diets and also determination of bacterial diversity in various environments is essential. Therefore a mixture sample of whole rumen content of 12 native goats from the north of Iran was collected. PCR amplification was done by universal primers of bacteria and 16S rRNA gene clone library was sequenced. Samples were collected from herds that fed by a mixture of forage and concentrate. Totally fifteen sequences of 16S rRNA were analyzed. From these 15 sequences, 4 sequences showed similarity (94%) to Streptococcus bovis and 3 sequences were similar (90%) to Selenomonas ruminantium. Also 2 sequences were similar (89%) to Megasphera elsdenii and two others resembled (96%) to Prevotella ruminicola. Two sequences had similarity (89%) to uncultured bacteria from the rumen of cow. One sequence from these 15 sequences showed similarity (90%) to Lactobacillus plantarum and one last sequence was similar (92%) to Prevotella multiformis. This study is the first identification of rumen bacterial population diversity from the goats in Iran. The results of this study showed that bacterial population diversity in the rumen of goats is similar to other ruminants in other parts of the world. Although we isolated some sequences which are not clustered to common bacteria in digestive tract. It may as results of individual differences during sample collection, types of rations, methods of DNA extraction or used primers.

C. jejuni and C. coli are considered as major causes of bacterial enteritis in human, worldwide. Birds are playing a substantial role in the transmission of infection. A relatively high prevalence of the infection in the rearing period leads to the cross-contamination during the slaughter processes which coincidently cause the contamination of poultry meat products. Investigation on the contamination of poultry carcasses at the end of rearing periods to C. jejuni and C. coli. In autumn 1388, total of 100 poultry farms were randomly selected, from which, 21 carcasses were chosen to collect a cecal specimen from each carcass. Each cecal contents of seven carcasses were then pooled, culture on a selective agar plate and enriched in a selective broth medium followed by culture on a selective agar plate. The isolates were finally confirmed using a multiplex-PCR assay. Total of 53.7% (70% of all the farms) were found positive using direct culture plating, whereas, 60% (76% of the farms) were positive when the samples were initially enriched in a selective broth. 31.3% to 35% and 36.7% to 40.0% of all the positives were respectively confirmed as C. jejuni and C. coli. A 6.3% increase in the isolation rate of Campylobacter, when the samples were initially enriched in the selective broth, was demonstrated. The present study revealed a considerable high prevalence of Campylobacter in poultry farms, and thus, a closer investigation on the possible risk factors involving in the contamination is essential. Furthermore, to reduce the risk of infection in human, improving the hygienic inspection in the poultry slaughter houses, is also recommended.

The aim of this experiment was consider of effects of replacing soybean meal (SBM) by different levels of sunflower meal (SFM) supplemented with protease enzyme on intestinal microflora, gizzard and cecum digeta pH and internal body organs characteristics in laying hens. In total, one hundred and twenty white Bovans laying hens were used from 77 to 86 weeks of age. This experiment including 6 treatments, 5 replicates and 4 hens in each. This study was conducted in a completely randomized design (CRD) as a factorial management 3×2 (0, 45 and 90% sunflower meal replaced by SBM) and protease enzyme (0 and 200 g/ton). Digesta cecum was taken for measurement of intestinal microflora of birds (lactic acid producing bacteria and Escherichia coli) at 86 weeks of age. Then, pH of the gizzard and cecum digesta and internal body organs characteristics were measured. Population of lactic acid producing bacteria and Escherichia coli, and gizzard and cecum pH were not affected by treatments (P>0.05). Duodenum length significantly increased by 45% replacing of SFM in the diet than other treatments (P

In this study a serological survey was carried out to detect group specific blue-tongue virus antibodies in goats serum collected in two regions of Chaharmahal-Va-Bakhtiari province of Iran in 2015. Blood samples were taken randomly. A competitive enzyme linked immunosorbent assay(C-ELISA) was conducted to test the serum samples for blue tongue virus (BTV) group specific antibodies. BTV seropositive reaction were obtained in 776 (57.48%) out of 1350 tested sera. From 520 serum samples in plain region and 820 serum samples in mountain region, the rate of positivity was 79.04% and 43.98%, respectively. Higher seroprevalence was observed in plain region (P

The predominant mode of growth of bacteria in the environment is, matrix-enclosed communities known as biofilms. This bacterial structure often complicate chronic and difficult-to-treat infections by protecting bacteria from the immune system, decreasing antibiotic efficacy, and dispersing planktonic cells to distant body sites. The purpose of the present study was to evaluate the biofilm producing ability of three standard strains of Escherichia coli and one standard strain of Staphylococcus aureus in different enrichment media and compared its structures.
For this study, Escherichia coli and staphylococcus aureus strains were cultured in four enrichment media (Brain-heart infusion broth contain 1% sucrose, Luria-Bertani, Tripti soy broth contain 3.5% glucose and Pepton Water) for phenotypic evaluation of biofilm production based on micro titer plate crystal violet method presented by stepanovic et al. (2007). Then, two Escherichia coli strains (1, 3) with strong and moderate biofilm producing ability, were selected for carbohydrate and protein extraction and compared with SDSPAGE by periodic acid shiff and methylene blue staining, respectively.
Based on results, BHI contain 1% sucrose, was selected for two strains. Protein profile of two studied strains were differences in protein components higher than 68 Kd, but, had a same carbohydrate and glycoprotein profiles. Finally, by observing the difference in protein structure of studied strains, identification and structural survey on different protein components can be useful in order to applying effective strategies to eradication of chronic infections.

In this survey, the microbiological control of poultry carcass slaughtered in industrial and semiindustrial slaughterhouses were compared (25 samples in each line). In industrial slaughter line, no process is performed manually. The visceral organs are removed automatically and chilling air is used to cool down the carcasses while in the semi-industrial line, the visceral organ removing is a manual procedure and chilling the carcasses is done by floating them in ice water. The bacterial contamination in each sample was studied using Total Count and Fecal Coli form by using pour plate count and MPN method, respectively and led to the isolation of E. coli, Salmonella, Staphylococcus aurous, Campylobacter, Listeria and Vibrio. The results showed that Total Count of%8 and%12 the industrial and semi-industrial lines ´samples were above the standard, respectively. In addition, Coli form count was above the standard in 24% of semi-industrial samples. None of the samples in the industrial line were Fecal Coli form & E. coli contaminated. In the semi-industrial line; Fecal Coliform & E, Coli were isolated from 44% and%52 of the sample, respectively. Staphylococcus aureus was not isolated from any of the samples in the industrial line while in the semi-industrial line, 8% of the samples had the contamination above the standard. No Campylobacter, Listeria, Staphylococcus aureus and Vibrio could be isolated from these 50 samples.

Chicken infectious anemia virus (CAV), cause severe anemia, and makes atrophy in lymphatic organs include thymus, bursa of fabricious, spleen and bone marrow. It suppresses immunity system and increase the susceptibility to other infections. Subclinical infection of CAV caused high economic losses in broiler flocks. Isolation of the virus in tissue culture, detection of genome by PCR based method and serological tests are diagnostic methods of chicken infectious anemia virus. The aims of the present study were detection and partial nucleotide sequencing of CAV genome by PCR and direct sequence method, respectively. Seventy seven liver samples from chickens belong to 8 different broilers flocks around the Sharekord city, in Chahrmahal VA Bakhtiari province, were tested. Results showed that 19 samples belong to 4 broiler flocks were positive. Amplicons of 4 PCR positive samples belong to 4 different flocks were sequenced and the results showed that partial gene sequences of chicken infectious anemia viruses had 100% homology. These sequences had more homology with nucleotide sequences of Indian, Malaysian, German and Chinese CIAV strains. With regards to phylogenic similarity, it seems that all of these isolates have a same origin.

Mastitis is an important disease that could turn up in both milking and dry off periods in dairy cattle. Considering the importance of Escherichia coli in causing microbial mastitis, this study in an Insilico way was undertaken to investigate the amount of biomass, toxin production and to identify genes affecting on Escherichia coli growth in both aerobic and anaerobic conditions in alveoli of mastitis dairy cow. The optimum rate of Escherichia coli growth or biomass production adopting aerobic condition was 2.0155 millimoles per gram dry weight per hour (mmol/gDW/h) but in anaerobic condition this value was 1.0152 mmol/gDW/h. The most toxin production was obtained in aerobic condition (1.6701 mmol/gDW/h). This substantial amount of biomass production could be a great indicative of rapid growth of Escherichia coli and therefore, inflicting the mastitis in dairy cow. Multiple gene deletion indicated that four genes ompF, ompN, PhoE and ompC brought about of stopping biomass production. Also, it was indicated that solA, LpxM ¡ ZnuA and znuB genes are suitable drug targets for reducing or stopping toxin production in Escherichia coli.

Foot& mouth disease (FMD) is a vesicular viral disease which cause severe damage in livestock industry. Today FMD is controlled by traditional inactivated vaccine. Due to short duration of immunity, using molecular adjuvant is recommended to increase efficiency of vaccine. Purpose of this study is synthesis fusion protein from VP1 protein of FMDV (Opanasia2) and flagelline of the Salmonella (fliC) in E. coli. 1D gene that is responsible for producing VP1 protein was produced by PCR (800bp)and cloned in PTZ57 vector. After cutting cloned fragment from the recombinant plasmid by HindIII&XhoI, inserted to expression vector (pET41b) that containsfliC of the Salmonella. The construct was transfected to competent E. coli (BL21DE3) cell. The expression was induced by IPTG (0.5mM) and purified by Ni-NTA Resin. Production of recombinant protein was resolved by SDS-PAGE in 12% acrylamide gel and western blotting. Recombinant fusion protein was evaluated by PCR and enzyme digestion. By molecular analysis, PCR product by length 700 and 1500 bp was observed. Ni-NTA Resin and gradient pH was used for purification. The most amount of protein recovering was observed in pH 4.5. Recombinant protein by molecular weight of 76 KDa determined in SDS-PAGE (12% acrylamide gel) and was confirmed by western blot by AntiHis. In this study, the fused gene of FMDV vp1 and flagelline of Salmonella (fliC/2200 bp) was cloned successfully in E.coliand recombinant protein (76 KDa) was expressed and purified.

Salmonellosis is a common disease among human, animal and poultry which is raised as a food-borne illness. One of the most common serotypes of this bacterium is salmonella infantis. The aim of this study was to investigate the antimicrobial resistance of 70 salmonella infantis isolates from poultry between 1389-1390 in Arak. In order to confirm the genus of Salmonella, all isolates showed 1.5 Kb band on agarose gel. The results of antibiogram that was performed by the Kirby-Bauer disk diffusion method according to Clinical Laboratory Standards Institute (CLSI) showed that all isolates (100%) were resistant to nalidixic acid and nitrofurantoin. Among 70 isolates, 2 (2.9%) cases were resistant to 11 antibiotic followed by 10 (14.3%) to 10 antibiotic, 8 (11.4%) to 9 antibiotic, 6 (8.6%) to 8 antibiotic, 22(31.4%) to 7 antibiotic, 12(17.1%) to 6 antibiotic and 8(11.4%) to 5 antibiotic. Also twenty-nine antibiotic resistance patterns were found.

The Mycobacteria grouped in the Mycobacterium tuberculosis complex are causes of Tuberculosis in animals and humans. This chronic disease also affects a wide range of other domestic and wildlife animals and may also cause disease in humans. It is very important to control and prevent the epidemic among human and animals because of zoonotic identity of the pathogen and several cattle flocks in suburb of cities, and it must be done better through more active surveillances. During this study, the causative agent of disease must be isolated from rodents found and hunted on the reactor or suspected farms. Finally, it must be distinguished that which isolates (species) are circulated in those areas. This study has been carried out by cooperation of Khuzestan Veterinary Office. By use of T.B test sample Farms have been selected. They consist of 16 mice from the same farms. All samples were referred to laboratory, then cultured in specific Lowenstein-Jensen slant media. After at least 8 weeks incubation (at 37ºc) DNA was extracted from 2 isolates of 16 mice to identify the isolates belonging to the Mycobacterium tuberculosis complex, by using PCR detection of insertion sequence 6110 (IS6110) and (16srRNA). According to the test 2 isolates of mice belong to the family of Mycobacterium tuberculosis complex.

The Sistani cattle are indigenous breed of Sistan region. The purpose of this research was the separation and studying the appearance morphology of anaerobic fungi in the Sistani cattle rumen in Sistan region. Sampling from the solid and liquid contents of 50 Sistani cattle was done randomly in Zabol slaughterhouse and these samples were used as the source of fungus to inoculation to culture. The semi-defined medium environment was used in this research to cultivation, separation and purification of anaerobic fungi. The roll bottle method was used for purification of rumen fungi and the antibiotic solution (ampicillin, penicillin, streptomycin, oxytetracycline and chloramphenicol) were used for inhibiting growth of bacteria. Samples of pure fungi were transferred to culture and were observed after growth in glass slide with light microscope. With regard of morphologic characteristics the genus of Neocallimastix and species of Orpinomyces joyonni, Piromyces mae, Piromyces communis, Piromyces minutus, Piromyces rhizinflata was isolated in rumen of Sistani cattle. With identification of these fungi species in rumen of Sistani cattle, it is recommended to perform molecular test and enzyme extraction for more survey characteristics in future research.