نشریه میکروبیولوژی دامپزشکی
سال سیزدهم شماره 2 (پیاپی 35، پاییز و زمستان 1396)

Melanoma is a malignancy that initiates from melanocytes. Zymosan is an insoluble compound prepared from yeast cell wall and consists of protein-carbohydrate complexes. In this study, we assessed the effects of zymosan on tumor cell viability, growth and apoptosis in melanoma cell line B16F10. Melanoma cells were cultured in RPMI1640 medium with 10% fetal bovine serum.
After one day, the cells were treated by zymosan with different concentrations for 48 and 72 hours. Cell proliferation was assessed by MTT assay. Apoptosis induction was evaluated by flow cytometric analysis using Annexin V/Propidium iodide apoptosis kit. Zymosan noticeably inhibited cancer cells growth in vitro. In addition, zymosan induced apoptosis in a dose dependent manner, mainly at doses of 25μg/ml and 50μg/ml, and reduced tumor growth in vivo. Since zymosan can induce apoptosis and reduce tumor growth, it seems to be an appropriate component for cancer biotherapy and further investigation on antitumor effects of this microbial element may be beneficial.

The study was conducted to evaluate the effect of one commercial probiotic in diet on growth performance and hygienic indices of broiler chickens with using 90 one-day old chicks reared under same condition in two groups with 3 replicates. Chickens in group 1 fed one commercial probiotic according to manufacture instruction. Chickens in group 2 received no supplement in diet. The accumulative growth indices at end of growing period were compared. In 42 days old, chickens slaughtered and breast muscles were sampled for E.coli detection. Cecal content were sampled for E.coli counting. Result showed that chickens received probiotic have significantly higher weight, and lower feed consumption and feed conversion ratio. The adding of probiotic in diet could reduce carcass contamination and microbial load of cecal content. It was concluded that continues administration of probiotic in broiler diet can improve growth indices and hygienic quality of carcass.

Cryptosporidiosis is one of the most important parasitic infection in sheep. The aim of the present study, was to identify species of Cryptosporidium isolated from sheep in Tehran province based on HSP70 gene by Nested PCR-RFLP assay. In the first step 1485 faecal samples were collected form sheep in Tehran province then the samples were examined for ý Cryptosporidium detection using modified acid fast staining. In the second step DNA of the ý positive samples were extracted, then gene of HSP70 was amplified by ýNested-PCR in order to differentiate between species. The PCR product was digested by Hind II restriction enzyme. ý According to the result, 22 positive ýsheep samples were detected by modified acid fast method. The results were confirmed by molecular technique. The 800 bp fragment of HSP70 digested ýby restriction enzymes. Twenty samples showed ýsimilar band on 2.5% agarose gel whereas 2 samples demonstrated different pattern. Based on sequence results, the first and second groups were identified as Cryptosporidium andersoni and C.parvum respectively. Generally, in spite of Cryptosporidium parvum introduced as the major agent of ýcryptosporidiosis in sheep but in our study Cryptosporidium andersoni was dominant.

Mycoplasma Synoviae is a major poultry pathogen which causes significant economic losses to the poultry industry. The aim of study was detection Mycoplasma synoviae infections in vaccinated broiler breeder flocks and their progeny. For this purpose, from 20 broiler breeder farms and their progeny 2 times in 40 and 60 weeks of age 20 tracheal swab and 20 blood samples from each broiler farm were collected. All 20 broiler breeder farms before vaccination against Myc.oplasma Synoviae were confirmed free from Mycoplasma synoviae infections. Tracheal swab transfered to cap tubes containing PPLO broth media and kept at 4°C less than 24 hrs send to laboratory for further proceeding. Antibody against Mycoplasma Synoviae was measured by rapid serum agglutination (RSA) and Elias tests. Tracheal swabs were examined by high resolution melting PCR(HRM PCR) for detection Mycoplasma Synoviae field isolate.. The results showed that vaccination broiler farm against Mycoplasma Synoviae upto 40 weeks could protect all broiler breeder farms from Mycoplasma Synoviae field infection and produce good antibody titers. With increasing age and at age 60 weeks one farms and its progeny was positive for Mycoplasma Synoviae which was differ from Mycoplasma Synoviae included in live vaccine. This study showed vaccination by present vaccine against Mycoplasma Synoviae in broiler breeder farms could not protect older broiler breeder farms in older ages and probably Mycoplasma Synoviae infection transferred to progeny. Therefore vaccinated farms should be monitor by molecular methods.

In recent years, the multicausal respiratory disease has caused severe economic losses in poultry industry, particularly in broiler flocks. A variety of viral and bacterial factors are involved in the incidence of this disease, one of the most prominent of which is infectious bronchitis virus (IBV). Despite the vaccination program, this disease is still regarded as the most prevalent virulence factor in poultry industry throughout the world. Recently, the outbreak of multicausal respiratory disease has led to an increase of considerable losses in broiler flocks in Bushehr province. In order to detect the infectious bronchitis virus in vaccinated and unvaccinated broilers with multicausal respiratory disease, 8 flocks, vaccinated against infectious bronchitis and 7 as unvaccinated flocks were selected as samples. For all vaccinated broilers, the Massachusetts Serotype Vaccine was used as bronchitis vaccine. During the study, representative samples showed respiratory problems including coughing, sneezing, and respiratory rales. In the present survey, 135 tracheal swabs and 150 blood samples of 15 flocks with respiratory infection and with high mortality rate were collected and examined using RT-PCR method and ELISA. Out of 15 sampled broilers, 12 (80%) were positive for IBV, and 3 were negative for molecular detection. All unvaccinated broilers were positive for IBV, while 3 among vaccinated ones were negative. In serological assay, it was elucidated that all unvaccinated flocks were positive, demonstrating correlation with molecular results. The findings revealed a high incidence of infectious bronchitis virus in broilers with multicausal respiratory disease; especially in unvaccinated flocks, showing an immediate action in controlling infectious bronchitis disease as to decrease the damages caused by multicausal respiratory disease.

Medicinal plants‚ which may have less adverse reactions‚ can be a suitable substitute for chemical drugs. In this comparative studies of antibacterial activity of extracts Nasturtium officinale and Coriandrum sativum used on Gram-positive (S. aureus and L. monocytogenes) and Gram-negative (S. Typhimurium and E . coli) bacteria.
We used Coriandrum sativum and Nasturtium officinale alcoholic extract this study. First‚ prepared the alcoholic extract of the plants. At concentration of 100 mg/ml, 50 mg/ml, 25 mg/ml, 12.5 mg/ml, 6.25 mg/ml, 3.12 mg/ml, 1.56 mg/ml and 0.78 mg/ml and evaluated its antibacterial effect. Well Diffusion and Microtitr Plate for determining the Minimum Bactericidal Concentration and Minimum Inhibitory Concentrations methods were used on Gram-positive (S. aureus and L. monocytogenes)
and Gram-negative (S. Typhimurium and E. coli) bacteria. Results showed that alcoholic extract of Coriandrum sativum have more antibacterial activity than alcohol extracts of Nasturtium officinale. Our results also showed that S. aureus and L. monocytogenes had more susceptibility and S. Typhimurium and E_coli were the resistant one. the extracts from Coriandrum sativum and Nasturtium officinale had MIC equal to 6.25 µg/ml and MBC equal to 12.5 µg/ml against S. aureus.
The antibacterial activity of Coriandrum sativum and Nasturtium officinale alcoholic extract was observed on gram positive bacteria and so may be beneficial in the treatment of infectious diseases.

The global popularity of probiotic products on one hand and their low viability in foods and digestive system on the other hand, has led researchers to innovate new techniques in order to enhance their viability. Chocolate is considered as a healthy food in societies due to its nutritional and delightful properties, ease of use and variety of forms. It is necessary to eliminate the limitations in order to use probiotics in chocolate. Therefore, to use probiotics in chocolate making, microencapsulation of probiotics can be an efficient method to protect their survival in unfavorable conditions of manufacturing process and digestive system and thus exploit the nutritional properties of chocolate. In this study, calcium alginate and resistant starch were used for making capsules by emulsion method and Bifidobacterium breveýwasýaddedýtoýchocolateýinýtwoýforms:ýfreeýandýmicroencapsulated;ýandýsurvival,ý acidity,ýwaterýactivity,ýsolidýcontentýandýsensoryýattributesýofýchocolateýwereýmonitoredý duringý30ýdaysýstorageýatý4ýͦC. the morphology and size of microcapsules were measured, SEM technique and particle size analyzer. Obtained results showed that probiotics in microencapsulated form had higher survival compared to those in free form. Acidity and water activity of chocolate with free form of probiotics were reported higher than those of samples with microencapsulated probiotics. Also chocolate contained microencapsulated probiotic had higher solid content than that of samples with free form. In addition, microencapsulation had no undesirable effect on sensory attributes of chocolate. microencapsulation helps to enhance the survival of probiotic bacteria in chocolateand manufacturing probiotic chocolate is now made possible.

This experiment was carried out to evaluate the effects of adding vinegar to drinking water (DW) and probiotic to starter diet (SD) on growth performance, gastrointestinal tract (GIT) contents pH, and ileal lactobacillus population of broiler chicks. A total of 330 one-day-old male Ross-308 broiler chicks were used in a completely randomized design (CRD) experiment consisted of 6 treatments with factorial arrangement 2×3 and 5 replicates of 11b each. Treatments consisted of 2 levels of adding probiotic zero and 0.1% “1×1010 CFU lactic acid/g of supplement” to SD and 3 levels of adding vinegar zero, 1 and 2% “5% acetic acid concentration” to DW. A corn-soybean meal based diet was formulated to meet the nutrient levels suggested by Ross-308 (2015). The study lasted from 110d of age. At the 10th day of age’s one bird from each experimental unit (5 for each treatment) were randomly selected and were slaughtered for evaluation GIT content pH, and ileal lactobacillus population. In the birds were fed supplemented diet with probiotic combination drunk supplemented water with 1 and 2% vinegar levels feed intake (FI) and feed conversion ratio (FCR) were significantly lower than birds fed diet and drunk water without any supplementation. In the birds were fed supplemented diet with probiotic and or were drunk supplemented water with vinegar, ileal lactobacillus population were significantly higher than birds fed diet and drunk water without any supplementation. The addition vinegar to DW and probiotic to SD, and their interaction effects were not significant on the pH of GIT contents. In conclusion, drinking water supplementation with vinegar and or starter diet supplementation with probiotic will improved ileal benefit microbial flora and feed efficiency of broiler chicks.

In order to survey on the causative agents and the occurrence of suspected disease to
Streptococcosis in some farms of rainbow trout in the areas of Dasht-e-Arzhan and Kohmareh in Shiraz province, 100 specimens of live fish with symptoms similar to Streptococcosis were caught and After dissection, bacterial sampling were taken from kidney, liver and brain then cultured on Blood Agar media. Samples were stored in the incubator at a temperature of 25 -30 ° C for 48 hours. After this time, at first gram negative samples were removed by using gram stain technique and catalase test. then the supplementary biochemical tests were performed after recultivating of the samples. The results showed that the samples isolated from the two species, Streptococcus iniae and Lactococcus garvieae. The occurrence of fish with clinical symptoms was 77% to S. iniae and 13% to L.garvieae and no Infection isolated from 10% of the samples.

E.coliO157:H7 is recognized globally as an important cause of diarrhea, hemorrhagic colitis (HC) and haemolytic uremic syndrome (HUS) in humans. The main and natural reservoirs of E. coliO157:H7 are feces of domestic and wild animals, which shed the bacteria with their feces into the environment.The aim of the present study was to isolation and the prevalence determination of E. coliO157:H7 in fecal samples of ostrich farms using culture and multiplex PCR in Lorestan province. In this cross sectional study, at all 100 fecal samples of ostrich were collected during March to May 2015. A 10-g of fecal sample was added to 90 ml of Trypticase soya broth containing novobiocin and all of enrichment samples were cultured on selective sorbitol Macconkey agar plates supplemented with Cefixime and tellurite (CT-SMAC) for screening test. All nonsorbitol fermenting isolates were evaluated to multiplex PCR using specific primers. From 100 fecal samples, after enrichment and selective plating, 15 (15%) nonsorbitol fermenting were isolated, but only 4 (4%) isolates were identified as E.coliO157:H7 using multiplex PCR. The present study is the first report on isolation of E. coliO157:H7 from ostrich feces sample in Iran. This study shows the importance of ostrich fecal as a reservoir of E. coliO157:H7.