نشریه میکروبیولوژی دامپزشکی
سال چهاردهم شماره 1 (پیاپی 36، بهار و تابستان 1397)

Every year, the outbreaks of highly pathogenic avian influenza occurs when migratory birds migrate to different countries.. Because of high economic losses and its effects on public health, increase in the outbreaks of this disease,have increased concern in global societies.Most of the non-waterfowl wild bird Species are susceptible to HPAI and for various reasons, including free-range flying, contact with domestic poultry and in some cases contact with industrial poultries, migrating some species and moving between different countries, international trade, in some cases smuggling of them, andclose contact with humans because of their maintenance as entertainment or as games bird, they arepotentialy important in the ecology HPAI viruses. Most reports of this disease in the nonwaterfowl wild birds are the H5N1 subtype. The least susceptibility to HPAI viruses has been observed in the pigeon and starling spicies, and probably these spicieshavethe least role in the epidemiology of these viruses. While other Passeriformes, Falconiformes and parrots are highly susceptible to HPAI. In the most reported outbreaks in the Falconiformes and crows, the main causes of the infection were using the carcasses of the affected birds or contact with them. The main cause of the susceptibility of these birds to the influenza viruses is the presences of the alpha 2 and 3 sialic acid receptors in the respiratory and gastrointestinal tracts of them. The most common signs that observed in these birds are neurologic signs and the highest viral concentration has been observed in the brain.Due to the susceptibility of non-waterfowl wild birds, and the large number of these birds and the close contact between migratory birds with domestic and industrial birds and the possibility of transmission of infection to other birds and humans, these birds should be considered in the designing of influenza surveillance programs and in the outbreaks of HPAI.

Lumpy skin disease virus (LSD), Goatpox virus and Sheeppox virus are members of genus Capripoxvirus of the Poxviridae, which have close genetic similarity. Therefore, the use of CaPV-vaccine strains derived from sheep and goat would be useful to protect cattle against LSD. The aim ofthis study was to evaluate the immune responses characteristics of available goat pox vaccine against LSD in cattle. All immunized calves were daily examined for any increase of rectal temperature and appearance of adverse reaction for 5 weeks following vaccination. Humoral immune responses were evaluated by serum neutralization test and cellular immune response were evaluated by MTT, Real-time PCR and ELISA tests. The results showed that theFever and localized swelling at the injection site were observed in vaccinated calves, in comparison to the control group. In assessing of humoral immune response, a significant increase in the neutralizing antibody titer was observed in the vaccinated group in comparison to control group, although this difference in days 35 was higher than the other days (p<0.05). In assessing the cellular immune response, a significant increase was shown for IFN-γ gene expression on day 7 and for IL-4 gene expression on day 21 postvaccination in comparison to day 0 and control group (P<0.05), While the highest cytokine production in cell culture supernatant for IFN-γ and IL-4 was observed at day 21 postvaccination (P<0.05). Also, the lymphocyte proliferation index in vaccinated calves showed a significant increase on day 7, 21, and 35 in comparison to day 0 and control group (P<0.05). According to the results of this study, the results of this study indicated, that goat pox-vaccine due to induction of high level of antibody titer, higher lymphocyte proliferation and IFN-γ and IL-4 production has a good immunogenic response, So It considered suitable vaccine to control of LSD.

Silver nanoparticles have been synthesized by several methods, but each of them has disadvantages. Chemical methods, have high toxicity and low stability, and physical methods, are costly and low-efficient. More recently, researchers have found a new technique for synthesizing AgNPs, using plants and microorganisms such as bacteria, fungi, and Actinomycetes. This method is called “eco-friendly”, “green synthesis” or “Biosynthesis”. The aim of this investigation was, biosynthesis of AgNPs using Saccharomyces cerevisiae, which is one of the safe microorganisms that the AgNPs synthesized by it, are suitable for using in pharmacology and medicine purposes. Saccharomyces cerevisiae was cultured in sabraud dextrose broth and incubated at 28-30°c for 24 h, then it was centrifuged, and its supernatant was added to the silver nitrate solution with the concentration of 0.001, 0.002 and 0.003 M at pH: 5.6, 7 and 9, then incubated at 28 -35 and 37°c. The formation of AgNPs, was monitored by color-changing, uv-vis spectroscopy, TEM, XRD and FTIR. Antifungal effect of AgNPs on Microsporumcanis investigated by disc diffusion method. MIC and MFC were also measured. The results, showed that Saccharomyces cerevisiae have had the great potential for synthesizing of AgNPs in the size of 27 nm and have shown the antifungal effect in the concentration of 33±0.29 μg/ml.In this study, we have reported a simple, biological, easy and nontoxic method for synthesis of silver nanoparticles, which have had suitable antifungal effect on Microsporumcanis, by using one of the nonpathogenic microorganisms.

Opportunistic fungi are scattered in aquatic and soil environments. Predisposing factors such as density, transporting, handling, water pollution, manual feeding and other stressors play role for fungal infections outbreak, especially in sensitive specie like as rainbow trout.This fish is one of the popular species in Iran that is one of the major trout producers in the world. Because of that study of bacterial, fungal and parasitical flora in such facilities are highlighted. For this reason, Skin, fins and gills samples of 150 rainbow trouts (with healthy appearance) with 100.98± 5.48 g body weight from 15 facilities of Alborz province, in autumn and winter, 2016-2017 were collected. All the farms water resources were ground water (well). Samples were transferred to lab in sterile condition and under the laminar flow hood were cultured in Sabouraud Dextrose Agar with chloramphenicol, Corn Meal Agar and Glucose Peptone Agar. Cultured media were incubated in 23±1 ºC for at last 2 weeks. Grown fungal colonies were evaluated by their both macroscopic and microscopic methods.Out of surface samples, 103 fungal isolates were identified that 87 of that are molds belong to 11 genera besides 16 yeast isolates and out of gill samples, 82 fungal isolates were identified that 70 of that are molds belong to 10 genera and others were yeast isolates.In gills and skin, Aspergillus was the most frequent fungal genus and flavus was the most one among Aspergillus species. In order to the Chi square statistical test, there was no more significant correlation between identified fungal isolates in body surface and gills (P>0.05). But in most cases, the quantity of each isolated species was more in body surface than the gills.Fungi are present everywhere, and it is normal to be in water resources. Environmental pollutants (e.g. industrial, agricultural and municipal wastewaters) can provide productivity of water and entering the development of non- water molds. Diversity of fungal isolates is the indicator of poor water quality resulting low production, fungal infections increase as well as treatment and medication costs. Consumption of fungal spore-polluted fish food in facilities can be another way of entering of such fungal pollution.

Mycoplasma bovigenitalium is one of the mastitiscausing pathogen in dairy cows. The complications of the disease arereduced feeding, changes in the quality and consistency ofits stiffness, swelling Cartier, arthritis, infection of the mucous membranes of the respiratory tract, urinary tract, ear infections and septicemia. The purpose of this study was to Isolation and Identification of Mycoplasma bovigenitalium from bovine milk suffering of clinical mastitis by culture and PCR methods. In this descriptive study, 156 samples of milk collected from cows with clinical signs of mastitis in dairies industry. Samples were cultured in PPLO broth and PPLO agar. The bacteria DNAs were extracted by phenol/ chloroform method. The PCR assey applied for detection of M. bovigenitalium by molecular method used MbgF, MbgR specific primers. In the culturing method from 156 samples, 57 (37%) samples were reported to be positive for Mycoplasmaand 5 (3%) were suspected to be positive.In the PCR test from 156 samples, 62 (40%) samples were positive for Mycoplasma. In the specific PCR test, 1 (2%) of these specimens were positive for Mycoplasma bovigenitalium. The use of the molecular method of PCR showed that it is a rapid diagnostic method with high sensitivity and specificity, which is used as a classical method.

Leishmaniasis is a protozoan parasitic infection that is reported in most parts of the world and is prevalent in different parts of the world. The disease is the third most important disease in the world transmitted through the biological vector. From 12 to 15 million people in many countries around the world are infected with leishmaniasis and every year there are 2 million new cases of leishmaniasis worldwide. Treatment with chemical drugs may have a lot of side effects for the patient. For herbs, garlic, walnut leaves, honey, thyme, and yarrow, there is a significant therapeutic effect. In the present study, products from medicinal herbs and honey for the treatment of cutaneous leishmaniasis were investigated in the Balb/C mice model. The results of this study showed that garlic extract at concentrations of 25, 50 and 100 mg/mldid not any statistically significant therapeutic effect on leishmania, and there was no statistically significant difference between the controls (p≥0.05). Walnut leaf extracts showed significant therapeutic effects. Also, honey extracts at different concentrations had a significant therapeutic effect and showed a lower effect on the extract of walnut leaves. Yarrow and thyme extracts also had good therapeutic effects on the walnut leaves and honey. Thyme extract with a 100mg/ml therapeutic effect had a favorable therapeutic effect. The results of this study might be effective in the treatment of leishmaniasis.

Clostridium perfringens is cause of enterotoxaemia disease in sheep. The purpose of this research was to select the best strain Clostridium perfringens type D isolated from Iran and their comparison with vaccinal strain by MLD and potency tests. In this research 15 strains Clostridium perfringens type D were studied. At first, microbiological and biochemical tests were done then specimens were confirmed by PCR test. Finally, the best strain was selected by MLD test and the enterotoxaemia vaccine was prepared by the selected and the vaccine strain separately. The vaccines were kept for 45 days at refrigerator then potency test was done according to standard protocols. The results showed that the conditions for epsilon toxin production by isolated strain and the vaccine strain was nearly similar.

Avian influenza (AI) is a serious infectious disease with world-wide significance causing considerable financial losses in poultry industry.Serodiagnosis based on detection of antibody to the nucleoprotein (NP) that is conserved among all influenza A virus, can be used to demonstrate immunity against all subtypes of influenza A virus. The aim of this study was to produce recombinant nucleoprotein (NP). For this purpose, the coding region of nucleoprotein (NP) gene of A/chicken/Iran/AH-1/06(H9N2) AI isolate was amplified by RTPCR and cloned into a prokaryotic expression vector (pMal-C2x) and transformed into E. coli BL21(DE3). The purified recombinant vector for the expression of recombinant MBP-NP was used.Target protein with a weight of about 97 kDa by SDS-PAGE and subsequent Western blotting was evaluated. Observationthe expressed protein with expected molecular weight in SDS-PAGE and positive reaction ofthis protein with bird ,serum infected avian influenza In western blot showed that the expression of NP gene in E.coli was successful.

Hepatitis E virus infection, including disease is emerging in recent years in most countries because of increased mortality and decreased egg production in laying hen flocks is about 20%. Due to the development of poultry farming laying hens and reduce economic losses for growers, pest control and waste reduction in egg production is very important. Since in our country no study has been done on this disease assessment and notification of the presence of the disease in cattle is evident laying. A total of 150 serum samples and 150 samples of fecal swabs of hens around the time of collection and Nested PCR method were investigated. A total of 150 serum samples and 150 fecal swab samples were taken. In 6 cases (4%) Since the virus genome was detected in 5 of 6 cases (3.33%) of the samples, but the virus was detected in fecal samples from the birds, the virus was detected in 1 case (0.66%) in both serum and swab samples were bird droppings containing the hepatitis E virus.

Causative agent of toxoplasmosis is an obligatory intracellular protozoa known as Toxoplasma gondii which is able to infect most of warm blooded vertebrates. This disease due to congenital infection and abortion is of significant importance in human and veterinary medicine. Aim of this study is seroepidemiologic evaluation of toxoplasmosis by latex agglutination method in poultry of Mazandaran province. In this experiment after taking blood samples and record of poultry charecteristics including age and gender, samples were transferred to laboratory and after centrifugation sera were seprated and stored in - 20ºC. Then for evaluation of toxoplasmosis infection, diagnostic latex aglutanision kit manufactured by ZistFaravaran Pars ( ZFP) was usd and obtained data and results were analysed.In this study done byserologic Latex agglutination method, toxoplasmosis infection rate in Mazandaran province domestic poultry was observed as fallows: hen ( 13.11%), rooster (40%), duck (16.82%), goose ( 10.41%), and turkey (21.56%).