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Differentiation of Infectious bursal disease viruses isolated from Iranian poultry flocks using real-time RT-PCR and high resolution melt curve analysis
2017
Peighambari, Seyed Mostafa | Cheraghchibashi, Mehdi | Hosseini, Hossein
BACKGROUND: Infectious bursal disease (IBD) is a highly contagious disease of young birds. Differentiation between classical virulent and very virulent IBDV (vvIBDV) isolate is very important for the poultry industry to choose the right vaccination program. Molecular and serological tests are time consuming and have variable sensitivity. However, the melting curve analysis is relatively fast method with high precision. OBJECTIVES: This study was designed to evaluate the efficacy of the melting curve analysis for differentiation of some Iranian IBDVs which their identity had been previously determined by RT-PCR/RFLP analysis. METHODS: In this study, after RNA extraction and reverse transcription and Real Time RT- PCR of IBDVs, high melting resolution at temperatures ranging from 81 to 92°C were performed. RESULTS: The findings of this study showed that in the high resolution melting curve analysis, the viruses were classified from A to D. Three vaccine strains of D78, Gumbokal, Bursa CE; IBD L; Bursine 2; and all field viruses were placed in groups A, B, C, and D, respectively. High resolution melting curve analysis after normalization also showed all viruses of this study were placed in 4 HRM genotypic group. Three strains, D78, Gumbokal, Bursa CE, produced similar and non-differentiable curve but were different from other vaccine and field strains. Two other vaccine strains, IBD L and Bursine 2, were different from each other and other viruses. CONCLUSIONS: We concluded that the real-time RT-PCR HRM technique is cost-effective and reliable among the currently used methods and can be used for differentiation of IBDV isolates.
Afficher plus [+] Moins [-]A survey on prevalence and molecular characteristics of Linguatula serrata isolated from slaughtered sheep and goat in Yazd slaughterhouse
2017
Eslami, Gilda | Khalatbari-limaki, Sepideh | Oryan, Ahmad | Zohortabar, Amin | Amiri, Asieh | Hajimohammadi, Bahador
Background: Linguatula serrata is a zoonotic parasite causing Halazoun syndrome in humans. Consumption of raw or semi-cooked infected edible offal induces the infection in human. Objectives: The main objective of this study was to investigate the outbreak and molecular characterization of Linguatula serrata in sheep and goat of Yazd slaughterhouse. Methods: To determine the prevalence and severity of Linguatula serrata, mesenteric lymph nodes of 200 slaughtered sheep and 200 slaughtered goats in the Yazd industrial slaughterhouse were examined. DNA extraction was performed using commercially DNA extraction kit as manufacturers’ protocol. In order to genetic evaluation, the partially 18srRNA gene as a target was amplified using the specific primer pair which was designed by Primer3 software.The PCR product sent for sequencing and the sequence was BLAST. Data were then analyzed using SPSS version 16.0 and by the Pearson correlation test and χ2 at a significance level of 0.01.Results: In the present study, prevalence of the infection of slaughtered goats and sheep was 25.5% and 22.5%, respectively. No statistically significant difference was observed between the prevalence of this parasite in different ages and sexes groups (goats and sheep). The results of genetic evaluation showed no variation between this isolate in comparison with the ones in GenBank. Conclusions: This study was the first report of molecular identification of Linguatula serrate in Iran. Considering high prevalence of infection in domestic animal and lack of knowledge and hygienic practice of the people about consumption of animal offal infection of the people to Linguatula serrata is probable. Therefore, in this context, using appropriate and reliable diagnostic methods for detection of infection in abattoirs as well as educating people on the proper use of animal offal is effective steps to prevent this disease.
Afficher plus [+] Moins [-]Molecular identification and antifungal susceptibility patterns of dermatophytes isolated from companion animals with clinical symptoms of dermatophytosis
2021
Katiraee, Farzad | Kouchak Kosari, Yasaman | Soltani, Minoo | Shokri, Hojjatollah | Hassan Minooieanhaghighi, Mohammad
Dermatophytosis is a common skin disease in cats and dogs caused by Microsporum and Trichophyton fungi. Species identification and knowledge of their antifungal susceptibility are therapeutically and epidemiologically important. This study assessed the prevalence of feline and canine dermatophytosis in Iran, identified the aetiological agents molecularly and tested their antifungal susceptibility. A total of 308 companion animals (134 dogs and 174 cats) with skin lesions were examined from March 2015 to March 2018. Hair and skin samples were examined by microscopy with 20% KOH and cultured on Sabouraud dextrose agar with cycloheximide and chloramphenicol. Fungal isolates were confirmed by sequencing of the internal transcribed spacer (ITS) r-DNA region. The antifungal susceptibility of dermatophytes was tested by broth microdilution assay using standard drugs. Dermatophytes were found in 130 (42.2%) samples, 62 of them feline and 68 canine. Based on sequencing of all strains, M. canis (78.5%, P<0.05), M. gypseum (10.7%), and T. mentagrophytes (10.7%) were the dermatophytes isolated. The non-dermatophyte species Nannizziopsis vriesii was also isolated from two feline dermatomycosis cases. Dogs and cats younger than one year (61.5%) showed a statistically significantly higher prevalence of infection (P<0.05). Caspofungin produced the lowest geometric mean MIC at 0.0018 μg/mL, followed by ketoconazole, terbinafine, itraconazole, miconazole, griseofulvin, clotrimazole and fluconazole, in a 0.038–1.53 μg/mL range. This is the first molecular study to identify the causes of pet dermatophytosis in north-western Iran. ITS-PCR was shown to be a useful and reliable method for the identification of closely related species of dermatophytes in clinical and epidemiological settings. The lowest MIC of caspofungin indicated that this drug was the most potent in vitro.
Afficher plus [+] Moins [-]Dogs’ gastrointestinal parasites and their association with public health in Iran
2017
Kohansal Mohammad Hasan | Fazaeli Asghar | Nourian Abbasali | Haniloo Ali | Kamali Koorosh
Introduction: Dogs harbour zoonotic parasites that cause serious infections in humans, such as visceral larva migrans, ocular larva migrans, cystic echinococcosis, and alveolar echinococcosis. Studies on dogs’ gastrointestinal parasites in different geographical locations are required to increase knowledge of the risk of canine zoonoses in human populations.
Afficher plus [+] Moins [-]Mixed infection zones may be important in the epidemiology of contagious agalactia
2016
Jafarizadeh, Amirreza | Pourbakhsh, Seyed Ali | Tadayon, Keyvan | Jamshidian, Mahmud | Ashtari, Abbas
Introduction: The current study was designed to detect Mycoplasma agalactiae (Ma), Mycoplasma mycoides subsp. capri (Mmc), Mycoplasma capricolum subsp. capricolum (Mcc) and Mycoplasma putrefaciens (Mp) in sheep and goats with clinical signs consistent with contagious agalactia.Material and Methods: A total of 299 samples were collected from 55 mixed herds in Azarbaijan-e-Sharghi province, Iran. Samples were examined using PCR and culture methods.Results: The findings showed that in 40 herds at least one sample was positive by PCR or culture method. Moreover, out of 274 sheep samples, 101 were proved to be positive using the PCR technique and 76 were found positive using the culture method. Out of 25 goat samples, 10 were found positive using PCR and 9 were positive through the culture method. Less than 20% of isolated mycoplasmas were Ma. Ma was detected from almost all studied regions in the province while Mmc, Mcc, and Mp were detected only in a very limited area that was deemed by the research group the mixed infection zone.Conclusion: In vaccination or eradication projects, it would be more economical to focus on mixed infection zones. Further investigation on mixed infection zones could facilitate better understanding of contagious agalactia epidemiology.
Afficher plus [+] Moins [-]Mixed infection zones may be important in the epidemiology of contagious agalactia
2016
Jafarizadeh Amirreza | Pourbakhsh Seyed Ali | Tadayon Keyvan | Jamshidian Mahmud | Ashtari Abbas
Introduction: The current study was designed to detect Mycoplasma agalactiae (Ma), Mycoplasma mycoides subsp. capri (Mmc), Mycoplasma capricolum subsp. capricolum (Mcc) and Mycoplasma putrefaciens (Mp) in sheep and goats with clinical signs consistent with contagious agalactia.
Afficher plus [+] Moins [-]Dogs’ gastrointestinal parasites and their association with public health in Iran
2017
Kohansal, Mohammad Hasan | Fazaeli, Asghar | Nourian, Abbasali | Haniloo, Ali | Kamali, Koorosh
Introduction: Dogs harbour zoonotic parasites that cause serious infections in humans, such as visceral larva migrans, ocular larva migrans, cystic echinococcosis, and alveolar echinococcosis. Studies on dogs’ gastrointestinal parasites in different geographical locations are required to increase knowledge of the risk of canine zoonoses in human populations.Material and Methods: The presence of parasites was examined in 450 faecal samples collected from eight zones of Zanjan province, northwest Iran from June to November 2015. The samples were examined using the sedimentation concentration method and modified Ziehl-Neelsen staining.Results: Gastrointestinal parasites were found in 86 (19.1%) faecal samples. Sarcocystis spp. (7.3%), Taenia/Echinococcus spp. (5.6%), Toxocara spp. (1.8%), and Cystoisospora spp. (1.6%) were the most common parasites observed. The other detected parasites consisted of Dicrocoelium dendriticum (0.7%), Eimeria spp. (0.7%), Cryptosporidium spp. (0.4%), Physaloptera spp. (0.4%), Giardia spp. (1.3%), and Spirocerca lupi (1.3%). The lowest parasite infection rates belonged to Trichuris vulpis and Acanthocephalans (0.2% each).Conclusion: This study provides current information on the infection rates in dog populations in Zanjan Province. Furthermore, the study shows a high prevalence of gastrointestinal parasitic infections, including zoonotic ones and particularly Taenia/Echinococcus spp., potentially transmissible to humans and thus relevant to public health.
Afficher plus [+] Moins [-]Pathological description and immunohistochemical demonstration of ovine abortion associated with Toxoplasma gondii in Iran
2013
Rassouli, M., Ferdowsi University of Mashhad, Mashhad 9177948974, Iran | Razmi, G.R., Ferdowsi University of Mashhad, Mashhad 9177948974, Iran | Movassaghi, A.R., Ferdowsi University of Mashhad, Mashhad 9177948974, Iran | Bassami, M.R., Ferdowsi University of Mashhad, Mashhad 9177948974, Iran | Sami, M., Ferdowsi University of Mashhad, Mashhad 9177948974, Iran
The obligatory intracellular protozoan parasite Toxoplasma gondii is a major world wide cause of infectious ovine abortion. In some different diagnostic techniques that are being used to detect this pathogen in ovine fetuses, immunohistochemistry (IHC) is a very sensitive and expensive one. Histopathology is not truly a specific and sensitive test for Toxoplasma infection but it can be helpful to choose some suspected tissues for IHC. In this study 9.5% of 200 samples (aborted ovine fetuses internal organs such as brain, liver, heart, lung, kidney, spleen) (4.6~14.4% with 95% CI) were positive in IHC with a very good logical agreement among different diagnostic techniques (κ = 0.73, 0.8) and with no significant difference among different fetal age groups (p 0.05).
Afficher plus [+] Moins [-]Molecular surveillance of <i>Theileria ovis, Theileria lestoquardi</i> and <i>Theileria annulata</i> infection in sheep and ixodid ticks in Iran
2013
Gholamreza Razmi | Saeed Yaghfoori
A molecular study was undertaken to detect Theileria ovis, Theileria lestoquardi and Theileria annulatain sheep and tick vectors. Investigation was conducted from 2010 to 2011 in the south of Khorasan Razavi Province, Iran. A total of 150 blood samples were collected from 30 different sheep flocks. In addition, ixodid ticks were sampled from the same flocks. The stained blood smears were microscopically examined for the presence of piroplasms and a semi-nested polymerase chain reaction-restriction (PCR) was used for subsequent molecular speciation. Salivary glands were isolated from the ticks and subsequently analysed by semi-nested PCR. polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to differentiate between T. lestoquardi and T. annulata from PCR-positive samples. Theileria species infection was microscopically detected in 18.6% of blood smears. The presence of T. ovis and T. lestoquardi or T. annulata was detected by semi-nested PCR in 58.6% and 6.6% of blood samples respectively. In total, 169 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 155; 91.7% of the total), followed by Hyalomma anatolicum anatolicum (n = 8; 4.7%) and Hyalomma marginatum turanicum (n = 6; 3.5%). From an organ pooling of 33 ticks, three pools of salivary glands from R. turanicus were positive for Theileria species by semi-nested PCR. Of the three R. turanicus samples testing positive for Theileria species, two (6.1%) were positive for T. ovis and one (3.0%) for T. lestoquardi or T. annulata. Amongst the 11 PCR-positive samples for T. lestoquardi or T. annulata, 10 were positive for T. lestoquardi and one sample was positive for both T. lestoquardi and T. annulata using PCR-RFLP. The results also demonstrated that PCR-RFLP could be used for the detection of T. ovis. Based on the results, it can be concluded that T. ovis has a higher prevalence than T. lestoquardi, and that R. turanicus could be a possible vector for T. ovis and T. lestoquardi. Finally, the PCR-RFLP based on Msp1 restriction enzyme is a simple method for differentiation of Theileria species in sheep and ixodid ticks.
Afficher plus [+] Moins [-]Determination of antibiotic resistance pattern in Aeromonas hydrophila isolated from Reared Oncorhynchus mykiss in Tabriz city, Iran
2019
Jafari-Sales, Abolfazl | Shadi-Dizaji, Azizeh
Aeromonas hydrophila is a Gram negative, positive oxidase, anaerobic, and opportunistic bacteria that, under certain conditions, become a pathogen (in humans and fish). This bacterium causes toxin and host infection in which different antibiotic resistance in isolated strains has been reported in different regions of the world. The aim of this study was to determine the prevalence of this bacterium and its susceptibility to common antibiotics in Tabriz city.50 samples from 5 Reared Oncorhynchus mykiss farms in Tabriz city (For each farm,10 numbers) were randomly assigned to suspected fish to the disease. By using biochemical tests, 14 samples (28%) from 2 Fish farms (40%) were identified as A.hydrophila. Antibiogram for these specimens showed that the bacterium had the highest resistance tovancomycin (100 %) and clindamycin (92.8%) antibiotics, and has the most sensitivity to the antibiotics ofsultrim, tetracycline and oxytetracycline with 71.4%.Considering the different antibiotic resistance pattern in this study and other similar studies,the necessity of examining the pattern of resistance in each region seems necessary.
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