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Open Access Article
1 - Comparison of PCR and histopathology methods for diagnosis of Yersiniosis infection (yersinia ruckeri) in Rainbow trout of Iran
عادل Haghighi Khiabanian Asl M.R Roozbahani بهرام KazemiYersinia ruckeri is the causative agent of Yersiniosis or Enteric Red mouth Disease that causeshuge amount of economic loss in mariculture industry worldwide. Definitive detection andeffective treatment for septicemia like Y. ruckeri that outbreak critically, requires r MoreYersinia ruckeri is the causative agent of Yersiniosis or Enteric Red mouth Disease that causeshuge amount of economic loss in mariculture industry worldwide. Definitive detection andeffective treatment for septicemia like Y. ruckeri that outbreak critically, requires rapid andspecific diagnosis. Despite the advantages of both PCR and Histopathology techniques in theterms of diagnosis and monitoring they have some defects, the aim of this study is thecomparison of PCR and histopathology methods for detection of Yersiniosis in trout fish inIran.In PCR-based detection, Extracted DNA from suspected Rainbow trout tissue utilized inPCR reaction and polymerase chain products were visualized by gel electrophoresis. Inpathological detection, all tissues from live or moribund fish were fixed in 10% formalin saline.Then Samples were embedded in paraffin block and sectioned with digital microtome at 5-7micrometer thickness. Slides were stained by haematoxillin & eosin, and then treated withmounting media. Differences in the number of positive samples in this comparison demonstratedthat each mentioned technique has its advantages so if the advantages of each method exploitand one method supplements the other one, the efficiency and accuracy of monitoring theexperiments would boost. In both mentioned methods, all positive results confirmed theexistence of yersiniosis infection in Iran, which their comparison is very useful for monitoringand diagnosis of pathogen in the country. Differences in the number of positive samples in thiscomparison demonstrate that although PCR technique would provide a more rapid and sensitivealternative to traditional diagnosis in detection of harmful pathogens but in some cases such asnecrotic and degenerated cells in tissue, PCR ability collapses in the comparison of otherdetection technique like histopathology. In this study, 20 suspected samples were tested by bothtechniques that 2 samples were positive and 18 samples were negative in PCR, and 5 sampleswere positive and 15 samples were negative in pathologic technique . Manuscript profile -
Open Access Article
2 - Distribution of staphylococcal enterotoxin A gene among Staphylococcus aureus isolates from traditional white–brined cheese
Kh Mohammadi,The consumption of food containing staphylococcal enterotoxins is regularly identified as thecause of intoxication. Enterotoxin A is considered as the most common toxin in staphylococcus–related food poisoning. The purpose of this study was to determine the preval MoreThe consumption of food containing staphylococcal enterotoxins is regularly identified as thecause of intoxication. Enterotoxin A is considered as the most common toxin in staphylococcus–related food poisoning. The purpose of this study was to determine the prevalence of S. aureusin traditional white–brined cheese and distribution of enterotoxin A gene (sea) among them. Atotal of 120 samples was examined and S. aureus was isolated from 11 (9.1%) of the samples.According to the results, load of S. aureus was estimated from 1.5×101 to 8.6×104 cfu/g. Nosample was in the critical cell density of >105 cfu/g. From each sample, five suspected colonieswere confirmed by biochemical tests. S. aureus isolates were further identified based on 23SrRNA, themonuclease and enterotoxin A genes using multiplex PCR. Based on multiplex PCRresults all 55 isolates were identified as S. aureus. The enterotoxin A gene (sea) was detected in6 (10.9%) of the isolates. In conclusion, S. aureus and sea gene was found in traditional white–brined cheese. It seems that if the favorable growth conditions are provided, S. aureus couldproliferate and produce enterotoxin and could be regarded as a potential risk for human health. Manuscript profile -
Open Access Article
3 - .Molecular identification of escherichia coli pathotypes EAEC and EPEC strains isolated from dairy cows with mastitis by Multiplex- PCR and determination of antibiotic resistance by disk diffusion method and E-test
Soleimanifard, N., Amini, K. . E.coli as normal intestinal tract flora of animals and humans are harmless bacteria. Although most strain of E.coli are not pathogenic, but some strains can cause a variety of enteritidis and non-enteritidis diseases. Antibiotic resistance microorganisms are imp More E.coli as normal intestinal tract flora of animals and humans are harmless bacteria. Although most strain of E.coli are not pathogenic, but some strains can cause a variety of enteritidis and non-enteritidis diseases. Antibiotic resistance microorganisms are important in treatment of infectious diseases. The aim of this study was to investigate the frequency of genes pathotypes EAEC, EPEC and antibiotic resistance Escherichia coli isolates from animal specimens.Collected 50 samples had undergone various biochemical and microbiological tests to identification or confirmation of microorganisms. Then antibiotic susceptibility tests were performed by disk diffusion method according to CLSI guidelines. E-test was performed with different antibiotics groups. Multiplex PCR assay was used to identify genes pathotypes.All of the clinically isolated E.coli was susceptible to erythromycin (100%), resistance to ampicillin (93%), sensitive to amikacin (100%) and susceptible to nitrofurantoin (96%). Many strains were multidrug resistance. The results of 50 samples of cattle milk Multiplex PCR have shown four samples (8%) carried gene bfPA (pathotypes EPEC).The results of the M-PCR in this study were inconsistent with the results of other study has done in different countries. This Inconsistency may raise due the difference source of isolation site thus in our study isolation source mastitis milk samples were examined, but in other research stool was the source. The difference in samples isolated from milk can be due to differences in geographical areas. Manuscript profile -
Open Access Article
4 - Survey on prevalence of main serogroups and F5 fimbria gene in Enterotoxigenic Escherichia coli isolates from diarrheic calves under five days in Alborz and Qazvin provinces
Gharabaghi, A., Lotfollahzadeh, S., Mokhber Dezfouli, M.R., Moosakhni, F., Yadegari, Z., Nikbakht Brojeni, G.R. .Escherichia coli is the natural inhabitant of gastrointestinal tract of animals and also human beings. However, some pathogenic strains of E. coli cause variety of intestinal and extra intestinal diseases. Enterotoxigenic E. coli (ETEC) is the most common cause of diarr MoreEscherichia coli is the natural inhabitant of gastrointestinal tract of animals and also human beings. However, some pathogenic strains of E. coli cause variety of intestinal and extra intestinal diseases. Enterotoxigenic E. coli (ETEC) is the most common cause of diarrhea syndrome in neonatal farm animals and thus is one of the significant causes of economic losses in herds. Fimbriae are the most important factor associated with pathogenicity of ETEC which carry adhesions and work as secretory enterotoxin. The most common fimbriae in calves are F5 (K99) and F41. K99 isolates most commonly belong to serogroups O8, O9 and O101. The objective of this study was to investigate the frequency of genes associated with different serogroups (O8, O9, O101, etc) and K99 fimbriae in fecal samples obtained from calves under 5 days of age. Samples were cultured on MacConkey’s agar, then from each palate, three colonies were cultured separately. Strains were screened for K99 genes and then for O101 by PCR assay, if negative results were obtained, Multiplex PCR was performed to identify genes associated with serogroups O8, O9 and O115. In this study, the frequency of genes associated with O8, O9, O101, O115 serogroups and K99 fimbriae in diarrheic calves were 18.7%, 9.3%, 56.3%, 7.8% and 9.3%, respectively. K99 fimbriae occurred at higher frequencies in O101 serogroup and were detected in 83.3% of those strains. Manuscript profile -
Open Access Article
5 - Registration and Identification of Toxic S. aureus genes Isolated from Tilapia Fish Using Multiplex PCR Technique
N. Niakhalili H. Ahari B. Nowruzi Introduction: Aquatic animal products are considered very important food items in the food basket regarding their high calorie, protein, and omega-3 unsaturated fat. Many aquatic animals, including fish, may always be infected with pathogenic microorganisms at var More Introduction: Aquatic animal products are considered very important food items in the food basket regarding their high calorie, protein, and omega-3 unsaturated fat. Many aquatic animals, including fish, may always be infected with pathogenic microorganisms at various stages, from hunting to purchasing. Eventually, these factors would cause severe poisoning in humans. The present study investigates the registration and identification of Staphylococcus aureus’s toxic genes isolated from tilapia using the Multiplex PCR technique.Materials and Methods: The sample size included 42 subjects (21 fresh fish samples and 21 frozen fish samples). First, after preparing the samples, Staphylococcus aureus was isolated and examined as surface culture in Baird-Parker medium. Next, a Coagulase test was performed, and the enterotoxin gene was identified by DNA extraction. Finally, gene sequencing was carried out automatically and systematically.Results: The results suggested that 92.9% of the total samples (21 fresh tilapia and 18 frozen fish) had no Staphylococcus aureus, and 7.1% of the samples were infected with Staphylococcus aureus. The coagulase test results also suggested that all three frozen fish samples were coagulase positive. Examining enterotoxin-producing genes (SEA, SEB, SEC, SED, SEE, and SEG) among three frozen samples infected with Staphylococcus bacteria revealed SEA enterotoxin gene only in one frozen sample. Conclusion: The results showed that due to the low cost and much shorter time required to identify toxic Staphylococcus aureus genes using Multiplex PCR, this method is highly effective in studying the genotypes of Staphylococcus isolates. Therefore, by using this method, one can improve the food health level in the society. Manuscript profile -
Open Access Article
6 - تفکیک و تایپینگ مولکولی گونه های بروسلا جداسازی شده از سقط جنین دام های اهلی در شمال و شرق ایران
علی نعمتی غلامرضا هاشمی تبار مهرناز راد -
Open Access Article
7 - Molecular characterization of quinolone resistance genes (qnr) in Salmonella typhimurium isolated from food samples
K. Amini T. Ghaseminezhad Raeini B. KheirkhahSalmonellosis is an important disease in animals and human which is caused by different serovars of Salmonella ‎enterica. Serovars Typhimurium is one of the most prevalent sorvars in humans. Quinolones and floroquinolones ‎are the family of extended-spectrum ant MoreSalmonellosis is an important disease in animals and human which is caused by different serovars of Salmonella ‎enterica. Serovars Typhimurium is one of the most prevalent sorvars in humans. Quinolones and floroquinolones ‎are the family of extended-spectrum antibiotics which are used in salmonellosis treatment. Qnr genes are the ‎plasmid-mediated quinolones resistance which leads to resistance in Enterobacteiacea. The aim of this study is ‎identification of quinolone resistance genes qnr in Salmonella Typhimurium isolated from food samples. In this ‎study, 60 Salmonella samples isolated from food was collected and confirmed by culture and biochemical tests. ‎Serotyping was done by O and H antisera. Multiplex-PCR‏ ‏‎ was performed to identify qnrA, qnrB and qnrS genes. ‎All of the 60 isolates were confirmed as Salmonella by culture and biochemical tests. The results of serotyping ‎showed all the 60 isolates were belonged to serogroup B and serovar Typhimurium. Multiplex-PCR test showed 5 ‎samples had the qnrB, 4 had the qnrS and 1 harboured qnrA gene. The results of this study shows the presence of ‎qnr genes in Salmonella Typhimurium isolated from food samples which has a specific public health importance. ‎Therefore, there should be sureveillance and montoring programs to prevent this quinolone resistance.‎ Manuscript profile -
Open Access Article
8 - Prevalence of six genes encoding enterotoxins production of Staphylococcus aureus isolated from white-brined-cheese by multiplex PCR
سامان MahdaviIsolation, identification and grouping of Staphylococcal enterotoxins in milk is of great importance since these toxins are considered as a major potential source of human illness. In this study, 22 coagulasepositive strains of Staphylococcus aureus isolated from white MoreIsolation, identification and grouping of Staphylococcal enterotoxins in milk is of great importance since these toxins are considered as a major potential source of human illness. In this study, 22 coagulasepositive strains of Staphylococcus aureus isolated from white brine cheese produced in rural areas of Maragheh, were assessed for the existence of six genes encoding enterotoxin synthesis using multiplex PCR. For this, extracted DNA was subjected to multiplex PCR for the presence of enterotoxin a, b, g, h, i and j genes. Amongst seg was the most frequent (8 strains) gene; meanwhile seb gene was not detected in any of the isolates. According to the results, 9 strains (40.9%) harbored the genes encoding enterotoxin production. Four strains (18.18%) contained more than one enterotoxin gene. The results of this study indicated that most of Staphylococcus aureus isolates in traditional cheeses have the potency to produce enterotoxin. Manuscript profile -
Open Access Article
9 - Contamination of Fresh Beef to Salmonella typhimurium and Salmonella enteritidis in Sanandaj during 2012
هیوا Karimi Darehabi فرزین Esmailneshad کیوان Ebrahimi mohammadi Salmonella infection is among the main food-borne gastrointestinal disease. Meat has been recognized as a major source of human illness caused by Salmonella serovars. The presence of Salmonella was detected in 60 samples of fresh beef from retailsof Sananda More Salmonella infection is among the main food-borne gastrointestinal disease. Meat has been recognized as a major source of human illness caused by Salmonella serovars. The presence of Salmonella was detected in 60 samples of fresh beef from retailsof Sanandaj. The presence of Salmonella was assessed by conventional culture method and confirmed by PCR assay. To confirm the identification of isolated colonies as Salmonella spp. and determining serovars as typhimuriumand enteritidisserovars, a multiplex PCR assay, using three pairs of primers were employed. S141 and S139 for InvAgene, specific for the genus of Salmonella, Fli15 and Tym for FliCgene, specific for typhimurium serovar and Prot6e-5 and Prot6e-6 for Prot6E gene, specific for Enteritidisserovar.12 samples 20% were determined as contaminated with Salmonella sppwith microbial culture method but with PCR method only seven samples 11.66% were confirmed. 4 samples (6.6%) of isolated colonies were confirmed as SalmonellaTyphimuriumand any number of isolated colonies were confirmed as Salmonella Enteritidis , the other isolated colonis were belong to other salmonella serovars. This study showed a relatively highprevalence of salmonella in fresh beef from Sanandaj. Manuscript profile -
Open Access Article
10 - Prevalence of genes encoding the classical enterotoxins of Staphylococcus aureus isolated from buffalo milk in Tabriz area by multipex PCR
مهرداد Esnaashari جلال Shayegh آیت الله Nasrollahi Omran Isolation, identification and grouping of staphylococcal enterotoxins in milk is of great importance, since these toxins are considered as a major potential source of humans illness. In this study, 75 staphylococcus aureus strains isolated from buffalo milk More Isolation, identification and grouping of staphylococcal enterotoxins in milk is of great importance, since these toxins are considered as a major potential source of humans illness. In this study, 75 staphylococcus aureus strains isolated from buffalo milk were assessed. For this, extracted DNA was analysed by multiplex-PCR for the presence and diversity of genes encoding the common classical staphylococcal enterotoxins. Results revealed that one of the isolates haboured both sec and seb genes; meanwhile, three other isolates contained merely sec gene. Moreover, the gene encoding the production of enterotoxin-A was not detected in any of the isolates. It was concluded that the diversity of genes encoding the classical enterotoxins of S. aureus isolated from Tabriz area was low. Manuscript profile -
Open Access Article
11 - Isolation of Salmonella from Iranian broiler breeder farms and feed
Mansour Mayahi Forough Talazade Ramezan Ali Jafari Vahid Keshavarz ZamanianContamination of poultry by salmonella spp. is an important issue both in the field of public health as well as in the poultry industry and poultry have a significant role in transmission and incidence of human salmonellosis. The aim of the present study was isolation a MoreContamination of poultry by salmonella spp. is an important issue both in the field of public health as well as in the poultry industry and poultry have a significant role in transmission and incidence of human salmonellosis. The aim of the present study was isolation and identification of Salmonella spp. in Iranian broiler breeder farms and their feed. Samples from Sixty two broiler breeder farms and their feed from 21 states of Iran were collected during one year. All samples were cultured in different conventional media, including pre-enrichment, enrichment, selective plating and 18 Salmonella isolates were identified. Salmonella identification was confirmed by multiplex PCR and 12 isolates were confirmed. Out of positive samples, seven samples (58.33%) were Salmonella enteritidis in Ghazvin, Mazandaran and Markazi provinces, three samples (25%) were Salmonella infantis in Kordestan and southern Khorasan provinces, and two samples (16.6%) were Salmonella typhimurium in Fars and Lorestan provinces. All feed samples were negative. The results of this study showed that some breeder farms in Iran are contaminated with Salmonella and the most prominent Salmonella inbroiler breeder farms are Salmonella enteritidis, Salmonella infantis, and Salmonella typhimurium respectively. Manuscript profile -
Open Access Article
12 - Evaluation of virulence and enterotoxin genes in Salmonella enteritidis strains isolated from Meat and Egg samples by Multiplex-PCR
Toktam Khodadadipour Kumarss Amini Razagh MahmoudiToday,food borne diseases are one of the most serious problems and occur as a result of consumption to contaminated food and water. The aim of the study was evaluation of virulence and enterotoxin genes in Salmonella enteritidis strains isolated from food samples by mul MoreToday,food borne diseases are one of the most serious problems and occur as a result of consumption to contaminated food and water. The aim of the study was evaluation of virulence and enterotoxin genes in Salmonella enteritidis strains isolated from food samples by multiplex-PCR. This descriptive, cross-sectional study was performed at 2014 on the 1250 no duplicative and non-repetitive food samples. M-PCR assay was done in order to detection of Stn،sopB،slyA،spvc and Phop/Q genes. Sixty Salmonella enteritidis strains were obtained from poultry meat (35 strains, 58.3%) and eggs (25 strains, 41.6%), respectively. molecular analysis distribution showed all isolates (100%) were absence for spvC gene. The highest and lowest prevalence of the genes were related to Phop/Q and SopB,33.3% and 1.6%, respectively. Evaluation of virulence genes and enterotoxin in the Salmonella enteritidis isolated from the food samples are useful because of presence of the genes and efficacy of M-PCR method in epidemiological investigation and assessment of intraspecies genes transfer in food samples. Manuscript profile -
Open Access Article
13 - Utilization of a 17 Microsatellites Set For Bovine Traceability in Czech Cattle Populations
L. Putnova I. Vrtkova P. Srubarova L. Stehlik -
Open Access Article
14 - تعیین تیپهای ایزولههای Clostridium perfringens به روش Multiplex PCR در شتر مرغ
E. Zandi M.R. Mohammadabadi M. Ezzatkhah A.K. Esmailizadehکلستریدیوم پرفرینجنس یک پاتوژن مهم است که بیماریهای گوناگون زیادی را ایجاد میکند. این باکتری دارای 5 تیپ مختلف است که هر تیپ قادر به ایجاد بیماری مشخصی میباشد. روشهای مختلفی برای شناسایی باکتریها وجود دارند که بیشتر آنها مشکل، زمان بر، پرهزینه و دارای حساسیت و اختصاصیت Moreکلستریدیوم پرفرینجنس یک پاتوژن مهم است که بیماریهای گوناگون زیادی را ایجاد میکند. این باکتری دارای 5 تیپ مختلف است که هر تیپ قادر به ایجاد بیماری مشخصی میباشد. روشهای مختلفی برای شناسایی باکتریها وجود دارند که بیشتر آنها مشکل، زمان بر، پرهزینه و دارای حساسیت و اختصاصیت پایینی هستند. هدف این پژوهش تعیین انواع نامشابه کلستریدیوم پرفرینجنس با روش PCR بود. در این مطالعه، نمونههای مدفوع 120 شترمرغ به صورت تصادفی از نواحی اطراف شهر کرمان واقع در جنوب شرقی ایران جمعآوری شدند. بعد از عملآوری و کشت نمونهها، کلونیهای تولید شده از نظر مورفولوژیکی مطالعه شدند، تست رنگ آمیزی گرم نیز انجام شد و جنس این باکتریها با استفاده از تستهای بیوشیمیایی تعیین گردید. DNA استخراج شده از باکتریهای جدا شده برای تعیین ژنوتیپ توسط multiplex PCR با آغازگرهای اختصاصی آزمون شد. بر اساس طول قطعات سنتز شده توسط PCR، تیپهای توکسین و استرینهای باکتریایی شناسایی شدند. تیپهای جدا شده کلستریدیوم پرفرینجنس عبارت بودند از: 100 درصد تیپ A، 0 درصد تیپ B، 0 درصد تیپ C و 0 درصد تیپ D. این نکته را میتوان مورد تأکید قرار داد که تا کنون تیپ A کلستریدیوم پرفرینجنس گزارش نشده است. Manuscript profile -
Open Access Article
15 - Used of Multiplex PCR for detection of gram-positive cocci causing farms rainbow trout (Oncorhynchus mykiss), Ilam
seyede saeedeh HeidarinejadIn the past five years, The disease that suspected streptococosis was epidemic in rainbow trout farms in Ilam, In this study, 60 rainbow trout with Clinical signs such as lethargie, erratic swimming, skin darkness and exophthalmia, were collected from rainbow trout farm MoreIn the past five years, The disease that suspected streptococosis was epidemic in rainbow trout farms in Ilam, In this study, 60 rainbow trout with Clinical signs such as lethargie, erratic swimming, skin darkness and exophthalmia, were collected from rainbow trout farms in Ilam in southwest of Iran in January 2010, for detection agent of disease samples were collected from spleen, kidney and liver of fish then cultured on blood agar and incubated at 22 ºC for 24 h. Based on biochemical tests, Lactococcus garvieae was detected and isolates were confirmed as L. garvieae by using a m-PCR targeting lctO-gene, 16S rRNA and 16S- 23S rRNA Internal Transcribed Spacer region(ITS) sequencing that detect L .garvieae, Streptococcus iniae and S. dysgalactiae,. In the present study we observed a single band of 1100 bp for L. garvieae for most of isolates and concluded that the main agent of septicemia was L. garvieae. In conclusion, Multiplex PCR was definite method for rapid detection of L. garvieae in mixed infection in rainbow trout. Manuscript profile -
Open Access Article
16 - Molecular characterization of serotypes and capsular virulence genes in cps gen group of Klebsiella pneumonia isolated from Tehran hospitals
Marzieh Tavakol Hassan MomtazBackground & Objectives: Klebsiella pneumoniae is one of the important clinical pathogens and responsible for some nosocomial infections; especially pneumonia, septicemia, and urinary tract infection (UTI). Study of the genotypic and phenotypic characteristics MoreBackground & Objectives: Klebsiella pneumoniae is one of the important clinical pathogens and responsible for some nosocomial infections; especially pneumonia, septicemia, and urinary tract infection (UTI). Study of the genotypic and phenotypic characteristics of virulence factors associated with pathogenicity of K. pneumoniae clinical isolates is of great importance. The aim of the present study was the molecular identification of serotypes and capsular virulence genes in cps gene group of K. pneumonia isolates. Material & Methods: This cross-sectional study was carried out on 50 K. pneumonia clinical isolates collected from patients admitted to the Payambaran and Baqiyatallah hospitals located in Tehran. Multiplex PCR was used for identification of K1, K2, K5, K54, K57 capsular types, and detection of rmpA and wcaG virulence factors from capsular polysaccharide genes group. Results: In this study, K54 (68%) was the most frequent capsular serotype, while K1 (8%) had the lowest frequency. The highest frequency of virulence gene rmpA was observed in capsular serotype K5 (60%), and the highest frequency of wcaG gene was observed in serotype K54 (17.64%). Conclusion: Multiplex PCR method can be used as a rapid tool to characterize and type the bacterial isolates causing nosocomial infections. Manuscript profile -
Open Access Article
17 - Comparison of Pap smear and Multiplex-PCR methods for detection of Human Papillomavirus isolated from women with cervical lesions
Fateme Zandnia Abbas Doosti Abbas Mokhtari-Farsani Mohammad Chehelgerdi Shahrzad SoleimaniBackground & Objectives: High-risk human papillomaviruses (HPV) is the main cause of cervical cancer in women. Early diagnosis and immediate treatment are very helpful. Detection of this infection by traditional methods, such as pap smear, are very inadequate. This MoreBackground & Objectives: High-risk human papillomaviruses (HPV) is the main cause of cervical cancer in women. Early diagnosis and immediate treatment are very helpful. Detection of this infection by traditional methods, such as pap smear, are very inadequate. This study was aimed to evaluate the HPV infection in women with cervical wounds using Multiplex PCR, and to compare the efficiency of this methods with pap smear. Materials & Methods: This cross-sectional study was carried out on 50 samples from patient with cervical wounds attended to Khanevadeh Hospital in Isfahan. After extraction and purification of DNA, 5 types of HPV viruses were identified using Multiplex PCR. At the end, the results of these technique were compared with the results obtained from Pop smear. Results: Our results showed that 41 samples were positive to different types of HPV infection. All of these strains were detected using Multiplex PCR method. However, only 14 cases were reported positive by pap smear technique. Conclusion: According to the results, pap smear has high error rate in HPV detection. But Multiplex PCR can provide a rapid, sensitive and affordable method for detection and screening of the viral infection. Manuscript profile -
Open Access Article
18 - Evaluation of virulance genes of Shiga toxin producing Escherichia coli from Juice Purchase and vegetables in Shiraz
Marjan Shah Illi Mohammad Kargar Abbasali Rezaeian Maryam Homayoon Mehdi Kargar Sadegh Ghorbani-DaliniBackground and Objective: Shiga Toxin-Producing Escherichia coli (STEC) strains can infect humans via vegetal and animal food consumption and cause food poisoning with diarrhoeas, hemorrhagic colitis, haemolytic uraemic syndrome that can lead to death. The purpose of th MoreBackground and Objective: Shiga Toxin-Producing Escherichia coli (STEC) strains can infect humans via vegetal and animal food consumption and cause food poisoning with diarrhoeas, hemorrhagic colitis, haemolytic uraemic syndrome that can lead to death. The purpose of this study is isolation and identification of Escherichia coli O157:H7 virulence genes from Juice Purchase and vegetables in Shiraz. Material and Methods: This cross- sectional study was performed on 89 samples of Juice Purchase and vegetables collected from Shiraz. Isolates were enriched in ECB with novobiocin medium in temperature37°C. After, in order to examine the fermentation of sorbitol and lactose the CT-SMAC and VRBA media and the activities of β-glucoronidase of separated bacteria were examined using the choromoagar medium. Then, the existence of E.coli O157:H7 was confirmed with the spesific antiserum. Finally, virulence genes stx1, stx2, eae and hly were tested by multiplex PCR and rfb O157 and fliC H7 were tested by single PCR. Results: Out of all examined samples 69 (77.53%) sorbitol negative bacteria were separated that after evaluation with specific antiserum 21 (23.60%) E.coli O157:H7 was detected. The molecular markers, stx1 genes in 3 samples and eae , stx1 in 1 sample were detected. Conclusion: Intensive studies of the sources, incidence, fate and transport of E.coli O157 near produce production are required to determine the mechanisms of pre-harvest contamination and potential risks for human. Manuscript profile -
Open Access Article
19 - The comparison between molecular and bacteriological detection for identification of abortion agents caused by Brucella and Salmonella in sheep in Shahrekord town
Ali Sharifzadeh Abbas Doosti Mohsen GaafarianBackground and Objective: Brucella spp and Salmonella abortus ovis are important causes of ovine abortion around the world. Both Bacteria can be serologically diagnosed, but many factors may cause false positive and negative results. Direct methods based on bacteriologi MoreBackground and Objective: Brucella spp and Salmonella abortus ovis are important causes of ovine abortion around the world. Both Bacteria can be serologically diagnosed, but many factors may cause false positive and negative results. Direct methods based on bacteriological isolation are usual, but they are difficult, time consuming and dangerous. Polymerase chain reaction (PCR) have been successfully described for the detection of Brucella spp. and Salmonella abortus ovis. Material and Methods: The detection of these agents in aborted ovine fetuses by multiplex PCR is described. The mPCR was applied to 38 fetal stomach contents. 5(13.1%) samples collected from ovine fetus were Brucella spp. Results: 19 (50%) samples collected were salmonella abortus ovis. 10 (26.3%) samples collected were negative and 4 (10.6%) samples collected were Brucella spp. and Salmonella abortus ovis .in Bacteriological examination 5(13.1%)samples collected from ovine fetus were Brucella spp. 9(23.7%)samples collected were salmonella abortus ovis and 24 (63.2%) samples collected were negative. Conclusion: Simplicity and the possibility of detection of both bactera in a single tube reaction support the use of the mPCR in the routine diagnosis. Manuscript profile -
Open Access Article
20 - Rapid and simultaneous identity of virulence factors and capsular typing of Pasteurella multocida isolated from sheep and goats by Multiplex PCR
Samaneh Danesh Lari Yahya Tahamtan Masoumeh Hayati Mohammad KargarBackground and objective: Pasteurella multocida exist commensally in respiratory track of a variety of wild and domestic animals. Serotypes A and D are responsible for pneumonia in goats and sheep, and Fimbriae, adhesions, capsule, toxin, iron uptake factor and outer me MoreBackground and objective: Pasteurella multocida exist commensally in respiratory track of a variety of wild and domestic animals. Serotypes A and D are responsible for pneumonia in goats and sheep, and Fimbriae, adhesions, capsule, toxin, iron uptake factor and outer membrane protein are its important virulence factors. The aim of this study was to design a multiplex PCR for fast and simultaneous detection of more important virulence factors of P. multocida isolated from sheep and goats as well as their capsular typing and identification. Materials and Methods: Totally 500 nasal swabs were collected from the goats and sheeps suspected to respiratory signs of pasteurellasis. After isolation abd detection of P. multosida by biochemical examination, the bacteria were assayed by specific primers for identification of major important virulence factors and their capsular typing. Result: Capsular typing of the isolates by PCR showed two capsular types A, D with 83.8% and 6.4% prevalence, respectively. The results also showed that 23 (74.19%), 21 (67.74%), 21(67.74%) and 24(77.4%) of the isolates were positive for presence of toxA, hgbA, ptfA and ompH genes, respectively. Conclusion: The most isolated P. multocida from goat and sheep were toxigenic, and capsular type A was the most common isolates in the Fars province. The remarkable high prevalence of toxA and ompH among the afflicted sheep and goats may imply to important role of these genes in epidemiology and virulence of P. multocida. Furthermore the high prevalence of P. multocida typeA harbor toxA gene can be attributed to its important role in the respiratory infection. Manuscript profile -
Open Access Article
21 - Genotyping of BVDV type 1 and 2 isolated from PI cattle in Tehran province by multiplex PCR
فرهاد Moosakhani آریا Badiei مهدی Loghmani علیرضا Shaghayegh محسن ZafariMucosal disease (MD) and bovine viral diarrhea (BVD) are diseases that have no clinical correlations but they have commonviral etiology. In this respect, genotyping of the virus is not performed in Iran yet, after genotyping, control of the disease byusing proper vaccin MoreMucosal disease (MD) and bovine viral diarrhea (BVD) are diseases that have no clinical correlations but they have commonviral etiology. In this respect, genotyping of the virus is not performed in Iran yet, after genotyping, control of the disease byusing proper vaccines could be more effective.In this study, serum samples of suspected calves were taken randomly from 20 farms around province of Tehran. Then, twoELISA tests were performed for detection of BVDV antigen. Twenty samples were chosen for RNA extraction as each samplebelonged to one farm. Then, a multiplex PCR was preformed on the basis of 5′ UTR of BVDV genome with positive samplesfor genotyping the virus.In conclusion, 3 samples (15%) were positive for BVD-2 and 20 samples (100%) were positive for BVD-1. All positive andnegative ELISA samples had equal results in RT-PCR. As for detection of BVD-2, phylogenetic analysis and molecular examination is recommended. Manuscript profile
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