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PCR Based Detection of Shiga Toxin Producing E. coli in Commercial Poultry and Related Environments
2015
Homaira Anzum Himi | Md. Shafiullah Parvej | M. Bahanur Rahman | K M Nasiruddin | Wahedul Karim Ansari | Md. Mostakin Ahamed
Shiga toxin (Stx)-producing E. coli (STEC) is the most important foodborne pathogen which is the causal agent of mild diarrhea, bloody diarrhea, hemolytic-uremic syndrome (HUS) in human. The present study was designed to determine the prevalence and identification of Shiga toxin (Stx)-producing E. coli in poultry, detection of its source of infection in poultry and transmission pattern to human. For this purpose a total of 150 samples (cloacal swab-60, feed -15, water-15 and egg -60) were collected and analyzed in bacteriology laboratory by cultured in different bacteriological media followed by gram’s staining, biochemical tests and Polymerase Chain reaction (PCR). The PCR was performed by targeting 16s rRNA gene and shiga toxin producing gene in E. coli. Out of 150 collected samples, E. coli was found in 81 (54%) samples. Presence of E. coli was 100% in both feed (n=15) and egg (n=60), whereas 10% in cloacal swab (n=6). Water samples were totally free of E. coli. The stx2 gene was detected in all samples whether all samples were negative for stx1 gene. The study revealed that, poultry feed acts as a source of E. coli infection in poultry, which may be transmitted to environment and human via meat or eggs. Antibiotic sensitivity test revealed that isolated bacteria were highly sensitive to Ciprofloxacin.
显示更多 [+] 显示较少 [-]Comparative Detection of Canine Parvovirus by Differential PCR, Conventional PCR and Nested PCR tests: Detecting Antigenic Variants by Differential PCR
2022
Sibel Hasırcıoğlu | Hatice Pelin Aslım
Canine parvovirus (CPV) is a pathogen causing hemorrhagic enteritis in puppies and mainly transmitting via feco-oral route. In this study, stool samples were collected from a total of 35 animals suspected of CPV. The samples were examined by Conventional PCR, Nested PCR and Differential PCR tests. 20 out of 35 dogs (57.1%) were detected positive by conventional PCR, 31 (88.6%) by nested PCR and 30 (85.7%) by differential PCR. CPV 2a was stated as the most common antigenic type, male animals and 0–3-month-olds had a high rate of becoming sick and vaccinated animals might also catch the disease, rarely. Accordingly, it is recommended to focus on studies providing molecular epidemiology surveillance in order to detect the existing subtypes and develop reliable diagnosis and vaccination methods.
显示更多 [+] 显示较少 [-]Detection of Sesame Allergen Traces with Two PCR Assays - The Challenge to Protect Food-Allergic Consumers
2015
Dimitra Panagiotis Houhoula | Vasilios Belsis | Leonidas Georgopoulos | Virginia Giannou | Vasiliki R. Kyrana | John Tsaknis | Vladimiros P. Lougovois | Stamatios Koussissis
The purpose of this study was to investigate the possible presence of sesame in commercial foods normally carrying no warning for the allergen, but which may have been subjected to contamination during processing. One hundred units of widely consumed goods with high potential to contain allergenic substances deriving from nuts were analyzed, using sensitive and capable PCR (C-PCR) and Real Time PCR (RT-PCR) methodologies. Of the products examined, 15 (15.0%) declared the presence of sesame, 36 (36.0%) carried no food allergy label, 44 (44.0%) were marked by the phrase “may contain traces of nuts” and 5 (5.0%) carried the indication “may contain sesame traces”. The sesame-positive products detected using the C-PCR method were 15 (100%), 12 (33.3%), 14 (31.8%) and 3 (60%), respectively. Using the RT-PCR technique, positive results were obtained for 15 (100%), 18 (50.0%), 18 (20.5%) and 5 (100%) samples, respectively. The results indicate that the PCR methods applied are highly sensitive and selective, which makes them suitable for the detection of sesame traces in food samples. In addition, they can be useful for monitoring the effectiveness of cleaning processes in the production units of the food industry.
显示更多 [+] 显示较少 [-]Screening of secondary metabolite biosynthesis genes of marine actinomycetes isolated from Trabzon (Black Sea) sea sediments
2017
Kadriye Özcan
In this study, active secondary metabolite production capacity of actinomycete isolates obtained from Trabzon (Black Sea) sea sediments was investigated by molecular techniques. Totaly 24 actinomycetes were investigated by PCR based on the presence of secondary metabolite biosynthesis genes PKS / NRPS. According to the PCR results, 25 and 58% of actinomycetes obtained from Trabzon sea sediments were found to contain PKS-NRPS and only NRPS gene regions, respectively. When PCR data were evaluated, it was found that the production of the peptide form active secondary metabolite of the isolates by non-ribosomal way was higher than that of the secondary metabolite production by the PKS pathway. In addition, it has been determined that Black Sea marine sediments have high potential for active secondary metabolite production.
显示更多 [+] 显示较少 [-]Molecular Investigation and Phylogenetic Analysis of Ehrlichia canis in Dogs in Siirt, Turkey
2022
Burçak Aslan Çelik | Özgür Yaşar Çelik | Ali Bilgin Yılmaz | Adnan Ayan | Özlem Orunç Kılınç | Ramazan Özdemir | Özge Oktay Ayan
Ehrlichia canis is the primary etiologic agent of canine monocytic ehrlichiosis, a tick-transmitted disease of dogs. The aim of this study is to molecularly investigate the presence of E. canis and to reveal its prevalence in dogs in Siirt province. The animal material of the study is consisted of a total of 82 dogs. A region of the 16S ribosomal RNA gene of E. canis was targeted for PCR amplification. As a result of the conducted Nested-PCR, positivity was detected at the rate of 10.53% (4/38) in male dogs and 13.64% (6/44) in females, and Ehrlichia canis specific bands of size 389 bp were obtained in 10 (12.20%) dogs in total. The phylogenetic tree was constructed with the Maximum Likelihood (MCL) method, The nucleotide sequence was registered in the NCBI GenBank database with access numbers OK331365.1-OK331366. Early detection of the disease by means of hematological, serological, or molecular tests is very important in terms of prognosis. More studies should be performed to determine vector-disease relationships in this region about ticks that vector the disease.
显示更多 [+] 显示较少 [-]A Systematic Detection for Brucellosis at Chronic Stage of Infection in Semen of Sheep and Saanen Goats
2019
Esra Buyukcangaz | Burcu Ustuner | Sevil Erdenlig | Selim Alcay | Huban Gocmen | Berk Toker | Engin Kennerman | Mihriban Ulgen
The study was conducted in a herd (n: 244) in which goats (n: 206) and sheep (n:38) had a history of brucellosis in Bursa which is located in Northwestern of Turkey between the years 2012-2014. For the detection of Brucella spp. and the other zoonotic bacterial agents, semen samples were taken from Saanen goats (n: 35) and rams (n: 8). Samples were tested by routine diagnostic procedures and PCR. The serum samples of male animals were also tested for Brucellosis by C-ELISA and I-ELISA. The culture results represented Trueperella pyogenes (n:2), Pasteurella pneumotopica (n: 5), Esherichia coli (n: 3), Aeromonas salmonicida subs. Salmonicida (1), Brevundimonas vesicularis (n: 2) and Mycoplasma bovigenitalium (n: 1) and Mycoplasma arginini (n: 1) from semen samples. Rams had no symptoms due to epididymitis or epididymoorchitis in clinical examination, but two bucks showed orchitis and they were serologically positive for brucellosis. Also, one seronegative buck showed epididymitis in a flock. There were no statistically significant differences between the serologically positive and negative animals in an examination of semen samples in terms of their volume, concentration, mass activity, motility and defectivity rate for acrosome. Although 20 of the serum samples were negative for anti-Brucella antibody, 23 of them were serologically positive for brucellosis. As a result of this study, Brucellae were not detected by bacteriologically and molecularly while there were some positive serum samples for brucellosis. This could be attributed that these samples might have been collected from chronically infected animals in which animals generally do not shed the organisms. Therefore, it was thought that sampling with regular intervals might help for the definitive incidence of brucellosis.
显示更多 [+] 显示较少 [-]Isolation and Characterization of Salmonella Enterica Serovar Typhimurium Circulating Among Healthy Chickens of Bangladesh
2016
Md. Shafiullah Parvej | Marzia Rahman | Md. Forhad Uddin | KHM Nazmul Hussain Nazir | Md. Sayduzzaman Jowel | Md. Ferdousur Rahman Khan | Md. Bahanur Rahman
Salmonella is considered as a global problem ranking first among food borne diseases. All motile Salmonella of poultry origin are zoonotic and readily transmit to human via meat and eggs but reports on non - typhoidal Salmonella serovars circulating in layer chickens is very limited in South-East Asian countries including Bangladesh. Salmonella serovars isolated from apparently healthy chickens were characterized in the present study. Of 170 samples (cloacal swab 150 and feed 20) collected from commercial layer farms, motile Salmonella was isolated 4% (6/150) and 50% (10/20) respectively by cultural, biochemical, motility test and by detection of hisJ gene. About 5% (8/170) samples possessed serovar-specific gene fimA, suggesting that isolates were Salmonella enterica serovar Typhimurium. Antimicrobial susceptibility testing demonstrated that the isolated serovars were multidrug resistant. Therefore apparently healthy layer chickens harbour and transmit S. Typhimurium to the environment, although little is alarming since it has zoonotic significance and the isolates were resistant to commonly used first line of antibiotic in Salmonella infection.
显示更多 [+] 显示较少 [-]Development of a PCR/RLB Test for Meat Source Authentication in Processed Meat and Meat Products
2015
Ibrahim Abbasi | Al-Shareef Abdel-Kareem | Matouk Imad | Akkawi Muataz
Concern over food authenticity has increased as a result of an increase in the consumption of processed foods containing meat or animal products. This raises a number of issues where the presence of pork in such foods is considered unacceptable in most Muslim and Jewish communities around the world. It also applied to the prohibition of beef consumption among Hindus. In order to ensure the absence of unwished meat products or mixing of meats from different sources in processed foods, a specific and sensitive test is essential. For this purpose we developed a molecular test based on DNA amplification by polymerase chain reaction (PCR) of the cytochrome b gene followed by reverse line blot analysis (RLB). Using this method many samples may be treated simultaneously and meat origins can easily be detected from processed foods or foods containing mixed meat sources; also, added pork components such as fat may be identified by this methodology. The PCR/RLB method is considered to be a sensitive and specific technique; it can detect one nucleotide change within the PCR-amplified DNA segment.
显示更多 [+] 显示较少 [-]Molecular Survey of Toxoplasma gondii Infection in Aborted Fetuses of Sheep in the Iğdır Province of Türkiye
2023
Davut Koca | Özlem Orunç Kılınç | Adnan Ayan | Fatma Ertaş Oğuz | Ali Osman Turgut | Özge Oktay Ayan
Toxoplasma gondii, an obligatory intracellular protozoan parasite, can infect a wide range of warm-blooded animals, including livestock species. T. gondii is a zoonotic protozoan parasite that affects both humans and other warm-blooded animals. The aim of this study was to detect T. gondii by using PCR in the brain tissues of 60 aborted sheep fetuses from the Iğdır Province in Türkiye. For this purpose, 60 brain tissue samples of sheep were collected within the lambing seasons of 2023 in Iğdır, Türkiye. The DNA extraction was performed using the PureLink™ Genomic DNA Mini Kit from brain samples. The PCR was performed with the appropriate primers from the obtained DNA samples. T. gondii was found in the brain (16.6%) samples of aborted sheep fetuses. According to the present study, T. gondii infection can be one of the causes of fetus abortion of sheep in Iğdır province, Türkiye. This result emphasizes the need for vigilance and preventive measures in managing this potential public and animal health concerns.
显示更多 [+] 显示较少 [-]Molecular Survey of the Toxoplasma gondii and Neospora caninum in brain tissue of aborted fetuses of Morkaraman sheep in Muş, Türkiye
2023
Davut Koca | Burçak Aslan Çelik | Özgür Yaşar Çelik | Adnan Ayan | Özlem Orunç Kılınç | Ali Osman Turgut | Özge Oktay Ayan
Toxoplasma gondii and Neospora caninum are obligate intracellular protozoan parasites that can affect different warm-blooded species worldwide. In this study, it was aimed to detect T. gondii and N. caninum using PCR method in brain tissues of aborted sheep fetuses. Brain specimens were collected from 50 Morkaraman sheep fetuses that had undergone abortion at various stages of pregnancy, within the lambing seasons of 2023 in Muş. Approximately 1 cm³ of brain tissue from the right cerebral hemisphere was excised and subsequently frozen at -20°C for DNA extraction. DNA extraction and PCR amplification were then performed. As a result of this study, 11 (22%) of 50 brain tissues were positive. All brain samples examined in this study were negative for Neospora caninum. Based on the results of this study, it is possible to say that T. gondii is an important abortion agent in sheep in this region. Although N. caninum was not detected in this study, larger scale studies are recommended. Moreover, this study provides important information to breeders and veterinarians in the field in the evaluation and management of abortion.
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