(2015) Phylo-typing of clinical Escherichiacoli isolates originating from bovine mastitis and canine pyometra and urinary tract infection by means of quadruplex PCR, Veterinary Quarterly, 35:4, 194-199, DOI: 10.1080/01652176.2015 Background: Escherichia coli is one of the major causative agents of bovine mastitis worldwide, and is typically associated with acute, clinical mastitis. Besides this, E. coli strains which belong to the extra-intestinal pathogenic group are also the major cause of urinary tract infections and pyometra in dogs. Objectives: In this study, it was aimed to investigate phylo-groups/subgroups in 155 E. coli isolates obtained from acute bovine mastitis, 43 from urinary tract infections of dogs and 20 from canine pyometra by a formerly described triplex PCR and recently described new quadruplex polymerase chain reaction (PCR) method. Results: Group A 1 (n D 118; 76%) and B1 (n D 71; 46%) were found to be the most prevalent groups by triplex and quadruplex PCR assays in mastitis isolates, respectively. Phylo-typing of 43 urinary tract isolates also revealed that most of the isolates belonged to A 1 (n D 23; 54%) by triplex and B2 (n D 36; 84%) by quadruplex PCR assays. The isolates assigned as group A 1 (n D 17; 85%) by triplex PCR could not be classified by quadruplex PCR in pyometra isolates. Conclusions:The results support the hypothesis that E. coli strains isolated from bovine mastitis cases are environmental. Also, groups C, E and F were identified as new phylo-groups for the first time in acute bovine mastitis cases. The comparison of triplex PCR with quadruplex PCR results revealed that most of the groups assigned in triplex PCR were altered by quadruplex PCR assay.

Quinolones have been extensively used for treatment of a variety of invasive and systemic infections of salmonellosis. Widespread use of these agents has been associated with the emergence and dissemination of quinolone-resistant pathogens. The quinolone resistance and plasmid-mediated quinolone resistance determinants (qnrA, qnrB, qnrS and aac(6')-Ib-cr) of 85 Salmonella isolates from chicken carcasses were investigated in this study. Isolates were serotyped according to the Kauffman-White-Le Minor scheme, and broth microdilution method was used to determine quinolone resistance. Plasmid-mediated quinolone resistance genes were investigated by real-time PCR and positive results were confirmed by sequencing. Among the Salmonella isolates, 30/85 (35%) and 18/85 (21%) were found to be resistant to enrofloxacin (MIC ≥ 2 mg/ml), and danofloxacin (MIC ≥ 2 mg/ml), respectively. All the isolates were negative for qnrA, qnrB and aac(6')-Ib-cr genes, nevertheless 2% (S. Brandenburg and S. Dabou) were positive for qnrS (qnrS1 determinant). This study is the first and unique investigating the plasmidmediated quinolone resistance determinants of Salmonella isolated from chicken carcasses in Turkey.

This study examined the incidence of Clostridium perfringens in raw, ready-to-cook (RTC), and ready-to-eat (RTE) meat and meat-based products (N = 306) collected from restaurants, supermarkets, and butcher shops in Bursa, Turkey. In addition, we investigated the presence of the C. perfringens enterotoxin (CPE), as well as cpe genes and their source (chromosomal or plasmid borne). In this study, tryptose sulfite cycloserine (TSC) agar for classic culture isolation and API and real-time polymerase chain reaction (RT-PCR) techniques were used to identify C. perfringens and detect cpa and cpe genes from these products, respectively. Seventeen C. perfringens isolates (5.6%) were isolated and identified with API 20A. In addition, 42 of 81 suspicious isolates (51.9%) were identified as C. perfringens using RT-PCR. Of the 81 suspicious isolates tested by RT-PCR, 22 (27.2%) carried the cpe gene either on the plasmid or chromosome. Twenty-one isolates were positive for chromosomal cpe (C-cpe), and one was positive for plasmid-borne cpe (P-cpe). CPE was detected in 31.8% (7/22) of the cpe positive isolates by the PET-RPLA test. In conclusion, C. perfringens and their CPEs were present in raw, RTC, and RTE meat and meat-based foods in this study. It is emphasized that the presence of C. perfringens and the cpe gene in these foods may be a potential risk for human health.

Birçok ülkede önemli halk sağlığı sorunu olan S. Enteritidis, hayvanlardan ve gıdalardan izole edilen en yaygın Salmonella serotiplerinden biridir. Hayvanların bakteriyel infeksiyonlarının kontrolünde bakteriyofaj kullanımı, son yıllarda üzerinde araştırma yapılan konular arasındadır. Bu çalışmada, ülkemizde kanatlılardan en çok izole edilen Salmonella serotiplerinden biri olan S. Enteritidis (15 adet), bakteriyofajların konak hedef hücreleri olarak kullanıldı. Virulent bakteriyofajların izolasyonu ve zenginleştirilmesi amacıyla overlay-agar yöntemi seçildi. İzole edilen bakteriyofajların hedef bakteri suşları üzerindeki litik aktiviteleri spot test uygulaması ile belirlenerek değerlendirildi. Türkiye'nin farklı bölgelerinde bulunan ticari broyler tesislerinden S. Enteritidis üzerinde litik etki gösteren fajların izolasyonu için alınan toplam 78 adet örnek (tavuk dışkı, kümes atık suları, kümes altlık) bakteriyofaj izolasyonu amacıyla test edildi. Bu örneklerden toplam 22 adet S. Enteritidis'e spesifik fajın purifikasyonu gerçekleştirildi. Yapılan bu çalışmada, Af1-Ka ve Af3-Ka fajları, test edilen 15 adet S. Enteritidis suşunu sırasıyla %78'ini ve %71'ini lize ederek geniş bir konak spektrumu göstermiştir. Bu iki bakteriyofajın ileri faj karakterizasyonu ve tiplendirilmesinin yapılmasıyla, kanatlı endüstrisindeki bakterileri kontrol etmede kullanılacak etkili bakteriyofajın seçilmesi için bir temel oluşturulmasında yardımcı olabileceği kanısına varılmıştır.

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This research was conducted to investigate the extended spectrum beta-lactamase activity and multidrug resistance of Salmonella serovars isolated from chicken carcasses. For this purpose, 99 Salmonella isolates from 930 chicken carcasses were tested against 12 different antimicrobials. The resistance rates of Salmonella isolates to antimicrobials were as follows: 35.3% (35/99) to ampicillin, 33.3% (33/99) to tetracycline, 29.2% to amoxicillin-clavulanic acid, 18.1% (18/99) to nalidixic acid, 17.1% (17/99) to chloramphenicol, 16.1% (16/99) to aztreonam, 12.1% (12/99) to trimethoprim-sulfamethoxazole, 4% (4/99) to gentamicin, 1.0% (1/99) to ceftazidime. Of the isolates 46.4% (46/99) were found to be resistant to two or more antimicrobials as a multidrug resistance. Extended spectrum beta-lactamase activity was detected in 1.0% (1/99) of the isolates. Furthermore, S. Typhimurium 26.2% (28/99), S. Infantis 16.1% (16/99), S. Hadar 12.1% (10/99) and S. Branderburg 9.0% (9/99) were found to be the predominant serovars. In conclusion, antimicrobial resistance and also multidrug resistance rates of Salmonella isolates in this study, indicated that monitoring of antimicrobial resistance profiles is important for Salmonella infections to plan treatment strategies.

Summary:This study aims to determine the presence of Salmonella in naturally contaminated grade A eggs by the standard culture method International Organization for Standardization Method 6579 (ISO) and a specific real-time PCR system (LightCycler PCR-LCPCR) to complement ISO. A total of 1635 eggs pooled into 101 samples were randomly collected within one year period from 20 different retail markets in Bursa, Turkey, carrying eggs of 16 large egg producers/suppliers of 5 cities with intensive layer production. Preparation of the egg and shell for analyses, Salmonella isolations and identifications, and detections were performed according to ISO 6887-4:2003, ISO 6579 and LCPCR, respectively. Overall Salmonella detection rate by ISO and LCPCR were 15.8 % (16/101) and 46.5 % (47/101), respectively. Out of 101 inner parts, Salmonella was detected in 11 (10.9 %) samples by ISO, and in 31 (30.7 %) samples by LCPCR. Six of 101 shell samples (5.9 %) were found to harbor Salmonella by ISO, while 18 (17.8 %) shells were positive by LCPCR. All isolates were determined as Salmonella enterica subsp. enterica serovar Enteritidis. These findings indicate considerably high Salmonella contamination in retail grade A eggs. This should be under routine monitoring by rapid methods such as PCR, complemented by standard culture to evaluate and assess the significance of risk for public health.

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