(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.
In this study, pathogenic mycobacteria were investigated in fecal samples of caged birds by PCR. A total of 47 feces samples collected from 4 different aviaries in Ankara. DNA extraction from fecal samples was performed with a commercial kit using spin column technology. PCR was performed with designed primers respectively amplifying 274 base pairs (bp), 128 bp, 102 bp and 219 bp nucleotide sequences of specific genes (16SrRNA, ISI245, IS901 and hypothetical 21kDa protein gene) of Mycobacterium spp., Mycobacterium avium complex (MAC), Mycobacterium avium subsp. Avium and Mycobacterium genavense, respectively. Five samples were positive and harbored the sequence for the Mycobacterium spp., of 4 of these 5 samples was identified as M. genavense by PCR. As a conclusion of this study, which is the first announcement of the detection of M. genavense DNA in fecal samples of caged birds in Turkey, PCR was seen to be a rapid, sensitive, and a reliable method in detection of avian mycobacteriosis.
Aim To investigate factors influencing Campylobacter spp. colonization of broiler chickens. Methods and Results Campylobacters were isolated from caeca from 319 flocks of two different breeds (199 Cobb and 120 Hubbard), reared as standard (199), Freedom Food/corn fed (57), free‐range (47) or organic (16). The standard category exclusively used Cobb birds slaughtered at 38‐41 days. The Freedom Food/corn‐fed and free‐range Hubbard birds were slaughtered at 49–56 days and the organic flocks at 70 days. Campylobacters were picked at random from direct plates. Both breed of chicken (Hubbard) and age at slaughter were independently associated with increased likelihood of colonization by Campylobacter coli rather than Campylobacter jejuni, but breed could not be separated from other aspects of husbandry with the data available. Conclusions Chickens are frequently colonized by C. jejuni and C. coli and most human infections originate from poultry. In most developed countries approximately 90% of human infections are caused by C. jejuni, but fewer than 10% by C. coli. This might be due to C. coli being less pathogenic than C. jejuni to humans, and/or to chicken meat carrying fewer C. coli than C. jejuni. More investigations are needed into these aspects before it can be concluded that slaughtering older birds from slower‐growing breeds would reduce the risk of human Campylobacter disease. Significance and Impact of the Study Meat from certain breeds of poultry are predominantly colonized by C. coli rather than C. jejuni. More research is needed to understand the impact this may have on the number and severity of human campylobacter infections.
SummaryMycoplasma bovis is one of the most pathogenic agents in the Mycoplasma species that cause disease in cattle. In particular, young calves at less than 4 months of age are a considerable risk from pneumonia caused by M. bovis. In this study, we investigated M. bovis from tracheal swabs and blood sera of cattle which showed respiratory symptoms. A total of 127 tracheal swab samples were collected from seven different farms in Turkey. In addition, a total 254 acute and convelance sera were collected from the same cattle at intervals 15 days. The materials were collected from cattle between 3-12 months of age that reported respiratory problems such as broncho-pneumonia with coughing, depression, lethargy and fever. Mycoplasma bovis was investigated in tracheal swab samples and sera collected from the cattle by using PCR and ELISA respectively. The PCR results showed that M. bovis infections were positive in 4 different farms. The rates ranged from 5.3% (1/19) to 37.5% (6/16). Out of the 127 cattle examined, 45 (35.4%) were positive for M. bovis antibodies, while 82 (64.6%) were found to be negative. All PCR positive cattle were also found to be positive by ELISA. However by using ELISA, M. bovis infections were positive in all farms and the ELISA positive rates ranged from 20% (2/10) to 68.8% (11/16). Considering these results, in especially chronic infections, ELISA is a more useful method than PCR to detect M. bovis infection. Keywords: Cattle, ELISA, Mycoplasma bovis, PCR Sığırlarda Mycoplasma bovis Infeksiyonunun ELISA ve PCR ile Teşhisi ÖzetMycoplasma bovis, Mycoplasma etkenleri içerisinde sığırlarda infeksiyona neden olan en patojen etkenlerden biridir. Özellikle, 4 aylık yaşın altındaki genç buzağılarda, M. bovis'in neden olduğu pnömonilerde artan bir risk bulunmaktadır. Bu çalışmada solunum sistemi infeksiyonu semptomları gösteren sığırların trachea svapları ve kan serumlarından M. bovis infeksiyonunun teşhisi ve M. bovis teşhisi için serolojik ve moleküler metodların karşılaştırılması amaçlanmıştır. Türkiye'de bulunan 7 farklı çiflikten gönderilen 127 tracheal svap örneği ile 15 gün arayla aynı sığırlardan alınan 254 adet akut ve konvelesans serum örneği PCR ve ELISA yöntemleriyle incelendi. Bu örnekler 3-12 aylık yaşlar da olan ve bronkopnömoni, öksürük, depresyon, halsizlik ve ateş gibi solunum sistemi infeksiyonu semptomu gösteren sığırlardan toplandı. Tracheal svap örneklerinin PCR sonuçlarına göre 4 farklı çiftik M. bovis infeksiyonu yönünden pozitif bulundu. Oranlar %5.3 (1/19) ile %37.5 (6/16) arasında bulundu. Mycoplasma bovis antikorları yönünden incelen 127 sığıra ait serumlarda, 45 (%35.4) adeti pozitif olarak saptandı; 82 (%64.6) serum ise negatif olarak saptandı. PCR'da pozitif olarak saptanan tüm sığırlar ELISA yöntemiyle de pozitif olarak saptandı. M. bovis infeksiyonu tüm çiftliklerde pozitif olarak saptandı ve ELISA oranları %20 (2/10) ile %68 (11/16) arasında değişkenlik gösterdi. Bu sonuçlar göz önüne alındığında, özellikle kronik infeksiyonlarda M. bovis infeksiyonunun teşhisinde ...
PurposeThere is currently an emerging need for developing improved approaches for preventing urinary tract infections (UTIs) occurring during diagnostic or interventional procedures of the lower urinary tract. We aimed to establish a rat model to assess the use of transurethral antibiotic administration and to provide evidence that this could be used as a preventive therapy.MethodsAnimals received fosfomycin trometamol (FOF) either urethrally or orally prior to the procedure. A third group was generated as treatment controls and did not receive any medication. Urethral dilation was conducted to recapitulate an interventional procedure prior to intravesical Escherichia coli administration in all three groups. Finally, sham-operated animals were introduced as a fourth group which did not receive antibiotics or E. coli. Colony counts of urine and tissue cultures for the identification of E. coli and histopathological examinations of the bladder and prostate were conducted.ResultsEvaluation of infection intensities in cultures as well as histopathological examination of the bladder and prostate demonstrated a preventative role of transurethral FOF administration. In terms of efficiency, local administration of FOF was similar to oral administration.ConclusionsThese results suggest that transurethral antibiotic administration is a promising alternative for preventing UTIs occurring during diagnostic or interventional procedures of the lower urinary tract.
In this study, virulence factors such as serum resistance, aerobactin iron uptake systems, adhesins (type 1 fimbria, P fimbria, S fimbria, afimbrial adhesins), haemolysine, uropathogenic spesific protein and cytotoxic necrotizing factor in 45 Escherichia coli isolates from dogs and cats with urinary tract and genital system infections, were investigated by both phenotypic methods and molecular methods. In PCR examinations of 45 E. coli, hlyA gene was detected in 73.3% of all isolates. fimH was found in 100%, papC in 71.1%, sfaDE in 82.2%, and afaBC in 0% of the isolates. iucD, traT, usp, cnf1 and cvaC genes were found in 20%, 71.1%, 84.4%, 75.5%, and 11.1% of the isolates, respectively. According to the isolation rate of hlyA and haemolytic activity on blood agar, this virulence factor is commonly seen in these strains and important in these infections especially pathogenesis. İsolates was found to be a high rate positive for usp gene. This result suggested that this strain may have higher infectivity. Cytotoxic necrotizing factor was common and important virulence factor in uropathogenic E. coli and prognosis of infection. However, colisin encoding gene (cvaC) only were detected from 5 of urine samples from dogs. This virulence factor could be important for the urinary tract infections in dogs rather than cats. Haemolysis, type 1 fimbriae, P fimbriae, S fimbriae, cnf, serum resistance, uropathogenic specific protein were detected at high rates by molecular method. In E. coli strains isolated from dogs, virulence factors were found to be higher than cat isolates. Kedi ve köpeklerin ürogenital sistem infeksiyonlarından izole edilen Escherichia coli'lerin virulens faktörlerinin belirlenmesi**
In this study, which was conducted for the first time in Balıkesir, it was purposed to determine the existense, virulence factors and antibiotic susceptibility of Escherichia coli O157, which is zoonotic in ruminant feces and feed, and the pathogen Listeria monocytogenes, which causes diseases in humans and animals, and to use these results as epidemiological data in our province, region and country. Feces and animal feed samples were analyzed simultaneously for in order of E. coli O157 and L. Monocytogenes according to ISO 16654: 2001 / Amd 1: 2017 and ISO 11290-1. 38 E.coli O157 was isolated from a total of one hunderd stool samples. 18 L. monocytogenes were isolated from a total of one hunderd stool samples. 6 L. monocytogenes were isolated from 50 silage samples. Three of these isolates were isolated from faeces and silage samples taken from the same farm with L. monocytogenes isolates isolated from sheep feces. E. coli O157 could not be isolated from a total of 100 silage and feed samples. All L. monocytogenes isolates were susceptible to sulfamethoxazole / trimethoprim, tetracycline, streptomycin, meropenem and erythromycin. The highest resistance was detected against Sulbactam / ampicillin. 3 E. coli O157 isolates were found resistant to Gentamicin and 7 isolates to Tobramycin. 21 isolates were resistant to erythromycin, and 12 isolates were intermediate. According to PCR results of fliCH7, Stx1, Stx2, eaeA and EhlyA genes, EhlyA gene was found in 20 E. coli O157 isolates. Of these isolates, 4 were isolated from sheep feces and 16 from calf feces. The stx1 gene was found in a total of 5 E. coli O157 isolates, one from sheep feces and four from calf feces. EhlyA gene was also found in all isolates with stx1 gene. The stx2 gene was found in a total of 3 E. coli O157 isolates, one from sheep dung and two from calf dung. Intimin gene was found in 8 E. coli O157 isolates, two of which are sheep faeces and six calf faecal isolates. EhlyA gene was found in all isolates with intimin gene. In this study, enterohaemolysin is the predominant virulence factor among the isolates. Epidemiologically, silage was thought to be the main source of L. monocytogenes contamination, and recently, silage contamination continued in Balikesir.
Listeria species lead to mastitis infection in cows. The aerobic mesophilic bacteria count (total bacteria count) is one of the most important factors affecting udder health and determining the milk quality. The aim of this study was to determine the aerobic mesophilic bacteria count, one of the most important factors affecting cow's milk quality, and presence and the antibiotic resistance profiles of Listeria spp., one of the factors causing mastitis in cows. As a result of isolation and identification for Listeria spp., totally 3 L. monocytogenes (n: 68, 4.41%), 7 L. innocua (n: 68, 10.29%) and 3 L. ivanovii (n: 68, 4.41%) were isolated from cow milk samples. According to results of the disc diffusion method performed to determine antibiotic susceptibility, it was found that L. monocytogenes, L. innocua, and L. ivanovii isolates were susceptive against sulfamethoxazole/ trimethoprim, meropenem, vancomycin, streptomycin, oxacillin and erythromycin. The aerobic mesophilic bacteria in the cow milk samples were detected 1.1x107 cfu/ml as the highest and 2.3x102 cfu/ml as the lowest. The average aerobic mesophilic bacteria count of milk samples was calculated 256623.971 cfu/ml. The total bacteria (aerobic mesophilic bacteria) count (cfu/ml) of milk samples in the study was found to be high based on the criteria stated in the national and international standards. Also, Listeria species were isolated from these samples. Since intermediate and resistant Listeria species were determined against the antibiotics used as a treatment option in these isolates, it is thought that Listeria species should also be considered in mastitis infections in terms of etiology and treatment. It is considered that a national mastitis control program is needed for preventing the mastitis infections and antibiotic resistance development causing economic losses in dairy cattle enterprises in order to provide milking hygiene completely.
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