Detection of AmpC and ESBL-producing Enterobacterales isolated from urinary tract infections in Tunisia

Urinary tract infections (UTIs) are the most frequent human infections in community and hospitals. This study aimed to determine the distribution of bacterial uropathogens among urinary tract infections diagnosed within the regional hospital Houcine Bouzaiene (Gafsa, South West Tunisia) during a survey of 54 days from the 8th of November to the 31st of December 2017. Enterobacterales strains were tested for antimicrobial resistance by disk diffusion method and extended-spectrum beta-lactamase (ESBL) production was tested by double-disc synergy test. Strains were further subjected to a molecular assessment of ESBL and AmpC beta-lactamase production by PCR. Overall, 173 bacterial isolates were studied, out of which 91.3% were Enterobacterales. Escherichia coli was the dominant pathogen, followed by Klebsiella pneumoniae. High to moderate resistance rates were observed, ranging from 66% to 90.7% for penicillin, from 6.7% to 18.6% for cephalosporins and from 16.2% to 25.4% for fluoroquinolones. Enterobacterales with decreased susceptibility to third-generation cephalosporins (3rd GC) carried several resistance genes: bIaCTX-M group 1 and group 9, and ACC and FOX AmpC beta-lactamase genes. Overall, ESBLs and AmpC beta-lactamases were detected in 57% and 14% of the 3rd GC-resistant isolates, respectively. This study proved the high potential of K. pneumaniae species to develop resistance against commonly used antibiotics. Thus, rigorous monitoring of the antibiotic resistance of clinical pathogens have to be implemented in Tunisia. Our results are very relevant to evaluate efficiency of the Tunisian therapeutic strategies against UTIs and adapt them to the emerging problem of antimicrobial resistance

Real time PCR gene profiling and detection of Salmonella using a novel target: The siiA gene

The objective of this study was to develop and evaluate a SYBR Green real time PCR method for the specificdetection of Salmonella spp using a novel target, the siiA gene. Primer specificity testing was done on a panel of76 Salmonella strains and 32 non-Salmonella strains. The primers directed against the siiA gene amplified allSalmonella strains tested, while non-Salmonella strains were not amplified. The melting temperatures of the107 bp amplicons were consistently specific as they gave melting peaks around 75.5 °C. The precision of theassay, based on intra and inter-run variations, was shown to be widely acceptable. In the second part of thisstudy, 45 Salmonella strains were screened for the presence of 6 virulence-associated genes (sopB, cat2, safC,sefB and SC1248) located in several Salmonella Pathogenicity Islands (SPIs) and the spvC gene from the Salmonellavirulence plasmid. The prevalence of these genes ranged from51% to 100%. Variable virulence gene profileswereobtained even within the same serotype

Effect of zeolite (clinoptilolite) as feed additive in Tunisian broilers on the total flora, meat texture and the production of omega 3 polyunsaturated fatty acid

Abstract Background Increasing consumer demand for healthier food products has led to the development of governmental policies regarding health claims in many developed countries. In this context, contamination of poultry by food-borne pathogens is considered one of the major problems facing the progress of the poultry industry in Tunisia. Result Zeolite (Clinoptilolites) was added to chicken feed at concentrations 0,5% or 1% and was evaluated for its effectiveness to reduce total flora in chickens and its effects on performance of the production. The broilers were given free and continuous access to a nutritionally non-limiting diet (in meal form)that was either a basal diet or a' zeolite diet' (the basal diet supplemented with clinoptilolite at a level of 0,5% or 1%). It was found that adding zeolite in the broiler diet significantly (p Conclusion This study showed the significance of using zeolite, as a feed additive for broilers, as part of a comprehensive program to control total flora at the broiler farm and to increase level of Omega 3 fatty acid on the chicken body.</p

Genetic diversity of food-isolated Salmonella strains through Pulsed Field Gel Electrophoresis (PFGE) and Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR).

All over the world, the incidence of Salmonella spp contamination on different food sources like broilers, clams and cow milk has increased rapidly in recent years. The multifaceted properties of Salomnella serovars allow the microorganism to grow and multiply in various food matrices, even under adverse conditions. Therefore, methods are needed to detect and trace this pathogen along the entire food supply network. In the present work, PFGE and ERIC-PCR were used to subtype 45 Salmonella isolates belonging to different serovars and derived from different food origins. Among these isolates, S. Enteritidis and S. Kentucky were found to be the most predominant serovars. The Discrimination Index obtained by ERIC-PCR (0.85) was slightly below the acceptable confidence value. The best discriminatory ability was observed when PFGE typing method was used alone (DI = 0.94) or combined with ERIC-PCR (DI = 0.93). A wide variety of profiles was observed between the different serovars using PFGE or/and ERIC-PCR. This diversity is particularly important when the sample origins are varied and even within the same sampling origin

The number of samples positive for each pair of pathogens.

<p>The number of samples positive for each pair of pathogens.</p

Representative ERIC-PCR fingerprint of different between <i>Salmonella</i> isolates on 2 per cent agarose gel.

<p>M: 1000 bp DNA marker; lane 1 to 13: F34′, F35′, C31′, C32′, C33′, C34′, C36′, L1, L2, L3, L4, L5 and L9.</p

Sequencing results of vaginal swab samples positive with the pan-<i>Chlamydiales</i> PCR.

<p>*These results were previously published by Barkallah et al. (2013).</p

The association of abortion with positive results for different agents and frequencies of positive results between vaginal and milk samples.

<p>The association of abortion with positive results for different agents by real-time PCR and frequencies of positive results between vaginal and milk samples were assessed using the Fisher's exact test. A p value<0.05 was taken as a level of significance. Cases: cows with clinical signs, controls: cows with normal pregnancies and normal parturitions, OR: odds ratio, CI: confidence interval.</p

Details of cases positive for <i>Brucella abortus</i>.

<p>+ positive/ − negative.</p

Sequences of primers and probes used in the study.

<p>Sequences of primers and probes used in the study.</p