Multidrug-resistant Escherichia coli isolated from cleaned and disinfected poultry houses prior to day-old chick placement.

The control and elimination of multidrug-resistant (MDR) Escherichia coli is an important challenge in the poultry industry. The aim of this study was to investigate the presence of MDR E. coli in cleaned and disinfected poultry houses before day-old chick placement to identify potential flock colonization sources. In this study, a total of 104 swab samples, collected from 104 cleaned and disinfected poultry houses, were analyzed for the presence of E. coli. Antimicrobial susceptibility profiles of E. coli isolates were determined using the disk diffusion method. Screening for extended-spectrum ß-lactamases-encoding genes was performed by polymerase chain reaction and Sanger sequencing. Out of the 104 samples analyzed, 92 (88.46%) were found to be colonized with E. coli. At least one poultry house per farm was found positive for the presence of E. coli, and one isolate per positive shed was subjected to antibiotic susceptibility testing. All isolates displayed high resistance rates to most of the antimicrobial agents tested, including commonly used frontline antibiotics in Algeria. All E. coli isolates (100%) exhibited MDR profiles. A single house on four different broiler farms was found to be contaminated with CTX-M-1-producing E. coli. This study suggests the need for the adoption of strict biosecurity measures and the implementation of improved or novel disinfection procedures in all the poultry houses, in order to avoid cross-contamination of day-old chicks by MDR E. coli.

Chemical Characterization, Antibacterial Activity, and Embryo Acute Toxicity of Rhus coriaria L. Genotype from Sicily (Italy).

This study reports a full characterization of the Sicilian sumac, Rhus coriaria L. This fruit represents a potential source of fiber (33.21 ± 1.02%) and unsaturated fatty acids, being the contents of linoleic and α-linolenic acids, 30.82 ± 1.21% and 1.85 ± 0.07%, respectively. In addition, the content of phenolic and total anthocyanin was 71.69 ± 1.23 mg/g as gallic acid equivalents, and 6.71 ± 0.12 mg/g as cyanidin-3-O-glucoside equivalents, respectively. The high content in mineral elements, consisting mainly of potassium, calcium, magnesium, and phosphorus, followed by aluminum, iron, sodium, boron, and zinc, was detected by inductively coupled plasma mass spectrometry (ICP-MS). Moreover, its antimicrobial activity was evaluated against multidrug resistant (MDR) microorganisms, represented by Escherichia coli and Klebsiella pneumoniae strains isolated from poultry. The activity of seven different sumac fruit extracts obtained using the following solvents-ethanol (SE), methanol (SM), acetone (SA), ethanol and water (SEW), methanol and water (SMW), acetone and water (SAW), water (SW)-was evaluated. The polyphenol profile of SM extract, which showed better activity, was analyzed by ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS). The major component identified was gallic acid, followed by quercetin, methyl digallate, pentagalloyl-hexoside, and kaempferol 3-O-glucoside. The non-toxicity of Sicilian R. coriaria was confirmed by testing the effect of the same extract on zebrafish embryos.

Comparison of sensitivity to a commercial Origanum vulgare essential oil between extended-spectrum ß-lactamases (ESBL-) and non-ESBL-producing Enterobacteriaceae isolates.

This study aimed to evaluate the antibacterial effect of Origanum vulgare essential oil (OVEO) against ESBL- and non-ESBL-producing multidrug-resistant (MDR) Enterobacteriaceae isolates. OVEO composition was determined using Gas chromatograph-mass spectrometer (GC/MS). In the current study, the effect of OVEO was studied on seven MDR Enterobacteriaceae isolates. Antibacterial activity of OVEO was investigated by the disc diffusion assay and twofold serial dilution method. GC/MS analysis identified thymol (78.21%) as the single major component present in the OVEO. This EO showed an unexpectedly high antibacterial activity against all the studied MDR Enterobacteriaceae isolates, with inhibition zone diameters and minimum inhibitory concentration (MIC) values ranging from 28 ± 0.6 to 35 ± 0.6 mm and 0.31 ± 0.0 to 5 ± 0.0 µL/mL, respectively. However, ESBL-producing isolates were more susceptible to OVEO than non-ESBL producing isolates. This study compared, for the first time, the sensitivity to OVEO between ESBL and non-ESBL-producing Enterobacteriaceae isolates.

Virulence, Antimicrobial Resistance and Biofilm Production of Escherichia coli Isolates from Healthy Broiler Chickens in Western Algeria.

The aim of this study was to assess the virulence, antimicrobial resistance and biofilm production of Escherichia coli strains isolated from healthy broiler chickens in Western Algeria. E. coli strains (n = 18) were identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. Susceptibility to 10 antibiotics was determined by standard methods. Virulence and extended-spectrum ß-lactamase (ESBL) genes were detected by PCR. The biofilm production was evaluated by microplate assay. All the isolates were negative for the major virulence/toxin genes tested (rfbE, fliC, eaeA, stx1), except one was stx2-positive. However, all were resistant to at least three antibiotics. Ten strains were ESBL-positive. Seven carried the ß-lactamase blaTEM gene only and two co-harbored blaTEM and blaCTX-M-1 genes. One carried the blaSHV gene. Among the seven strains harboring blaTEM only, six had putative enteroaggregative genes. Two contained irp2, two contained both irp2 and astA, one contained astA and another contained aggR, astA and irp2 genes. All isolates carrying ESBL genes were non-biofilm producers, except one weak producer. The ESBL-negative isolates were moderate biofilm producers and, among them, two harbored astA, two irp2, and one aggR, astA and irp2 genes. This study highlights the spread of antimicrobial-resistant E. coli strains from healthy broiler chickens in Western Algeria.

Potentially Toxic Elements in Xiphias gladius from Mediterranean Sea and risks related to human consumption.

Swordfish is a widely consumed fish worldwide. The aim of this work was to evaluate levels of As, Zn, Mn, Fe, Cu, Al, Cr, Ni, Se, Cd, Pd and Hg in 28 samples of swordfish muscles, coming from zone of Food and Agriculture Organization (FAO) namely 37.1 and 37.2, in order to assess potential public health risks. 100% of the samples had Cd levels not detectable, 32.14% of samples exceeded European maximum limits set for Hg. Health risk assessment showed a not dangerous condition for Al (0.65% of Tolerable Weekly Intake), Ni (0.27% of Tolerable Daily Intake) and Pb (0.68% of Provisional Tolerable Weekly Intake), but very hazardous states for Hg (159.69% of Tolerable Weekly Intake) and As (1099.45% for minimum value of Benchmark Dose Limit). Target Hazard Quotient (THQ) showed dangerous values only for As and Hg. Analytical data of the present study suggest a swordfish prudent consumption.

Resistance to multiple first-line antibiotics among Escherichia coli from poultry in Western Algeria.

BACKGROUND AND AIM: Escherichia coli can cause a number of serious infections both in human and veterinary medicine. Their management is increasingly complicated by the emergence and dissemination of multiresistance to various first-line antimicrobial agents. This study aimed to evaluate the resistance level to the commonly used antibiotics, with a focus on the first-line antimicrobial agents, in E. coli strains isolated from poultry in Western Algeria. MATERIALS AND METHODS: E. coli culture was done on MacConkey agar and their identification was determined by AP20E system. For susceptibility testing, disk diffusion method to 14 antimicrobials, including first-line antibiotics, was used according to Kirby-Bauer disk diffusion method in Mueller-Hinton agar and the results were interpreted according to the Clinical and Laboratory Standards Institute guidelines. E. coli isolates were considered as multidrug resistance (MDR) when found resistant to at least one antimicrobial agent of three different families of antibiotics. Double-disk synergy and combination disk tests were used for initial screening and confirmation for extended-spectrum ß-lactamases (ESBLs) production, respectively. RESULTS: A total of 145 E. coli strains were isolated in this study. High resistance levels to various antibiotics, including commonly used first-line antimicrobial agents, were recorded in this study. The highest resistance level was observed against nalidixic acid (90.34%, n=131), followed by tetracycline (86.89%, n=126), ampicillin (82.75%, n=120), enrofloxacin (80.68%, n=117) and neomycin (80.68%, n=117), trimethoprim/sulfamethoxazole (73.79%, n=107), norfloxacin (72.41%, n=105) and cephalothin (72.41%, n=105), amoxicillin/clavulanic acid (51.72%, n=75), chloramphenicol (22.75%, n=33), nitrofurantoin (17.24%, n=25), gentamicin (13.10%, n=19), and ceftiofur (3.44%, n=5). Moreover, resistance to multiple first-line antibiotics was also demonstrated in the present study. Overall, 139 out of 145 isolates (95.86%) demonstrated MDR (resistant to at least three antibiotics). In addition, five E. coli isolates (3.44%) were confirmed to be ESBL producers. CONCLUSION: The alarming rate of E. coli resistant to multiple first-line antibiotics in poultry demands intensified surveillance. These results call for taking drastic measures to preserve antibiotic effectiveness and reduce the emergence risks of extensively drug-resistant and pandrug-resistant E. coli isolates.

Polyphenols contents, heavy metals analysis and in vitro antibacterial activity of extracts from Cladanthus arabicus and Bubonium imbricatum of Moroccan Origin.

Aim of this study was to evaluate polyphenols and major and trace elements of Cladanthus arabicus and Bubonium imbricatum, along with their in vitro antibacterial activity against six multidrug resistant Enterobacteriaceae (Escherichia coli S33/16, E. coli S34/16, Proteus mirabilis S32/16, Klebsiella pneumoniae S12/16, Enterobacter cloacae S5/16, and Salmonella sp S12/14). UV spectrophotometry, ultra-high-performance liquid chromatography coupled to mass spectrometry and inductively coupled plasma mass spectrometry were used to evaluate total polyphenol content, quali-quantitative profile of single polyphenols and inorganic elements of the extract. The antibacterial activity was investigated by standard methods. Twelve polyphenols were identified in both plants and these were more concentrated in B. imbricatum than C. arabicus extracts. High levels of minerals, essential trace elements and tolerable levels of heavy metals (Cd, As and Pb) were found. Furthermore, the extracts showed also a strong in vitro antibacterial activity, particularly versus E. coli S33/16 (MIC, 0.125 mg ml-1).

Detection of multidrug resistant Escherichia coli in the ovaries of healthy broiler breeders with emphasis on extended-spectrum ß-lactamases producers.

In the last few years, antimicrobial resistant (AMR) Escherichia coli have been detected in newborn chickens suggesting their vertical transmission from breeding birds to their offspring. However, little is known about the presence of AMR E. coli in the reproductive organs of broiler breeders. The aim of this study was to investigate the presence of E. coli in the ovaries of healthy broiler breeders and to study their antimicrobial resistance. Samples from broiler breeders (n = 80) collected from 80 different broiler breeder flocks were included in this study. Antibiotic susceptibility testing was performed using disk diffusion method according to Clinical and Laboratory Standards Institute guidelines. Minimal inhibitory concentrations (MICs) of five antimicrobial agents were determined by Etest. PCR and sequencing were used to detect the blaESBL genes. E. coli were detected in the ovaries of thirty seven out of 80 (46.25%) sampled flocks. High levels of resistance to various first-line antimicrobial agents were recorded in E. coli isolates. This study showed that 89.18% of E. coli isolates were multidrug resistant (MDR). Furthermore, MDR extended-spectrum ß-lactamases (ESBL)-producing E. coli were detected in the ovaries of four different broiler breeder flocks. Molecular characterization revealed that three isolates harboured blaCTX-M-1 gene and one isolate expressed blaSHV-12 gene. In addition, one blaCTX-M-1 -producing E. coli co-harboured the blaTEM-1 gene. These findings would contribute to a better epidemiological understanding of MDR E. coli for improve existing preventive strategies in order to reduce the dissemination of antimicrobial resistance in the broiler production system.

Antibacterial activity of Thymus vulgaris essential oil alone and in combination with cefotaxime against blaESBL producing multidrug resistant Enterobacteriaceae isolates.

The aim was to evaluate the susceptibility of blaESBL producing Enterobacteriaceae to Slovakian Thymus vulgaris essential oil (TVEO) alone and in combination with cefotaxime (CTX). TVEO composition was determined by gas chromatograph-mass spectrometer (GC/MS). Susceptibility to 21 antibiotics was determined by disc diffusion assay. Genes characterization for resistance to ß-lactams was accomplished by polymerase chain reaction (PCR). The antibacterial activity was investigated by standard methods. The synergistic interaction was determined by checkerboard test. Thymol (34.5%), p-cymene (22.27%) and linalool (5.35%) were the major components present in the TVEO. The identified strains were multi-drug resistant (MDR). TVEO showed high activity against all MDR strains, including blaESBL producing isolates, with inhibition zones and MIC values in the range of 24-40 mm/10µL and 2.87-11.5 µg/mL, respectively. TVEO in combination with CTX showed a synergistic action against blaSHV-12 producing Escherichia coli (FICI 0.28) and an additive effect vs ESBL producing Enterobacter cloacae (FICI 0.987).

Isolation of Escherichia coli carrying the blaCTX-M-1 and qnrS1 genes from reproductive organs of broiler breeders and internal contents of hatching eggs.

This study aimed to characterize two third-generation cephalosporins- and quinolone-resistant Escherichia coli (TGCs- and Q-R-Ec) isolates recovered from the ovaries of a broiler breeder flock and the internal contents of hatching eggs produced by the broiler breeder flock. Clonal relatedness was determined by multilocus sequence typing (MLST). The isolates displayed the same multidrug resistance profile, with resistance to ampicillin, ticarcillin, piperacillin, cefazollin, cephalothin, cefotaxime, nalidixic acid, tetracycline and sulfonamides. Double disk synergy test demonstrated that the two isolates presented an ESBL phenotype. PCR and sequencing results showed that both the isolates harbored the blaCTX-M-1 and qnrS1 genes. MLST revealed a novel allele combination, designated as ST461, in these isolates. This study would contribute to the molecular epidemiological understanding of TGCs- and/or Q-R-Ec.

Characterization of quinolone-resistant Enterobacteriaceae strains isolated from poultry in Western Algeria: First report of qnrS in an Enterobacter cloacae.

AIM: Multidrug-resistant (MDR) Enterobacteriaceae have frequently been reported, in both human and veterinary medicine, from different parts of the world as a consequence of antibiotic usage. However, there is a lack of published data regarding antimicrobial resistance in non-Escherichia coli (E. coli) Enterobacteriaceae from animals in Algeria. This study aimed to evaluate the frequency of resistance to antibiotics with a focus on quinolones and to investigate the presence of qnr genes inEnterobacteriaceaeof poultry origin. MATERIALS AND METHODS: A total of 310 samples of poultry origin were collected from 2010 to 2014 from broiler and layer farms and hatcheries located in different geographic areas of Western Algeria (including Mostaganem, Oran, Mascara, Relizane, Chlef, Tiaret, and Tissemsilt). Antimicrobial susceptibility testing was performed using disc diffusion assay. Polymerase chain reaction and sequencing accomplished the characterization of qnr genes (qnrA, qnrB, and qnrS). RESULTS: A total of 253 Enterobacteriaceaestrains were isolated in this study. These isolates exhibited high levels of resistance to quinolones and other families of antibiotics. All the strains isolated in this study were resistant to at least one antibiotic. Among them, 233 (92.09%) were considered MDR. Among the 18 randomly selected nalidixic acid (NA)-resistant Enterobacteriaceaeisolates, one E. coli and one Enterobacter cloacae were carrying qnrS1. By contrast, qnrA and qnrB were not detected in this study. CONCLUSION: This is the first report on the identification of the qnrS gene in E. cloacae isolated from animal source in Algeria. Further studies have to be conducted to determine the real prevalence of qnr genes.

Valorization of raw materials from agricultural industry for astaxanthin and ß-carotene production by Xanthophyllomyces dendrorhous.

Waste deriving from food and agro-industries is a growing problem in our modern society. In order to reduce food waste and its associated impacts over the environment and public health, innovative strategies for the conversion of food wastes in added value products are studied. Whey is one of the prominent waste deriving from Sicilian diary industry. The growth and the carotenoid biosynthesis of the yeast Xanthophyllomyces dendrorhous by its cultivation in pre-treated whey were studied. The study showed the possibility of increasing the astaxanthin productivity from X. dendrorhous in a batch fermentative process under continuous illumination. Our work is focused on the importance of carotenogenesis by X. dendrorhous on whey for the economic and ecological aspect. The results obtained in the study provide useful information about the possible use of this waste material for carotenogenesis from X. dendrorhous.