Molecular Characterization of Staphylococcus aureus Isolated from Human and Food Samples in Northern Algeria.

Staphylococcus aureus is a commensal resident of the skin and nasal cavities of humans and can cause various infections. Some toxigenic strains can contaminate food matrices and cause foodborne intoxications. The present study aimed to provide relevant information (clonal complex lineages, agr types, virulence and antimicrobial resistance-associated genes) based on DNA microarray analyses as well as the origins and dissemination of several circulating clones of 60 Staphylococcus aureus isolated from food matrices (n = 24), clinical samples (n = 20), and nasal carriers (n = 16) in northern Algeria. Staphylococcus aureus were genotyped into 14 different clonal complexes. Out of 60 S. aureus, 13 and 10 isolates belonged to CC1-MSSA and CC97-MSSA, respectively. The CC 80-MRSA-IV was the predominant S. aureus strain in clinical isolates. The accessory gene regulator allele agr group III was mainly found among clinical isolates (70.4%). Panton-Valentine leukocidin genes lukF/lukS-PV were detected in 13.3% of isolates that all belonged to CC80-MRSA. The lukF/S-hlg, hlgA, and hla genes encoding for hemolysins and leucocidin components were detected in all Staphylococcusaureus isolates. Clinical and food isolates harbored more often the antibiotic resistance genes markers. Seventeen (28.3%) methicillin-resistant Staphylococcus aureus carrying the mecA gene localized on a SCCmec type IV element were identified. The penicillinase operon (blaZ/I/R) was found in 71.7% (43/60) of isolates. Food isolates belonging to CC97-MSSA carried several antibiotic resistance genes (blaZ, ermB, aphA3, sat, tetM, and tetK). The results of this study showed that all clones were found in their typical host, but interestingly, some nasal carriers had isolates assigned to CC705 thought to be absent in humans. The detection of MRSA strains among food isolates should be considered as a potential public health risk. Therefore, controlling the antibiotics prescription for a rational use in human and animal infections is mandatory.

Short communication: Diversity of staphylococci isolated from sheep mastitis in northern Algeria.

Mastitis in ruminants is an important disease with major effects on both the economy and animal welfare. It is caused by major pathogens such as Staphylococcus aureus and minor pathogens such as coagulase-negative staphylococci. The objective of this study was to identify and characterize staphylococci as a cause of sheep mastitis in Algeria. In this study, 123 milk samples were collected directly from the udder of sheep suffering from clinical mastitis in 2 provinces in Algeria. Recovered isolates were identified using MALDI-TOF mass spectrometry. Virulence-associated and antimicrobial resistance genes as well as clonal complexes (CC) of S. aureus were determined using microarray-based analysis. A total of 45 staphylococci isolates were cultivated from sheep milk samples, and 28 S. aureus were identified as methicillin susceptible (62.2%). Seventeen other Staphylococcus isolates of different species were identified using MALDI-TOF mass spectrometry. Subsequent microarray analysis typed the methicillin-susceptible S. aureus to 6 CC: CC8-MSSA, CC97-MSSA, CC130/521-MSSA, CC479-MSSA, CC522-MSSA, and CC705-MSSA. The accessory gene regulator agrIII and the ruminant leukocidin genes lukF-P83 and lukM were found in all isolates of CC130/521, CC479, CC522, and CC705. The toxic shock syndrome toxin gene tst1 was detected exclusively in CC130/521. Additionally, virulence-associated genes (sea, sed, sak, hld, hlgA, edinB, and others) were detected. The presence of antibiotic resistance genes [blaZ, erm(B), and tet(K)] was detected in small numbers of staphylococci. Staphylococci possessing these genes are considered potential hazards for farm animals, farmers, and consumers. Data concerning the prevalence and diversity of staphylococci causing mastitis in sheep from Algeria are lacking. Presented results on different aspects about staphylococci in Algerian sheep populations should at least partially close that gap. However, further extensive studies covering more geographical regions are needed to assess the epidemiological risk.

Influence of some parameters on the ability of Listeria monocytogenes, Listeria innocua, and Escherichia coli to form biofilms.

AIM: The present study was conducted to evaluate the capacity of Listeria monocytogenes (L.m), Listeria innocua (L.i), and Escherichia coli to form biofilms on polystyrene support under different parameters by performing crystal violet (CV) staining technique. MATERIALS AND METHODS: Different suspensions were prepared with single strains and with multiple combinations of strains including two serogroups of L.m (IIa and IIb), L.i, and E. coli strains at different microbial load. Selected strains and combinations were grown in biofilms for 6 days attached to polystyrene microplates under aerobic and microaerophilic conditions. The evaluation of the power of adhesion and biofilm formation was determined by CV staining followed by the measurement of optical density at 24 h, 72 h, and 6 days incubation time with and without renewal of the culture medium. RESULTS: All the strains tested, presented more or less adhesion power depending on the variation of the studied parameters as well as the ability to form multispecies biofilms. Their development is more important by renewing the culture medium and increasing the initial load of bacteria. The ability to adhere and form biofilms differs from one serogroup to another within the same species. In bacterial combination, strains and species of bacteria adopt different behaviors. CONCLUSION: The ability to form biofilms is a key factor in the persistence of tested strains in the environment. Our study showed that L.m, L.i, and E. coli could adhere to polystyrene and form biofilms under different conditions. More researches are necessary to understand the mechanisms of biofilm formation and the influence of different parameters in their development.

Staphylococcus aureus isolated from selected dairies of Algeria: Prevalence and susceptibility to antibiotics.

AIM: The objectives of this study were to assess the prevalence of Staphylococcus aureus in raw milk in Algerian dairies, to study the effect of seasons on the contamination of milk and the susceptibility of isolated strains to antibiotics, and to estimate the risk on the health consumer. MATERIALS AND METHODS: The ISO method 6888-1 (1) was used for Staphylococcus screening. Antimicrobial susceptibility to the 11most used antibiotics in veterinary medicine was assessed using the disk diffusion assay. RESULTS: The overall prevalence was 31.56% (95/301); 34.84% (85/244) from raw milk collectors cisterns (MCC), 22.73% (5/22) from mixing tank milk before pasteurization, and 14.29% (5/35) from pasteurized tank milk (p<0.05). A significant difference (p<0.001) of contamination on MCC was observed between dairies without season influence (p≥0.05). It was observed that 49.47% of S. aureus isolates were resistant to penicillin, 5.26% to tetracycline, 4.21% to erythromycin, 3.15% to neomycin, 2.10% to cefoxitin, 2.10% to clindamycin, and 1.05% to ofloxacin. No resistance was observed for vancomycin, gentamicin, chloramphenicol, and trimethoprim-sulfamethoxazole. CONCLUSION: A high prevalence of S. aureus from MCC was observed without significant effect of season. The pasteurization does not ensure the elimination of bacteria in all samples. Half of the isolates were resistant to penicillin. These findings emphasize the importance of S. aureus control in Algerian milk industry at different levels to improve public health.

Combined impacts of oregano extract and vacuum packaging on the quality changes of frigate tuna muscles stored at 3±1°C.

AIM: The combined effects of oregano extract with vacuum packing (VP) on the quality enhancement of dark and white muscles of frigate tuna (Auxis thazard) stored as intact fillet at refrigerated (3±1°C) conditions were studied. MATERIALS AND METHODS: About 35 kg of fish were filleted without skin removal and randomly divided into two groups. One group without treatment (control) and the remaining group were dipped in a sterilized oregano extract solution for 5 min. Chemical, microbiological, sensorial, and textural analyses were carried out in each of dark and white muscles of frigate tuna fillets during storage. RESULTS: Several quality indexes were higher in dark muscle than white muscle. The sensory assessment indicated that both muscles from control had a shelf life of 12 days. Quality parameters of both muscles had the same tendency and were significantly affected by time and also by the presence of plant extract in VP. Although VP alone was sufficient to delay lipid oxidation on fish fillets, especially on dark muscle but cannot enhance the textural deterioration in both muscles. CONCLUSION: Consequently, the employment of such combination had a cumulative effect on preservation, resulting in prolonging the shelf life of both frigate tuna muscles.

Serotype diversity and slaughterhouse-level risk factors related to Salmonella contamination on poultry carcasses in Algiers.

INTRODUCTION: In Algeria, the latest studies on Salmonella demonstrated warning contamination rates in farms and slaughterhouses. This pathogen can contaminate poultry meat and put humans at risk especially that such product is nowadays widely consumed. METHODOLOGY: a cross-sectional study was conducted in Algiers to evaluate prevalence, determine serotypes and quantify risk for Salmonella contamination in broiler chickens and turkeys at the post-chill stage of slaughter process. RESULTS: batch prevalence was 63.1% for chickens and 34.9% for turkeys. Eleven serotypes were isolated from chickens and five from turkeys. The most predominant at both sample and batch levels was S. Kentucky either in chicken (65.1%) or in turkey carcasses (63.2%). Univariate analysis screened 3 variables for chickens and 5 variables for turkeys. Final multivariate regression models provided one potential risk factor for Salmonella contamination in each poultry species. Presence of less than 6 broilers simultaneously in the traditional scalding tank of small scale slaughterhouses had a significantly reduced contamination risk (OR = 0.31; p < 0.05). Slaughtering turkeys in sites processing only this specie than in mixed poultry slaughterhouses increased significantly the contamination probability (OR = 4.44; p < 0.05). CONCLUSIONS: Our study indicates a high prevalence of Salmonella-contaminated poultry carcass with wide diversity of serotypes. Moreover, two potential risk factors identified for the first time in Algeria are found to be associated with the lack in hygienic management on production sites. A real threat for consumers exists highlighting the imperative need for improved safety throughout the local poultry meat supply chain.

Emerging of antimicrobial resistance in staphylococci isolated from clinical and food samples in Algeria.

OBJECTIVE: The antimicrobial resistance of staphylococci rose worldwide. In total, 96 Staphylococcus isolates from food and clinical samples were collected from two provinces in Algeria. The antimicrobial susceptibility testing was performed and resistance-associated genes were detected. RESULTS: Fifty-one strains were isolated from food samples and differentiated into 33 Staphylococcus aureus and 18 coagulase-negative staphylococci. Forty-five staphylococci were collected from hospital and community-acquired infection cases. All S. aureus isolated from food were resistant to penicillin and 45.5% were resistant to tetracycline. The resistance rates of 45 clinical Staphylococcus isolates were 86.7%, 48.9%, 37.8% and 20.0% to penicillin, tetracycline, erythromycin and kanamycin, respectively. Nine isolates were confirmed as MRSA from food and clinical isolates. One S. aureus originated from food was confirmed as vancomycin-resistant. Multidrug-resistance was observed among 25.5% and 53.3% of food and clinical staphylococci, respectively. The tetM/K, blaZ, aacA-aphD, ermC and mecA genes were detected in food and clinical isolates. ermA gene was not found. This study provided insight into the status of antimicrobial resistance of staphylococci isolated from food and clinical samples in Algeria. Further investigations and surveillance programmes are mandatory.

Sources of contamination, prevalence, and antimicrobial resistance of thermophilic Campylobacter isolated from turkeys.

AIM: Sources of contamination, prevalence, and antimicrobial susceptibility of thermophilic Campylobacter isolated from turkey samples were determined. MATERIALS AND METHODS: A total of 300 samples were collected from 3 farms (fecal droppings) and 4 poultry slaughterhouses (neck skins and ceca) located in the middle area of Algeria (Algiers, Boumerdès, and Bouira). After detection, an antibiogram was realized only for slaughterhouses samples. RESULTS: Samples from cecum (90.0%, 90/100; 95% confidence interval (CI)=84.1-95.9%), fecal dropping (68.0%, 68/100; 95% CI=58.9-77.1%), and neck skin (55.0%, 55/100; 95% CI=45.2-64.8%) were positive for thermophilic Campylobacter (p<0.05). Contamination rate of turkey carcasses was higher in modern slaughterhouse (96.7%) than in traditional slaughterhouses (37.1%) (p<0.05). Isolated strains were resistant to nalidixic acid (NA) (87.5%), tetracycline (TE) (81.3%), ciprofloxacin (CIP) (75.0%), ampicillin (AM) (65.6%), and erythromycin (25.0%) (p<0.05). 96.9% (124/128) of the isolates were multiresistant and 18 drug resistance patterns were registered. The predominant one (43.0%) was AM, NA, CIP, and TE. CONCLUSIONS: Potential sources of contamination of this fastidious bacterium were noticed in farms and slaughterhouses. Modern slaughterhouse allowed contamination of turkey carcasses more than a traditional slaughterhouse. However, the scalding step could not represent a source of contamination. The most tested strains exhibited resistance to erythromycin and/or CIP. It is worrisome because these molecules are considered as first-choice antibiotics for human campylobacteriosis.

Prevalence and antimicrobial resistance of Salmonella isolated from meat and meat products in Algiers (Algeria).

This study was conducted in order to estimate the proportion of raw meat and processed meat products contaminated by Salmonella in the region of Algiers, Algeria, to identify serovars and to determine the antimicrobial resistance patterns of isolates. Out of the total 314 samples (144 of raw red meat and meat products, 128 of raw poultry meat and poultry products, and 42 of processed meat products) collected from various retail outlets, 61 (19.43%) were tested positive for Salmonella. The most significant occurrences were recorded for the categories of red meat (23.61%, n=34) and poultry (17.97%, n=23). Among the 64 isolates recovered, 21 different serovars were identified and two strains were nontypable. The most prevalent serovars were Salmonella Anatum (14.6%, n=9), Salmonella Altona (12.50%, n=8), Salmonella Corvallis (7.81%, n=5), Salmonella Enteritidis (7.81%, n=5), and Salmonella Typhimurium (7.81%, n=5). Sixty-two Salmonella isolates were tested for their susceptibility to 32 selected antimicrobial agents. Fifty-six (90.32%) isolates were resistant to at least one antimicrobial, of which 20 (32.26%) showed multidrug resistance. Resistance to sulphonamides (87.10%, n=54) was the most common. Resistance rates were lower to nalidixic acid (16.13%, n=10), streptomycin (16.13%, n=10), and tetracycline (12.90%, n=8), while resistance to pefloxacin was estimated at 4.84% (n=3). Fourteen different resistance patterns were observed. The "ACSSuT" pentaresistance pattern was observed in three of the Salmonella Typhimurium strains. The obtained results show that these foodstuffs are a potential source of antimicrobial-resistant Salmonella for human infections.

Identification and molecular characterization of Listeria monocytogenes isolated in raw milk in the region of Algiers (Algeria).

Listeria monocytogenes was isolated from raw milk, whey and curdled milk produced and collected in the region of Algiers and Blida between September 2003 and July 2004. Four out of 153 (2.61%) farm milk samples and 6 out of 80 (7.50%) tankers' samples tested positive for L. monocytogenes. All samples of whey and curdled milk (n=12) tested negative for L. monocytogenes, but 2 of 22 (9%) samples of whey were contaminated by L. innocua. L. monocytogenes isolates were grouped by a multiplex PCR assay; all isolates belonged to the PCR-group IVb, which corresponds to serovars 4b, 4d and 4e. L. monocytogenes isolates were characterized by Pulsed-Field Gel Electrophoresis (PFGE). The combination of AscI and ApaI macrorestriction patterns yielded five different pulsovars (I to V). The results indicate that raw milk, and raw milk products are potential sources of the L. monocytogenes and represent a potential risk for consumers.