Occurrence and antibiogram of multidrug-resistant Salmonella enterica isolated from dairy products in Libya.

Background and Aim: Foodborne illnesses are a serious challenge to human health and the economic sector. For example, salmonellosis remains a burden in developed and developing nations. Rapid and reliable molecular methods to identify Salmonella strains are essential for minimizing human infection. This study aimed to identify Salmonella spp. in raw milk and dairy products using conventional and molecular techniques and to test the antibiotic susceptibility of the isolated strains. Materials and Methods: One hundred and thirty-one milk and dairy product samples were randomly collected from different localities in Libya. Samples were examined for the presence of Salmonella by conventional culture techniques, including cultivation in Rappaport-Vassiliadis broth and streaking on xylose lysine deoxycholate agar. Identification also used polymerase chain reaction and partial sequencing of 16S rDNA. Twenty-four antibiotics were used for the examination of antimicrobial resistance of Salmonella spp. isolates with the agar disk diffusion method (Kirby-Bauer technique). Multi-antibiotic resistance index and antibiotic resistance index (ARI)for Salmonella enterica isolates were calculated. Results: Twenty-one of 131 samples (16%) were positive for Salmonella spp. recovered from 9 (16%), 2 (11%), 4 (22.2%), and 6 (46%) samples of raw cow milk, fermented raw milk, and fresh locally made soft cheeses, Maasora and Ricotta), respectively. Samples of ice cream, milk powder, and infant formula showed no Salmonella spp. contamination. Only 9 of 21 (42.8%) isolates were confirmed as S. enterica by partial sequence 16S rDNA analysis. All isolates were resistant to amoxycillin, bacitracin, penicillin G, lincomycin, vancomycin, clindamycin, and cloxacillin with an ARI of 0.042. In contrast, all tested strains were sensitive to levofloxacin, doxycycline, and ciprofloxacin. In addition, all of the tested isolates (100%) were resistant to more than one antibiotic. Conclusion: This study demonstrated the applicability of molecular techniques, compared with conventional methods, as preferable for the identification of Salmonella in milk and dairy products and thus reduction of milk-borne transmission to the consumers. From the view of public health, isolation and identification of Salmonella multidrug-resistant strains from raw cow`s milk and locally prepared dairy products sold in the Libyan markets indicate the need to improve the handling and processing of milk and dairy products to minimize the prevalence of Salmonella, one of the most important foodborne microorganisms that cause food poisoning.

Occurrence, characterization, and antibiogram of Staphylococcus aureus in meat, meat products, and some seafood from Libyan retail markets.

AIM: The aim of the current investigation was to screen the presence of Staphylococci spp., especially S. aureus in meat, meat products of different animal species, and some seafood sold in some retail markets in Libya using cultural and molecular techniques, and to study their antibiotics resistance profiles. MATERIALS AND METHODS: A total of 139 samples from red meat, meat products, and seafood were collected from many areas in Libya. Enumeration and isolation of Staphylococci spp. and S. aureus by normal cultural methods followed by molecular identification using molecular techniques by bacterial DNA extraction and partial sequencing of 16S rDNA. RESULTS: Out of 139 samples, 112 (80.6%) were contaminated with different species of Staphylococci based on cultural characteristics of Staphylococci on Baird-Parker medium, for which S. aureus was detected in only 32 samples (23%). However, only six out of 18 (33.3%) isolates sent for sequencing were confirmed to be S. aureus using the molecular technique. The six identified isolates of S. aureus were tested for antimicrobial resistance against 24 most commonly used antibiotics. All isolates were resistant to only two antibiotics (cefotaxime and clindamycin). Among these six isolates, only one confirmed to be Methicillin-resistant Staphylococcus aureus. CONCLUSION: Results of this study suggest that food of animal origin could be a source of S. aureus with antimicrobial resistance characteristics that can be spread through the food chain, and raise the importance of these results for public health.

Enterohemorrhagic Escherichia coli O157 in milk and dairy products from Libya: Isolation and molecular identification by partial sequencing of 16S rDNA.

AIM: The aim of this work was to isolate and molecularly identify enterohemorrhagic Escherichia coli (EHEC) O157 in milk and dairy products in Libya, in addition; to clear the accuracy of cultural and biochemical identification as compared with molecular identification by partial sequencing of 16S rDNA for the existing isolates. MATERIALS AND METHODS: A total of 108 samples of raw milk (cow, she-camel, and goat) and locally made dairy products (fermented cow's milk, Maasora, Ricotta and ice cream) were collected from some regions (Janzour, Tripoli, Kremiya, Tajoura and Tobruk) in Libya. Samples were subjected to microbiological analysis for isolation of E. coli that was detected by conventional cultural and molecular method using polymerase chain reaction and partial sequencing of 16S rDNA. RESULTS: Out of 108 samples, only 27 isolates were found to be EHEC O157 based on their cultural characteristics (Tellurite-Cefixime-Sorbitol MacConkey) that include 3 isolates from cow's milk (11%), 3 isolates from she-camel's milk (11%), two isolates from goat's milk (7.4%) and 7 isolates from fermented raw milk samples (26%), isolates from fresh locally made soft cheeses (Maasora and Ricotta) were 9 (33%) and 3 (11%), respectively, while none of the ice cream samples revealed any growth. However, out of these 27 isolates, only 11 were confirmed to be E. coli by partial sequencing of 16S rDNA and E. coli O157 Latex agglutination test. Phylogenetic analysis revealed that majority of local E. coli isolates were related to E. coli O157:H7 FRIK944 strain. CONCLUSION: These results can be used for further studies on EHEC O157 as an emerging foodborne pathogen and its role in human infection in Libya.

ß4-integrin-mediated cytotoxic activity of AexU in human prostate cancer PC3 cells.

The present study aimed to characterize the cytotoxic activity of AexU, an effector-mediating type three secretion system (TTSS) of gram-negative bacteria, in human prostate cancer cells, focusing on the association with ß4-integrin expression. The cytotoxic effects of AexU either alone or in combination with chemotherapeutic agents were evaluated using several human prostate cancer cell lines. Human prostate cancer PC3 cells, in which an expression vector containing siRNA targeting ß4-integrin had been introduced, were established (PC3/sh-In), and the cytotoxic effects of AexU on the PC3/sh-In cells were compared with the PC3 cells that were transfected with a control vector (PC3/C). The expression levels of ß4-integrin in the PC3 cells were markedly higher compared with those in the LNCaP or DU145 cells, and the cytotoxic effects of AexU in the PC3 cells were more pronounced compared with those in the LNCaP or DU145 cells. The sensitivity of the PC3 cells to docetaxel and cisplatin was significantly enhanced following treatment with AexU, resulting in a decrease in the IC50 of the two agents by ~90%. The cytotoxic effect of AexU in the PC3/C cells was more marked compared with that in the PC3/sh-In cells, and the phosphorylation of Akt in the PC3/C cells appeared to be significantly more inhibited by the treatment with AexU compared with the PC3/sh-In cells. In conclusion, treatment with AexU may be a useful therapeutic option for prostate cancer when ß4-integrin is overexpressed. The treatment appears to exert its effects through growth inhibition and by enhancing the sensitivity of the cancer cells to chemotherapeutic agents.