Dataset Information

Prevalence and Characterization of PVL-Positive Staphylococcus aureus Isolated from Raw Cow's Milk.

ABSTRACT: The present study aimed to investigate the prevalence, antibiotic susceptibility profiles, and some toxin genes of Panton-Valentine leukocidin (PVL)-positive Staphylococcus aureus (S. aureus) in unpasteurized raw cow's milk collected from retail outlets located at Mansoura, Dakahliya governorate, Egypt. In that context, a total of 700 raw cow's milk samples were investigated for the presence of S. aureus, which was identified in 41.1% (288/700) of the samples. Among the S. aureus isolates, 113 PVL-positive S. aureus were identified and subjected for further analysis. The PVL-positive S. aureus were investigated for the existence of toxin-related genes, including hemolysin (hla), toxic shock syndrome toxin-1 (tst), and enterotoxins (sea, seb, sec, see, seg, sei, and selj). Genotypic resistance of PVL-positive strains was performed for the detection of blaZ and mecA genes. Among the PVL-positive S. aureus, sea, seb, and sec were detected in 44.2, 6.2%, and 0.9%, respectively, while the hla and tst genes were identified in 54.9% and 0.9%, respectively. The blaZ and mecA genes were successfully identified in 84.9 (96/113) and 32.7% (37/113) of the total evaluated S. aureus isolates, respectively. PVL-positive S. aureus displayed a high level of resistance to penicillin, ampicillin, and trimethoprim-sulfamethoxazole. Multidrug resistance (resistant to ≥3 antimicrobial classes) was displayed by all methicillin-resistant S. aureus (MRSA) and 38.2% of methicillin-sensitive S. aureus (MSSA) isolates. The obtained findings are raising the alarm of virulent PVL-positive MRSA clones in retail milk in Egypt, suggesting the requirement for limiting the use of β-lactam drugs in food-producing animals and the importance of implementing strong hygiene procedures in dairy farms and processing plants.

SUBMITTER: Sadat A

PROVIDER: S-EPMC8876356 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature

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Prevalence and Characterization of PVL-Positive Staphylococcus aureus Isolated from Raw Cow's Milk.

Sadat Asmaa A Shata Radwa Reda RR Farag Alshimaa M M AMM Ramadan Hazem H Alkhedaide Adel A Soliman Mohamed Mohamed MM Elbadawy Mohamed M Abugomaa Amira A Awad Amal A

Toxins 20220125 2

The present study aimed to investigate the prevalence, antibiotic susceptibility profiles, and some toxin genes of Panton-Valentine leukocidin (PVL)-positive Staphylococcus aureus (S. aureus) in unpasteurized raw cow's milk collected from retail outlets located at Mansoura, Dakahliya governorate, Egypt. In that context, a total of 700 raw cow's milk samples were investigated for the presence of S. aureus, which was identified in 41.1% (288/700) of the samples. Among the S ...[more]

PMID: 35202125

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Project description:Staphylococcus aureus is one of the most important food-borne pathogens globally. It produces various toxins and invasive enzymes and can be found in numerous food products. Milk is an important source of staphylococcal food poisoning. After pasteurization, this microorganism or its enterotoxins might still remain in pasteurized milk. Therefore, this study was to investigate the contamination of S. aureus in 258 pasteurized milk from 39 cities of China. The prevalence and levels of S. aureus in these samples as well as antibiotic susceptibility profiles, virulence genes, biofilm formation, and biofilm related genes, spa typing and MLST were used to determine the characterization among the isolates. It was found 3.9% of samples were detected S. aureus in 8 of 39 cities in China. The contaminated level were not very excessive which showed the MPN values of the most positive samples (9/10) were less than 1 MPN/g. All pasteurized milk-related S. aureus isolates have ability to produce biofilm and harbored icaA, icaD, eno, clfA, clfB, fnbA, fnbB, fib genes, other biofilm related genes icaC were showed in 91.7% of isolates and cna gene were showed in 50%, except bap gene which were free in all isolates. The antibiotic susceptibility test showed that all isolates were resistant or intermediate-resistant to different concentrations of the antibiotics. Furthermore, 75.0% of the isolates were resistant to three or more antibiotic classes, which indicated multidrug resistance. The isolates had virulence potential, which showed 66.7% (8/12) of the isolates carried one or more virulence-associated genes. Molecular typing by MLST and spa typing enabled classification of these isolates into a total of 11 sequence types (STs) and spa types, which indicated high genetic diversity. Most of these types were related to various clinical S. aureus infections. Thus, the findings of this study reflect the potential risk of S. aureus infection in China. Our study also provides comprehensive analysis of the prevalence of S. aureus in pasteurized milk and helps ensure more accurate treatment of human infection with effective antibiotics.

Project description:Staphylococcus aureus is a common causative agent of bovine mastitis in dairy herds. The emergence of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals as well as the community is a significant and costly public health concern. S. aureus-related bovine mastitis is a common reason for therapeutic and/or prophylactic use of antibiotics on dairy farms. In this study, herd prevalence of S. aureus, including MRSA, was estimated from bulk tank milk (BTM) from Minnesota farms. A total of 150 pooled BTM samples from 50 farms, collected over 3 seasons (spring, summer, and fall of 2009), were assessed. Herd prevalence of methicillin-susceptible S. aureus (MSSA) was 84%, while MRSA herd prevalence was 4%. A total of 93 MSSA isolates and 2 MRSA isolates were recovered from 150 BTM samples. Antibiotic susceptibility testing of S. aureus isolates showed pansusceptibility in 54 isolates, resistance to a single antibiotic class in 21 isolates, resistance to two antibiotic classes in 13 isolates, and resistance to ?3 antibiotics classes and thus multidrug resistance in 5 isolates. The two MRSA isolates displayed resistance to ?-lactams, cephalosporins, and lincosamides and were multiresistant. Staphylococcal protein A gene (spa) typing identified spa types t529 and t034 most frequently among methicillin-susceptible isolates, while t121 was observed in MRSA isolates. Seven isolates, including the two MRSA isolates, produced staphylococcal enterotoxins B, C, D, and E on overnight culture. MRSA isolates were further genotyped using multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Of the 2 MRSA isolates, one had a composite genotype profile of MLST ST 5-PFGE USA100-unknown spa type, which has been reported among hospital-associated MRSA isolates, while the second isolate carried the MLST ST 8-PFGE USA300-spa type t121 genotype, commonly identified among community-associated MRSA isolates. These results suggest that MRSA genotypes associated with hospitals and community can be isolated from milk at very low rates.

Project description:Staphylococcus aureus (S. aureus) is one of the main pathogens in different raw milk and dairy products, which may lead to economic losses. Staphylococcus aureus is a significant and costly public health concern because it may enter the human food chain and contaminate milk causing foodborne illness. This study aimed to investigate the prevalence, antimicrobial susceptibility and virulence genes of S. aureus in raw milks. In total, 125 raw milk samples collected from goat (n = 50), buffalo (n = 25), camel (n = 25), and yak (n = 25) were collected from 5 provinces in China in 2016. Out of 125 samples, 36 (28.8%) S. aureus were isolated (16 from goat, 9 from buffalo, 6 from camel, and 5 from yak). Out of 36 S. aureus, 26 strains (26/36, 72.2%) showed antibiotics resistance, and 6 strains isolated from goats were identified as methicillin-resistant S. aureus (MRSA). The antimicrobial resistance against Penicillin G, tetracycline and gentamicin was 50% (18/36), 41.7% (15/36), and 36.1% (13/36), respectively. 19 S. aureus (52.8%) were considered as multidrug resistant. The highest prevalence of resistant S. aureus was observed in goat milk (13/36, 36.1%). Among the 36 strains, 16 isolates harbored three or more resistant genes. The resistance genes were detected in 25 S. aureus, including 13 strains in goat, 5 strains in buffalo, 4 strains in camel, and 3 strains in yak. Among the 26 resistant strains, 61.5% of isolates harbored three or more resistant genes. The resistance genes were detected in 25 S. aureus, including 13 strains in goat milk, 5 strains in buffalo milk, 4 strains in camel milk, and 3 strains in yak milk. The most predominant resistance genes were blaZ (18/26, 69.2%), aac6'-aph2″ (13/26, 50.0%), and tet(M) (10/26, 38.5%). The mecA, ant(6)-Ia and fexA gene were only detected in S. aureus from goat milk. The most predominant toxin gene were sec (8/26, 30.8%). The majority of S. aureus were multidrug resistant and carried multiple virulence genes, which may pose potential risk to public health. Our findings indicated that the prevalence and antimicrobial resistance of S. aureus was a serious concern in different raw milks in China, especially goat milks.

Project description:BackgroundSome Staphylococcus aureus strains produce Panton-Valentine leukocidin (PVL), a bi-component pore-forming toxin, which causes leukocyte lysis and tissue necrosis. Currently, there is very limited information on the molecular epidemiology of PVL-encoding S. aureus strains in Iran. This study aimed to determine the molecular epidemiology and genetic background of PVL-positive S. aureus clinical strains isolated from Iranian patients.MethodsA total of 28 PVL-positive S. aureus strains were detected from 600 S. aureus isolates between February 2015 and March 2018 from different hospitals in Tehran, Iran. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Molecular genotyping was performed using SCCmec and accessory gene regulator (agr) typing, PVL haplotyping, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE).ResultsThe highest antibiotic resistance rate was found to be against erythromycin (57.1%), followed by ciprofloxacin (42.8%) and clindamycin (35.7%). Moreover, 19 (67.9%) out of 28 S. aureus isolates were identified as MRSA, including CA-MRSA (14/19, 73.7%) and HA-MRSA (5/19, 26.3%). SCCmec type IVa was detected as the predominant type (10/19, 52.6%), followed by type III (5/19, 26.3%) and type V (4/19, 21.1%). The agr type I was identified as the most common type (14/28, 50%), and H and R haplotype groups were observed at frequencies of 67.9 and 32.1%, respectively. Among H variants, the predominant variant was H2 (78/9%). The isolates encompassed 21 different sequence types (STs), including 16 new STs (ST5147 to ST5162). Based on eBURST analysis, the isolates were clustered into five CCs, including CC30, CC22, CC1, CC8, and CC5 (ST5160), and nine singletons. PFGE typing showed that 24 isolates were clustered into A (4 pulsotypes), B (9 pulsotypes), and C (11 pulsotypes) clusters.ConclusionsA high prevalence of PVL-positive CA-MRSA strains was detected in Iran. The majority of PVL-positive isolates were of H (mostly H2) variant, while R variant was harbored by 100% of PVL-positive MRSA strains. Also, CC8, CC22, and CC30 were identified as the dominant clones among PVL-encoding S. aureus strains. This study promotes a better understanding of the molecular epidemiology and evolution of PVL-positive S. aureus strains in Iran.

Project description:Staphylococcus aureus and staphylococcal enterotoxins are a serious public health concern associated with hospital and community-acquired illnesses. Dairy animals frequently shed S. aureus into the milk supply which can lead to food poisoning in humans. This study aims to investigate the prevalence and genetic diversity of S. aureus and staphylococcal enterotoxins in raw milk from the main dairy region of mainland Portugal. S. aureus was found in 53.0% (95% CI: 40.6-65.4%) of 100 raw cow's milk samples collected from bulk cooling tanks. The highest contamination level was 3.4 log10 CFU.mL-1, and in some samples more than one S. aureus strain was identified. Staphylococcal enterotoxins (SEA-SEE) were detected in one sample. Spa typing revealed 62 distinct S. aureus isolates, being t529 (17.7%, 95% CI: 8.2-27.3%) and t1403 (16.1%, 95% CI: 7.0-25.3%) the predominant types, commonly associated with livestock infection or carriage. The antimicrobial susceptibility test showed that 35.5% of the S. aureus isolates were resistant to at least one antimicrobial agent, with resistance to penicillin being the highest (32.3%, 95% CI: 20.6-43.9%) followed by tetracycline (24.2%, 95% CI: 13.5-34.9%), ciprofloxacin (16.1%, 95% CI: 7.0-25.3%) and chloramphenicol (16.1%, 95% CI: 7.0-25.3%). Moreover, five isolates (8.1%, 95% CI: 1.3-14.8%) were identified as methicillin-resistant S. aureus (MRSA, cefoxitin resistant). Regarding virulence/resistance genes, 46,8% (95% CI: 34.4-59.2%) isolates harbored at least one enterotoxin-encoding gene, and the seg gene was the most frequently detected (41.9%, 95% CI: 29.7-54.2%) followed by the sei (40.3%, 95% CI: 28.1-52.5%), sec (6.5%, 95% CI: 0.3-12.6%), seh (4.8%, 95% CI: 0.0-10.2%), and sea (1.6%, 95% CI: 0.0-4.7%) genes. Five (8.1%, 95% CI: 1.3-14.8%) non-enterotoxigenic isolates carried the mecA gene (corresponding to isolates phenotypically classified as MRSA), and 4.8% (95% CI: 0.0-10.2%) enterotoxigenic strains also had the tsst-1 gene. Our study confirm that raw milk can be a zoonotic source of S. aureus, including enterotoxigenic and MRSA strains. Furthermore, the majority of enterotoxigenic isolates were found to contain genes encoding SEs (SEG, SEH and SEI) not routinely screened. This shows the need for a broader SE screening in food safety control, as well as the relevance of risk mitigation measures to control S. aureus transmission along the food chain in Portugal.

Dataset Information

Prevalence and Characterization of PVL-Positive Staphylococcus aureus Isolated from Raw Cow's Milk.

Publications

Prevalence and Characterization of PVL-Positive <i>Staphylococcus aureus</i> Isolated from Raw Cow's Milk.

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