The occurrence of Staphylococcus aureus in fermented milk products (fura and manshanu) in Nigeria

Ninety-three samples of fermented milk cereal (Fura) and 79 of local butter (Manshanu) were collected from four different markets around Zaria. For Fura the mean content of Staphylococci for each of the four markets ranged from 4.5 × 10³ to 4.3 × 10⁴ cfu/ml and the mean aerobic mesophilic plate count from 5.6 × 10⁵ to 2.7 × 10⁶ cfu/ml. For Manshanu the mean staphylococcal count and aerobic mesophilic plate count ranged from 3.4 × 10² to 2.2 × 10³ cfu/ml and 6.7 × 10⁴ to 1.1 × 10⁶ cfu/ml respectively. Significant differences were seen between the different markets.     

Occurrence and characterization of Staphylococcus aureus isolated from raw milk for cheesemaking

Fifty-four samples of raw milk for cheesemaking were tested for the presence of Staphylococcus aureus . Multiplex polymerase chain reactions were performed to identify presumptive isolates and the presence of enterotoxin-coding genes sea-see . The strains were tested for antibiotic resistance. Eighty strains were identified as S. aureus and 31 of these carried one or more enterotoxin genes ( sea-see ). Resistance to eritromycin, penicillin and ampicillin was widespread among isolates . Staphylococcus aureus in raw milk for cheesemaking may constitute a risk with respect to staphylococcal food poisoning from raw milk products.     

云南乳饼中金黄色葡萄球菌污染状况及其耐药性分析

目的了解云南省乳饼中主要致病菌的污染状况及耐药情况。方法采集云南省乳饼主产地区样品62件,用传统方法结合基质辅助激光解吸电离飞行时间质谱进行分离鉴定,采用微量肉汤稀释法对金黄色葡萄球菌进行12种抗生素敏感性分析。结果共检出18株金黄色葡萄球菌,检出率为29.03%,沙门氏菌和单增李斯特氏菌均未检出。农贸市场、加工店和奶牛养殖户检出率分别为44.44%、26.67%和16.67%(P=0.321),超市和网店的样品均未检出;散装和包装样品检出率分别为34.38%、23.33%(P=0.433)。所有菌株的耐药性如下:青霉100%,红霉素44.44%,复方新诺明33.33%,苯唑西林、克林霉素和头孢西丁的耐药率均为27.78%,四环素22.22%,多重耐药率为72.22%。18株菌对环丙沙星、达托霉素、万古霉素、氯霉素和庆大霉素均为敏感。结论金黄色葡萄球菌为乳饼中的主要食源性致病菌,其对抗生素耐药性较普遍,需加强畜禽养殖及乳饼加工卫生条件监管。     

A note on the incidence and antibiotic resistance of Staphylococcus aureus isolated from meat and chicken samples

One hundred and fifty samples of raw calf/lamb meat samples (mince and chunks) and chicken parts (giblets, carcass) were analysed for the presence of Staphylococcus aureus. Eighty S. aureus strains were isolated and identified. Resistance of the strains to methicillin and other antibiotics was determined by the Kirby-Bauer disc diffusion test. The overall methicillin resistance rate for S. aureus was 67.5%. Of S. aureus strains, 87.5% were resistant to bacitracin. A high prevalence of penicillin G resistance was detected for S. aureus (53.8%). Few of the strains were resistant to erythromycin (7.5%). All strains were susceptible to vancomycin, sulbactam–ampicillin, ciprofloxacin and cefaperazone–sulbactam. This study confirmed the presence of S. aureus, especially antibiotic-resistant strains, in the foods examined, indicating poor sanitary conditions during processing which may create a health risk for consumers.     

免疫磁球捕获-PCR检测牛奶中金黄色葡萄球菌的研究

对免疫磁球捕获-PCR（IMS-PCR）检测牛奶中金黄色葡萄球菌进行了初步研究。优化了免疫磁球制备参数,确定了金黄色葡萄球菌免疫磁球对目标菌的结合时间。研究结果显示,当纯菌浓度为101¹04CFU/m L水平时,金黄色葡萄球菌免疫磁球对目标菌的捕获率大于80%。通过对目标菌和非目标菌的检测,IMS-PCR检测方法显示了很强的特异性;在纯培养、无需增菌情况,IMS-PCR检测方法检测限为104CFU/m L;牛奶中的金黄色葡萄球菌经磁分离后增菌2h用PCR检测,可检测出104CFU/m L的金黄色葡萄球菌。      

Prevalence of multidrug resistant Staphylococcus aureus in milk from large- and small-scale producers in Kenya

This study evaluated the prevalence of multidrug resistant Staphylococcus aureus in Kenyan milk and investigated any differences in antimicrobial resistance between large- (>200 L/d) and small- (<50 L/d) scale producers. Susceptibility profiles for penicillin G, tetracycline, erythromycin, trimethoprim/sulfamethazine, and chloramphenicol were determined for Staph. aureus (n=402) isolated from cows with subclinical mastitis. There was a significant difference in the overall mean resistance profile between large- (7.1%) and small-scale farm (14.7%) isolates. The overall prevalence of multidrug resistance (> or =2 antibiotics) differed significantly between isolates from small farms (34.3%) and those from large farms (18.0%). Additionally, the producers were interviewed about their usage of antimicrobial drugs and their attitudes toward education in related fields. There was an evident difference between the producer types in their documentation of the use of antimicrobial drugs. Small-scale farms were less inclined to documentation, and treatment records were available from 22% of small-scale farms, compared with 73% of large-scale farms. Farmers expressed a need for more information in 5 areas, ranking preventive management highest (34.0%); followed by affordable tests to control residues in milk (22.8%); preparation of antimicrobial drugs (20.0%); public health concerns (11.2%); disposal of surplus antimicrobial drugs (7.8%); and antimicrobial drug residue persistence in milk (4.2%). It was concluded that herd size might be an indirect risk factor in the development of antimicrobial resistance in Staph. aureus within the region.     

A large meta-analysis of the global prevalence rates of S. aureus and MRSA contamination of milk

ABSTRACT

Past reports have indicated a high prevalence of milk contaminated with Staphylococcus aureus (S. aureus) and methicillin-resistant Staphylococcus aureus (MRSA), but the pooled prevalence rates of S. aureus and MRSA in pasteurized and boiled cow's milk, raw cow's milk, and raw Caprinae milk (raw sheep's milk and raw goat's milk) and across different periods, continents, economic conditions and purchase locations remain inconsistent. We searched relevant articles published in PubMed, EMBASE, and Web of Science before July 2016. The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement was used to evaluate the quality of 93 included studies. We observed that the pooled prevalence rates of S. aureus contamination in pasteurized and boiled cow's milk, raw cow's milk, and raw Caprinae milk were 15.4% (95% CI, 6.1–27.5%), 33.5% (95% CI, 29.5–37.7%) and 25.8% (95% CI, 17.5–35.0%), respectively. The pooled prevalence rates of MRSA contamination were 4.9% (95% CI, 0.0–15.7%), 2.3% (95% CI, 1.3–3.6%), and 1.1% (95% CI, 0.5–1.8%), respectively. The prevalence of S. aureus contamination in raw cow's milk increased over time. However, the pooled prevalence of raw cow's milk contaminated with S. aureus was lowest in European studies. These findings give an indication of the consequence of better milk regulation in Europe. High S. aureus prevalence rates in raw milk collected from farms and processing companies pose a potential threat to consumers. The implementation of good hygiene practices, appropriate health knowledge, and food safety principles at the farm level, as well as the prudent use of antibiotics in veterinary medicine and heat treatment before drinking, are necessary to reduce the potential risk of S. aureus and MRSA contamination.     

利用水牛乳资源优势,开发南方特色乳制品

介绍了我国奶水牛产业的发展状况，水牛乳的理化、加工特性，水牛乳的加工状况，干酪在我国的发展状况，分析了我国南方发展水牛乳干酪的条件、优势和前景，提出了南方发展水牛乳干酪的设想。     

Antimicrobial Effect of Bacteriocin KU24 Produced by Lactococcus lactis KU24 against Methicillin‐Resistant Staphylococcus aureus

Bacteriocin KU24 produced by Lactococcus lactis KU24 exhibited an inhibitory effect against methicillin‐resistant Staphylococcus aureus (MRSA). Bacteriocin KU24 was inactivated by protease XIV, showing that it has a proteinaceous nature on S. aureus ATCC 33591. Also, bacteriocin KU24 exhibited a strong heat stability (121 °C for 15 min) and pH stability (pH 3 to 9). The mode of inhibition was determined for S. aureus ATCC 33591 by treatment of 0, 250, and 500 AU/mL of bacteriocin KU24. S. aureus ATCC 33591 was inhibited by added bacteriocin KU24, while control was increased. The cell membranes of S. aureus ATCC 33591 were damaged with treatment of 500 AU/mL of bacteriocin KU24. Also, bacteriocin KU24 inhibited the occurrence of mecA gene, the methicillin resistance gene in S. aureus ATCC 33591. Bacteriocin KU24 was purified by C₁₈ Sep‐Pack column, cation exchange chromatography, and gel filtration chromatography, and molecular mass is approximately 6.5 kDa by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. These results demonstrate that bacteriocin KU24 can be used as an alternative antimicrobial agent for the treatment of infection of MRSA in the food industry.     

Occurrence and antibiotic resistance of Escherichia coli,Staphylococcus aureus and Bacillus cereus in raw milk and dairy products in Turkey

In this survey, 150 samples of raw milk, white cheese and ice cream from three different dairy‐processing plants in Ankara were analysed to find out if they were contaminated with Escherichia coli, Staphylococcus aureus or Bacillus cereus. The highest contamination percentages were found in raw milk samples as follows: B. cereus (90%), E. coli (74%) and S. aureus (56%) followed by cheese (70% B. cereus, 60% E. coli, and 48% S. aureus) and ice cream (56% E. coli, 36% S. aureus and 20% B. cereus). The survey showed that 2% of cheese samples were contaminated with E. coli O157. It was also found that the numbers of S. aureus and E. coli in raw milk, cheese and ice cream samples exceeded the numbers permitted under the Turkish Food Codex (TFC). The number of B. cereus in raw milk, cheese and ice cream samples was lower than the limit given in the TFC standards. The study also showed that E. coli and S. aureus exhibit resistance to ampicillin, penicillin, tetracycline, erythromycin, gentamicin and trimethoprim/sulfamethoxazole. Escherichia coli isolates also showed resistance to chloramphenicol and ciprofloxacin but none of them exhibited resistance to cefotaxime. All S. aureus isolates were found to be susceptible to cefotaxime, chloramphenicol, and ciprofloxacin. Bacillus cereus isolates were found to be resistant to ampicillin, penicillin and trimethoprim/sulfamethoxazole and sensitive to cefotaxime, chloramphenicol, ciprofloxacin erythromycin, gentamicin and tetracycline.     

2016—2020年上海地区乳源金黄色葡萄球菌耐药性及最低抑菌浓度变迁情况分析

分析上海地区乳源金黄色葡萄球菌对常见抗菌药物的耐药性及其MIC变迁情况,加强致病菌耐药性检测。方法 采用微量肉汤稀释法对2016-2020年间采集的349株金黄色葡萄球菌进行10种常见抗菌药物敏感性测试,通过分析药敏数据了解5年间上海地区乳源金黄色葡萄球菌耐药性变迁的情况。结果 349株金黄色葡萄球菌对青霉素（74.2%）耐药率较高,对其余9种抗菌药物的耐药率较低,均在20%以下。五年间,上海地区乳源金黄色葡萄球菌对青霉素、苯唑西林及阿莫西林/克拉维酸的抑制50%微生物生长的抑菌浓度（minimal inhibitory concentration,MIC）MIC50值及抑制90%微生物生长的抑菌浓度MIC90值均呈下降趋势;对头孢西丁、头孢噻呋、恩诺沙星、氧氟沙星及庆大霉素的MIC50值五年来无明显变化,MIC90值呈下降趋势;对万古霉素和利奈唑胺的MIC50值和MIC90值五年来无明显变化,均处于较低水平。结论 金黄色葡萄球菌的耐药情况近年来有好转趋势,但仍需继续加强乳源金黄色葡萄球菌耐药性监测,为科学合理使用抗菌药物提供技术支撑。     

金黄色葡萄球菌在生乳与灭菌乳中生长状况分析

目的 了解并比较金黄色葡萄球菌(S.aureus)在生乳与灭菌乳中的生长状况.方法 采用GB/T4789.37-2008<食品微生物学检验金黄色葡萄球菌检验>,测定不同温度下金黄色葡萄球菌在生乳和灭菌乳中的生长情况,利用MicroFit 1.0软件对所测数据进行分析,利用SPSS统计软件对该菌在生乳与灭菌乳中生长参数进行统计分析.结果 金黄色葡萄球菌在生乳中的最大生长速率明显低于在灭菌乳中的最大生长速率,二者的迟滞期、代时无明显差异.结论 金黄色葡萄球菌在生乳与灭菌乳中的生长情况差异显著.有统计学意义(P<0.05).     

Comparison of the antipathogenic effect toward Staphylococcus aureus of N-linked and free oligosaccharides derived from human,bovine, and goat milk

N-linked oligosaccharides (N-glycans) derived from milk were recently found to be antipathogenic. This study compares the antimicrobial activity of N-linked glycans and free oligosaccharides from human, bovine, and goat milk against Staphylococcus aureus. Milk N-glycans showed a bactericidal/bacteriostatic effect on the pathogen when compared to free milk oligosaccharides, evidenced by the clear zone from the halo assay, with the order of human milk >goat milk >bovine milk. None of the free milk oligosaccharide samples were bactericidal/bacteriostatic, despite its positive results in growth curve and minimum inhibitory concentration (MIC) assays which are believed to be related to hyperosmosis. Both N-glycans and free milk oligosaccharides can reduce the adhesion of Staphylococcus aureus to Caco-2 cells, however, N-glycans worked significantly more effective than free milk oligosaccharides. Structural analysis of all free oligosaccharide and N-glycan samples showed the obvious interspecies differences, and the structure/function relationship of the respected N-glycans is of interest for future study. The significant bactericidal/bacteriostatic activity possessed by human, bovine, and goat milk N-linked glycans holds great potential as a novel substitute for antibiotics.     

Species Diversity and Pheno‐ and Genotypic Antibiotic Resistance Patterns of Staphylococci Isolated from Retail Ground Meats

The presence and species diversity of staphylococci in 250 ground beef and lamb meat samples obtained from Diyarbakir, Turkey were investigated. The presence of the 16S rRNA gene, mecA, nuc, pvl, and femA was analyzed by multiplex PCR. Pheno‐ and genotypic antibiotic resistance profiles of 208 staphylococci isolates were established. Of the ground beef and ground lamb samples, 86.4% and 62.4% were positive for staphylococci, respectively. Staphylococcus aureus, S. saprophyticus, S. hominis, S. lentus, S. pasteuri, S. warneri, S. intermedius, and S. vitulinus made up 40.8%, 28.8%, 11%, 3.8%, 3.8%, 2.4%, 2.4%, and 2.4% of isolates, respectively. Of the 85 S. aureus isolates, 40%, 47%, and 5.8% carried femA, mecA, and pvl, respectively, whereas the corresponding rates for the 118 coagulase‐negative staphylococci (CoNS) were 0%, 10.1%, and 0%, respectively. We determined from the 208 isolates, the highest antibiotic resistances were to tetracycline and oxytetracycline (85.5%), followed by penicillin (51.4%), novobiocin (45.6%), ampicillin (39.9%), and doxycycline (31.7%), using the Clinical and Laboratory Standards Inst. (CLSI) method. All isolates were sensitive to gentamycin, ofloxacin, and tobramycin, but 2.3% of the S. aureus isolates had resistance to vancomycin. The staphylococci isolates carried tet(K), blaZ, tet(L), tet(W), cat, tet(S), tet(M), ermB, ermA, and ermC antibiotic resistance genes at rates of 59%, 51.7%, 36.9%, 31.8%, 27.2%, 27.2%, 24.4%, 18.1%, 7.9%, and 3.9%, respectively.     

Short communication: Space allocation in intensive Mediterranean buffalo production influences the profile of functional biomolecules in milk and dairy products

The aim of the present study was to determine if space allocation influenced the concentration of biomolecules in buffalo milk and dairy products. Intensively housed buffaloes (n = 96) were randomly assigned to 2 groups according to days in milk, parity, and milk yield: group S10 had a space allocation of 10 m² per buffalo and group S15 had a space allocation of 15 m² per buffalo. Individual milk yield was recorded daily. Twice a month, a bulk milk sample was collected for each group, as well as whey, ricotta, and mozzarella cheese, to assess cheese yield and to conduct HPLC-electrospray ionization-tandem mass spectrometry, milk antioxidant activity, and cell viability analyses. We tested milk extracts from the 2 groups in vitro to evaluate their efficacy in counteracting endothelial oxidative damage induced by high glucose. We evaluated reproductive function in 28 buffaloes from each group using the Ovsynch-timed artificial insemination program. We observed no differences in milk quantity or quality in terms of fat, protein, or lactose, and reproductive function did not differ between the 2 groups. Compared with group S10, group S15 had higher concentrations of carnitine (56.7 ± 1.1 vs. 39.8 ± 0.7 mg/L in milk and 40.9 ± 0.8 vs. 31.7 ± 0.7 mg/L in whey), acetyl-l-carnitine (51.9 ± 0.3 vs. 39.7 ± 0.7 mg/L in milk and 41.1 ± 1.7 vs. 28.7 ± 2.6 mg/L in whey), propionyl-l-carnitine (34.8 ± 1.0 vs. 21.0 ± 0.9 mg/L in milk and 26.9 ± 0.8 vs. 17.6 ± 1.2 mg/L in whey), glycine betaine (23.1 ± 1.9 vs. 13.5 ± 1.6 mg/L in milk and 10.7 ± 0.4 vs. 7.9 ± 0.5 mg/L in whey), and δ-valerobetaine (24.2 ± 0.5 vs. 16.7 ± 0.5 mg/L in milk and 22.0 ± 0.9 vs. 15.5 ± 0.7 mg/L in whey). Group S15 also had higher total antioxidant activity than group S10 (56.7 ± 1.9 vs. 46.4 ± 1.13 mM Trolox equivalents). Co-incubation of high-glucose-treated endothelial cells with milk extracts from group S15 improved cell viability compared with cells treated with high glucose only; it also reduced intracellular lipid peroxidation (144.3 ± 0.4 vs. 177.5 ± 1.9%), reactive oxygen species (141.3 ± 0.9 vs. 189.3 ± 4.7 optical density units), and cytokine release (tumor necrosis factor-α, IL-1β, IL-6). Greater space allocation was associated with higher levels of biomolecules in buffalo milk. This could have been the result of improved welfare in buffaloes that were allocated more space.     

多重实时荧光定量PCR快速检测婴幼儿奶粉中沙门氏菌、克罗诺杆菌和金黄色葡萄球菌

目的建立多重实时荧光定量PCR(multiplex quantitative real-time PCR,multiplex qPCR)快速检测奶粉中金黄色葡萄球菌、沙门氏菌和克罗诺杆菌3种常见致病菌的方法。方法筛选目标菌株的特异性引物与探针,优化反应体系,建立稳定的多重q PCR反应体系。通过阳性菌株加标的方式验证体系的特异性,并确定人工污染奶粉的检出限。结果各对引物探针对目标菌株均能扩增,多重实时荧光PCR未发现交叉反应,对17株非目标菌进行检测均未检出,人工污染奶粉中克罗诺杆菌和沙门氏菌的检出限均为10~3 CFU/mL,金黄色葡萄球菌的检出限为10~4 CFU/mL。结论本研究方法可实现婴幼儿奶粉样品中3种致病菌qPCR高效率检测。