Phenotypic and genotypic methods for identification of slime layer production, efflux pump activity, and antimicrobial resistance genes as potential causes of the antimicrobial resistance of some mastitis pathogens from farms in Menoufia, Egypt

El-Sayed, Mohamed; Roshdey, Tamer; Salah, Ahmed; Tarabees, Reda; Younis, Gamal; Eldeep, Doaa

doi:10.1007/s11033-019-05099-6

Cited by 10 publications

(4 citation statements)

References 75 publications

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“…Staphylococcus aureus has various implications within nosocomial diseases and foodborne illnesses in terms of public health and economic effect. S. aureus has caused 1,681 illnesses and 86 hospitalizations reported in the foodborne-associated outbreaks in the United States (Dewey-Mattia et al, 2018); decreased animal production and milk production caused by clinical and subclinical mastitis in dairy animals; and increased use of antimicrobials for the treatment and prevention of mastitis and numerous diseases, including abscesses, septicemia, and pneumonia, which could lead to the emergence of antimicrobial resistance (Kumar et al, 2010;Akanbi et al, 2017;Abdi et al, 2018;Elsayed et al, 2019;Zhang et al, 2020). In addition, S. aureus can form biofilm and could be involved in 65-85% of microbial and chronic infections that are associated with biofilm formation reported by the National Institutes of Health (Flemming et al, 2016;Jamal et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Genetic and compositional analysis of biofilm formed by Staphylococcus aureus isolated from food contact surfaces

et al. 2022

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IntroductionStaphylococcus aureus is an important pathogen that can form biofilms on food contact surfaces (FCS) in the dairy industry, posing a serious food safety, and quality concern. Biofilm is a complex system, influenced by nutritional-related factors that regulate the synthesis of the components of the biofilm matrix. This study determines the prevalence of biofilm-associated genes and evaluates the development under different growth conditions and compositions of biofilms produced by S. aureus.MethodsBiofilms were developed in TSB, TSBG, TSBNaCl, and TSBGNaCl on stainless-steel (SS), with enumeration at 24 and 192 h visualized by epifluorescence and scanning electron microscopy (SEM). The composition of biofilms was determined using enzymatic and chemical treatments and confocal laser scanning microscopy (CLSM).Results and discussionA total of 84 S. aureus (SA1–SA84) strains were collected from 293 dairy industry FCS (FCS-stainless steel [n = 183] and FCS-polypropylene [n = 110]) for this study. The isolates harbored the genes sigB (66%), sar (53%), agrD (52%), clfB/clfA (38%), fnbA/fnbB (20%), and bap (9.5%). 99. In particular, the biofilm formed by bap-positive S. aureus onto SS showed a high cell density in all culture media at 192 h in comparison with the biofilms formed at 24 h (p < 0.05). Epifluorescence microscopy and SEM revealed the metabolically active cells and the different stages of biofilm formation. CLSM analysis detected extracellular polymeric of S. aureus biofilms on SS, such as eDNA, proteins, and polysaccharides. Finally, the level of detachment on being treated with DNase I (44.7%) and NaIO 4(42.4%) was greater in the biofilms developed in TSB compared to culture medium supplemented with NaCl at 24 h; however, there was no significant difference when the culture medium was supplemented with glucose. In addition, after treatment with proteinase K, there was a lower level of biomass detachment (17.7%) of the biofilm developed in TSBNaCl (p < 0.05 at 24 h) compared to that in TSB, TSBG, and TSBGNaCl (33.6, 36.9, and 37.8%, respectively). These results represent a deep insight into the composition of S. aureus biofilms present in the dairy industry, which promotes the development of more efficient composition-specific disinfection strategies.

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Section: Introductionmentioning

confidence: 99%

Genetic and compositional analysis of biofilm formed by Staphylococcus aureus isolated from food contact surfaces

et al. 2022

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“…The tested isolates contained resistance genes, including ermA (26.7%), ermB (10.8%), ermC (11.7%). Also, The study ofElsayed et al, (2019) demonstrated the high antimicrobial resistance of the investigated isolates. A total of 20 methicillin-resistant S. aureus (MRSA) isolates.…”

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confidence: 75%

Phenotypic and genotypic characterization of macrolide resistance in Staphylococcus aureus isolates from wound infection

Ali

Al-Amara²

2022

ijhs

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Microbiologists are increasingly concerned about the rise in S. aureus MLSB (macrolide, lincosamide, streptogramin B) drug resistance. Clindamycin has been effective in treating infections by S. aureus, and the variations in clindamycin sensitivity patterns cause treatment to fail. Inducible clindamycin resistance in S. aureus is expressed via erythromycin ribosome methylase genes. In the current study, 25 S. aureus isolates were identified by conventional chemical tests and the Vitek®2 system. Specific primers were used for the amplification of Macrolide genes by PCR. Among 25 S. aureus isolates, 23(92%) isolates were methicillin resistant and 2(8%) isolates were methicilin sensitive. The 5(20%) isolates showed resistance to Erythromycin and sensitivity to Clindamycin with a positive D test which was identified as inducible MLSB, while 2(8%) isolates showed resistance criteria for both Erythromycin and Clindamycin which identity as a constitutive MLSB and 18(92%) isolates were given the sensitivity for both Erythromycin and Clindamycin. The erm resistance genes (ermA, ermB, ermC, ermF, and ermG) were detected in 5(20%), 17(68%), 25(100%), 24(96%),11(44%) respectively. The D-test, and Vitek ®2 system should be routinely done to avoid treatment failure due to clindamycin resistance.

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“…For this reason, the detection of antibioresistance genes directly from biological samples could be a useful tool in Tunisia and other countries where the bla CTX-M gene clusters were predominant. The CTX-M, TEM, and SHV enzymes were among the most common variants in Tunisia [ 27 ], in Palestine, in Egypt [ 28 ], and in European region [ 29 ] with varying prevalence rates. These results are similar to our qPCR data.…”

Section: Discussionmentioning

confidence: 99%

Rapid Detection of Beta-Lactamases Genes among Enterobacterales in Urine Samples by Using Real-Time PCR

Yengui

Trabelsi

Khannous

et al. 2022

BioMed Research International

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The objective of this study was to develop and evaluate newly improved, rapid, and reliable strategies based on real-time PCR to detect the most frequent beta-lactamase genes recorded in clinical Enterobacterales strains, particularly in Tunisia (blaSHV12, blaTEM, blaCTX-M-15, blaCTX-M-9, blaCMY-2, blaOXA-48, blaNDM-1, and blaIMP) directly from the urine. Following the design of primers for a specific gene pool and their validation, a series of real-time PCR reactions were performed to detect these genes in 78 urine samples showing high antibiotic resistance after culture and susceptibility testing. Assays were applied to DNA extracted from cultured bacteria and collected urine. qPCR results were compared for phenotypic sensitivity. qPCR results were similar regardless of whether cultures or urine were collected, with 100% sensitivity and specificity. Out of 78 multiresistant uropathogenic, strains of Enterobacterales (44 E. coli and 34 K. pneumoniae strains) show the presence of the genes of the bla group. In all, 44% E. coli and 36 of K. pneumoniae clinical strains harbored the bla group genes with 36.4%, 52.3%, 70.5%, 68.2%, 18.2%, and 4.5% of E. coli having blaSHV-12, blaTEM, blaCTX-M 15, blaCTX-M-9, blaCMY-2, and blaOXA-48 group genes, respectively, whereas 52.9%, 67.6%, 76.5%, 35.5%, 61.8, 14.7, and 1.28% of K. pneumoniae had blaSHV-12, blaTEM, blaCTX-M 15, blaCTX-M-9, blaCMY-2, blaOXA-48, and blaNDM-1 group genes, respectively. The time required to have a result was 3 hours by real-time PCR and 2 to 3 days by the conventional method. Resistance genes of Gram-negative bacteria in urine, as well as cultured bacteria, were rapidly detected using qPCR techniques. These techniques will be used as rapid and cost-effective methods in the laboratory. Therefore, this test could be a good candidate to create real-time PCR kits for the detection of resistance genes directly from urine in clinical or epidemiological settings.

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Phenotypic and genotypic methods for identification of slime layer production, efflux pump activity, and antimicrobial resistance genes as potential causes of the antimicrobial resistance of some mastitis pathogens from farms in Menoufia, Egypt

Cited by 10 publications

References 75 publications

Genetic and compositional analysis of biofilm formed by Staphylococcus aureus isolated from food contact surfaces

Genetic and compositional analysis of biofilm formed by Staphylococcus aureus isolated from food contact surfaces

Phenotypic and genotypic characterization of macrolide resistance in Staphylococcus aureus isolates from wound infection

Rapid Detection of Beta-Lactamases Genes among Enterobacterales in Urine Samples by Using Real-Time PCR

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