scholarly journals Molecular detection of methicillin heat-resistant Staphylococcus aureus strains in pasteurized camel milk in Saudi Arabia

2020 ◽  
Vol 40 (4) ◽  
Author(s):  
Amani H. Aljahani ◽  
Khaloud M. Alarjani ◽  
Zeinab K. Hassan ◽  
Manal F. Elkhadragy ◽  
Elsayed A. Ismail ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Saudi Arabia ◽  
Sodium Dodecyl ◽  
Meca Gene ◽  
Camel Milk ◽  
Resistant Bacteria ◽  
Local Alignment ◽  
Nucleotide Sequencing ◽  
Methicillin Resistant ◽  
Heat Resistant

Abstract Antibiotic- and heat-resistant bacteria in camel milk is a potential public health problem. Staphylococcus aureus (S. aureus) is an opportunistic pathogen in humans, dairy cattle and camels. We characterized the phenotype and genotype of methicillin-resistant staphylococcal strains recovered from pasteurized and raw camel milk (as control) distributed in the retail markets of Saudi Arabia. Of the 100 samples assessed between March and May 2016, 20 S. aureus isolates were recovered from pasteurized milk, 10 of which were resistant to cefoxitin, and as such, were methicillin-resistant. However, raw camel milk did not contain methicillin-resistant S. aureus (MRSA). Antimicrobial susceptibility tests showed that the resistance ratio for other antibiotics was 60%. We performed a polymerase chain reaction (PCR) assay using primers for the methicillin-resistant gene mecA and nucleotide sequencing to detect and verify the methicillin-resistant strains. Basic local alignment search tool (BLAST) analysis of the gene sequences showed a 96–100% similarity between the resistant isolates and the S. aureus CS100 strain’s mecA gene. Ten of the methicillin-resistant isolates were heat-resistant and were stable at temperatures up to 85°C for 60 s, and three of these were resistant at 90°C for 60 or 90 s. The mean decimal reduction time (D85-value) was 111 s for the ten isolates. Sodium dodecyl sulfate (SDS)/polyacrylamide gel electrophoresis (PAGE) showed that there was no difference in the total protein profiles for the ten methicillin heat-resistant S. aureus (MHRSA) isolates and for S. aureus ATCC 29737. In conclusion, a relatively high percentage of the tested pasteurized camel milk samples contained S. aureus (20%) and MHRSA (10%).

scholarly journals Prevalence of methicillin-resistant (mecA gene) and heat-resistant Staphylococcus aureus strains in pasteurized camel milk

2020 ◽  
Vol 103 (7) ◽  
pp. 5947-5963
Author(s):  
Hany M. Yehia ◽  
Abdulrahman H. Al-Masoud ◽  
Khaloud M. Alarjani ◽  
Mohamed S. Alamri
Keyword(s):  
Staphylococcus Aureus ◽  
Meca Gene ◽  
Camel Milk ◽  
Methicillin Resistant ◽  
Heat Resistant

scholarly journals Occurrence of Methicillin-Resistant Coagulase-Negative Staphylococci (MRCoNS) and Methicillin-Resistant Staphylococcus aureus (MRSA) from Pigs and Farm Environment in Northwestern Italy

Antibiotics ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 676
Author(s):  
Miryam Bonvegna ◽  
Elena Grego ◽  
Bruno Sona ◽  
Maria Cristina Stella ◽  
Patrizia Nebbia ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Human Health ◽  
Environmental Samples ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Bacterial Species ◽  
Meca Gene ◽  
Nucleotide Sequencing ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant ◽  
Nasal Swabs

Swine farming as a source of methicillin-resistant Staphylococcus aureus (MRSA) has been well documented. Methicillin-resistant coagulase-negative staphylococci (MRCoNS) have been less studied, but their importance as pathogens is increasing. MRCoNS are indeed considered relevant nosocomial pathogens; identifying putative sources of MRCoNS is thus gaining importance to prevent human health hazards. In the present study, we investigated MRSA and MRCoNS in animals and environment in five pigsties in a high farm-density area of northwestern Italy. Farms were three intensive, one intensive with antibiotic-free finishing, and one organic. We tested nasal swabs from 195 animals and 26 environmental samples from three production phases: post-weaning, finishing and female breeders. Phenotypic tests, including MALDI-TOF MS, were used for the identification of Staphylococcus species; PCR and nucleotide sequencing confirmed resistance and bacterial species. MRCoNS were recovered in 64.5% of nasal swabs, in all farms and animal categories, while MRSA was detected only in one post-weaning sample in one farm. The lowest prevalence of MRCoNS was detected in pigs from the organic farm and in the finishing of the antibiotic-free farm. MRCoNS were mainly Staphylococcus sciuri, but we also recovered S. pasteuri, S. haemolyticus, S. cohnii, S. equorum and S. xylosus. Fifteen environmental samples were positive for MRCoNS, which were mainly S. sciuri; no MRSA was found in the farms’ environment. The analyses of the mecA gene and the PBP2-a protein highlighted the same mecA fragment in strains of S. aureus, S. sciuri and S. haemolyticus. Our results show the emergence of MRCoNS carrying the mecA gene in swine farms. Moreover, they suggest that this gene might be horizontally transferred from MRCoNS to bacterial species more relevant for human health, such as S. aureus.

Assessment of the Multifactorial Effect on Antimicrobial Sensitivity in Positive Staphylococcus Aureus Clinical Isolates from Assir Region, Saudi Arabia

2012 ◽  
Vol 13 (2) ◽  
pp. 152-159 ◽  
Author(s):  
Nazar M Abdalla ◽  
Waleed O Haimour ◽  
Amani A Osman ◽  
Hassan Abdul Aziz
Keyword(s):  
Staphylococcus Aureus ◽  
Saudi Arabia ◽  
Culture Media ◽  
Meca Gene ◽  
Laboratory Data ◽  
Age Groups ◽  
Clinical Isolates ◽  
Diabetic Patients ◽  
Staph Aureus ◽  
Antimicrobial Sensitivity

General objectives: This study aimed at assessment of factors affecting antimicrobial sensitivity in Staphylococcus aureus clinical isolates from Assir region, Saudi Arabia. Materials and Methods: In this study, eighty one patients presented with Staph. aureus infections either nosocomial or community acquired infections were involved by collecting nasal swabs from them at Aseer Central Hospital General Lab. These patients were from all age groups and from males and females during the period of Jan 2011- Jun 2011. These samples were undergone variable laboratory procedures mainly; bactech, culture media, antibiotics sensitivity test using diffusion disc test (MIC) and molecular (PCR) for detection of mec A gene. Clinical and laboratory data were recorded in special formats and analyzed by statistical computer program (SPSS). Results: Showed that; Descriptive and analytical statistical analysis were performed and final results were plotted in tables. In Staph aureus MecA gene positive cases (50) showed: Oxacillin/ Mithicillin, Ciprofloxacin and Fusidin resistant in diabetic patients were 13, 26.0%, 9, 18% and 7, 14% respectively and in non diabetic patients were 37, 74.0%, 22, 44% and 20, 40% respectively. While no sensitivity in diabetic and non diabetic patients using Oxacillin/ Mithicillin. In Staph aureus MecA gene negative cases (31) showed: Oxacillin/ Mithicillin, sensitivity in diabetic patients (5, 16.1%) and in non diabetic were (26, 83.9%). While no resistant in diabetic and non diabetic patients. In Ciprofloxacin and Fusidin resistant in diabetic patients were 1, 3.2% and 1, 3.2% respectively and in non diabetic patients were 12, 38.7% and 7, 22.6%respectively. Erythromycin in Staph aureus ( MecA gene) positive cases (50) showed: resistant in age (0-15) years were (5, 10%), (16-50) years were (16, 32%) and ( ›50 years) were (12, 24%). Erythromycin in Staph aureus (MecA gene) negative cases (31) showed: resistant in age (0-15) years were (6, 19.3%), (16-50) years were (5, 16.1%) and ( ›50 years) were (3, 9.7%). Conclusion: Drugs resistance is a major progressive multifactorial problem facing the treatment of Staph aureus infections. DOI: http://dx.doi.org/10.3329/jom.v13i2.12750 J Medicine 2012; 13 : 152-159

scholarly journals Mutation-Based Antibiotic Resistance Mechanism in Methicillin-Resistant Staphylococcus aureus Clinical Isolates

2021 ◽  
Vol 14 (5) ◽  
pp. 420
Author(s):  
Tanveer Ali ◽  
Abdul Basit ◽  
Asad Mustafa Karim ◽  
Jung-Hun Lee ◽  
Jeong-Ho Jeon ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Active Site ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Resistance Mechanism ◽  
Ligand Docking ◽  
Clinical Samples ◽  
Allosteric Site ◽  
Methicillin Resistant

β-Lactam antibiotics target penicillin-binding proteins and inhibit the synthesis of peptidoglycan, a crucial step in cell wall biosynthesis. Staphylococcus aureus acquires resistance against β-lactam antibiotics by producing a penicillin-binding protein 2a (PBP2a), encoded by the mecA gene. PBP2a participates in peptidoglycan biosynthesis and exhibits a poor affinity towards β-lactam antibiotics. The current study was performed to determine the diversity and the role of missense mutations of PBP2a in the antibiotic resistance mechanism. The methicillin-resistant Staphylococcus aureus (MRSA) isolates from clinical samples were identified using phenotypic and genotypic techniques. The highest frequency (60%, 18 out of 30) of MRSA was observed in wound specimens. Sequence variation analysis of the mecA gene showed four amino acid substitutions (i.e., E239K, E239R, G246E, and E447K). The E239R mutation was found to be novel. The protein-ligand docking results showed that the E239R mutation in the allosteric site of PBP2a induces conformational changes in the active site and, thus, hinders its interaction with cefoxitin. Therefore, the present report indicates that mutation in the allosteric site of PBP2a provides a more closed active site conformation than wide-type PBP2a and then causes the high-level resistance to cefoxitin.

scholarly journals Burden of methicillin-resistant Staphylococcus aureus pneumonia among hospitalized patients in Lebanon and Saudi Arabia

2017 ◽  
Vol Volume 10 ◽  
pp. 49-55 ◽  
Author(s):  
Abdulhakeem O Althaqafi ◽  
Madonna J Matar ◽  
Rima Moghnieh ◽  
Adel F Alothman ◽  
Thamer H Alenazi ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Saudi Arabia ◽  
Hospitalized Patients ◽  
Methicillin Resistant

Methicillin-Resistant Staphylococcus aureus in Two Tertiary-Care Centers in Jeddah, Saudi Arabia

2001 ◽  
Vol 22 (4) ◽  
pp. 211-216 ◽  
Author(s):  
Tariq A. Madani ◽  
Nabeela A. Al-Abdullah ◽  
Ali A. Al-Sanousi ◽  
Tawfik M. Ghabrah ◽  
Shadia Z. Afandi ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Saudi Arabia ◽  
Tertiary Care ◽  
Age Groups ◽  
Clinical Manifestations ◽  
Total Mortality ◽  
Control Measures ◽  
Methicillin Resistant ◽  
Mrsa Infection ◽  
Tertiary Care Hospitals

AbstractObjective:To review clinical experience with methicillin-resistant Staphylococcus aureus (MRSA) in tertiary-care hospitals in Jeddah, Saudi Arabia.Design:Retrospective review for the year 1998.Setting:Two tertiary-care hospitals.Methods:Results of MRSA-positive cultures of clinical specimens obtained as part of investigations for suspected infections were retrieved from the microbiology laboratories' records. Charts of patients were reviewed, with standardized data collection.Results:Of 673 S aureus isolates identified, 222 (33%, or 6.8 isolates/1,000 admissions) were MRSA Overall MRSA prevalence was 2% in 1988. Nosocomial acquisition occurred in 84.2% of cases. All age groups were affected, and 52% of patients had at least one comorbidity. MRSA prevalence was highest in the intensive care units (26.6% of all isolates), the medical wards (24.8%), and the surgical wards (19.8%). Seventy-three percent of isolates caused infection; the rest represented colonization. Surgical wounds (35.2%), the chest (29%), and central venous catheters (13%) were the most common sites of infection. Bacteremia occurred in 15.4% of patients. Local signs (84%) and fever (75.9%) were the most common clinical manifestations. Respiratory distress and septic shock occurred in 30.2% and 13.6% of cases, respectively. Of 162 patients with MRSA infection and 60 patients with MRSA colonization, 95.7% and 70% received antibiotics in the preceding 6 weeks, respectively (P<.0001). The total mortality of patients with MRSA infection was 53.7%: 36.4% as a result of MRSA infection and 17.3% as a result of other causes.Conclusions:The prevalence of MRSA is high and rapidly increasing in the two hospitals, as it is worldwide. Control measures to prevent die spread of MRSA in hospitals should continue, with reinforcement of hygienic precautions and development of policies to restrict the use of antibiotics.

scholarly journals Detection of mecA gene by latex agglutination and its molecular analysis by PCR in methicillin resistant staphylococcus aureus.

2020 ◽  
Vol 27 (07) ◽  
pp. 1363-1370
Author(s):  
Aneela Khawaja ◽  
Iffat Javed ◽  
Sohaila Mushtaq ◽  
Saeed Anwar ◽  
Faiqa Arshad ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Latex Agglutination ◽  
Medical Institute ◽  
Cross Sectional ◽  
Methicillin Resistant ◽  
Agglutination Method ◽  
Resistant Strains

Antimicrobial resistance (AMR) is a devastating question that is threatening the health globally. The extensive and indiscriminative use of antibiotics has evolved a notorious resistance in Staphylococcus aureus.  This resistance developed through possession of mecA gene, which codes for modified penicillin binding protein (PBP2a) and the emergent strain being labeled “methicillin resistant Staphylococcus aureus”. Conventional phenotypic techniques for detection of MRSA rely on standardization of cultural characteristics. The drawbacks of diagnostic error to report MRSA include: poor prognosis, expensive treatment, dissemination of multi-drug resistant strains and even treatment failure. Latex agglutination method can be adopted as a more accurate and quick strategy for rapid detection of methicillin resistance. Objectives: To compare detection of mecA gene in methicillin resistant isolates of Staphylococcus aureus by latex agglutination and PCR; by assessing the sensitivity and specificity of both methods. Study Design: Descriptive Cross-Sectional study. Setting: Pathology Department, Post Graduate Medical Institute, Lahore. Period: From January 2015 to December 2015; according to standard operating procedures at Microbiology laboratory. Material & Methods: A total 713 consecutive, non-duplicate isolates of Staphylococcus aureus were processed. Methicillin resistance was determined using cefoxitin (30mg) by Kirby-Bauer method using CLSI guideline (2016), latex agglutination method; and PCR for mecA gene. Results: The results showed that out of 713 Staphylococcus aureus isolates, 92 (12.90%) isolates were resistant to cefoxitin and were labelled as MRSA. majority MRSA isolates recovered from pus (44.57%) and wound swab (20.65%), followed by blood (13.04%), fluid (8.70%), CSF (4.35%), CVP (3.26%), HVS (3.26%) and tracheal secretion (2.17%). By latex agglutination method, 87 (94.50%) were positive for PBP2a; while on PCR mecA gene was detected only in 82 (89.10%) MRSA isolates. When assessed with PCR (gold standard) the sensitivity and diagnostic accuracy of latex agglutination was 100% and 94.57%, respectively. Conclusion: Latex agglutination test can be employed as rapid and reliable diagnostic technique in MRSA isolates for mecA gene detection, where resources for molecular methods are inadequate. This can effectually lessen the misdiagnosis of resistant strains, and over/ ill-use of antibiotics.

scholarly journals An outbreak of methicillin resistant Staphylococcus epidermidis among neonates in a hospital in Saudi Arabia

2011 ◽  
Vol 5 (10) ◽  
pp. 692-699 ◽  
Author(s):  
Maha Abd El Hafez ◽  
Noha G. Khalaf ◽  
Mohamed El Ahmady ◽  
Ahmed Abd El Aziz ◽  
Abd El Gawad Hashim
Keyword(s):  
Saudi Arabia ◽  
Antibiotic Resistance ◽  
Staphylococcus Epidermidis ◽  
Neonatal Intensive Care ◽  
Meca Gene ◽  
Neonatal Intensive Care Units ◽  
Methicillin Resistant ◽  
Resistance Patterns ◽  
Clonal Origin ◽  
Resistance Rates

Introduction: Staphylococcus epidermidis is a pathogen associated with nosocomial infection in neonatal intensive care units (NICU). This study investigates an outbreak of methicillin resistant S. epidermidis in an NICU in a hospital in Saudi Arabia. Methodology: A total of 41 isolates identified as Gram-positive cocci were obtained from blood culture, umbilical wound swabs and endotracheal aspirate specimens of neonates, of which 29 were identified as S. epidermidis. Bacterial identification at the species level and determination of antibiotic resistance were performed by MicroScan (Dade Behring, USA). Genotyping was completed using randomly amplified polymorphic DNA (RAPD) and the mecA gene was detected by PCR. Results: All 29 S. epidermidis isolates were found to be resistant to oxacillin and were positive for the mecA gene. The isolates showed several multidrug-resistance patterns; the resistance rates to gentamicin, erythromycin, clindamycin, and trimethoprim/sulfamethoxazole were 89.7%, 86.2%, 75.9% and 72.4%, respectively. All isolates were susceptible to vancomycin, teicoplanin, rifampin, synercid, and ciprofloxacin. Several genotypic and phenotypic patterns were detected among the S. epidermidis isolates: antibiogram typing showed seven different patterns, one of which was shared by 65% of the isolates, whereas the most prevalent RAPD genotype was shared by only five S. epidermidis isolates, and did not correlate with antibiotic resistance phenotype. Conclusion: The diverse clonal origin of tested isolates indicates the presence of multiple S. epidermidis strains among neonates in the NICU setting

Evaluation of antibiotic resistance pattern and mecA gene frequency in methicillin-resistant Staphylococcus aureus strains collected from clinical specimens in Marand hospital and treatment centers, Iran

2021 ◽  
Author(s):  
Abolfazl Jafari-Sales ◽  
Zahra Sadeghi Deylamdeh ◽  
Afsoon Shariat
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Medical Centers ◽  
Wide Range

Introduction: Staphylococcus aureus causes a wide range of infections and as a multivalent pathogen is one of the causative agents of nosocomial and community infections. Therefore, the aim of this study was to identify and determine the pattern of antibiotic resistance of methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients in hospitals and medical centers in Marand city and also to evaluate the presence of mecA gene. Materials and Methods: In this cross-sectional descriptive study, 385 samples of S. aureus were collected from different clinical samples of patients in hospitals and medical centers of Marand city. S. aureus was identified using standard biochemical methods.  Methicillin resistance was determined by disk diffusion method in the presence of oxacillin and cefoxitin. The pattern of antibiotic resistance of the strains was determined by disk diffusion method and according to CLSI recommendation and also PCR method was used to evaluate the frequency of MecA gene. Results: In the present study, out of 385 samples of S. aureus, 215 (55.84%) samples were methicillin resistant. PCR results for mecA gene showed that 110 samples had mecA gene.  The highest antibiotic resistance was observed against penicillin (100%) and erythromycin (83.63%). Most MRSA were isolated from urine and wound samples. Conclusion: The results of this study indicate the prevalence of methicillin-resistant species and also the increase in antibiotic resistance of MRSA to various antibiotics.  Therefore, in order to prevent increased resistance to other antibiotics, it is recommended to avoid inappropriate use of antibiotics.

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