scholarly journals Inhibition of Staphylococcus aureus LC 554891 by Moringa oleifera Seed Extract either Singly or in Combination with Antibiotics

Molecules ◽  
2020 ◽  
Vol 25 (19) ◽  
pp. 4583
Author(s):  
Gamal Enan ◽  
Abdul-Raouf Al-Mohammadi ◽  
Samir Mahgoub ◽  
Seham Abdel-Shafi ◽  
Eman Askar ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Essential Oils ◽  
Inhibitory Activity ◽  
Moringa Oleifera ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Resistance Index ◽  
Aureus Strain ◽  
Rrna Gene ◽  
Bacterial Strains

Bacterial outbreaks caused by Staphylococcus aureus (S. aureus) are interesting due to the existence of multidrug resistant (MDR) isolates. Therefore, there is a need to develop novel ways to control such MDR S. aureus. In this study, some natural agents such as honey bee (HB), extracts of either Moringa oleifera seeds (MSE), or leaves (MLE) and essential oils of garlic, clove, and moringa were studied for their inhibitory activity against this S. aureus pathogen. About 100 food samples including beef luncheon (n = 25), potato chips (n = 50), and corn flakes (n = 25) were investigated for possible pollution with the S. aureus bacteria. The isolated bacteria suspected to belong S. aureus that grew well onto Baird–Parker agar (Oxoid) and shiny halo zones and positive coagulase reaction were selected and identified by API-Kits; all of them that were approved belong to S. aureus (18 strains). The sensitivity of the obtained 18 S. aureus bacterial strains to 12 antibiotics were evaluated; all of them were resistant to ofloxacin; however, other antibiotics tested showed variable results. Interestingly, the S. aureus No. B3 isolated from beef luncheon was resistant to 10 antibiotics out of 12 ones tested. Multiple antibiotic resistance index (MAR) of this S. aureus strain was about 83.3%. Therefore, its identification was confirmed by sequencing of a 16S rRNA gene which approved a successful biochemical identification carried out by API Kits and such strain was designated S. aureus LC 554891. The genome of such strain appeared to contain mecA gene encoding methicillin resistance; it was found to contain hla, hlb, tsst-1, and finbA that encode α-blood hemolysis, β-blood hemolysis, toxic shock syndrome gene, and fibrinogen-binding protein gene, respectively. In addition, the virulence factors viz. sea; seb; sec encoding enterotoxins were detected in the DNA extracted from S. aureus B3 strain. Aqueous extract of Moringa oleifera seeds (MSE) showed inhibitory activity against S. aureus LC 554891 better than that obtained by tetracycline, essential oils or HB. Minimum inhibitory concentration (MIC) of MSE was 20µg/mL. Instrumental analysis of MSE showed 14 bioactive chemical compounds. Combinations of both MSE and tetracycline showed distinctive inhibitory activity against S. aureus LC 554891 than that obtained by either tetracycline or MSE singly.

scholarly journals Mannitol Fermenting Methicillin-Resistant Coagulase Negative Staphylococci Isolated From Diabetic Foot Infections

2020 ◽  
Vol 11 (4) ◽  
pp. 6095-6101
Author(s):  
Samira Fattah Hamid ◽  
Aza Bahadeen Taha
Keyword(s):  
Staphylococcus Aureus ◽  
Diabetic Foot ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Rrna Gene ◽  
Human Pathogens ◽  
Coagulase Negative Staphylococci ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant ◽  
Diabetic Foot Infections

Detection of mannitol fermenting coagulase-negative staphylococci is frequently unnoticed when Staphylococcus aureus is screening in the laboratory. On the other hand, the emergence of coagulase-negative staphylococci as critical human pathogens need dependable methods for the identification of clinically significant coagulase-negative staphylococci to understand the epidemiology of infections caused by these bacteria. The study aimed to identify mannitol fermenting coagulase-negative staphylococci that assumed to be Staphylococcus aureus as they formed yellow colonies on Mannitol Salt agar plates. Samples were taken from eighty-four patients with diabetic foot infections. The specimen was cultured on Blood agar and Mannitol Salt agar. Mannitol fermenting coagulase-negative staphylococci isolates diagnosed through Vitek2 system then confirmed by detecting 16S rRNA gene and absence of the nuc gene. Antibiotic sensitivity and methicillin resistance were detected by Vitek2 system, then methicillin resistance was confirmed by Oxacillin Salt Agar Screen test and detection of the mecA gene. Out of 81 Staphylococcus isolated from foot and nose of diabetic foot patients, twenty isolates were mannitol fermenting coagulase-negative staphylococci, they related to following species; Staphylococcus haemolyticus, staphylococcus lentus, Staphylococcus xylosus, Staphylococcus lugdunensis, Staphylococcus hominis, Staphylococcus galinarum and Staphylococcus saprophyticus). The majority of them (85%) were phenotypically methicillin-resistant and genotypically harbouring mecA gene. 80% were resistant to Erythromycin, 70% to Clindamycin, 35% to Trimethoprim-Sulphamethoxazole, 30% to Gentamicin and Rifampicin, 15% to Levofloxacin and Teicoplanin. 30% expressed inducible clindamycin resistance.

scholarly journals Molecular detection and typing of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci isolated from cattle, animal handlers, and their environment from Karnataka, Southern Province of India

2019 ◽  
Vol 12 (11) ◽  
pp. 1760-1768 ◽  
Author(s):  
Nimita Venugopal ◽  
Susweta Mitra ◽  
Rituparna Tewari ◽  
Feroze Ganaie ◽  
Rajeswari Shome ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Animal Health ◽  
Sccmec Type ◽  
Meca Gene ◽  
Molecular Characteristics ◽  
Rrna Gene ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant

Background and Aim: Methicillin-resistant staphylococci are among the emerging pathogens which have become a threat to both human and animal health. The present investigation intended to examine the occurrence and the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) recovered from cattle, its handlers, and their environment. Materials and Methods: A total of 666 specimens were subjected to culture method and genus-specific polymerase chain reaction (PCR) for the identification of Staphylococcus. Methicillin resistance was substantiated by PCR identification of mecA and mecC resistance determinants. Species-specific identification of mecA positive isolates was conducted by multiplex PCR. The unidentified species were deciphered by 16S rRNA gene sequencing approach. The mecA positive isolates were further characterized by staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). Results: Duplex PCR identified 728 Staphylococcus isolates, of which 66 (9%) were positive for mecA gene. MRSA constituted 24% of the total mecA positive isolates. Among MRCoNS, Staphylococcus epidermidis (42%), and Staphylococcus haemolyticus (11%) were the most common species identified. Overall, 47% of the mecA positive isolates belonged to SCCmec type V. MLST analysis showed eight different sequence types (STs) among MRSA isolates of which five were novel STs. Among methicillin-resistant S. epidermidis, 19 different STs were found, of which nine novel STs were detected. Conclusion: The increase in the prevalence of mecA positive staphylococci, especially MRCoNS in cattle is a great concern in view of their transmission potential. Hence, continuous monitoring and molecular characterization of methicillin-resistant staphylococci should be elucidated in human and animal sectors so as to prevent the spread of these resistant pathogens.

Panton-Valentine Leukocidin Gene Detected in a Staphylococcus Aureus Strain Isolated from a Knee Arthroprosthesis Infection

2009 ◽  
Vol 32 (9) ◽  
pp. 630-634 ◽  
Author(s):  
Lucio Montanaro ◽  
Lucilla Baldassarri ◽  
Tolmino Corazzari ◽  
Roberta Creti ◽  
Stefano Ravaioli ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Research Unit ◽  
Aureus Strain ◽  
Toxin Gene ◽  
Strain Collection ◽  
Rizzoli Orthopaedic Institute ◽  
Panton Valentine Leukocidin ◽  
Valentine Leukocidin

This report focuses on the molecular characterization of a Staphylococcus aureus strain isolated from a knee arthroprosthesis infection and recognized retrospectively as a carrier of the Panton-Valentine leukocidin gene. The stored microbiological isolate, which belonged to the strain collection of the Research Unit on Implant Infections of the Rizzoli Orthopaedic Institute, was retrieved for molecular analysis. Genotyping was carried out, revealing an interesting profile. In addition to the positivity for the Panton-Valentine toxin gene, the results indicated that the isolate belonged to the agr III group and was endowed with bbp and cna genes, both encoding for staphylococcal adhesins that bind bone proteins. The strain had the mecA gene for methicillin resistance, even though it was unable to resist any of the β-lactam or other antibiotics. Its gene configuration matched that of other community-acquired methicillin-resistant and methicillin-susceptible Staphylococcus aureus (CA-MRSA and CA-MSSA) strains which have recently been reported worldwide. As far as we know, this is the first report on a PVL-positive S. aureus strain associated with an orthopedic implant (knee arthroprosthesis) infection.

scholarly journals Detection of Methicillin-Resistant Staphylococcus aureus in Ready-to-Eat Shellfish (Corbiculid heterodont) in Bayelsa State, Nigeria

2020 ◽  
pp. 22-35
Author(s):  
Seighazi Regina Egege ◽  
Nedie Patience Akani ◽  
Chidiebele Emmanuel Ikechukwu Nwankwo
Keyword(s):  
Staphylococcus Aureus ◽  
Multidrug Resistance ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Resistance Index ◽  
Multidrug Resistant ◽  
Coagulase Negative Staphylococcus ◽  
Methicillin Resistant ◽  
Awareness Campaigns

Aims/Objective: Methicillin-Resistant Staphylococcus aureus (MRSA) is a dominant cause of severe healthcare-associated (HA) infections but has recently emerged as Community-Associated (CA-MRSA) and Livestock-Associated MRSA (LA-MRSA). This study was carried out to detect the presence of Methicillin-Resistant S. aureus in Ready-to-Eat (RTE) Corbiculid heterodont in Bayelsa state, Nigeria. Place of Study: This study was conducted in Yenagoa, Southern Ijaw and Sagbama Local Government Areas, Bayelsa State, Nigeria. Methodology: Fifty-three samples (dried, fried and stewed) of the RTE C. heterodont were collected and subjected to standard microbiological procedures. Results: Seventy Staphylococcus spp. were isolated, consisting of 65 (92.86%) S. aureus and 5 (7.14%) coagulase negative Staphylococcus (CONS). Out of the 65 S. aureus isolates, 53.8% were MRSA. Susceptibility pattern of MRSA showed a decreasing trend of resistance in the order: Amoxacillin, Ampicillin-Cloxacillin, Cefoxitin, and Oxacillin (100%) > Streptomycin (82.9%) >Ceftriaxone (40%) >Cefuroxime (31.4%), > Ciprofloxacin and Trimethoprim-Sulfamethoxazole (17.1%) > Gentamycin and Pefloxacin (8.6%) > Erythromycin (2.9%).Out of the 65 S. aureus isolates, 62 (95.4%) had Multidrug Resistance Index > 0.2 while 3 (4.6%) had 0.2 as their Multidrug Resistance Index. The MRSA isolates were 100% Multidrug Resistant (MDR) while the Methicillin-Sensitive S. aureus (MSSA) were 90% MDR. Molecular characterization of ten S. aureus isolates confirmed 80% S. aureus isolates and 20% CONS (S. sciuri and S. warneri).  Polymerase chain reaction (PCR) and Agarose gel-electrophoresis were used to detect the presence of mecA gene, a gene that confers methicillin resistance. The mecA gene with size 500bp was identified in 50% of MRSA. This study reported presence of MRSA and MDR S. aureus in C. heterodont sold in Bayelsa state, Nigeria. This could be the avenue for the spread of MRSA. Public health awareness campaigns should be advocated to enhance hygienic practices in the handling and distribution of food.

scholarly journals Suppression of methicillin resistance in a mecA-containing pre-methicillin-resistant Staphylococcus aureus strain is caused by the mecI-mediated repression of PBP 2' production.

1996 ◽  
Vol 40 (12) ◽  
pp. 2680-2685 ◽  
Author(s):  
K Kuwahara-Arai ◽  
N Kondo ◽  
S Hori ◽  
E Tateda-Suzuki ◽  
K Hiramatsu
Keyword(s):  
Staphylococcus Aureus ◽  
Gene Transcription ◽  
Culture Medium ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Aureus Strain ◽  
Methicillin Resistant ◽  
Gene Transcripts ◽  
Regulator Genes

The mechanism of methicillin susceptibility was studied in Staphylococcus aureus N315P, a pre-methicillin-resistant S. aureus strain that is susceptible to methicillin, despite the presence of mecA in the chromosome. In the presence of mec regulator genes mecI and mecR1, transcription of the mecA gene was not inducible by the addition of methicillin to the culture medium. Inactivation of the mecI gene function by replacing it with tetL made N315P express heterogeneous-type methicillin resistance. The subclone, in which the mecI gene was replaced, subclone P delta I, produced 12 times greater amounts of mecA gene transcripts and 8.5 times more PBP 2' protein than N315P. These data indicate that the mecI gene-encoded repression of mecA gene transcription is responsible for the apparent methicillin susceptibility phenotype of pre-methicillin-resistant S. aureus N315P.

scholarly journals Isolation of Biofilm Producing Methicillin-Resistant Staphylococcus aureus from Hospitalized Orthopaedic Patients in Kano State, Nigeria

2021 ◽  
Vol 28 (1) ◽  
pp. 66-74
Author(s):  
D.A. Oche ◽  
U. Abdulrahim ◽  
A.S. Oheagbulem ◽  
B.O. Olayinka
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Community Settings ◽  
Potential Threat ◽  
Orthopaedic Patients ◽  
Nasal Swabs ◽  
Public Health Challenge

Biofilm formation and resistance to methicillin are among the factors that makes Staphylococcus aureus a very important human pathogen in both health-care and community settings. This study investigated methicillin-resistance among biofilm-producing S. aureus isolated from 49 orthopaedic in-patients within a 3 months period. Wound swabs, nasal swabs, bed swabs and urine samples were collected from each patient. The samples were cultured and screened for presence of S. aureus while the micro-titre plate method was used to detect biofilm producing isolates. PCR technique was finally used to detect the presence of mecA gene in methicilin resistant S. aureus (MRSA) isolates. Findings reveal 14.8% of bacterial isolates were Staphylococcus aureus of which 96.4% were biofilm-producers. However, strong biofilm producers constitute 11.1%. The mecA gene was detected in 15.8% of the MRSA isolates. Therefore, MRSA among biofilm-producing S. aureus is a potential threat primarily to the community of National Orthopaedic Hospital Dala and a major public health challenge. Keywords: Biofilm, Methicillin-resistance Staphylococcus aureus (MRSA), mecA gene, Orthopaedic patient

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 Effect of exogenous glycine on peptidoglycan composition and resistance in a methicillin-resistant Staphylococcus aureus strain.

1996 ◽  
Vol 40 (6) ◽  
pp. 1498-1503 ◽  
Author(s):  
B L de Jonge ◽  
Y S Chang ◽  
N Xu ◽  
D Gage
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Aureus Strain ◽  
Methicillin Resistant ◽  
Resistance Phenotype ◽  
Concomitant Decrease

A highly homogeneously methicillin-resistant Staphylococcus aureus strain was grown in the presence of various concentrations of exogenous glycine. Increasing concentrations of glycine in the medium resulted in a decrease in methicillin resistance and the appearance of a heterogeneous resistance phenotype. Parallel to the gradual changes in resistance was an alteration in the muropeptide composition of peptidoglycan. Increasing concentrations of glycine in the medium resulted in peptidoglycan in which muropeptides with a D-alanyl-D-alanine terminus were replaced with D-alanyl-glycine-terminating muropeptides. The disappearance of D-alanyl-D-alanine-terminating muropeptides in peptidoglycan and the concomitant decrease in resistance indicate a central role for D-alanyl-D-alanine-terminating precursors in methicillin resistance.

scholarly journals Single and Synergistic Effects of Solar Radiation, Water guard and Moringa oleifera Treatments on Bacterial Loads and Physicochemical Parameters of Surface Water in Umur and Bele Streams of Benue State

2018 ◽  
Vol 6 (3) ◽  
pp. 244-251
Author(s):  
G.M. Gberikon ◽  
I.I. Adeoti ◽  
I.O. Ogbonna
Keyword(s):  
Surface Water ◽  
Solar Radiation ◽  
Water Sample ◽  
Water Samples ◽  
Moringa Oleifera ◽  
Synergistic Effects ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Ph Values ◽  
Significant Difference

Bacteriological profiles of surface water samples treated with solar radiation, Moringa oleifera powder and water guard was carried out to ascertain their effectiveness in water treatment. Bacterial loads of the surface water collected from Umur and Bele streams in Gwer East Local Government area samples were determined before and after treatments and characterised using biochemical and molecular methods. Single and synergistic effects of these treatments on water quality were also examined. Bacteriological analysis showed that Umur stream had the highest bacterial loads of 4.47 x 103 cfu/mL while Bele had the lowest counts. There was significant reduction in the mean viable counts recorded for all the water samples (p ˂ 0.05). In the daily bacteria counts, the control water samples gave extremes values. There were no bacteria count recorded following combine treatment in water samples from Bele stream on the fifth day. Molecular analysis based on the 16S rRNA gene sequence showed bacterial strains to be phylogenetically close to bacterial strains which are capable of causing infectious diseases to man. Normal pH values were recorded in Umur stream while low pH values were recorded in Bele streams. Treatment impacted significantly on the pH of the water samples from Umur stream (p ˂ 0.05) while no significant difference was observed with water samples from Bele stream (p > 0.05). Treatment impacted significantly on turbidity in water sample from Umur stream (p ˂ 0.05). Sulphate was found to be within the permissible limit except for water sample from Bele stream which gave mean values within the range of 51.00 to 68.00 mg/L. Treatments impacted no significant difference on sulphate (p > 0.05). Surface water in these rural areas should be thoroughly treated before use. Int. J. Appl. Sci. Biotechnol. Vol 6(3): 244-251

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