scholarly journals Randomly amplified polymorphic DNA analysis of Staphylococcus chromogenes isolated from bovine and bubaline mastitis in Karnataka

2021 ◽  
Vol 14 (1) ◽  
pp. 285-291
Author(s):  
P. Sheela ◽  
Malathi Shekar ◽  
Shrikrishna Isloor ◽  
D. Rathnamma ◽  
B. M. Veeregowda ◽  
...  
Keyword(s):  
Dna Analysis ◽  
Methicillin Resistance ◽  
Subclinical Mastitis ◽  
Image Correlation ◽  
Diffusion Method ◽  
Randomly Amplified Polymorphic Dna ◽  
Base Pairs ◽  
Disk Diffusion Method ◽  
Dairy Animals ◽  
Intraspecies Diversity

Background and Aim: In recent times, non-aureus staphylococci (NAS) have emerged as the major organisms isolated from mastitis cases in dairy animals, with a predominance of Staphylococcus epidermidis and Staphylococcus chromogenes. As compared to Staphylococcus aureus, much less is known about the molecular types or the spatiotemporal epidemiology of these NAS species. In the present study, randomly amplified polymorphic DNA (RAPD) was employed to detect genetic polymorphisms, intraspecies diversity, and epidemiology of S. chromogenes strains (n=37) isolated from bovine and bubaline mastitis cases in the state of Karnataka. Materials and Methods: Thirty-seven S. chromogenes isolates (14 from bovines and 23 from bubaline) isolated from subclinical mastitis cases, from organized and unorganized sectors, were subjected to RAPD typing. Further, methicillin resistance was determined by cefoxitin disk diffusion method. Results: The amplified DNA fragments ranged from 150 to 3000 base pairs and yielded several RAPD profiles. Further analysis using Digital Image Correlation Engine correlation coefficient and UPGMA method showed that the 37 isolates could be classified into 12 distinct RAPD types (A to L) at 62% similarity (D=0.889). Four of the most predominant RAPD types, B, A, C, and E, in that order, and together, represented 65% of the isolates. High diversity was observed among the isolates both within farms and between geographic locations. Most of the isolates exhibited methicillin resistance. This is the first such report from India. Conclusion: In the absence of defined multilocus sequence type protocols or sufficient sequences available in the public domain, RAPD can be employed to determine genetic diversity of S. chromogenes isolates.

scholarly journals Methicillin-resistant Staphylococcus aureus bloodstream infection: risk factors and clinical outcome in non-intensive-care units

2012 ◽  
Vol 45 (2) ◽  
pp. 189-193 ◽  
Author(s):  
Karinne Spirandelli Carvalho Naves ◽  
Natália Vaz da Trindade ◽  
Paulo Pinto Gontijo Filho
Keyword(s):  
Risk Factors ◽  
Staphylococcus Aureus ◽  
Bloodstream Infection ◽  
Antimicrobial Agents ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Staphylococcus Aureus Bloodstream Infection

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is spread out in hospitals across different regions of the world and is regarded as the major agent of nosocomial infections, causing infections such as skin and soft tissue pneumonia and sepsis. The aim of this study was to identify risk factors for methicillin-resistance in Staphylococcus aureus bloodstream infection (BSI) and the predictive factors for death. METHODS: A retrospective cohort of fifty-one patients presenting bacteraemia due to S. aureus between September 2006 and September 2008 was analysed. Staphylococcu aureus samples were obtained from blood cultures performed by clinical hospital microbiology laboratory from the Uberlândia Federal University. Methicillinresistance was determined by growth on oxacillin screen agar and antimicrobial susceptibility by means of the disk diffusion method. RESULTS: We found similar numbers of MRSA (56.8%) and methicillin-susceptible Staphylococcus aureus (MSSA) (43.2%) infections, and the overall hospital mortality ratio was 47%, predominantly in MRSA group (70.8% vs. 29.2%) (p=0.05). Age (p=0.02) was significantly higher in MRSA patients as also was the use of central venous catheter (p=0.02). The use of two or more antimicrobial agents (p=0.03) and the length of hospital stay prior to bacteraemia superior to seven days (p=0.006) were associated with mortality. High odds ratio value was observed in cardiopathy as comorbidity. CONCLUSIONS: Despite several risk factors associated with MRSA and MSSA infection, the use of two or more antimicrobial agents was the unique independent variable associated with mortality.

scholarly journals Evaluation of the CLSI cefoxitin 30-µg disk-diffusion method for detecting methicillin resistance in staphylococci

2006 ◽  
Vol 12 (10) ◽  
pp. 1039-1042 ◽  
Author(s):  
L.-X. Zhu ◽  
Z.-W. Zhang ◽  
C. Wang ◽  
H.-W. Yang ◽  
Q. Zhang ◽  
...  
Keyword(s):  
Methicillin Resistance ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method

scholarly journals Correlation between mazEF Toxin-Antitoxin System Expression and Methicillin Susceptibility in Staphylococcus aureus and Its Relation to Biofilm-Formation

2021 ◽  
Vol 9 (11) ◽  
pp. 2274
Author(s):  
Aya Abd El rahman ◽  
Yasmine El kholy ◽  
Rania Y. Shash
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Methicillin Resistance ◽  
Diffusion Method ◽  
P Value ◽  
Sequence Motifs ◽  
University Hospitals ◽  
Specific Sequence ◽  
Disk Diffusion Method ◽  
Cellular Mrnas

Methicillin resistance in Staphylococcus aureus has become prevalent globally. Moreover, biofilm-formation makes it more difficult to eradicate bacteria by antibiotics. The mazEF toxin-antitoxin system encodes for mazF, which acts as an endoribonuclease that cleaves cellular mRNAs at specific sequence motifs (ACA), and mazE, which opposes the mazF action. Our goal was to detect mazEF expression in methicillin-resistant S. aureus (MRSA) isolates compared with methicillin-sensitive S. aureus (MSSA) isolates and determine its relation to methicillin susceptibility as well as biofilm-formation. According to their susceptibility to cefoxitin disks, 100 S. aureus isolates obtained from patients admitted to Cairo University Hospitals were categorized into 50 MSSA and 50 MRSA according to their susceptibility to cefoxitin disks (30 µg). Antimicrobial susceptibility and biofilm-formation were investigated using the disk diffusion method and tissue culture plate method, respectively. Finally, using real-time PCR, mazEF expression was estimated and correlated to methicillin susceptibility and biofilm formation. Both MRSA and MSSA isolates showed the best sensitivity results with linezolid and gentamicin, where about 88% of MRSA isolates and 96% of MSSA isolates were sensitive to linezolid while 76% of MRSA isolates and 84% of MSSA isolates were sensitive to gentamicin. MRSA isolates were significantly more able to form biofilm than MSSA isolates (p-value = 0.037). The mazEF expression was significantly correlated to methicillin resistance in S. aureus (p-value < 0.001), but not to biofilm-formation.

scholarly journals Department of Microbiology, Bangladesh Agricultural University, Mymensingh

2016 ◽  
Vol 2 (2) ◽  
pp. 3-6
Author(s):  
Nahida Akther Zahan ◽  
Md. Akram Hossain ◽  
AKM Shamsuzzaman ◽  
AKM Musa ◽  
Md. Chand Mahamud ◽  
...  
Keyword(s):  
Clinical Laboratory ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Toxin Production ◽  
Latex Agglutination ◽  
Diffusion Method ◽  
National Committee ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method

The study was done to detect different exotoxins among the strains of Staphylococcus aureus isolated in the department of Microbiology, Mymensingh Medical College in collaboration with the Department of Medicine under the Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh between the periods from July, 2006 to June, 2007. A total of 40 S. aureus isolates investigated in this study were identified by standard microbiological techniques. Antimicrobial susceptibility of the isolates to Oxacillin was carried out by disk diffusion method as per recommendation of the National Committee for Clinical Laboratory Standards. Any isolate showing resistance to Oxacillin was tested again by agar dilution method to determine minimum inhibitory concentration (MIC) of Methicillin. All strains were also tested for mecA gene by Polymerase Chain Reaction (PCR) for confirmation of Methicillin resistance. Enterotoxin (A-D) and Toxic Shock Syndrome Toxin-1 (TSST-1) were detected by Reverse Passive Latex Agglutination (RPLA) test. Out of 40 S. aureus isolates, 7 (17.5%) Methicillin Resistant S. aureus (MRSA), 1 (2.5%) Methicillin Sensitive S. aureus (MSSA) produced Staphylococcal Enterotoxin A (SE-A) and 1 MRSA isolate was positive for TSST-1. In case of combined toxin production among the S. aureus isolates, 2 (5%) MSSA were found to produce SE-A and SE-B, 2 (5%) MSSA produced SE-C and SE-D, and 1 (2.5%) MRSA, 1 (2.5%) MSSA produced SE-C and TSST-1.Bangladesh J Med Microbiol 2008; 02 (02): 3-6

scholarly journals Characteristics of coagulase positive staphylococci isolated from milk in cases of subclinical mastitis

2014 ◽  
Vol 64 (1) ◽  
pp. 115-123 ◽  
Author(s):  
Nataša Rajić Savić ◽  
Vera Katić ◽  
Branko Velebit
Keyword(s):  
Meca Gene ◽  
Subclinical Mastitis ◽  
Penicillin G ◽  
Diffusion Method ◽  
Disc Diffusion ◽  
Disk Diffusion Method ◽  
Phenotypic Resistance ◽  
High Virulence ◽  
Pcr Method ◽  
Coagulase Positive Staphylococci

Abstract Coagulase-positive staphylococci are the most common pathogen causing subclinical mastitis in cows. Their main characteristic is a high virulence which leads to chronic infection. A total of 213 isolates of coagulase-positive staphylococci were tested. The majority of isolates (58%) formed a gold pigment, then light gold (28%), white gold (8%), golden gray, creamy white and white (2%). The majority of isolated coagulase-positive staphylococci produce beta hemolysis on esculin blood agar (50%), alpha and beta hemolysis (36%), beta and delta hemolysis (8%), delta hemolysis (4%), and alpha hemolysis (2%). Biochemical and molecular identification was performed by APISTAPH and multiplex PCR method. The majority of isolates were identified as S. aureus (88%), S. chromogenes (4%), and 2% of the isolates were identified as S. lentus, S. sciuri, S. xylosus, S. intermedius by APISTAPH. Antimicrobial susceptibility to penicillin G, was found by the Kirby Bauer disk diffusion method to be resistant (62.44%). For disc diffusion penicillin G sensitive isolates the minimal inhibitory concentration (MIC) was established for MIC50 and MIC90 as 0.003 mg/ml and 2μg/ml, respectively. For disc diffusion penicillin G resistant isolates MIC50 and MIC90 was 1.0μg/ml and 16μg/ ml, respectively. The study of phenotypic resistance to methicillin, as recommended by CLSI, established resistance to oxacillin in 5.26% of the isolates, while no resistance was found to cefoxitine. None of the tested isolates have the mecA gene.

scholarly journals Childhood Septicemia in Nepal: Documenting the Bacterial Etiology and Its Susceptibility to Antibiotics

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Shamshul Ansari ◽  
Hari Prasad Nepal ◽  
Rajendra Gautam ◽  
Sony Shrestha ◽  
Puja Neopane ◽  
...  
Keyword(s):  
Antibiotic Susceptibility ◽  
Methicillin Resistance ◽  
Care Center ◽  
Tertiary Care ◽  
Diffusion Method ◽  
Coagulase Negative Staphylococci ◽  
Gram Positive ◽  
Disk Diffusion Method ◽  
Gram Negative ◽  
Microbiological Methods

Introduction. Children are among the most vulnerable population groups to contract illnesses. The varying microbiological pattern of septicemia warrants the need for an ongoing review of the causative organisms and their antimicrobial susceptibility pattern. Therefore, the objective of this study was to document the bacterial etiology of childhood septicemia and its antibiotic susceptibility profile.Methods.Cross-sectional type of study in 1630 suspected patients was conducted at CMCTH from January 2012 to December 2013. Blood samples were collected aseptically for culture. The organisms grown were identified by standard microbiological methods recommended by American Society for Microbiology (ASM) and subjected to antibiotic susceptibility testing by modified Kirby-Bauer disk diffusion method. Methicillin resistance was confirmed using cefoxitin and oxacillin disks methods.Results.Septicemia was detected in 172 (10.6%) cases. Among Gram-positive organisms, coagulase negative staphylococci (CoNS) were leading pathogen andAcinetobacterspp. were leading pathogen among Gram-negative isolates. Vancomycin, teicoplanin, and clindamycin were the most effective antibiotics against Gram-positive isolates while amikacin was effective against Gram-positive as well as Gram-negative isolates. Methicillin resistance was detected in 44.4% ofStaphylococcus aureus.Conclusions. This study has highlighted the burden of bacterial etiology for septicemia among children in a tertiary care center of central Nepal.

scholarly journals A study of inducible clindamycin resistance in erythromycin resistant clinical isolates of Staphylococcus species

2015 ◽  
Vol 6 (6) ◽  
pp. 48-52
Author(s):  
Fatima Khan ◽  
Sana Ali ◽  
Asfia Sultan ◽  
Meher Rizvi ◽  
Abida Khatoon ◽  
...  
Keyword(s):  
Methicillin Resistance ◽  
Clinical Isolates ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Clinical Failure ◽  
Coagulase Negative Staphylococci ◽  
Sensitivity Testing ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Inducible Clindamycin Resistance

Introduction: Erythromycin resistant Staphylococcus isolates with inducible resistance appear sensitive to clindamycin in in?vitro sensitivity testing. If clindamycin is used for treatment of such isolates, selection for constitutive mutants may lead to clinical failure. Current study was conducted to detect the presence of inducible clindamycin resistance in erythromycin resistant Staphylococcus isolates by disk diffusion method (D test). To correlate clindamycin resistance phenotypes with minimum inhibitory concentrations (MICs) of clindamycin, erythromycin, oxacillin and vancomycin among the isolates. To correlate various resistance phenotypes with methicillin resistance. Material and Methods: 150 non duplicate isolates of Staphylococcus species were identified and antibiotic susceptibility testing was done using Kirby Bauer’s disc diffusion method. MICs were determined using E?test for oxacillin, vancomycin, clindamycin and erythromycin using E?test strips (Himedia) Results: Among 150 staphylococcus clinical isolates, 96 were of S. aureus and 54 were coagulase negative Staphylococci (CONS). About 81.2% of the S.aureus isolates and 72.2% of the CONS were found to be methicillin resistant. Inducible clindamycin resistance was found in 39.3% of the isolates, constitutive resistance phenotype in 48% while 12.7% demonstrated MS phenotype. 18% and 11.3% of all the isolates had MICs for clindamycin between 0.01?0.06 μg/ml and 0.06?0.1 respectively. 12.5% had MIC ranging from 4?8 μg/ml while 58% had MIC > 8 μg/ml. Constitutive resistant phenotype (cMLS) was the predominant phenotype in methicillin resistant isolates. MS phenotype was the predominant among MSSA (methicillin sensitive S. aureus) while MSCNS (methicillin sensitive CONS) cMLS (46.7%) predominated. MIC of all erythromycin resistant isolates were ≥ 240 μg/ml. Nearly 16.7% of the cMLS and 57.9% of MS isolates were found to be oxacillin sensitive and 83% of iMLS and 83.3% of MS phenotype isolates were oxacillin resistant on MIC testing. 47.2% of cMLS and 73.6% of MS isolates had MIC ≤ 2 μg/ml for vancomycin and 52.7% of cMLS and 26.3% of MS isolates had MICs in intermediate range for vancomycin. Conclusions: D?testing might help clinicians to decide whether to use clindamycin in Staphylococcal infections when erythromycin resistance is present. Determination of MICs help to identify exact sensitivity profile of isolates in cases where clinical failure occurs due to misleading disk diffusion tests.DOI: http://dx.doi.org/10.3126/ajms.v6i6.11811 Asian Journal of Medical Sciences Vol.6(6) 2015 48-52

scholarly journals Genes conferring anti­microbial resistance in cattle with subclinical mastitis

2021 ◽  
Vol 24 (1) ◽  
pp. 67-85
Author(s):  
N. H. Youssif ◽  
N. M. Hafiz ◽  
M. A. Halawa ◽  
H. M. Aziz
Keyword(s):  
Resistance Genes ◽  
Methicillin Resistance ◽  
Subclinical Mastitis ◽  
Cow Milk ◽  
In Vitro Susceptibility ◽  
Milk Samples ◽  
Dairy Animals ◽  
Susceptibility Tests ◽  
Moderate Resistance

This study was carried out to evaluate the antimicrobial resistance (AMR) as a risk factor associated with some microorganisms isolated from subclinical mastitis (SCM) milk samples from Holstein Friesian dairy animals in Fayoum area, Egypt. The percentage of the SCM in the farm was found to be 41.18% and 63.88% at quarter and cows level respectively, with mean somatic cell count (SCC) of 8.8×105 ± 9.2×103 cells/mL and electrical conductivity (EC) 6.27 ± 0.066 mS/cm for SCM quarter milk samples. Out of the total 444 SCM cow milk samples, the most often isolated microorganisms were Staphylococcus aureus: 296 (66.6%), Enterococcus spp.: 230 (51.80%), Escherichia coli: 210 (47.29%) and Streptococcus agalactiae: 106 (23.87%). AMR was determined by disc diffusion test and the corresponding resistance genes were detected by PCR. Results of the in vitro susceptibility tests performed and the phenotypes indicated that the highest resistance to antibiotics for isolated microorganisms was against penicillin followed by amoxicillin + clavulanic acid, oxacillin and tetracycline, whereas moderate resistance was exhibited to oxytetracycline, ampicillin, sulfamethazole/trimethoprim, cefotaxime and erythromycin. However the most effective antibiotics against most isolates were nitrofurantoin and gentamicin followed by enrofloxacin, norfloxacin and cefoxitin. It was shown that the resistance to tetracyclines was due to the tetK or tetA(A) genes, the resistance to β-lactams (penicillins) – to blaZ and blaTEM genes, to macrolides (erythromycin): to ermB and ermC genes. Methicillin resistance genes were mecA, mec1 and mecC, glycopeptides (vancomycin) resistance gene was vanA, and norfloxacin resistance was attributed to norA gene.

scholarly journals Comparison of rapid and conventional methods for investigating of mecA presence in Staphylococcus Species

2021 ◽  
Vol 37 (5) ◽  
Author(s):  
Selim Görgün ◽  
Hacer İşler ◽  
Mehmet Cenk Turgut
Keyword(s):  
Methicillin Resistance ◽  
Meca Gene ◽  
Latex Agglutination ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
Chromogenic Agar ◽  
Pcr Method ◽  
Conventional Methods

Objectives: Taking the determination of mecA gene by polymerized chain reaction (PCR) method as a reference in determining methicillin resistance in Staphylococcus species, we aimed at comparing the reliability levels of disk diffusion, latex agglutination test and chromogenic agar use methods. Methods: This prospective study was conducted on 228 Staphylococcus strains isolated between January 2020 and December 2020 in Samsun Training and Research Hospital. Disk diffusion, latex agglutination and chromogen agar medium methods were applied along with the polymerized chain reaction (PCR) method. Results: The mecA gene was detected in 47 of the isolates (20.6%) by the PCR method, and these isolates were accepted as methicillin-resistant. When the PCR result was taken as a reference, the sensitivity of the disk diffusion method became 100%, and specificity became 45.9%; sensitivity of latex agglutination was determined as 80.9%, and specificity as 70.2%; sensitivity of chromogenic agar as 85.1% and its specificity was found to be 95%. Only in S. aureus isolates, the highest sensitivity and specificity rate (100% and 88%, respectively) belonged to chromogenic agar. Conclusion: Chromogenic agar provides more reliable data for S. aureus isolates, and the combined use of all three methods does not significantly increase reliability. doi: https://doi.org/10.12669/pjms.37.5.4274 How to cite this:Gorgun S, Isler H, Turgut MC. Comparison of rapid and conventional methods for investigating of mecA presence in Staphylococcus Species. Pak J Med Sci. 2021;37(5):---------. doi: https://doi.org/10.12669/pjms.37.5.4274 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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