scholarly journals Isolation and Identification of Methicillin Resistance Staphylococcus aureus and Detection their Ability to the Production of Virulence Factors

2018 ◽  
Vol 26 (8) ◽  
pp. 100-111
Author(s):  
Angham Najah Al-Khafaji
Keyword(s):  
Staphylococcus Aureus ◽  
Virulence Factors ◽  
Bacterial Resistance ◽  
Methicillin Resistance ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Frequent Use ◽  
Slime Layer ◽  
Antibiotics Susceptibility ◽  
Disc Assay

* Objectives: Isolation and Identification of S.aureus and study their susceptibility to the production of virulence factors. * Methods: A total of 50 clinical specimens of Staphylococcus aureus isolates were collected in AL- Najaf AL-Asharaf governorate during the period of (10/10/2013-20/1/2014) by the cultural characteristic colonies, microscopic for cells, biochemical tests, antibiotics susceptibility test to Staphylococcus aureus using an antibiotic diffusion disc assay, in addition to studying the virulence factors produced from S.aureus . * Results: The results show that the Staphylococcus aureus resistant to methicillin formed 70% and 30% of isolates were sensitive to the same antibiotics. Tested 10  isolates to methicillin-resistant for their efficiency in forming the highest inhibitory areas for testing their sensitivity to other antibiotics used, the results show S.aureus resistant to penicillin about 100%, while there is no resistance to CO-Trimethoprim because it was sensitive completely 100%.       The susceptibility investigated of Staphylococcus aureus resistance methicillin isolates to the production of biofilm and slime layer with using both tube method and growth on Congo red agar respectively. Ten isolates of S.aureus  were tested based on resistance to methicillin and their high susceptibility to the production of the biofilm and the slime layer. * Conclusion: So we conclude, they have Staphylococcus aureus is highly resistant to β- Lactam antibiotics and the CO-trimethoprim is the most effective in the inhibiting growth of this bacteria. In addition to its ability to produce virulence factors. The increase cause bacterial resistance to our local isolates may be due to the frequent use of antibiotics, which allowed for increased bacterial resistance to various antibiotics.  

scholarly journals Molecular characterization of virulence factors in Staphylococcus aureus isolated from bovine subclinical mastitis in central Ethiopia

2021 ◽  
Vol 71 (1) ◽  
Author(s):  
Desiye Tesfaye Tegegne ◽  
Gezahegne Mamo ◽  
Hika Waktole ◽  
Yohannes Equar Messele
Keyword(s):  
Staphylococcus Aureus ◽  
Dairy Cows ◽  
Virulence Factors ◽  
Virulence Genes ◽  
Animal Health ◽  
Subclinical Mastitis ◽  
Biochemical Tests ◽  
Milk Samples ◽  
Lactating Dairy Cows ◽  
Genomic Study

Abstract Purpose Staphylococcus aureus (S. aureus) is the most important pathogen involved in bovine mastitis in dairy production. S. aureus produces a spectrum of extracellular protein toxins and virulence factors which are thought to contribute to the pathogenicity of the organism. The aim of this work was to isolate and molecular characterize S. aureus associated with bovine subclinical mastitis in the central part of Ethiopia. Methods A total of 265 lactating dairy cows from various dairy farms in four different geographical locations were screened by the California mastitis test (CMT) for bovine subclinical mastitis. One hundred thirty CMT-positive milk samples were collected and transported to the laboratory. Different biochemical tests and polymerase chain reaction (PCR) were used for the identification of S. aureus isolates. Finally, PCR was performed for molecular detection of virulence genes. Results From a total of 265 lactating dairy cows screened, 49% (n = 130) were positive for bovine subclinical mastitis. One hundred thirty mastitic milk samples were subjected to bacterial culturing, and one hundred (76%) S. aureus isolates were identified based on phenotypic characters. Sixty-eight confirmed S. aureus isolates were obtained using PCR. The confirmed S. aureus isolates were tested for six virulence genes (tsst-1, hlb, eta, sea, clfA, and icaD) using PCR. Of the six virulence genes screened from all the isolates, only two (clfA and eta) were detected in the isolates. Out of 68 isolates, 25% and 22% were possessed the eta and clfA genes, respectively. Conclusion The presence of Staphylococcus aureus having virulence genes (eta and clfA) revealed that mastitis is a major concern nowadays affecting animal health, milk quality, and yield. Further genomic study of these isolates will provide broad new insights on virulence.

scholarly journals Bacterial Etiology of Wound Exudates in Tertiary Care Cancer Patients and Antibiogram of the Isolates

2020 ◽  
Vol 13 ◽  
pp. 117863372095207
Author(s):  
Kiran Duwadi ◽  
Sujan Khadka ◽  
Sanjib Adhikari ◽  
Sanjeep Sapkota ◽  
Pabitra Shrestha
Keyword(s):  
Staphylococcus Aureus ◽  
Pathogenic Bacteria ◽  
Methicillin Resistance ◽  
Tertiary Care ◽  
Antibiotic Sensitivity ◽  
Cross Sectional Study ◽  
Diffusion Method ◽  
Disc Diffusion ◽  
Biochemical Tests ◽  
Cross Sectional

Introduction: Patients with malignancies frequently develop infections as a result of surgical procedures and fungating wounds leading to pus formation. This cross-sectional study was conducted to explore the bacteriological spectra of infections of various cancer sites and their antibiotic sensitivity patterns among the patients visiting minor operation theatre (OT) of B.P. Koirala Memorial Cancer Hospital (BPKMCH), Chitwan, Nepal. Methods: Over a period of 3 months from September to November 2018, a total of 183 wound exudates and pus samples were collected and analyzed by standard microbiological procedures. Isolates were identified based on the colony characters, Gram staining and an array of biochemical tests. Antibiotic susceptibility testing was performed by Kirby-Bauer disc diffusion technique according to criteria set by CLSI, 2016. Methicillin resistance in Staphylococcus aureus was tested with the help of cefoxitin using disc diffusion method. Results: Out of the 183 samples, 149 (81.4%) were culture positive. Among 13 different isolates identified, S. aureus (43.0%) was predominant followed by E. coli (14.0%). Higher incidence of bacteria was seen among the males (52.3%), in the age group 51 to 60 years (26.8%) and among the patients undergoing surgical intervention to deal with cancer (34.2%). The prevalence of wound infection was significantly affected by gender, age, and treatment regimen ( P < .01). Out of the total 68 S. aureus isolates, 38 (44.1%) were deemed as Methicillin-resistant Staphylococcus aureus (MRSA). Among the 158 isolates, 85 (53.8%) were multi-drug resistant (MDR). Cefepime was the most effective antibiotic for Gram positive isolates whereas both imipenem and meropenem were found to be equally more effective for Gram negative isolates. Conclusion: This study suggests that patients with malignancies harbor pathogenic bacteria; therefore, prudent use of antibiotics is essential to prevent the emergence of MDR pathogens.

scholarly journals Carriage Methicillin-Resistant Staphylococcus aureus in Poultry and Cattle in Northern Algeria

2018 ◽  
Vol 2018 ◽  
pp. 1-5 ◽  
Author(s):  
Saliha Bounar-Kechih ◽  
Mossadak Taha Hamdi ◽  
Hebib Aggad ◽  
Nacima Meguenni ◽  
Zafer Cantekin
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Broiler Chickens ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Laying Hens ◽  
Methicillin Resistance ◽  
Public Health Problem ◽  
Cross Resistance ◽  
Isolation And Identification ◽  
Methicillin Resistant

Multiresistant and especially Methicillin-Resistant Staphylococcus aureus (MRSA) poses a serious public health problem that requires their immediate identification and antibiotic resistance characteristics. In order to determine antibiotic resistance S. aureus poultry and bovine origin, 8840 samples were collected from slaughterhouses in the northern region of Algeria between years 2009 and 2014. 8375 samples were from an avian origin (1875 from laying hens and 6500 from broiler chickens) and the rest was from bovine origin. Bacteriological isolation and identification were made by classical culture method and antibiotic resistance patterns were determined by disc diffusion test. The prevalence of S. aureus was 42% in laying hens, 12% in broilers, and 55% in bovine samples. The prevalence of MRSA was 57%, 50%, and 31% in laying hens, broiler chickens, and bovine, respectively. While MRSA strains isolated from poultry showed cross-resistance to aminoglycosides, fluoroquinolones, macrolides, sulphonamides, and cyclins, those isolated from bovine also revealed similar multiresistance except for sulphonamide. This high percentage of methicillin resistance and multidrug resistance in S. aureus poultry and bovine origin may have importance for human health and curing of human infections.

scholarly journals Detection of icaA Gene Expression in Clinical Biofilm-Producing Staphylococcus Aureus Isolates

2020 ◽  
pp. 3154-3163
Author(s):  
Shaimaa W. Mohammed ◽  
Hala M. Radif
Keyword(s):  
Gene Expression ◽  
Staphylococcus Aureus ◽  
Bacterial Infections ◽  
Pcr Assay ◽  
Biochemical Tests ◽  
Slime Layer ◽  
Tract Infections ◽  
Strip Test ◽  
Icaa Gene ◽  
Quantitative Pcr Assay

The pathogenicity resulting from Staphylococcus aureus infection has remarkable importance as one of the community-associated bacterial infections, due to the virulent ability of these bacteria to produce biofilms. This study was designed to detect biofilm production in clinical isolates from samples of wounds and urinary tract infections. The expression levels of the icaA gene that is responsible of slime layer production in biofilms was compared in isolates with different biofilm producing capabilities. Fifty seven samples that included 32 samples from urine and 25 samples from wounds were collected from Alwasti Hospital, Al-Kindi Teaching Hospital, and Alzahraa Clinic, Baghdad, Iraq. The bacteria was identified according to biochemical tests, API20 strip test, and PCR assay. The results of 16S rRNA PCR detection revealed that nine isolates were identified as S. aureus. The biofilm assay showed that 46.15% of the isolates were strong biofilm producers, 46.15% had moderate ability to produce biofilm, and 7.70% were weak producers. Quantitative PCR assay was carried out on three isolates with different biofilm-producing abilities. The results demonstrated  that the strong biofilm-producing isolates had significantly higher (P ≤0.01) gene expression level (6.508) compared with the moderate (1.624) and the weak (1.231) isolates.

Isolation and Identification of Staphylococcus Aureus from Dairy Farms in Bishoftu Town, Ethiopia

2021 ◽  
Author(s):  
Sara Amanuel Bude ◽  
Abdi Kidane Mengesha
Keyword(s):  
Staphylococcus Aureus ◽  
Dairy Farms ◽  
Nutrient Agar ◽  
Blood Agar ◽  
Swab Sample ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Mannitol Salt Agar ◽  
Milk Samples ◽  
Golden Yellow

Abstract A cross-sectional study was conducted from February, 2020 to March, 2020 in selected dairy farms in Bishoftu Town to isolate and identify Staphylococcus aureus from milk samples, swab (teat swab, tank swab and bucket swab) samples. The samples were transported to microbiology laboratory and, isolation and identification of an organism was based on morphological, cultural and biochemical characteristics. Accordingly a total of 120 samples of them 58 milk sample, 58 polled teat swab sample, 2 polled tank swab sample and 2 polled bucket swab samples. Those milk samples and swab sample were collected and cultured parallel on both Blood agar and nutrient agar. Out of total samples 68/120 were well grown on both Medias and sub cultured on nutrient agar for farther identification using Primary and secondary biochemical tests. Accordingly, 68 milk samples were shown typical large, round, golden yellow colonies with hemolysis when grown on blood agar, round shaped grape like clusters are seen under microscope after stained with grams stain, and catalase positive Staphylococcus aureus with bubble formation is observed and typical yellow pigmentation on mannitol salt agar which is selective media for genus Staphylococcus. Finally, 68(56.67%) Staphylococcus aureus were identified with coagulase test with the clumping result observed. Hence, implementing hygiene conditions, creation of awareness on control and prevention of subclinical mastitis in dairy farms and conducting drug sensitivity test for Staphylococcus aureus is recommended.

scholarly journals Detection of Antibiotics Sensitivity at Different PH Levels for Proteus Mirabilis Isolated from Patients with Urinary Tract Infections

2018 ◽  
Vol 26 (5) ◽  
pp. 132-144
Author(s):  
Ehsan F. Hussein
Keyword(s):  
High Activity ◽  
Proteus Mirabilis ◽  
Penicillin G ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Neutral Ph ◽  
Antibiotics Susceptibility ◽  
Antibiotics Sensitivity ◽  
Tract Infections

One hundred and sixty-nine samples of urine have been collected through the period from December/2016 to May/2017. Were for isolation and identification of Proteus mirabilis. Isolated from urine in patients with UTI. Identification was done by grow-  ing on different media and biochemical tests as well as the antibiotics susceptibility were studied by using twenty types of antibiotics at acidic pH, neutral pH and alkaline or basic pH by disc diffusion method. These antibiotics were involved Ciprofloxacin, Amikcin, Meromenem, Imipenem, Ampicillin, Rifampin, Gentamicin, Trimethoprim, Tetracycline, Amoxicillin, Sulfamethoxazole, Carbnicillin, Rifaximin, Penicillin-G, Ox- olinic acid, Bacitracin, Clindamycin, Erthromycin, Novamicin and Aztreomycin. The Ciprofloxacin and Aztreomycin have high activity at both acidic and neutral pH, while the Amikcin, Meromenem, Imipenem and Ciprofloxacin have high activity at basicity pH against these bacteria.

scholarly journals Prevalence and Antibacterial Susceptibility Pattern of Bacteria Contaminating the Wards and Operating Room of Mama Lucy Kibaki Hospital, Kenya

2021 ◽  
Author(s):  
Johnstone Amulioto ◽  
Margaret Muturi ◽  
Scholastica Mathenge ◽  
Gideon M. Mutua
Keyword(s):  
Staphylococcus Aureus ◽  
Operating Room ◽  
Hospital Environment ◽  
Health Concern ◽  
Coagulase Negative Staphylococcus ◽  
Susceptibility Pattern ◽  
Biochemical Tests ◽  
Cross Sectional ◽  
Antibiotics Susceptibility ◽  
Susceptibility Tests

Abstract Background Nosocomial infections are of public health concern globally. These infections are frequently caused by microbes residing in the healthcare environment, including contaminated medical equipment, wards, and operating theaters. Objectives To determine the prevalence and susceptibility pattern of bacteria that contaminated the wards and operating rooms of Mama Lucy Kibaki Hospital. MethodsA cross-sectional descriptive study was done from October - November 2018. Surface swabs taken from predefined areas within the wards and operating room were assessed for bacterial contamination. The samples were processed through gram reaction, culture, and an array of biochemical tests. Antibiotics susceptibility tests were done for each swab by the Kirby Bauer disc diffusion technique. Statistical work was done through a statistical package for the social sciences version 20. ResultsThe present work revealed an isolation rate of 64.7% (44/68) for culture-positive surface swabs while 35.3% (24/68) of the swabs failed to yield any organism. A total of 59 bacteria were isolated from the processed swabs. Of the isolates, gram-positive bacteria were more compared to gram-negative rods. Overall, Staphylococcus aureus 55.9% (n=33) was the prevalent bacteria followed by Bacillus species 30.5% (n=18), Citrobacter species 3.4% (n=2), Enterobacter species 3.4% (n=2), Pseudomonas aeruginosa 1.7% (n=1), Acinetobacter species 1.7% (n=1), Escherichia coli 1.7% (n=1) and Coagulase-negative staphylococcus 1.7% (n=1). Methicillin-resistant Staphylococcus aureus (MRSA) accounted for 61.8% of all staphylococci species. The bacteria were resistant to Ampicillin/Cloxacillin at 96.6% (n=57/59). Conclusion The infection of hospitalized patients by resident flora of the hospital environment may worsen the clinical condition of the patient. Therefore, it's paramount for all facilities to ensure that they assess periodically their healthcare environments to identify potential bacterial pathogens of nosocomial significance.

scholarly journals HOSPITALIZED PATIENTS

2016 ◽  
Vol 23 (07) ◽  
pp. 795-801
Author(s):  
Muhammad Saeed ◽  
Farhan Rasheed ◽  
Shahida Hussain ◽  
Maqsood Ahmad ◽  
Mizna Arif ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Clinical Laboratory ◽  
Methicillin Resistance ◽  
Nasal Carriage ◽  
Cross Sectional Study ◽  
Hospitalized Patients ◽  
Coagulase Negative Staphylococcus ◽  
Biochemical Tests ◽  
Carriage Rate ◽  
Cross Sectional

Objectives: An alarming rise in Methicillin resistant Staphylococcus aureus(MRSA) associated hospital based infections has been reported in recent decades.Prolongedhospital stay, unhygienic health services are confined to this challenging serious problem.Anterior nares of humans are a natural reservoir for Staphylococcus aureus with asymptomaticcolonization. This study was designed to determine the prevalence of nasal carriage rate ofMRSA among hospitalized patients. Study Design: Cross sectional study. Setting: PunjabInstitute of Cardiology, Lahore, Pakistan. Period: January 2013- 2014. Materials & Methods:Nasal swabs were collected from hospitalized patients. Total 2,440 patients were screened forthe nasal carriage of MRSA. Microbiologically, identification of S. aureus was done on the basisof colony morphology, Gram staining and biochemical tests. Identified isolates of S. aureus werefurther tested for cefoxitin susceptibility testing, following the technique of modified Kirby Bauer discdiffusion using Mueller-Hinton agar as per Clinical Laboratory Standards Institute (CLSI) guidelines2013. Results: Out of total 2,440 nasal swabs, 60.45% (1475/2440) were Staphylococcusspecies. Among Staphylococcus group, 86.10% (1270/1475) were Staphylococcus aureus.The frequency of MRSA and MSSA (Methicillin sensitive Staphylococcus aureus) was 5.20%(66/1270) and 94.8% (1204/1270) respectively. Overall nasal carriage of MRSA is 2.70%(66/2440). The coagulase negative Staphylococcus were found in 205 (13.90%) cases whileMRSE (Methicillin resistance Staphylococcus epidermdis) were found in only 7 (3.41%) cases.Conclusion: Overall nasal carriage rate of MRSA was 2.70%.

scholarly journals VraSR and Virulence Trait Modulation during Daptomycin Resistance in Methicillin-Resistant Staphylococcus aureus Infection

mSphere ◽  
2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Agustina Taglialegna ◽  
Maria C. Varela ◽  
Roberto R. Rosato ◽  
Adriana E. Rosato
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Cell Membrane ◽  
Virulence Factors ◽  
Bacterial Resistance ◽  
Methicillin Resistant ◽  
Content Type ◽  
Bacterial Physiology ◽  
Two Component ◽  
Mrsa Infection

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) threatens human health in hospital and community settings. The lipopeptide antibiotic daptomycin (DAP) is a frequently used treatment option for MRSA infection. DAP exposure can cause bacterial resistance because mutations are induced in genes implicated in cell membrane and cell wall metabolism. Adaptations aimed at surviving antimicrobial pressure can affect bacterial physiology and modify in vivo aptitude and pathogenesis. In this study, clinical DAP-susceptible (DAPs) and DAP-resistant (DAPr) MRSA isolates were used to investigate associations between DAP resistance and staphylococcal virulence. We previously found that VraSR is a critical sensor of cell membrane/wall homeostasis associated with DAP acquisition during MRSA infection. The present study found that DAPr CB1634 and CB5014 MRSA strains with vraSR upregulation were less virulent than their susceptible counterparts, CB1631 and CB5013. Differential gene-transcription profile analysis revealed that DAPr CB1634 had decreased agr two-component system expression, virulence factors, and highly suppressed hemolysis activity. Functional genetic analysis performed in DAPr CB1634 strains using vraSR inactivation followed by gene complementation found that vraSR acted as a transcriptional agrA regulator. These results indicated that VraSR has a broad range of regulatory functions. VraSR also appeared to affect DAPr adherence to epithelial cells, which would affect DAPr strain colonization and survival in the host. The correlation between DAP resistance and decreased virulence was also found in the CB5013 (DAPs) and CB5014 (DAPr) pair. Taken together, these findings are the first evidence that DAP resistance and MRSA virulence are tightly connected and involve compromised expression of regulatory and virulence determinants. IMPORTANCE Methicillin-resistant S. aureus continues to develop resistance to antimicrobials, including those in current clinical use as daptomycin (DAP). Resistance to DAP arises by mutations in cell membrane and cell wall genes and/or upregulation of the two-component VraSR system. However, less is known about the connection between the pathogen and virulence traits during DAP resistance development. We provide new insights into VraSR and its regulatory role for virulence factors during DAP resistance, highlighting coordinated interactions that favor the higher persistence of MRSA DAP-resistant strains in the infected host.

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