Coagulase Test Research Articles

Isolasi Staphylococcus aureus dari swab tangan penjamah makanan di kantin Universitas Mataram

One of the main qualities of food is seen from the biological safety aspect, such as being free from Pathogenic microorganisms. Staphylococcus aureus is a pathogenic bacterium that can contaminate food ingredients come from the skin of food handlers. S. aureus produces enterotoxins that have a poisoning in consumers.The preliminary observationstated that there were still many food handlers in the Mataram University canteen who did not pay attention to hand hygiene and personal sanitation. This study aims to determine the bacterial contamination of S. aureus swabbed from the hands of food handlers in the Mataram University cafetaria. The method was used in this research is descriptive observational. The research consisted of two stages, namely isolation and biochemical identification. The isolation stage resulted 10 culturable bacterial isolates. All bacteria were able to ferment mannitol on Mannitol Salt Then, the biochemical test identification resulted that all isolates had a positive catalase test, a positive slide coagulase test, Gram-positive, and staphylococci cell. The TSIA test resulted A/A or (acid)/(acid) which indicated that all isolates were able to use diverse carbon sources from sucrose, lactose, glucose. Moreover, other test as sulfur, indole and motility tests were negative. Therefore, all isolates in this study are classified as S. aureus. This research is also a promotive and preventive effort for the campus in implementing hygiene and sanitation for food handlers in the cafetaria.

Characterization and Prevalence of Antibiotic Resistance Staphylococcus aureus in Street Food: A Public Health Concern

Food samples containing Staphylococcus aureus pose a serious health risk. This study aimed to examine the prevalence of S. aureus strains in various food samples sourced from Mailsi and Multan. Many food samples including yogurt, bakery products, and raw and cooked food were examined for S. aureus. The isolated strains were confirmed through Mannitol agar fermentation, catalase, coagulase, and urease tests. Hemolysis on blood agar and biofilm formation were also assessed to determine toxin production. Antibiotic sensitivity testing was conducted using the Kirby Bauer method on MH agar, and multiple antibiotics were tested. Out of all samples, a total of 50 S. aureus strains were obtained, mainly from milk and milk-based products. Yellow colonies on mannitol salt agar confirmed S. aureus presence, with all isolates testing positive for coagulase, catalase, and urease. The presence of hemolysins: beta, gamma, and alpha were revealed by hemolysis assays. Biofilm assay results showed variation among the strains, with some categorized as strong, moderate, or weak biofilm formers. Regarding antibiotic sensitivity, most strains exhibited multidrug resistance, particularly against certain antibiotics. Vancomycin showed varying susceptibility patterns, some strains showed susceptibility and intermediate resistance, whereas only milk samples showed resistance. This study emphasizes the prevalence of MDR S. aureus strains in food samples. The study underscores the significance of antimicrobial stewardship programs and stringent food safety measures in preventing the spread of antibiotic-resistant strains and reducing foodborne illnesses. Further research is needed to explore the mechanisms behind antibiotic resistance and toxin production in S. aureus strains from food samples.

Biofilm Formation among Staphylococcus aureus Strains from Food Contact Surfaces in Households: A Public Health Concern

Staphylococcus aureus is a foodborne pathogen of serious public health concern due to its high biofilm formation capability. One of the contributing factors to foodborne illnesses is improper cleaning and sanitization of food contact surfaces This study aimed to determine the prevalence of S. aureus with biofilm formation potential (BFP) from various food contact surfaces. A total of 100 samples were obtained, 20 from each of stainless steel spoons, plastic plates, plastic eating bowls, plastic cutting boards and glass cups. Samples were obtained by surface swabbing. The cotton swabs were streaked onto nutrient agar plates and incubated overnight. Colonies obtained were subjected to Gram staining, coagulase, catalase and DNAse tests for the identification of S. aureus. The identified S. aureus strains were subjected to biofilm formation potential assay using crystal violet assay (CV-assay) in polystyrene 96-well microtitre plates. Out of the 100 samples obtained, 26 were found to harbour S. aureus. Moreover, out of the 20 isolates, 2 (10%) were from stainless steel spoons, 8 (40%) from plastic plates, 5 (25%) from plastic-eating bowls, 10 (50%) from plastic cutting boards and 1 (5%) from glass cups. Additionally, 11 (42%) of the strains had strong BFP, 11 (42%) had moderate BFP, 3 (12%) and 1 (4%) had no BFP. Presence of S. aureus with BFP on food contact surfaces should be checked to avoid foodborne outbreaks due to this organism.

Developing Field Effect Transistor Sensors Based on DNA Probes to Detect Staphylococcus Lugdunensis Bacteria

The goal of this research is to work with an Electrical Double Layer (EDL)-gated Field Effect Transistor (FET) to detect the bacteria Staphylococcus lugdunensis, which can cause a variety of illnesses, including infections of the skin and soft tissues. In rare cases, Staphylococcus lugdunensis can be successfully identified using the combination of a positive catalase test, a negative tube coagulase test, a positive PYR test, and a positive ornithine decarboxylase test. We are employing an Electrical DoubleLayer (EDL)-gated Field Effect Transistor (FET) to speed up the procedure because this test is time consuming and requires a highly competent individual to perform. An ssDNA probe was developed, and effectively immobilized on a gold surface electrode sensor with a square surface, and due to hybridization with the cDNA at different concentrations, electrical signals were generated in the form of current.Our sensor was fabricated using photolithography where the gold electrode surface having 500 μm x 500 μm gate sensing area consisting of each individually addressable sensor arranged in a 1 x 8 array which is the extent gate was expose for immobilization purpose. The sensor was then subjectedto O2 plasma & HCl solution for surface cleaning and fluorescence image of baregold were taken Figure1(b).The Single-Stranded DNA comprising of fluorescent dye (FAM) at 3’ and thiol modifier at 5’ was drop-casted on the gold surface and was incubated for 24 hours at 24°C. After, it went through a washing process in order to remove the unbound ssDNA and fluorescence images were taken. We observed that intensity was higher than that of baregold which indicates that our probe has successfully immobilized on the gold surface Figure 1(b).Figure 1(a) represents the device that we designed. The system works with N-Channel Enhancement-Mode DMOS FET (VN10LP) in which a continuous drain voltage (3.5 V) was applied and the drain current which is the signal, were measured at two distinct gate bias voltage (2V and 3V). cDNA having quencher (BHQ1) at 5’ was diluted in different concentration namely 1fM, 10fM, 100fM and 1000fM. These cDNA with different concentrations were drop-casted on our sensor with Tris-EDTA (TE) buffer as the baseline and starting with lower concentration to higher concentration every after 12 seconds periodically where we record the change in signal as shown in Figure 1(c).After the electrical measurement, fluorescence image was taken and we can observe that it quenched which indicates that the cDNA and the immobilized ssDNA has successfully hybridized which give the signal change. Figure 1: (a) Surface Immobilization Principal Display in Extended Gate EDL BioFETs. (b)Fluorescence value comparison. (c) Plot showing the change in drain current over time for aptamersensors exposed to various cDNA concentrations. Figure 1

Panton-Valentine leucocidin gene in methicillin resistant Staphylococcus aureus isolated from tertiary care hospital in Nepal.

Methicillin-resistant Staphylococcus aureus (MRSA) expresses the Panton-Valentine leukocidin (PVL) virulence gene, which is associated with community and hospital-acquired severe MRSA infections. The objective of this study was to determine the prevalence and antibiotic susceptibility profile with a focus on the presence of the PVL gene among MRSA isolates in healthcare settings. A total of 1,207 clinical specimens and 304 hospital environment swabs were collected in a tertiary care hospital in Nepal, and investigated following basic microbiological techniques. S. aureus was confirmed with the coagulase test. An antibiotic susceptibility test (AST) was performed by the Kirby-Bauer method and screening for MRSA was carried out by the cefoxitin disc diffusion method guided by the Clinical and Laboratory Standards Institute (CLSI), 2020. DNA was extracted and used in a polymerase chain reaction (PCR) to detect mecA and PVL genes. Of the 1,511 samples, 45 (2.9%) S. aureus (23 clinical and 22 environmental) were isolated. Among them, 69.6% (16/23) and 27.3% (6/22) were MRSA in clinical and environmental isolates, respectively. Twelve (52.2%) clinical isolates and seven (31.8%) environmental isolates were multidrug resistant. The majority of isolates were susceptible to vancomycin and linezolid. The PVL gene was detected in 18.2% (n = 4/22) of the MRSA isolates, of which three were from clinical sources and one was from an environmental swab. The prevalence of MRSA, and PVL-producing S. aureus were higher in the hospital setting. Hence, immediate and urgent implementation of infection control and sanitation measures are needed in the hospital.

KOLONISASI BAKTERI METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) PADA UANG KERTAS YANG BEREDAR DI KANTIN FAKULTAS KEDOKTERAN UNIVERSITAS UDAYANA

Money serves as a widely utilized medium of transactional value in society. The risk of infection resulting from bacterial dissemination on paper currency is notably high. Infections caused by microorganisms on paper currency are largely caused by bacteria that resist antibiotics, exemplified by Methicillin-Resistant Staphylococcus aureus (MRSA). The resistance exhibited by these bacteria to antibiotic treatments has claimed millions of lives and can lead to substantial global health repercussions. Therefore, this study aims to ascertain the presence of Methicillin-Resistant Staphylococcus aureus (MRSA) bacteria contaminating paper currency. This research was conducted with observational methodology and a descriptive cross-sectional approach. Thirty samples of paper currency are swabbed, and specimens are subsequently inoculated onto Staphylococcus medium. Colonies of Staphylococcus are re-identified using gram staining, catalase tests, followed by coagulase tests to ascertain the presence of Staphylococcus aureus bacteria. Isolates are classified as Methicillin-Resistant Staphylococcus aureus (MRSA) if the inhibition zone measures less than or equal to 21 mm. The research findings indicate the colonization of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus (MRSA) bacteria on the paper currency circulating in the canteen of the Medical Faculty at Udayana University. The percentage of Staphylococcus aureus colonization is 56.66% and Methicillin-Resistant Staphylococcus aureus (MRSA) colonization is 53.33%. Keywords : Paper Currency, Methicillin-Resistant Staphylococcus aureus(MRSA).

Antimicrobial susceptibility patterns of urinary tract infections causing bacterial isolates and associated risk factors among HIV patients in Tigray, Northern Ethiopia

BackgroundUrinary tract infections, a prevalent global infectious disease, are clinical issues not well studied in HIV-positive individuals. UTIs have become a global drug resistance issue, but the prevalence and antibiotic susceptibility patterns of UTI-causing bacteria among HIV patients in Tigray, Ethiopia, are poorly understood. This study aims to identify the prevalence of UTI-causing bacteria, their antibiotic susceptibility patterns, and associated risk factors in HIV patients attending ART clinics at Mekelle General Hospital and Ayder Comprehensive Specialized Hospital in Tigray, Northern Ethiopia.MethodClean-catch midstream urine samples (10–15 mL) were collected from HIV patients who are attending ART clinics at Mekelle General Hospital and Ayder Comprehensive Specialized Hospital. Samples were analyzed based on standard microbiological protocols using cysteine-lactose electrolyte deficient (CLED) agar. Pure colonies of bacterial isolates were obtained by sub-culturing into Mac-Conkey, Manitol Salt agar and blood agar plates. The bacterial isolates were then identified using macroscopic, microscopic, biochemical, and Gram staining methods. Gram-negative bacteria were identified using biochemical tests like triple sugar iron agar, Simon’s citrate agar, lysine iron agar, urea, motility test, and indol test, whereas Gram-positive isolates were identified using catalase and coagulase tests. The Kirby-Bauer disk diffusion technique was used to analyze the antimicrobial susceptibility pattern of bacterial isolates. Data was analyzed using SPSS version 25.0.ResultsAmong the 224 patients, 28 (12.5%) of them had been infected by UTIs-causing bacteria. E. coli was the dominant bacterium (16 (57%)) followed by K. pneumoniae (4 (14%)), and S. aureus (3 (11%)). Of the total bacterial isolates, 22 (78.6%) of them developed multi-drug resistance. All Gram-positive (100%) and 75% of Gram-negative bacterial isolates were found to be resistant to two or more drugs. Patients with a history of UTIs, and with CD4 count < 200 cells/ mm3, were more likely to have significant bacteriuria. Compared to male patients, female patients were more affected by the UTIs-causing bacteria. More than 93% of the UTIs-causing bacterial isolates were susceptible to nitrofurantoin, ceftriaxone, ciprofloxacin, and gentamycin; whereas they are highly resistant to ampicillin (96%), cotrimoxazole (82%) and tetracycline (71%).ConclusionsMost of the bacterial isolates were highly resistant to ampicillin, cotrimoxazole, and tetracycline. Female patients were more affected by the UTIs causing bacteria. The highest prevalence (12.5%) of UTIs in HIV patients needs special attention for better management and monitoring. Previous UTI history and immune suppression are predictors of UTIs, highlighting the need for intervention measures involving molecular studies to identify resistant bacteria genes and promote patient immune reconstitution.

Occurrence, antimicrobial susceptibility, and resistance genes of Staphylococcus aureus in milk and milk products in the Arsi highlands of Ethiopia.

In Ethiopia, milk production and handling practices often lack proper hygiene measures, leading to the potential contamination of milk and milk products with Staphylococcus aureus (S. aureus), including methicillin-resistant strains,posing significant public health concerns. This study aimed to investigate the occurrence, antimicrobial susceptibility profiles and presence of resistance genes in S. aureus strains isolated from milk and milk products. A cross-sectional study was conducted in the Arsi highlands, Oromia, Ethiopia from March 2022 to February 2023. A total of 503 milk and milk product samples were collected, comprising 259 raw milk, 219 cottage cheese, and 25 traditional yogurt samples. S. aureus isolation and coagulase-positive staphylococci enumeration were performed using Baird-Parker agar supplemented with tellurite and egg yolk. S. aureus was further characterized based on colony morphology, Gram stain, mannitol fermentation, catalase test, and coagulase test. Phenotypic antimicrobial resistance was assessed using the Kirby-Bauer disc diffusion method, while the polymerase chain reaction (PCR) was employed for confirming the presence of S. aureus and detecting antimicrobial resistance genes. S. aureus was detected in 24.9% of the milk and milk products, with the highest occurrence in raw milk (40.9%), followed by yogurt (20%), and cottage cheese (6.4%). The geometric mean for coagulase-positive staphylococci counts in raw milk, yogurt, and cottage cheese was 4.6, 3.8, and 3.2 log10 CFU/mL, respectively. Antimicrobial resistance analysis revealed high levels of resistance to ampicillin (89.7%) and penicillin G (87.2%), with 71.8% of the isolates demonstrating multidrug resistance. Of the 16 S. aureus isolates analyzed using PCR, all were found to carry the nuc gene, with the mecA and blaZ genes detected in 50% of these isolates each. This study revealed the widespread distribution of S. aureus in milk and milk products in the Arsi highlands of Ethiopia. The isolates displayed high resistance to ampicillin and penicillin, with a concerning level of multidrug resistance. The detection of the mecA and blaZ genes in selected isolates is of particular concern, highlighting a potential public health hazard and posing a challenge to effective antimicrobial treatment. These findings highlight the urgent need to enhance hygiene standards in milk and milk product handling and promote the rational use of antimicrobial drugs. Provision of adequate training for all individuals involved in the dairy sector can help minimize contamination. These measures are crucial in addressing the threats posed by S. aureus, including methicillin-resistant strains, and ensuring the safety of milk and its products for consumers.

Frequency and antimicrobial susceptibility profile of Methicillin-resistant Staphylococcus aureus isolated from Pus and Swab samples in a Teaching Hospital of Lahore

Background: Staphylococcus is a ubiquitous bacterium and well-known pathogen causing a variety of infections. The global spread of MRSA constitutes one of the most prevailing challenges to the management of infections caused by this bug.&#x0D; Aim: To determine the frequency of MRSA and antibiogram profile of S. aureus recovered from various clinical samples.&#x0D; Setting: Microbiology Laboratory of Shalamar Hospital Lahore.&#x0D; Methods: The data of the Staphylococcus isolates including MRSA from various pus and swab samples was collected through Electronic Medical Record of the Shalamar Hospital from 1st Jan to 31st Dec 2021. S. aureus was identified by standard protocol including Gram stain, catalase, coagulase, and DNase tests. Antimicrobial susceptibility was carried out by modified Kirby Bauer method. MRSA frequency was determined by the result of sensitivity to cefoxitin.&#x0D; Results: Out of 885 samples submitted for culture, 517 (58.4%) were reported for microbial growth of a known pathogen. The most frequently isolated pathogen was S. aureus (37.9%), followed by E. coli (22.4%), other members of Enterobacteriaceae family (17.8%), Pseudomonas (15.5%), Enterococcus (3.5%), Candida (2.1%), and Streptococcus (0.8%). Amongst S. aureus, MRSA was documented in 46.9% cases. Substantial difference was detected in the susceptibility pattern of methicillin-sensitive and resistant staphylococci. All staphylococci were resistant to ampicillin while no vancomycin resistance was encountered.&#x0D; Conclusion: MRSA was seen in the local population with a high frequency of 46.9% and such MRSA also showed marked resistance against other commonly used antibiotics. Fortunately no vancomycin-resistant S. aureus was reported.

Antibiotic Susceptibility and Biofilm Production among Coagulase Negative Staphylococci Isolated from Clinical Samples at Tertiary Care Hospital.

Coagulase Negative Staphylococci have been widely associated with medical device implant treatment and immune-compromised patients. Despite having increasing interest in Coagulase Negative Staphylococci, few studies from Nepal have reported the association of these organisms with urinary tract infections, conjunctivitis, high vaginal swabs, and cerebrospinal fluid. This study was carried out to determine antibiotic susceptibility pattern and biofilm production among Coagulase Negative Staphylococci isolated from clinical samples at tertiary care hospital. This study was a hospital based cross-sectional study in which 3690 clinical samples were included. Isolation and identification of isolates was done following standard microbiological protocol. Coagulase Negative Staphylococci were identified phenotypically on the basis of gram staining, slide and tube coagulase test and by various sugar fermentation tests. Antibiotic susceptibility test was done following Kirby Bauer disk diffusion method (Clinical and Laboratory Standards Institute 2020). Biofilm production was determined by Tissue Culture Plate technique. A total of 113 isolates of Coagulase Negative Staphylococci were detected. Among them S. epidermidis (45.1%), S. saprophyticus (23.9%), S. haemolyticus (16.8%), S. hominis (5.3%), S. capitis (2.7%), -----S. cohini (1.8%), S. lugdunensis (1.8%) and S. sciuri (2.7%) were identified phenotypically. All isolates were found to be resistant against Ampicillin and 111 (98.2%) were sensitive against Linezolid.23.9% of CoNS were strong biofilm producers, 19.5% moderate and 56.6 % were non/weak biofilm producers. It requires susceptibility test for prescribing antibiotics against Coagulase Negative Staphylococci in hospital and the misuse of antibiotics should be prevented.

Antibacterial activities of honey bee against gram-positive bacteria isolated from urine

The medicinal benefits of honey have long been recorded in ancient days as a solution for various ailments and infections. This study x-rayed the effects of honey on common gram-positive bacteria isolated from urine. The study evaluated the in vitro susceptibility of two gram positive bacteria strains, isolated from urine of students of Nnamdi Azikiwe University, Awka Anambra state, Nigeria. The end point was achieved via macroscopic, microscopic methods and biochemical examinations which includes: gram staining, urease, catalase, coagulase testing to a hsoney sample. The evaluation of the antibacterial activity was ascertained by the agar diffusion well method. The results obtained in this study proved that the honey sample exhibited potent antibacterial activity against the tested strains, the zone of inhibition was at a mean value of 20.5 mm and 29.5 mm respectively at 100% concentration of honey. Staphylococcus epidermidis was found to be less susceptible than Enterococcus faecalis with a mean inhibition zones being 29.5 mm. This contribution has provided a broad overview of the antibacterial activity of honey and shown that honey bee has great potential for therapeutic use as an alternative therapy for infections caused by the isolates.

Resistencia a antibióticos de Staphylococcus aureus aislado de la cavidad bucal de participantes paraguayos de Coronel Oviedo, año 2021

A cross-sectional study was carried out to identify the resistance profile of Staphylococcus aureus colonizing the oral cavity of patients. Sampling was carried out with sterile swabs. Mannitol-positive colonies were cultured on blood agar, identifying S. aureus by conventional biochemical tests that included positive results for catalase, coagulase, DNAse and latex agglutination test for S. aureus. The resistance to 11 antibiotics was tested. Forty-three subjects were included; 83.7% presented oral pathologies, and 6.9% had a history of hospitalization, 18.6% mentioned having used antibiotics in the last three months. S. aureus was isolated in 3 patients; the 3 isolates presented resistance to penicillin, while 2 of the three isolates were resistant to oxacillin and cefoxitin. Resistance to erythromycin and clindamycin was also present in one patient. Two of the cases presented resistance to various drugs and didn´t report any risk factors.

Prevalence of Staphylococcus aureus on Noncritical Surfaces of the Laboratories of a Medical Laboratory Training Institution

Staphylococcus aureus is a Gram positive bacterium that is frequently encountered on surfaces such as benches and skin. This organism being a normal flora of the skin is usually nonpathogenic but becomes pathogenic when found outside its normal flora where it can cause varying number of infections such as nosocomial infections and sepsis which may even lead to death if left untreated. This research was therefore carried out to determine the prevalence of Staphylococcus aureus on noncritical surface areas including: Furniture, Doors and Windows of the laboratories of Federal College of Medical Laboratory Science &amp; Technology Jos, Nigeria. Samples were processed, examined and analyzed accordingly. It was observed that out of the 120 samples examined, 8 were gram positive cocci in clusters under Gram staining examination Coagulase and catalase tests were positive indicating specific biochemical tests to identify the organisms in the 8 organisms as Stapylococcus aureus showing a prevalence rate of 6.67%. The 17 working Benches examined, 3(2.50%) were positive. Out of 75 Chairs examined, 5(4.17%) were positive. The 18 Windows examined, 10 Doors examined showed no growth for Staphylococcus aureus. Distribution according to laboratory sections showed that, out of the 95 samples in main laboratory, 6(5.00%) were positive. Out of the 14 samples examined in side laboratory one, 2(1.67%) were positive while none 0(0.0%) was positive out of 11 samples examined in side laboratory two. The isolation of Stapylococcus aureus from these locations stresses the urgent need for public enlightenment campaigns by the appropriate authorities to educate the laboratory workers as well as the masses on the etiological agents, the possible risk factors, routes of transmission and health implication of Stapylococcus aureus infection and ways of preventing these bacteria in the Laboratories especially in Federal College of Medical Laboratory Science and Technology Jos Nigeria.

Distinguishing characteristics of Staphylococcus schleiferi and Staphylococcus coagulans of human and canine origin.

Staphylococcus schleiferi and Staphylococcus coagulans are opportunistic pathogens of animals and humans. They were previously classified as Staphylococcus schleiferi subs. schleiferi and Staphylococcus schleiferi subs. coagulans, respectively, and recently reclassified as separate species. S. coagulans, is frequently associated with dogs, whereas S. schleiferi is more commonly isolated from humans. Coagulase activity status is a defining characteristic of the otherwise closely related species. However, the use of coagulase tests originally developed to distinguish S. aureus from non-coagulase-producing staphylococci, for this purpose is questionable and the basis for their host preference has not been elucidated. In the current study, a putative coa gene was identified and correlated with coagulase activity measured using a chromogenic assay with human and bovine prothrombin (closely related to canine prothrombin). The results of the tests performed with human prothrombin showed greater reactivity of S. coagulans isolates from humans than isolates obtained from dogs with the same substrate. Our data suggest that unlike S. coagulans isolates from humans, isolates from dogs have more coagulase activity with bovine prothrombin (similar to canine prothrombin) than human prothrombin. Differences in nuc and 16s rRNA genes suggest a divergence in S. coagulans and S. schleiferi. Phenotypic and genotypic variation based on the number of IgG binding domains, and the numbers of tandem repeats in C-terminal fibronectin binding motifs was also found in protein A, and fibronectin-binding protein B respectively. This study identified a coa gene and associated phenotypic activity that differentiates S. coagulans and S. schleiferi and identified key phylogenetic and phenotypic differences between the species.

Study of Methicillin-resistant Staphylococcus aureus Isolated from Various Clinical Specimens from Tertiary Care Hospital

ABSTRACT Introduction: Staphylococci are Gram-positive bacteria, with a diameter of 0.5–1.5 μm and individual cocci, which are divided in more than one plane from the grape-like cluster. In the genus staphylococci, 32 species and 8 subspecies are present in many of which are present as a commensal in the human body. Staphylococci are frequently found as a normal flora of skin mucosal, surface of respiratory, upper elementary, and urogenital tract of the human body. In immunocompromised condition, the infection can be spread by skin-to-skin contact and also by excretion such as saliva aerosol releasing during sneezing and coughing. Infection of staphylococci may also spread from the animal product such as nonpasteurized milk and milk products. Over the last few decades, Staphylococcus aureus has emerge as the most dangerous organism, which is known as methicillin-resistant S. aureus (MRSA). Some strains show vancomycin resistance by S. aureus in hospitalized patients. The present study was conducted to study the prevalence and antibiogram of MRSA isolates in a clinical sample from Acharya Vinoba Bhave Rural Hospital. Materials and Methods: This study was conducted in the Department of Microbiology at Jawaharlal Nehru Medical College (Datta Meghe Institute of Higher Education and Research), Wardha, from August 2020 to December 2021. The samples received in the microbiology laboratory were inoculated on blood agar and MacConkey’s agar, and incubated for 24 h at 37°C for staphylococcal isolation. A total of 145 Staphylococcal aureus were isolated and confirmed by Gram stain and various biochemical tests such as catalase test, oxidase test, coagulase test, and mannitol salt agar. Antibiotic susceptibility test was examined on the Mueller–Hinton agar plate using the modified Kirby–Bauer disc diffusion method. Screening of methicillin resistant was performed by cefoxitin (30 μg) disc diffusion method and interpreted according to CLSI guideline. Results: The prevalence of MRSA was 76 (52.41%) to methicillin-susceptible S. aureus. The highest MRSA was found in the age group &gt;50 years (26.31%). Out of 76 MRSA isolates from different departments, the medicine and orthopedic departments show maximum MRSA as 19.74% each followed by pediatrics (17.11%) and surgery (13.16). Out of 76 MRSA isolates, vancomycin showed 93.42% sensitivity followed by linezolid (85.52%), Gentamicin (71.05%), tetracycline (56.57%), clindamycin (46.05%), and erythromycin (31.57%). Conclusion: With the advancement of new therapeutic agent, MRSA is also being easily resistant. Due to increasing cases of MRSA, there is an increase in the rate of morbidity and mortality. Moreover, except vancomycin, other drugs are resistant, as their MIC is not in the susceptible range. Thus, the results of this study indicate that vancomycin is a suitable medication to take when MRSA cases rise. In such cases, MRSA can be controlled by regular monitoring and checking the irrational use of antibiotics.

Alarming multidrug resistance in Staphylococcus aureus isolated from raw milk of cows with subclinical mastitis: Antibiotic resistance patterns and occurrence of selected resistance genes.

Bovine mastitis is a widespread and costly disease that affects dairy farming globally, characterized by mammary gland inflammation. Bovine intramammary gland infection has been associated with more than 135 different pathogens of which Staphylococcus aureus is the main etiology of sub-clinical mastitis (SCM). The current study was designed to investigate the prevalence, antibiotic resistance pattern, and the presence of antibiotic resistance genes (mecA, tetK, aacA-aphD and blaZ) in S. aureus isolated from the raw milk of cows with subclinical mastitis. A total of 543 milk samples were collected from lactating cows such as Holstein Friesian (n = 79), Sahiwal (n = 175), Cholistani (n = 107), and Red Sindhi (n = 182) from different dairy farms in Pakistan. From the milk samples microscopic slides were prepared and the somatic cell count was assessed to find SCM. To isolate and identify S. aureus, milk was streaked on mannitol salt agar (MSA) plates. Further confirmation was done based on biochemical assays, including gram staining (+ coccus), catalase test (+), and coagulase test (+). All the biochemically confirmed S. aureus isolates were molecularly identified using the thermonuclease (nuc) gene. The antibiotic resistance pattern of all the S. aureus isolates was evaluated through the disc diffusion method. Out of 543 milk samples, 310 (57.09%) were positive for SCM. Among the SCM-positive samples, S. aureus was detected in 30.32% (94/310) samples. Out of 94 isolates, 47 (50%) were determined to be multidrug resistant (MDR). Among these MDR isolates, 11 exhibited resistance to Cefoxitin, and hence were classified as methicillin-resistant Staphylococcus aureus (MRSA). The S. aureus isolates showed the highest resistance to Lincomycin (84.04%) followed by Ampicillin (45.74%), while the least resistance was shown to Sulfamethoxazole/Trimethoprim (3.19%) and Gentamycin (6.38%). Polymerase chain reaction (PCR) analysis revealed that 55.31% of the isolates carried blaZ gene, 46.80% carried tetK gene, 17.02% harbored the mecA gene, whereas, aacA-aphD gene was found in 13.82% samples. Our findings revealed a significant level of contamination of milk with S. aureus and half (50%) of the isolates were MDR. The isolated S. aureus harbored various antibiotic resistance genes responsible for the absorbed phenotypic resistance. The alarmingly high prevalence of MDR S. aureus isolates and MRSA strains in these cases possess a serious risk to public health, emphasizes the urgent need to address this issue to protect both human and animal health in Pakistan.

The Antibacterial Effect of Aloe Vera Gel Extract against Escherichia coli, Salmonella typhi and Staphylococcus aureus Isolated from Gastrointestinal Tract of Poultry Birds.

The Antibacterial effect of Aloe vera gel extract against Escherichia coli, Salmonella typhi and Staphylococcus aureus isolated from the gastrointestinal tract of poultry birds was investigated. Standard microbiological methods were used for the isolation of Escherichia coli, Salmonella typhi and Staphylococcus aureus. The prevalence of Escherichia coli, Salmonella typhi and Staphylococcus aureus in the droppings of the poultry birds was 89%. The organisms were identified following various biochemical tests, like Gram Staining, catalase test, Coagulase test, Citrate test and Indole test. The antibacterial activity of Aloe vera gel extracts were standardized by plating out 1.05×105 cfu/ml, while serial dilutions of 10-1-10-10 were used to evaluate efficacy of the inhibitory effect using the Agar Gel diffusion method on different concentration on the test organisms. The result shows that Aloe vera gel extract at different concentrations inhibited the growth of the microorganisms. Though there was a higher inhibitory effect on Staphylococcus aureus at 10-1, 13.5 mm and 14.5 mm for Ist and 2nd agar well respectively and a lower inhibitory growth of 8 mm and 8.3 mm at 10-8. There was no inhibition on 10-9 and 10-10, followed by Escherichia coli at 10-1, 11.5 mm and 12.5 mm for Ist and 2nd agar well respectively and a lower inhibitory growth of 10 mm at 10-6. There was no inhibition on 10-7 to 10-10 and lastly Salmonella typhi at 10-1, 10.5 mm and 11.5 mm for Ist and 2nd agar well respectively and a lower inhibitory growth of 9 mm at 10-7. There was no inhibition on 10-8 to 10-10. The study revealed that Aloe vera gel extract possessed antibacterial properties. Therefore, it can be added to the feed of poultry birds as a prophylactic to reduce bacterial infections and would be used to test the natural efficacy of Herbal medicine.

COMPARATIVE STUDY OF STAPHYLOCOCCUS AUREUS ISOLATES IN PUS AND NASAL SWABS AMONG LAHORE PATIENTS

Staphylococcus aureus predominately found in nasal and pus samples is one of the major causes of skin infections and invasive diseases like endocarditis, pneumonia, and osteomyelitis. The current study was performed over the 3 month period, from April 2022 to June 2022. A total of 368 participants’ nasal and pus swab specimens were included in this study and were collected from the different hospitals in Lahore. The objective of the study was to discern the relative prevalence of Staphylococcus aureus isolates in Pus and Nasal Swabs among Patients in Lahore. All the collected samples were cultured in sterile conditions on Blood agar, Chocolate agar, MacConkey agar, and Mannitol salt agar according to the standard laboratory guidelines. S. aureus culture plates were identified based on the colonial morphology after 48hrs. Furthermore, isolated bacteria were characterized based on Gram stain, and differential biochemical analyses like the Coagulase test, Oxidase test, Catalase test, Triple sugar iron, Urease test, Indole test, Motility agar test that was performed to reconfirm S. aureus presence in the given pus and nasal specimens. The modified Kirby Bauer disc diffusion method was employed for antimicrobial susceptibility testing and Zone sizes were interpreted according to the Clinical and Laboratory Standard Institute guidelines (2020). From a total of 368 bacterial samples, 283 were pus swabs and 85 were nasal swabs, whereas the most frequently isolated organism was gram-positive Staphylococcus aureus which accounts for 90% (330) of the total samples. Our results showed a 90 % MRSA prevalence rate in Lahore and in our study oxacillin represents high resistance against MRSA in nasal samples, whereas, clindamycin represents high resistance in pus samples respectively. The reason for this increased prevalence of MRSA is due to the irrational use of antibiotics without prior cultural reports, the absence of antimicrobial situation programs in hospitals, and the failure to look at the infection management practices which may be some of the highlighted reasons for this drawback in the hospital and clinical setup.

Species distribution and antimicrobial susceptibility pattern of coagulase-negative staphylococci isolated from clinical specimens at a tertiary care hospital

Background and objectives: Coagulase-negative staphylococci (CoNS) are considered important causative agents of hospital acquired infection. These organisms are found in various clinical specimens from hospitalized patients. Present study was carried out to determine the species distribution and antimicrobial susceptibility pattern of CoNS isolated from clinical specimens at a tertiary care hospital. Materials and methods: CoNS isolated from various clinical samples were included in this study. The isolates were identified by colony morphology, Gram’s staining, catalase and coagulase tests. Further differentiation of species was performed by susceptibility to novobiocin, urease activity and ornithine decarboxylase test. Antibiotic susceptibility testing was performed according to the Clinical and Laboratory Standard Institute (CLSI) guidelines. Results: Total 108 isolates of CoNS were included and analysed. Out of 108 CoNS, S. epidermidis was the most common species (36.1%) followed by S. saprophyticus (23.1%), S. hemolyticus (17.6%), S. hominis (13%) and S. lugdunensis (10.2%). Most of the isolates showed resistance to penicillin, oxacillin, amoxycillin, erythromycin, ciprofloxacin and ofloxacin. All the isolates were sensitive to vancomycin. Conclusions: CoNS emerged as an important nosocomial pathogen and should not be neglected as contaminant. High rate of antimicrobial resistance warrants susceptibility testing prior to the treatment of CoNS. IMC J Med Sci. 2024; 18(1):010. DOI: https://doi.org/10.55010/imcjms.18.010 *Correspondence: Sabiha S Tamboli, Department of Microbiology, Parbhani Medical College and RP Hospital Research Institute, Pathri Road, Parbhani, Maharashtra, India. Email: sabihatamboli77@gmail.com

Prevalence of livestock associated methicillin resistant Staphylococcus aureus (La-mrsa) in domestic livestock of Dharan, Nepal

Introduction: Methicillin Resistant Staphylococcus aureus (MRSA) infection has drawn a lot of attention since studies suggested that animals may act as reservoirs for human infection. Over the past 20 years, MRSA infections have become more prevalent worldwide. MRSA was initially only discovered in humans, but later it was discovered in animals as well. Objectives: The study aimed to determine the prevalence of LA-MRSA in domestic livestock of Dharan, Nepal. Methods: Overall, 320 skin swab samples of cattle (cows) were collected by swabbing the skin of cattle aseptically with a sterile cotton swab and the samples were transported to the laboratory in a cold chain. The samples were streaked in Mannitol salt agar(MSA) containing oxacillin concentration of 6mg/L and incubated at 37°C for 24 hours. Staphylococcus aureus colonies were identified based on cultural characteristics on MSA plates (golden yellow colonies), Gram’s reaction, and positive results for coagulase and catalase test. The purified isolates of MRSA were subjected to antibiotic susceptibility tests and Biofilm formation. Results: MRSA was found to be prevalent in 10% of cattle. Antibiotic-resistant tests reported that MRSA was found to be sensitive to chloramphenicol 12(37.5%) followed by ceftriaxone 12 (37.25%), gentamycin 9(28.12%), cefotaxime 4(12.5%), while 100% of MRSA were resistant to penicillin and vancomycin. The moderate biofilm-forming MRSA was 5(15.625%), followed by 3(9.375%) weak biofilm producers and 24(75%) were negative towards biofilm formation. Conclusion: The prevalence of MRSA (10%) and Vancomycin-Resistant Staphylococcus aureus (100% VRSA) in cattle shows the need for regular surveillance. Chloramphenicol can be a treatment of choice for MRSA infections. However, emerging VRSA is a serious epidemiological issue that needs to be addressed properly. Therefore, Healthcare organizations must adopt precise criteria to control and prevent MRSA infection..