Detection of blaTEM, blaCTX-M, blaCMY, and blaSHV Genes Among Extended-Spectrum Beta-Lactamase-Producing Escherichia coli Isolated from Migratory Birds Travelling to Bangladesh.

Extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli cause severe health hazards. Migratory birds are reservoirs and transmitters of many pathogens including ESBL-producing E. coli. To examine migratory birds as potential carriers of ESBL-producing E. coli and E. coli-carrying antibiotic resistance genes, 55 PCR-positive E. coli isolates were screened using the disk diffusion method, double-disk synergy test, and further polymerase chain reaction (PCR) tests. Genes encoding resistance to tetracycline [tetA, 100% (35/35); tetB, 31.43% (11/35)], fluoroquinolone [qnrA, 35.71% (10/28); qnrB, 25% (7/28)], and streptomycin [aadA1, 90.24% (37/41)] were detected in the isolated E. coli. Of the 55 E. coli isolates, 21 (38.18%) were ESBL producers, and all of them were multidrug resistant. All the ESBL-producing E. coli isolates harbored at least two or more beta-lactamase genes, of which blaTEM, blaCMY, blaCTX-M, and blaSHV were detected in 95.24%, 90.48%, 85.71%, and 42.86% of isolates, respectively. All the beta-lactamase genes were present in four of the ESBL-producing E. coli isolates. Furthermore, 95.24% of ESBL-producing E. coli isolates were positive for one or more antibiotic resistance genes. To the best of our knowledge, this is the first study to detect E. coli-carrying antibiotic resistance genes including beta-lactamase blaCMY and blaSHV originating from migratory birds in Bangladesh. These results suggest that migratory birds are potential carriers of ESBL-producing E. coli along with other clinically important antibiotic resistance genes which may have detrimental impacts on human health.

Draft genome sequencing of a multidrug-resistant Salmonella enterica subspecies enterica serovar Typhimurium strain isolated from chicken in Bangladesh.

Herein this study, we sequenced the genome of a multidrug-resistant Salmonella enterica serovar Typhimurium strain MBR-MFRK-23 isolated from the liver tissue of a diseased layer chicken. The 4,964,854-bp draft genome comprises 50 contigs with 50.5× coverage and 52.1% GC content and is typed as S. enterica sequence type 19.

Complete genome sequence of Pseudomonas aeruginosa lytic bacteriophage PA1O which resembles temperate bacteriophage D3112.

A novel Pseudomonas aeruginosa lytic bacteriophage (phage), PA1Ø, was isolated, and its genome was sequenced completely. This phage is able to lyse not only P. aeruginosa but also Staphylococcus aureus. Genome analysis of PA1Ø showed that it is similar to a P. aeruginosa temperate phage, D3112, with the exception of the absence of a c repressor-encoding gene, which is known to play a critical role in the maintenance of the lysogenic state of D3112 in P. aeruginosa.

Draft genome sequence of Staphylococcus gallinarum BAU_KME002 strain isolated from egg surface in Bangladesh.

This report describes the genome sequence of the Staphylococcus gallinarum BAU_KME002 strain isolated in Bangladesh in 2021 from a chicken egg surface. Our assembled genome had 50 contigs, an estimated genome length of 2,866,882 bp (with coverage of 90.0×), 36 predicted antibiotic resistance genes, and 28 predicted virulence factor genes.

Domestic cats are potential reservoirs of multidrug-resistant human enteropathogenic E. coli strains in Bangladesh.

Companion animals serve as our best friends, confidants, and family members. Thus, disease and antibiotic resistance gene transmission in pets and humans must be sought out. The study aimed to identify the common pathogenic Escherichia coli (E.coli) in pet cats and the antibiotic resistance patterns and resistant gene distribution. Samples (n = 210) were collected from different veterinary clinics in Bangladesh's cities of Mymensingh and Dhaka. Pathogenic E. coli was identified using conventional and molecular approaches. The disc diffusion method assessed the resistance profile against 12 antibiotics, and PCR was used to identify the beta-lactam resistance genes. The prevalence of the stx-1 gene was found to be 2.86%, whereas the rfbO157 prevalence was found to be 1.90% in cats. The stx-1 gene (n = 6) was 100% resistant to erythromycin and imipenem, whereas 100% sensitive to chloramphenicol. In turn, the rfbO157 gene (n = 4) exhibited 100% resistance to erythromycin, imipenem, cefixime, and azithromycin. In addtion, we identified genes that exhibit resistance to beta-lactam antibiotics (100% blaTEM, 40% blaCTX-M, 40% blaSHV2). This study found shiga-toxin producing and extended-spectrum beta-lactamase (ESBL) producing E. coli for the first time in pet cats of Bangladesh. Furthermore, the antimicrobial resistance (AMR) profile of the isolated strains refers to the occurrence of multidrug, which concerns cats and their owners. The existence of these genes in non-diarrheic pet animal isolates indicates that domestic pets may serve as a reservoir for human infection. Thus, one health strategy comprising animal and human health sectors, governments, together with stakeholders is needed to confront multidrug-resistant E. coli infections in Bangladesh.

Physico-chemical properties of the association of cetyltrimethylammonium bromide and bovine serum albumin mixture in aqueous-organic mixed solvents.

Herein, we have investigated the association behavior of bovine serum albumin (BSA) and cetyltrimethylammonium bromide (CTAB) using the conductivity method in H2O and H2O + organic mixed solvents at different temperatures. The association phenomenon was detected from the deviation of the conductivity changes with enhancing the surfactant concentration and changes of numerous physico-chemical properties, such as CMC, α, ß and thermodynamic variables (∆G0m, ∆H0m and ∆S0m). The values of CMC for the CTAB + BSA system in 10 % (v/v) solvents follow the trend: CMCwater < CMCwater+DMSO < CMCwater+AN < CMCwater+DX < CMCwater+DMF. The interaction of BSA with CTAB is notably influenced due to a change of temperature and extent of hydration of BSA and surfactant. The obtained values of -∆G0m manifest that the association of BSA and CTAB mixture is a spontaneous process, while the values of -∆G0m in presence of 10 % (v/v) aq. organic solvents come out in the given sequence: -∆Gmo (H2O + DMSO) > ∆Gmo (H2O + DMF) > -∆Gmo (H2O + DX) > -∆Gmo (H2O + AN). The H-bonding, ion-dipole, along with the hydrophobic interactions, are believed to be the binding interactions between BSA and CTAB in the study media.

Antimicrobial resistance patterns of Staphylococcus aureus isolated from apparently healthy pet cats of Bangladesh.

Objective: This study sought to determine the occurrence, molecular identification, antimicrobial-resistant trends, and gene distribution of Staphylococcus aureus in pet cats and their owners' hand swabs. Materials and Methods: From different places and clinics in Mymensingh and Dhaka, 168 pet cat samples and 42 hand swab samples from cat owners were obtained. The organisms were scrutinized by assessing the outcomes using conventional and molecular techniques. The disc diffusion technique was applied to find the resistance pattern against 12 antibiotics, and genes were discovered by targeting specific genes using PCR. Results: The occurrence of pathogenic S. aureus in pet cats was 7.74%, while it was 9.50% in pet owners' hand swabs, and 25.0% of the pet owner's hand swabs contained these genes. Staphylococcus aureus was utterly resistant to amoxicillin, ampicillin, cefixime, erythromycin, and imipenem in both pet cat and hand swabs of pet owner samples. All S. aureus isolates had a multidrug-resistant phenotype, and 1 from pet cats (O19) and 1 from pet owner hand swabs (H9) were resistant to all 12 antibiotics in the 7 antimicrobial classes. Several antibiotic-resistance genes were detected by PCR. Conclusion: The study confirmed multidrug-resistant pathogenic S. aureus in pet cats and their owners in Bangladesh, indicating a major health risk to both people and cats. Thus, a holistic and integrated one-health approach between veterinary and medical specialists is needed to mitigate the global distribution of these zoonotic antibiotic-resistant S. aureus strains.

Draft genome sequence of biofilm-forming methicillin-resistant Staphylococcus aureus MTR_V1 strain isolated from a ready-to-eat food in Bangladesh.

This announcement provides the genome sequence of the biofilm-forming methicillin-resistant Staphylococcus aureus MTR_V1 strain isolated from a ready-to-eat food sample in Bangladesh. Our assembled genome had a length of 2.8 Mb, 27 contigs, two CRISPR arrays, 38 predicted antibiotic resistance genes, and 66 predicted virulence factor genes.

Pseudomonas aeruginosa bacteriophage PA1Ø requires type IV pili for infection and shows broad bactericidal and biofilm removal activities.

We isolated a new lytic Pseudomonas aeruginosa phage that requires type IV pili for infection. PA1Ø has a broad bactericidal spectrum, covering Gram-positive and Gram-negative bacteria, and can eradicate biofilm cells. PA1Ø may be developed as a therapeutic agent for biofilm-related mixed infections with P. aeruginosa and Staphylococcus aureus.

Wound healing effects and antibacterial properties of heterologous platelet-rich plasma on Staphylococcus aureus induced septic wounds in rabbits.

Objective: This study has been designed to determine the effectiveness of heterologous platelet-rich plasma (hPRP) against infected wounds in rabbits. Materials and Methods: Staphylococcus aureus contamination was present in 24 surgical incisions, each 8 mm in diameter. The animals were then divided into two groups as follows: group A, also known as the hPRP group, received topically applied, freshly manufactured hPRP twice weekly, and group B, also known as the control group, only received sterile saline. Evaluations of the histological architecture of wounds, posttherapeutic morphology, morphometry, and in-vitro and in-vivo antimicrobial potentials of hPRP were made. Results: Rabbits that were given hPRP exhibited quicker rates of wound contraction and shorter healing times. The samples from day 7 in the hPRP group showed less inflammation and more structured fibroblasts than those from the control wounds, according to histological analysis. On day 21 of the histological examination, the hPRP group's epidermis showed notable thickening. As demonstrated by in-vitro antibacterial activity, undiluted hPRP successfully suppressed S. aureus growth. A serum biochemical analysis showed that hPRP had no harmful effects on the liver or kidneys. Conclusions: Based on the findings of the histological features, antibacterial properties, and wound morphology, it can be said that hPRP gel holds promise as an alternative to antibiotics for the treatment of wound infections.

Phenotypic and Genotypic Detection of Biofilm-Forming Staphylococcus aureus from Different Food Sources in Bangladesh.

Staphylococcus aureus is a major foodborne pathogen. The ability of S. aureus to produce biofilm is a significant virulence factor, triggering its persistence in hostile environments. In this study, we screened a total of 420 different food samples and human hand swabs to detect S. aureus and to determine their biofilm formation ability. Samples analyzed were meat, milk, eggs, fish, fast foods, and hand swabs. S. aureus were detected by culturing, staining, biochemical, and PCR. Biofilm formation ability was determined by Congo Red Agar (CRA) plate and Crystal Violet Microtiter Plate (CVMP) tests. The icaA, icaB, icaC, icaD, and bap genes involved in the synthesis of biofilm-forming intracellular adhesion compounds were detected by PCR. About 23.81% (100/420; 95% CI: 14.17−29.98%) of the samples harbored S. aureus, as revealed by detection of the nuc gene. The CRA plate test revealed 20% of S. aureus isolates as strong biofilm producers and 69% and 11% as intermediate and non-biofilm producers, respectively. By the CVMP staining method, 20%, 77%, and 3% of the isolates were found to be strong, intermediate, and non-biofilm producers. Furthermore, 21% of S. aureus isolates carried at least one biofilm-forming gene, where icaA, icaB, icaC, icaD, and bap genes were detected in 15%, 20%, 7%, 20%, and 10% of the S. aureus isolates, respectively. Bivariate analysis showed highly significant correlations (p < 0.001) between any of the two adhesion genes of S. aureus isolates. To the best of our knowledge, this is the first study in Bangladesh describing the detection of biofilm-forming S. aureus from foods and hand swabs using molecular-based evidence. Our findings suggest that food samples should be deemed a potential reservoir of biofilm-forming S. aureus, which indicates a potential public health significance.

Virulence Determinants and Methicillin Resistance in Biofilm-Forming Staphylococcus aureus from Various Food Sources in Bangladesh.

The eradication of staphylococcal infections has become more difficult due to the development of antibiotic resistance and virulence in biofilm-forming Staphylococcus aureus. The presence of the life-threatening zoonotic pathogen, methicillin-resistant S. aureus (MRSA), in foods indicates a public health issue. This study, therefore, aimed to determine virulence factors and methicillin resistance in biofilm-forming S. aureus isolates from different foods and food handlers. A total of 100 PCR-positive S. aureus isolates (97 biofilm formers and three non-biofilm formers) were screened using the disk diffusion method and PCR assay. By PCR, genes encoding virulence factors, e.g., enterotoxin (sea, 30%, 95% CI: 21.90−39.59%), toxic shock syndrome toxin (tst, 20%, 95% CI: 13.34−28.88%), and Panton−Valentine leukocidin toxin (PVL, 15%, 95% CI: 9.31−23.28%), were detected in the S. aureus isolates. By the disk diffusion method, 100% (95% CI: 96.30−100.00%) of S. aureus isolates were phenotypically MRSA in nature, showing 100% resistance to oxacillin and cefoxitin. Moreover, the methicillin-resistant gene mecA was found in 61 (61%, 95% CI: 51.20−69.98%) MRSA isolates. Furthermore, all the S. aureus isolates were phenotypically resistant to ampicillin and penicillin, 30% to erythromycin, and 11% to gentamycin. Among them, 51% (95% CI: 41.35−60.58%) of S. aureus isolates were phenotypically multidrug-resistant in nature, and the multiple antibiotic resistance index varied from 0.33 to 0.55. Genes encoding resistance to beta-lactams (blaZ, 100%, 95% CI: 96.30−100.00%) and tetracyclines (tetA and tetC, 3%, 95% CI: 0.82−8.45%) were found positive in the S. aureus isolates. Genes encoding virulence determinants and MRSA were significantly (p < 0.05) higher in strong biofilm-forming S. aureus than in moderate and non-biofilm-forming isolates. To our knowledge, this is the first study in Bangladesh to incorporate preliminary data on the occurrence of virulence determinants and methicillin resistance, including resistance to clinically important antibiotics, in biofilm-forming S. aureus isolates from different foods and food handlers in Bangladesh, emphasizing a potential threat to human health.

Characterization of induced Staphylococcus aureus bacteriophage SAP-26 and its anti-biofilm activity with rifampicin.

Lytic bacteriophages (phages) have been investigated as treatments for bacterial infectious diseases. An induced phage, SAP-26, was isolated from a clinical isolate of Staphylococcus aureus. It belongs to the family Siphoviridae and its genome consists of double-stranded 41,207 bp DNA coding for 63 open reading frames. The phage SAP-26 showed a wide spectrum of lytic activity against both methicillin-resistant S. aureus and methicillin-susceptible S.aureus. Furthermore, combined treatment with a phage and antimicrobial agents showed a strong biofilm removal effect which induced structural changes in the biofilm matrix and a substantial decrease in the number of bacteria. Such a broad host range in S. aureus and biofilm removal activity of the phage SAP-26 suggests the possibility of its use as a therapeutic phage in combination with appropriate antimicrobial agent(s). Among the three antimicrobial agents combined with phage, the combination of rifampicin showed the best biofilm removal effect. To the authors' knowledge, this study showed for the first time that S. aureus biofilm could be efficiently eradicated with the mixture of phage and an antimicrobial agent, especially rifampicin.

Antibiotic resistance patterns of Staphylococcus spp. isolated from fast foods sold in different restaurants of Mymensingh, Bangladesh.

OBJECTIVE: Fast foods are often responsible for staphylococcal foodborne illness. The present study was carried out to isolate Staphylococcus spp. from various fast foods sold in Mymensingh and to determine their antibiogram. MATERIALS AND METHODS: Overall, 60 samples of fast foods sold in different restaurants were screened by culture, biochemical tests, and polymerase chain reaction (PCR) to isolate and identify Staphylococcus spp., followed by employing of disk diffusion method to determine their antibiotic resistance patterns. RESULTS: Among these 60 samples, 8 [13.33%, 95% confidence interval (CI): 6.91%-24.17%] were positive for Staphylococcus spp. by cultural and biochemical properties. By PCR, four (6.67%, 95% CI: 2.62%-15.93%) isolates were confirmed as Staphylococcus aureus by targeting nuc gene. Additionally, all the S. aureus isolates were coagulase-positive. By antibiogram profiles, all the Staphylococcus isolates exhibited resistance to azithromycin and erythromycin (95% CI: 67.56%-100.00%), and frequently resistance to cefixime (75%, 95% CI: 40.93%-95.56%), ampicillin (50%, 95% CI: 21.52%-78.48%), and amoxicillin (37.5%, 95% CI: 13.68%-69.43%); moderate to lower resistance was found against cefotaxime, gentamicin, and doxycycline. In addition, all the isolates were sensitive to ciprofloxacin and chloramphenicol. Interestingly, 75% (6/8; 95% CI: 40.93%-95.56%) isolates were multidrug-resistant (MDR) in nature. Furthermore, the indices of multiple antibiotic resistance (MAR) were ranged from 0.2 to 0.6. CONCLUSION: This study revealed that fast foods sold in restaurants were contaminated with MDR and MAR Staphylococcus isolates, having potential public health significance.

Standard feeding strategies with natural insemination improved fertility in repeat breeding dairy cows.

OBJECTIVE: The experiment was designed to establish suitable management strategies through the different feeding and breeding approaches on fertility improvement in the experimental repeat breeding (RB) cows. MATERIALS AND METHODS: 80 RB cows were selected for this experiment. Before grouping, all cows were deworming and then divided into four equal groups, namely Group-TF1 [traditional feeding practice and natural insemination (NI)], Group-TF2 [traditional feeding practice and Artificial insemination (AI)], Group-SF1 [standard (STD) feeding practice and NI], and Group-SF2 (STD feeding practice and AI). These allocated RB cows were fed by traditional and STD feeding methods for 90 days and then inseminated by AI and NI breeding systems. The dominant follicle (DF) diameter, hemato-biochemical elements, and estrogen (E2) hormone were estimated during the insemination of cows. Estimation of the pregnancy rate was carried out at days 45-90 post-insemination in the cows. RESULTS: The pregnancy rate was significantly (p < 0.05) higher in STD feeding practice with NI when compared to traditional feeding practice irrespective of breeding systems, and it was also significantly (p < 0.05) higher in NI than in AI breeding system, irrespective of feeding strategies. The results also showed that the diameter of DF, serum E2, total erythrocyte count, hemoglobin, packed cell volume, total cholesterol, total protein, glucose, calcium, phosphorus, ferric iron, copper, zinc, and magnesium at the time of insemination were significantly (p < 0.01) elevated in the experimental RB cows with STD feeding practice. The diameter of DF and serum E2 were significant (p < 0.01) and positively correlated with all hemato-biochemical elements in the cows at the time of insemination. CONCLUSION: The results suggest that NI with STD feeding practice may increase fertility in RB cows by improving general health status. Finally, it could support the veterinarians and researchers to define the management strategies using feeding and breeding strategies to prevent repeat breeding syndrome in dairy cows.

Bacterial loads and antibiotic resistance profile of bacteria isolated from the most popular street food (Phuchka) in Bangladesh.

OBJECTIVE: Phuchka is one of the most common street foods in Bangladesh. It is served with salad, sweet and sour tamarind dispersed water, and minced eggs as topping at places where people usually gather. This makes these foods susceptible to bacterial contamination. Therefore, assessing the bacterial load and antimicrobial profile of organisms isolated from phuchka and other foodstuffs served with it was the focus of this study. MATERIALS AND METHODS: Bacterial isolates were isolated and identified from the samples after the bacterial loads were assessed as total viable count (TVC), total coliform count (TCC), and total staphylococcal count (TSC). The antibiotic resistance profile of the isolates was obtained using the disk diffusion method. Molecular detection of Escherichia coli isolates and the presence of gene responsible for tetracycline resistance was confirmed by polymerase chain reaction. RESULTS: According to the recommendations, the TVC value of 70% phuchka and egg samples was safe, whereas TSC value illustrated that 80% of both phuchka and egg samples were at safety level. For the TCC value, 80% egg and 70% phuchka samples were found to be safe for consumption. Among all the samples, the microbial loads of the vendors' hand wash were least in the safety level. Antibiotic sensitivity tests revealed that both Staphylococcus spp. and E. coli isolates were sensitive to gentamicin and ciprofloxacin but showed resistance to ampicillin. CONCLUSION: The data of this study indicate that phuchka can pose a public health problem as foodborne bacterial isolates which are antibiotic-resistant are found in it.

Challenges and Opportunities in the Pharmacological Treatment of Acute Venous Thromboembolism in Children.

Venous thromboembolism (VTE) is an important but historically under-recognized problem in pediatrics, with an incidence concentrated in hospitalized children. A number of specific VTE diseases with discrete triggers have been described, but the most common pediatric trigger is the presence of central venous access devices. VTE diseases, though heterogenous in etiology, are linked by the common therapeutic strategies shared by their management. Historically, the most commonly used drug therapies have been unfractionated heparin, low-molecular-weight heparins, and vitamin K antagonists, based on extrapolation from adult data rather than any specific pediatric trials. Although these widely used drugs appear safe and effective in expert hands, the historical lack of pediatric data is problematic in view of the recognized significant differences between children and adults with regards to hemostatic physiology, VTE etiology, and drug pharmacokinetics. The increasing adult usage of novel VTE pharmacotherapies such as direct oral anticoagulants (DOACs) has led to considerable interest in exploring the pediatric applications of these newer drugs. This review summarizes the advantages and disadvantages of existing VTE pharmacotherapies and outlines emerging novel pediatric VTE therapies, particularly DOACs, within the context of the current pediatric trial landscape.

Hot topics in coagulation testing: Important considerations for testing children for bleeding/thrombotic disorders.

The accurate use and interpretation of diagnostic investigations are essential for safe and effective patient care. Appropriate application and interpretation of coagulation testing can be challenging, and many controversies exist relating to the standardization of testing procedures, the application of relevant tests to different patient populations and the interpretation of test results. We present a list of the most prominent controversies in coagulation testing and have selected three specific examples (age-appropriate reference ranges, therapeutic anticoagulation monitoring and tests of thrombin generation) for closer discussion, highlighting examples with a paediatric framework. We discuss the limitations of discrete age-partitioned reference intervals, given the established principle of developmental haemostasis; the difficulties in establishing normative data across different laboratories; important pre-analytical variables affecting coagulation testing; the challenges in interpreting APTT and anti-Xa assays for monitoring unfractionated heparin therapy in different clinical situations; and the limitations in interpreting tests of thrombin generation due to current available thrombin-specific substrates and the complicating factor of variable alpha2-macroglobulin levels. These controversies are demonstrated using paediatric examples, but raise important implications for coagulation testing in patients of all ages and highlight the pressing need for further research in these areas.

Molecular Detection of Multidrug Resistant Salmonella Species Isolated from Broiler Farm in Bangladesh.

Multidrug resistant (MDR) Salmonella are a leading cause of foodborne diseases and serious human health concerns worldwide. In this study we detected MDR Salmonella in broiler chicken along with the resistance genes and class 1 integron gene intl1. A total of 100 samples were collected from broiler farms comprising 50 cloacal swabs, 35 litter and 15 feed samples. Overall prevalence of Salmonella was 35% with the highest detected in cloacal swabs. Among the Salmonella, 30 isolates were confirmed as S. enterica serovar Typhimurium using molecular methods of PCR. Disk diffusion susceptibility test revealed that all the Salmonella were classified as MDR with the highest resistance to tetracycline (97.14%), chloramphenicol (94.28%), ampicillin (82.85%) and streptomycin (77.14%). The most prevalent resistance genotypes were tetA (97.14%), floR (94.28%), blaTEM-1 (82.85%) and aadA1 (77.14%). In addition, among the MDR Salmonella, 20% were positive for class 1 integron gene (intl1). As far as we know, this is the first study describing the molecular basis of antibiotic resistance in MDR Salmonella from broiler farms in Bangladesh. In addition to tetA, floR, blaTEM-1, aadA1 and intl1 were also detected in the isolated MDR Salmonella. The detection of MDR Salmonella in broiler chicken carrying intl1 is of serious public health concern because of their zoonotic nature and possibilities to enter into the food chain.

Antibacterial efficacy of ethanolic extract of Camellia sinensis and Azadirachta indica leaves on methicillin-resistant Staphylococcus aureus and shiga-toxigenic Escherichia coli.

OBJECTIVE: This study aims at investigating the antibacterial potential of ethanolic extract of Camellia sinensis (common name: Green tea) and Azadirachta indica (common name: Neem) leaves on methicillin-resistant Staphylococcus aureus (MRSA) and shiga-toxigenic Escherichia coli (STEC). MATERIALS AND METHODS: Fresh leaves were processed and extracted by 99% ethanol and reconstituted with 50% ethanol before testing. Disk diffusion and broth microdilution methods were used to determine zone diameter of inhibition (ZDI) and minimum inhibitory concentration (MIC), respectively. Nutrient agar plate was used to estimate the minimum bactericidal concentration (MBC). RESULTS: Maximum ZDI value was observed for green tea against MRSA (7.5 mm) and minimum for neem (4.9 mm). Moreover, the highest ZDI against STEC was also for green tea and the combination of green tea and neem (4.5 mm). The MIC values of green tea extract were 15.625 and 31.25 mg/ml against MRSA and STEC, respectively, whereas the MIC of neem was 31.25 and 125 mg/ml, respectively. The combination had similar MIC (46.87 mg/ml) against both organisms. Green tea showed the lowest MBC values, 31.25 and 62.5 mg/ml, against MRSA and STEC, respectively. However, MBC of neem and the combination against MRSA and STEC were found >250 mg/ml, >500 mg/ml and 93.75 mg/ml, >375 mg/ml, respectively. CONCLUSION: Green tea and neem leaves showed good antimicrobial effects and can be used to explore novel antimicrobial compounds against MRSA and STEC.