Antimicrobial resistance pattern of methicillin-resistant Staphylococcus aureus isolated from sheep and humans in Veterinary Hospital Maiduguri, Nigeria.

Background and Aim: Methicillin-resistant Staphylococcus aureus (MRSA), an important opportunistic pathogen, is a Gram-positive coccus known to be resistant to ß-lactam antibiotics. Its virulence depends on a large range of factors, mainly extracellular proteins, such as enzymes and exotoxins, that contribute to causing a wide range of diseases in human and animal species. The major reasons for the success of this pathogen are its great variability, which enables it to occur and thrive at different periods and places with diverse clonal types and antibiotic resistance patterns within regions and countries. Infections caused by antibiotic-resistant S. aureus bring about serious problems in the general population (humans and animals). Infections with these pathogens can be devastating, particularly for the very young, adults and immunocompromised patients in both humans and animals. This study aimed to determine the presence of MRSA in both apparently healthy and sick sheep brought to the veterinary hospital as well as veterinary staff and students on clinical attachment in the hospital. Materials and Methods: A total of 200 nasal swab samples were collected aseptically from sheep and humans (100 each) for the isolation of MRSA. The samples were processed by appropriately transporting them to the laboratory, then propagated in nutrient broth at 37°C for 24 h followed by subculturing on mannitol salt agar at 37°C for 24 h, to identify S. aureus. This was followed by biochemical tests (catalase and coagulase tests) and Gram staining. MRSA was isolated using Clinical Laboratory Standard Institute (CLSI) guideline and confirmed by plating onto Oxacillin (OX) Resistance Screening Agar Base agar. The antimicrobial susceptibility pattern of the MRSA isolates was determined using the disk diffusion method against 12 commonly used antimicrobial agents. Results: The total rate of nasal carriage of S. aureus and MRSA was found to be 51% and 43% in sheep and humans, respectively. The MRSA prevalence in male and female sheep was 18% and 8%, while 9% and 8% were for male and female human samples, respectively. The antimicrobial susceptibility test showed 100% resistance to OX, cefoxitin, oxytetracycline, cephazolin, and penicillin-G (Pen) by MRSA isolates from humans. Conversely, there was 100% susceptibility to ciprofloxacin, imipenem, and gentamicin; for linezolid (LZD), it was 87.5%, norfloxacin (NOR) (71%), and erythromycin (ERY) (50%) susceptibility was recorded. The MRSA isolates from sheep recorded 100% resistance to the same set of drugs used for human MRSA isolates and were equally 100% susceptible to gentamicin, imipenem, LZD, ciprofloxacin, NOR (92%), and ERY (50%). Conclusion: This study determined the presence of MRSA in sheep and humans from the Veterinary Hospital, Maiduguri. It appears that certain drugs such as ciprofloxacin, imipenem, and gentamicin will continue to remain effective against MRSA associated with humans and sheep. Reasons for the observed patterns of resistance must be explored to reduce the burdens of MRSA resistance. Furthermore, the present study did not confirm the MRSA resistance genes such as mecA and spa typing to ascertain the polymorphism in the X-region using appropriate molecular techniques. Hence more studies need to be conducted to elucidate these findings using robust techniques.

Identification of potent inhibitors of ATP synthase subunit c (AtpE) from Mycobacterium tuberculosis using in silico approach.

ATP synthase subunit c (AtpE) is an enzyme that catalyzes the production of ATP from ADP in the presence of sodium or proton gradient from Mycobacterium tuberculosis (MTB). This enzyme considered an essential target for drug design and shares the same pathway with the target of Isoniazid. Thus, this enzyme would serve as an alternative target of the Isoniazid. The three dimensional (3D) model structure of the AtpE was constructed based on the principle of homology modeling using the Modeller9.16. The developed model was subjected to energy minimization and refinement using molecular dynamic (MD) simulation. The minimized model structure was searched against Zinc and PubChem database to determine ligands that bind to the enzyme with minimum binding energy using RASPD and PyRx tool. A total of 4776 compounds capable of bindings to AtpE with minimum binding energy were selected. These compounds further screened for physicochemical properties (Lipinski rule of five). All the compounds that possessed the desirable property selected and used for molecular docking analysis. Five (5) compounds with minimum binding energies ranged between ─8.69, and ─8.44 kcal/mol, less than the free binding energy of ATP were selected. These compounds further screened for the absorption, distribution, metabolism, excretion, and toxicity (ADME and toxicity) properties. Of the five compounds, three (ZINC14732869, ZINC14742188, and ZINC12205447) fitted all the ADME and toxicity properties and subjected to MD simulation and Molecular Mechanics Generalized Born and Surface Area (MM-GBSA) analyses. The results indicated that the ligands formed relatively stable complexes and had free binding energies, less than the binding energy of the ATP. Therefore, these ligands considered as prospective inhibitors of MTB after successful experimental validation.