Investigation of methicillin-resistant Staphylococcus aureus among clinical isolates from humans and animals by culture methods and multiplex PCR

Abstract

BackgroundStaphylococcus aureus is responsible for large numbers of hospital-related and community-acquired infections. In this study, we investigated the presence of S. aureus and methicillin-resistant S. aureus (MRSA) in 100 samples from animals (55 cattle, 36 dogs, and 9 cats) and 150 samples from hospitalized human patients. The samples were collected from healthy and diseased animals and from diseased humans and included milk, wound swab, pus, exudates, nasal swab and diabetic ulcer. Initially, S. aureus was isolated and identified by colony morphology, Gram staining, and biochemical tests (catalase and coagulase tests). The S. aureus-positive samples were examined by polymerase chain reaction (PCR) to determine their MRSA status.ResultsOf the 100 animal samples, 29 were positive for S. aureus. Four samples (13.8%) from dogs were MRSA-positive, but samples from cattle and cats were MRSA-negative. Of the 150 human samples we collected, 64 were S. aureus-positive and, of these, 34 (53.1%) were MRSA-positive. Most (28%) of the MRSA samples were isolated from surgical wound swabs, followed by the pus from skin infections (11%), exudates from diabetic ulcers (6%), exudates from burns (4%), and aural swabs (3%). By contrast, a low MRSA detection rate (n = 4) was seen in the non-human isolates, where all MRSA bacteria were isolated from nasal swabs from dogs. The antimicrobials susceptibility testing results showed that S. aureus isolates with mecA genes showed resistance to penicillin (100%), oxacillin (100%), erythromycin (73.5%), ciprofloxacin (70.6%), and gentamicin (67.7%). The lowest resistance was found against ceftazidime, and no vancomycin-resistant isolates were obtained.ConclusionsWe detected S. aureus and MRSA in both human and canine specimens. Isolates were found to be resistant to some of the antimicrobials available locally. MRSA carriage in humans and animals appears to be a great threat to effective antimicrobials treatment. The prudent use of antimicrobials will reduce the antimicrobial resistance. Our findings will help to find the most appropriate treatment and to reduce antimicrobial resistance in the future by implementing prudent use of antimicrobials. Further studies are required to better understand the epidemiology of MRSA human–animal inter-species transmission in Bangladesh.Electronic supplementary materialThe online version of this article (10.1186/s12917-018-1611-0) contains supplementary material, which is available to authorized users.

Occurrence of multidrug-resistant methicillin-resistantStaphylococcus aureusamong healthy farm animals: a public health concern

“…Nowadays, emergence of antibiotic-resistant bacteria, especially MRSA is not only a global public health challenge but also an emerging veterinary pathogen throughout the world [21]. After the introduction of β-lactam antimicrobials, the prevalence of MRSA infections and colonization in foodproducing animals has steadily increased over time [22,23]. In this study, the overall occurrence rate of MRSA among examined animals was 3.6% with the following rates among different animal species (sheep: 3.8%, goats: 3.9%, cattle: 4.3% and buffalo: 2.2%).…”

Nasopharyngeal Carriage of Methicillin-Resistant Staphylococcus aureus (MRSA) among Sickle Cell Disease (SCD) Children in the Pneumococcal Conjugate Vaccine Era

“…The emulsion was vortexed and heated in a heat block compartment at 100 °C for 10 min, frozen at −20 °C for 10 min, and centrifuged at 3000× g for 10 min. The supernatant containing the bacterial DNA was aliquoted into a new, labelled Eppendorf tube, stored at −20 °C, and used as a DNA template for multiplex PCR [ 53 ].…”

Prevalence of Panton-Valentine leukocidin in methicillin-resistant Staphylococcus aureus clinical isolates at a university hospital in Northern Cyprus: a pilot study

“…Multiplex PCR assay included 2 µl of the DNA template added to a 25 µl final reaction mixture containing: 2X PCR Master Mix (Thermo Fischer Scientific, Waltham, MA USA) containing reaction buffer, Taq DNA polymerase (0.05 U/µL), 4 mM MgCl 2 , 0.4 mM of each dNTP and 10 pmol of each primer. The primer sets were used for amplification are shown in Additional file 1: Table S1 [26] and amplification was performed as described before [27] with a modification of an initial denaturation step at 94 °C for 10 min. PCR products were analyzed by agarose gel electrophoresis using 1.5% agarose gel.…”