Wild Animals Are Reservoirs and Sentinels of Staphylococcus aureus and MRSA Clones: A Problem with “One Health” Concern

Background: The availability of comprehensive data on the ecology and molecular epidemiology of Staphylococcus aureus/MRSA in wild animals is necessary to understand their relevance in the “One Health” domain. Objective: In this study, we determined the pooled prevalence of nasal, tracheal and/or oral (NTO) Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA) carriage in wild animals, with a special focus on mecA and mecC genes as well as the frequency of MRSA and methicillin susceptible S. aureus (MSSA) of the lineages CC398 and CC130 in wild animals. Methodology: This systematic review was executed on cross-sectional studies that reported S. aureus and MRSA in the NTO cavities of wild animals distributed in four groups: non-human primates (NHP), wild mammals (WM, excluding rodents and NHP), wild birds (WB) and wild rodents (WR). Appropriate and eligible articles published (in English) between 1 January 2011 to 30 August 2021 were searched for from PubMed, Scopus, Google Scholar, SciElo and Web of Science. Results: Of the 33 eligible and analysed studies, the pooled prevalence of NTO S. aureus and MRSA carriage was 18.5% (range: 0–100%) and 2.1% (range: 0.0–63.9%), respectively. The pooled prevalence of S. aureus/MRSA in WM, NHP, WB and WR groups was 15.8/1.6, 32.9/2.0, 10.3/3.4 and 24.2/3.4%, respectively. The prevalence of mecC-MRSA among WM/NHP/WB/WR was 1.64/0.0/2.1/0.59%, respectively, representing 89.9/0.0/59.1/25.0% of total MRSA detected in these groups of animals.The MRSA-CC398 and MRSA-CC130 lineages were most prevalent in wild birds (0.64 and 2.07%, respectively); none of these lineages were reported in NHP studies. The MRSA-CC398 (mainly of spa-type t011, 53%), MRSA-CC130 (mainly of spa types t843 and t1535, 73%), MSSA-CC398 (spa-types t571, t1451, t6606 and t034) and MSSA-CC130 (spa types t843, t1535, t3625 and t3256) lineages were mostly reported. Conclusion: Although the global prevalence of MRSA is low in wild animals, mecC-mediated resistance was particularly prevalent among MRSA isolates, especially among WM and WB. Considering the genetic diversity of MRSA in wild animals, they need to be monitored for effective control of the spread of antimicrobial resistance.

Diversity and Distribution of Spa types among Methicillin Resistant Staphylococcus Aureus Isolated from Humans and Livestock in Kabale District - South Western Uganda

Background: S. aureus is a skin and mucosal bacterial commensal of both humans and animals which has evolved as an important pathogen implicated to cause various infections. High levels of antibiotic use have resulted into multi-drug resistance MRSA, especially among HA-MRSA, CA-and LA - MRSA. Awareness on coexistence and diversity of MRSA clones among humans and household Livestock particularly cattle and swine in our region is limited. We used spa typing method to determine spa diversity, distribution and coexistence in outpatients, household contacts and respective livestock (cattle and swine) in Kabale region, south western Uganda. Methods: This was a cross sectional study by design consisting of outpatients, household contacts and livestock. Outpatients (n =100) colonized with MRSA were traced back to their respective homesteads where household members, domestic cattle, and, swine were tested for S. aureus and subsequently MRSA colonization. High-resolution DNA melting analysis was used to determine spa types among MRSA isolates. Overlap of MRSA isolates among humans and livestock was based on the presence of similar spa types.Results: A total of 3371 S.aureus isolates were collected from outpatients (n =376), household contacts (n = 1531), Cattle (n = 1159) and Swine (n = 305), among which 482 had mecA gene where 27% (100/376) and 8% (123/1531) were outpatients and household contacts respectively while 11% (132/1159) and 42% (127/305) were cattle and swine respectively. Twenty different spa types were identified; t034, t4677, t108, t1451, t9377, t1081, t040, t701, t041, t002, t044, t037,t121, t127, t922, t032, t019, t018, t012 and t030, among which t034 (109/482), t4677 (53/482), t9377 (63/482) and t1081 (53/482) were most prevalent and distributed among human and livestock. All the MRSA isolates were multidrug resistant to antibiotics tested. Conclusion: In Kabale region, there is high diversity of spa types among MRSA. Presence of similar spa types was found circulating among humans and their respective livestock which demonstrates a possible bidirectional transmission. Presence of MDR - MRSA highlights the need for effective prevention and control of MRSA among livestock and in the community using One Health approach.

Molecular Epidemiology, Virulence Factors, Antibiotic Resistance and Risk Factors for Nasal Carriage of Staphylococcus aureus in A Teenage Student Population: High Prevalence of Oxacillin Susceptible MRSA Isolates

Background: Asymptomatic carriage of Staphylococcus aureus can lead to endogenous infections and cross-transmission to other individuals. Objectives: The prevalence, molecular epidemiology, antibiotic resistance, and risk factors for nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) were studied in school children in Ardabil, Iran. Methods: Totally, 510 nasal samples were collected during 2017. Isolates were identified and subjected to antimicrobial susceptibility testing, identification of oxacillin resistance, and molecular typing. Results: Totally, 13.5% of volunteers were positive for methicillin-susceptible Staphylococcus aureus (MSSA) and 17.5% colonized with mecA positive S. aureus strains, including 6.07% oxacillin-resistant MRSA (OR-MRSA) and 11.56% oxacillin-susceptible MRSA (OS-MRSA). Excluding β-lactam antibiotics, high resistance rate was observed for erythromycin (71%), tetracycline (25.8%), clindamycin (35%) in our isolates. Surprisingly, 11% of the isolates [OR-MRSA (25.8%), OS-MRSA (10.1%), and MSSA (5.7%) isolates] were resistant to mupirocin. Moreover, 18 (58%), 29 (49%), and 29 (42%) of OR-MRSA, OS-MRSA, and MSSA isolates were multidrug-resistant (MDR), respectively. Overall, 97.48% of isolates carried ≥ 3 toxin encoding genes. The pvl gene was found in 46 (29%) isolates. In comparison, 25.50% of MRSA (9.60% OR-MRSA and 34% OS-MRSA) and 33% of MSSA isolates carried pvl gene. SCCmec type IV had the highest rate among OR-MRSA (87%) and OS-MRSA (74.5%) isolates, which indicates CA-MRSA phenotype. Eleven and 21 spa types were identified in OR-MRSA, and OS-MRSA isolates, respectively. The most common spa types were t11332 (14.3%) and t012 (11.4%) in OS-MRSA isolates. ERIC-PCR revealed high genetic diversity among isolates. The number of students in classroom and incomplete antibiotic course were associated with OS-MRSA nasal carriage. Conclusions: This study showed a high proportion of MDR CA-MRSA nasal carriage among Iranian healthy school children community.

1168. Are the Staphylococcus aureus Isolates that Recolonize Infants in a NICU Following Successful Decolonization the Same or Different from the Initial Colonizing Isolate?

Background Staphylococcus aureus is an important pathogen of infants in a neonatal intensive care unit (NICU). Colonization precedes infection and decolonization may prevent infection. The origin of colonizing organisms may be the NICU environment or personnel or visitors. We have observed infants who became recolonized after successful decolonization. The purpose of this study was to determine the proportion of infants who become recolonized with the same strain or a different strain. Methods Eligible infants were consecutive infants who 1. were colonized with methicillin-susceptible S. aureus and were successfully decolonized with topical mupirocin ointment (nares and umbilicus) as evidenced by 2 or more consecutive negative weekly surveillance cultures (in the absence of a course of systemic antibiotics with activity against MSSA), 2. subsequently became recolonized, and 3. the pair of isolates was available for analysis. Isolates were analyzed by staphylococcal protein A (spa) typing and pairs with concordant spa types were subjected to whole genome sequencing (WGS; Illumina MiSeq) and phylogenetic analyses. Pairs of isolates with fewer than 25 single nucleotide polymorphism differences were considered closely related. Results There were 19 occurrences of MSSA recolonization in 17 infants following 2-6 (median, 2) negative weekly intervening surveillance cultures. Based upon spa typing (that identified 19 spa types), in 11 (58%) there was a concordant spa type and in 8 (42%) there was a discordant spa type. Of the 11 pairs of isolates with concordant spa types that were compared after WGS, 10 were closely related resulting overall in recolonization with a closely related strain in 53% of episodes. Conclusion Among MSSA colonized infants who become recolonized after successful decolonization, the recolonizing strain is the same as the original strain in over half of cases. In such cases the source is more likely to be a visitor than the NICU environment or staff. The possibility that some cases classified as recolonization were in fact persistent low level colonization or carriage in another body site not detected by surveillance cultures cannot be excluded. Disclosures Anne-Catrin Uhlemann, MD, PhD, Merck (Grant/Research Support)

Prevalence and genotypic characteristics of Panton-Valentine Leukocidin-producing Staphylococcus aureus isolates obtained in Jos, North Central Nigeria

Background: Staphylococcus aureus is an opportunistic pathogen that colonizes and causes infections in humans. Panton Valentine Leukocidin (PVL) is a cytolytic toxin produced by some strains of S. aureus and are mostly associated with skin and soft tissue infections and necrotizing pneumonia. Aim: To investigate the prevalence and genotypic characteristics of PVL-positive S. aureus strains cultured from patients in three tertiary hospitals in Jos, Nigeria. Methods: Two hundred and fourteen clinical S. aureus isolates were obtained from three tertiary hospitals in Jos. Polymerase chain reaction was used to detect lukSF-PV gene that encodes PVL, and sensitivity to antimicrobial agents was performed on PVL-positive S. aureus. Genotypic characteristics of the PVL-positive S. aureus was determined by spa typing and multilocus sequence typing (MLST). Results: The genes for PVL were detected in 67/214 (31.3%) of S. aureus isolates. Majority of the PVL-positive isolates were obtained from wound (n=37; 55.2%), blood (n=11; 16.4%) and urine (n=10; 14.9). Most of PVL-positive isolates (n=58; 34.7%) were methicillin sensitive S. aureus (MSSA) while nine isolates (19.1%) were methicillin resistant S. aureus (MRSA). Spa typing identified 14 different spa types, dominated by t355 (n=33; 49.3%), followed by t174 (n=7; 10.4%), t019 and t159 (n=5; 7.5%). MLST revealed six sequence types (ST) namely, ST152 (n=35), ST121 (n=9), ST1 (n=8), ST30 (n=8), ST772 (n=6) and ST15 (N=1). Conclusion: This study revealed that 31.3% of S. aureus isolated in Jos hospitals carried genes for PVL, belonged to six sequence types and 14 spa types with t355-ST152-MSSA as the dominant genotype.

Genotypic Characterization of Clinical Isolates of Staphylococcus aureus from Pakistan

In this study, we compared pulsed-field gel electrophoretic (PFGE), multilocus sequence typing (MLST), Staphylococcal cassette chromosome mec (SCCmec), spa typing, and virulence gene profiles of 19 Panton–Valentine leucocidin (PVL)-positive, multidrug-, and methicillin-resistant clinical Staphylococcus aureus (MRSA) isolates obtained from a hospital intensive care unit in Pakistan. The isolates exhibited 10 pulsotypes, contained eight adhesin genes (bbp, clfA, clfB, cna, fnbA, fnbB, map-eap, and spa), 10 toxin genes (hla, hlb, hld, hlg, pvl, sed, see, seg, seh, and tst), and two other virulence genes (cfb, v8) that were commonly present in all isolates. The spa-typing indicated seven known spa types (t030, t064, t138, t314, t987, t1509, and t5414) and three novel spa types. MLST analysis indicated eight ST types (ST8, ST15, ST30, ST239, ST291, ST503, ST772, and ST1413). All isolates belonged to the agr group 1. Most of the isolates possessed SCCmec type III, but some isolates had it in combination with types SCCmec IV and V. The presence of multidrug-resistant MRSA isolates in Pakistan indicates poor hygienic conditions, overuse of antibiotics, and a lack of rational antibiotic therapy that have led to the evolution and development of hypervirulent MRSA clones. The study warrants development of a robust epidemiological screening program and adoption of effective measures to stop their spread in hospitals and the community.

Antimicrobial Resistance and Genetic Lineages of Staphylococcus aureus from Wild Rodents: First Report of mecC-Positive Methicillin-Resistant S. aureus (MRSA) in Portugal

The frequent carriage of Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), by wild animals along with its zoonotic potential poses a public health problem. Furthermore, the repeated detection of the mecA gene homologue, mecC, in wildlife raises the question whether these animals may be a reservoir for mecC-MRSA. Thus, we aimed to isolate S. aureus and MRSA from wild rodents living in port areas and to characterize their antimicrobial resistance and genetic lineages. Mouth and rectal swab samples were recovered from 204 wild rodents. The samples were incubated in BHI broth with 6.5% of NaCl and after 24 h at 37 °C the inoculum was seeded onto Baird-Parker agar, Mannitol Salt agar and ORSAB (supplemented with 2 mg/L of oxacillin) plates. Species identification was confirmed by MALDI-TOF MS. The antimicrobial susceptibility testing was performed by the Kirby–Bauer disc diffusion method against 14 antibiotics. The presence of virulence and resistance genes was performed by PCR. The immune evasion cluster (IEC) system was investigated in all S. aureus. All isolates were characterized by MLST, spa- and agr typing. From 204 samples, 38 S. aureus were isolated of which six MRSA were detected. Among the six MRSA isolates, three harbored the mecC gene and the other three, the mecA gene. All mecC-MRSA isolates were ascribed to sequence type (ST) 1945 (which belongs to CC130) and spa-type t1535 whereas the mecA isolates belonged to ST22 and ST36 and spa-types t747 and t018. Twenty-five S. aureus were susceptible to all antibiotics tested. S. aureus isolates were ascribed to 11 MLST and 12 spa-types. S. aureus presents a great diversity of genetic lineages in wild rodents. This is the first report of mecC-MRSA in Portugal.

Antibiotic Resistance, spa Typing and Clonal Analysis of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolates from Blood of Patients Hospitalized in the Czech Republic

Staphylococcus aureus is one of the major causes of bloodstream infections. The aim of our study was to characterize methicillin-resistant Staphylococcus aureus (MRSA) isolates from blood of patients hospitalized in the Czech Republic between 2016 and 2018. All MRSA strains were tested for antibiotic susceptibility, analyzed by spa typing and clustered using a Based Upon Repeat Pattern (BURP) algorithm. The representative isolates of the four most common spa types and representative isolates of all spa clonal complexes were further typed by multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. The majority of MRSA strains were resistant to ciprofloxacin (94%), erythromycin (95.5%) and clindamycin (95.6%). Among the 618 strains analyzed, 52 different spa types were detected. BURP analysis divided them into six different clusters. The most common spa types were t003, t586, t014 and t002, all belonging to the CC5 (clonal complex). CC5 was the most abundant MLST CC of our study, comprising of 91.7% (n = 565) of spa-typeable isolates. Other CCs present in our study were CC398, CC22, CC8, CC45 and CC97. To our knowledge, this is the biggest nationwide study aimed at typing MRSA blood isolates from the Czech Republic.