Neutralization of the Staphylococcus aureus Panton-Valentine leukocidin by African and Caucasian sera.

BACKGROUND: The prevalence of Staphylococcus aureus isolates carrying the Panton-Valentine leukocidin (PVL) gene is higher in Africa (≈50%) compared to Europe (< 5%). The study aimed to measure anti-PVL-antibodies in Africans and Germans in a multi-center study and to test whether detected antibodies can neutralize the cytotoxic effect of PVL on polymorphonuclear leukocytes (PMNs). METHODS: Sera from asymptomatic Africans (n = 22, Nigeria, Gabon) and Caucasians (n = 22, Germany) were used to quantify antibody titers against PVL and α-hemolysin (in arbitrary units [AU]) by ELISA. PMNs from one African and German donor were exposed to 5 nM recombinant PVL to measure the neutralizing effect of serial dilutions of pooled sera from African and Caucasian participants, or donor sera at 0.625 and 2.5% (v/v). RESULTS: Anti-PVL-antibodies were significantly higher in Africans than in Germans (1.9 vs. 0.7 AU, p < 0.0001). The pooled sera from the study participants neutralized the cytotoxic effect of PVL on African and German PMNs in a dose dependent manner. Also, neutralization of PVL on PMNs from the African and German donors had a stronger effect with African sera (half-maximal inhibitory concentration (IC50) = 0.27 and 0.47%, respectively) compared to Caucasian sera (IC50 = 3.51 and 3.59% respectively). CONCLUSION: Africans have higher levels of neutralizing anti-PVL-antibodies. It remains unclear if or at what level these antibodies protect against PVL-related diseases.

Antibiotic-resistant staphylococci from the wastewater treatment plant and grey-water samples in Obafemi Awolowo University, Ile-Ife, Nigeria.

This study examined the occurrence and molecular basis for antibiotic-resistant staphylococci from the wastewater treatment plant and grey-water samples in Obafemi Awolowo University, Nigeria. Standard microbiological techniques and molecular methods were utilized. The species identified (MALDI score >1.7) comprised S. saprophyticus (19), S. cohnii (8), S. sciuri (7), S. aureus (4), S. epidermidis (3), S. warneri (2), S. equorum (1), S. haemolyticus (1), S. nepalensis (1), S. condimenti (1), and S. pasteuri (1). Resistance to trimethoprim, tetracycline and cefoxitin were observed in 78.3% (47/60), 36.7% (22/60) and 25% (15/60) of the isolates, respectively. The rate of multidrug resistance was 53.3% (32/60) and observed in eight species from different sampling sites. Seven (S. sciuri; n = 5; S. aureus; n = 1; S. warneri; n = 1) of the 20 selected (representing the various staphylococcal species and antibiotypes) isolates were mecA-positive. Furthermore, the tetK gene was detected in nine isolates, six with dfrA, and four were positive for the dfrG gene. One S. aureus was mecA, tetK and dfrG gene positive. The study provides insights on antibiotic-resistant staphylococci from a non-clinical setting and highlights the need for active surveillance to understand the burden of antimicrobial resistance in Nigeria. This is key to improve synergy across the human, animal and environmental health sectors in Nigeria.

A 6-Year Update on the Diversity of Methicillin-Resistant Staphylococcus aureus Clones in Africa: A Systematic Review.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of hospital-associated (HA) and community-associated (CA) infections globally. The multi-drug resistant nature of this pathogen and its capacity to cause outbreaks in hospital and community settings highlight the need for effective interventions, including its surveillance for prevention and control. This study provides an update on the clonal distribution of MRSA in Africa. Methods: A systematic review was conducted by screening for eligible English, French, and Arabic articles from November 2014 to December 2020, using six electronic databases (PubMed, EBSCOhost, Web of Science, Scopus, African Journals Online, and Google Scholar). Data were retrieved and analyzed according to the Preferred Reporting Items for Systematic Review and Meta-Analysis guidelines (registered at PROSPERO: CRD42021277238). Genotyping data was based primarily on multilocus sequence types (STs) and Staphylococcal Cassette Chromosome mec (SCCmec) types. We utilized the Phyloviz algorithm in the cluster analysis and categorization of the MRSA STs into various clonal complexes (CCs). Results: We identified 65 studies and 26 publications from 16 of 54 (30%) African countries that provided sufficient genotyping data. MRSA with diverse staphylococcal protein A (spa) and SCCmec types in CC5 and CC8 were reported across the continent. The ST5-IV [2B] and ST8-IV [2B] were dominant clones in Angola and the Democratic Republic of Congo (DRC), respectively. Also, ST88-IV [2B] was widely distributed across the continent, particularly in three Portuguese-speaking countries (Angola, Cape Verde, and São Tomé and Príncipe). The ST80-IV [2B] was described in Algeria and Egypt, while the HA-ST239/ST241-III [3A] was only identified in Egypt, Ghana, Kenya, and South Africa. ST152-MRSA was documented in the DRC, Kenya, Nigeria, and South Africa. Panton-Valentine leukocidin (PVL)-positive MRSA was observed in several CCs across the continent. The median prevalence of PVL-positive MRSA was 33% (ranged from 0 to 77%; n = 15). Conclusion: We observed an increase in the distribution of ST1, ST22, and ST152, but a decline of ST239/241 in Africa. Data on MRSA clones in Africa is still limited. There is a need to strengthen genomic surveillance capacity based on a "One-Health" strategy to prevent and control MRSA in Africa.

First Report of a Methicillin-Resistant, High-Level Mupirocin-Resistant Staphylococcus argenteus.

We describe the identification of a methicillin-resistant, high-level mupirocin-resistant Staphylococcus argenteus. The isolate (1801221) was characterized as t6675-ST2250-SCCmecIVc, and whole-genome sequencing revealed that the isolate possessed two plasmids. One plasmid (34,870 bp), designated p1_1801221 with rep23, harboured the mupirocin resistance (mupA) gene. The second plasmid (20,644 bp), assigned as p2_1801221 with rep5a and rep16, carried the resistance determinants for penicillin (blaZ) and cadmium (cadD). Phylogenetic analysis revealed that the isolate clustered with the European ST2250 lineage. The overall high similarity of both plasmids in S. argenteus with published DNA sequences of Staphylococcus aureus plasmids strongly suggests an interspecies transfer. The pathogenic potential, community and nosocomial spread, and acquisition of antibiotic resistance gene determinants, including the mupA gene by S. argenteus, highlight its clinical significance and the need for its correct identification.

DNA microarray analysis of Staphylococcus aureus from Nigeria and South Africa.

Staphylococcus aureus is an important human pathogen with an arsenal of virulence factors and a propensity to acquire antibiotic resistance genes. The understanding of the global epidemiology of S. aureus through the use of various typing methods is important in the detection and tracking of novel and epidemic clones in countries and regions. However, detailed information on antibiotic resistance and virulence genes of S. aureus, and its population structure is still limited in Africa. In this study, S. aureus isolates collected in South Africa (n = 38) and Nigeria (n = 2) from 2001-2004 were characterized by spa typing and DNA microarray. The combination of these two methods classified the isolates into seven spa types and three clonal complexes (CCs) i.e. t064-CC8 (n = 17), t037-CC8 (n = 8), t1257-CC8 (n = 6), t045-CC5 (n = 5), t951-CC8 (n = 1), t2723-CC88 (n = 1), t6238-CC8 (n = 1), and untypeable-CC8 (n = 1). A high percentage agreement (>95%) and kappa coefficient (>0.60) was largely observed with antibiotic susceptibility testing and DNA microarray, indicating substantial agreement. Some antibiotic and virulence gene markers were associated with specific clones. The detection of the collagen-binding adhesion (cna) gene was unique for t037-CC8-MRSA while the enterotoxin gene cluster (egc) and staphylococcal complement inhibitor (scn) gene were identified with t045-CC5-MRSA. Moreover, the combination of genes encoding enterotoxins (entA, entB, entK, entQ) was noted with most of the CC8 isolates. The t045-CC5-MRSA clone was positive for the mercury resistance (mer) operon. DNA microarray provides information on antibiotic resistance and virulence gene determinants and can be a useful tool to identify gene markers for specific S. aureus clones in Africa.

Genomic analysis of Staphylococcus aureus from the West African Dwarf (WAD) goat in Nigeria.

BACKGROUND: Staphylococcus aureus can colonize various host species, and human-animal interaction is a significant factor for cross-species transmission. However, data on S. aureus colonization in animals, particularly on ruminants in close contact with humans, is limited. The West African Dwarf (WAD) goat is among the earliest domesticated ruminant associated with rural dwellers and small-holder farmers in sub-Saharan Africa. This study aimed to investigate the population structure, antibiotic resistance, and virulence gene determinants of S. aureus from the WAD goat in Nigeria. METHODS: Nasal samples were obtained from the WAD goat in five markets in Osun State, South-West Nigeria. S. aureus was characterized by antibiotic susceptibility testing, detection of virulence determinants, spa typing, and multilocus sequence typing (MLST). Representative isolates were selected for whole-genome sequencing, biofilm, and cytotoxicity assay. RESULTS: Of the 726 nasal samples obtained from the WAD goat, 90 S. aureus (12.4%) were recovered. Overall, 86 isolates were methicillin-susceptible, and four were mecA-positive (i.e., methicillin-resistant S. aureus [MRSA]). A diverse S. aureus clonal population was observed (20 sequence types [STs] and 37 spa types), while 35% (13/37) and 40% (8/20) were new spa types and STs, respectively. Eleven MLST clonal complexes (CC) were identified (CC1, CC5, CC8, CC15, CC30, CC45, CC97, CC121, CC133, CC152, CC522). The MRSA isolates were designated as t127-ST852-CC1-SCCmec type VII, t4690-ST152-CC152-SCCmec type Vc, and t8821-ST152-CC152-SCCmec type Vc. Phylogenetic analysis revealed that 60% (54/90) of all isolates were associated with ruminant lineages (i.e., CC133, CC522). Panton-Valentine Leukocidin (PVL)-positive S. aureus was identified in CC1, CC30, CC121, and CC152. For the CC522 isolates, we illustrate their pathogenic potential by the detection of the toxic shock syndrome gene and hemolysins, as well as their strong cytotoxicity and ability to form biofilms. CONCLUSIONS: This is the first detailed investigation on the genomic content of S. aureus from the WAD goat in Nigeria. The S. aureus population of the WAD goat consists mainly of ruminant-associated lineages (e.g., CC133, CC522), interspersed with human-associated clones, including PVL-positive MRSA CC1 and CC152.

Molecular characterization of Staphylococcus aureus complex from fomites in Nigeria.

Fomites serve as a potential route for the transmission of pathogens including community-acquired methicillin-resistant Staphylococcus aureus to humans. Phylogenetic and taxonomic analyses have established the Staphylococcus aureus complex (S. aureus, S. argenteus and S. schweitzeri), however, phenotypic characteristics are insufficient in the delineation of these species. In this study, we describe the S. aureus complex from inanimate surfaces in Nigeria. Fomite samples in Obafemi Awolowo University were initially screened for S. aureus and species differentiation was determined by MALDI-TOF, PCR of the S. aureus specific thermonuclease and the nonribosomal peptide synthetase genes. Characterization of the isolates was based on antimicrobial susceptibility, spa typing, multilocus sequence typing and virulence gene detection (lukS/lukF-PV, chp, sak, scn). Whole-genome sequencing was done for selected isolates. Of the 239 fomites samples, 14  S. aureus and two S. schweitzeri isolates were identified including three MRSA. Genotyping classified the S. aureus isolates into ST8/CC8, ST30/CC30, ST15/ST5875/CC15, ST508/ST5876/CC45, ST121/CC121, ST152/CC152 and ST3961. All the isolates in CC30, CC121, and CC152 were lukS/lukF-PV positive. The MRSA (PVL+) were assigned with CC152. Phylogenetic analysis revealed that the S. schweitzeri isolates were closely related with those from fruit bats (Eidolon helvum) in Nigeria. The differentiation of S. aureus from S. schweitzeri was clearly achieved through MALDI-TOF and PCR. Fomites are not only a reservoir for S. aureus but also for S. schweitzeri that was so far recovered primarily in African wildlife.

Persistent Staphylococcus aureus nasal colonization in ambulatory human immunodeficiency virus-infected patients in Nigeria: Risk factors and molecular features.

This longitudinal study on Staphylococcus aureus colonization in Nigerian human immunodeficiency virus patients (n = 187) found a trend towards a higher proportion of persistent S. aureus carriage in patients with advanced human immunodeficiency virus infection, low CD4+ cell counts, and a predominance of isolates belonging to ST8/spa-CC064 in persistent carriers.

The discovery of a multi-resistant Staphylococcus haemolyticus clone in the hospital and community environment in south western Nigeria.

Among clinically significant isolates of coagulase negative staphylococci, Staphylococcus haemolyticus is ranked second after Staphylococcus epidermidis. It has been associated with septicemia in newborns and various infections in persons with compromised host defenses and implanted foreign bodies. The existence of a multi-resistant Staphylococcus haemolyticus clone was discovered during a study on patients with skin and soft tissue infections at two local health clinics and in a referral hospital in South Western Nigeria. The clonal nature of these strains was determined by antibiotic susceptibility profile and pulsed field gel electrophoresis. This represents the first report of what appears to be a hospital-acquired and transmitted Staphylococcus haemolyticus clone in South Western Nigeria. Careful infection control measures and strain typing are urgently needed to understand species epidemiology and to limit the spread of multi-resistant strains within and beyond healthcare facilities.

Wound infections in two health institutions in Ile-Ife, Nigeria: results of a cohort study.

The control of wound infections is increasingly complicated, yet treatment is not always guided by microbiological diagnosis. To describe the distribution of wound infections and causative agents, a prospective, 6-month cohort study involving 102 outpatients was conducted at the University Teaching Hospital and the Health Center in Ile-Ife, Nigeria. Location and type of infected wounds were recorded and bacterial isolates were identified by standard microbiological techniques. Almost half (40%) of all infected wounds were attributed to trauma and, in most cases, located on the extremities. Of the 162 bacterial isolates obtained from wound cultures, 39 were monomicrobial and 55 were polymicrobial; no bacterial isolate was obtained in eight cases. Staphylococcus aureus was the predominant micro-organism (25%), followed by Escherichia coli (12%), Pseudomonas aeruginosa (9%), and Staphylococcus epidermidis (9%). The diversity of micro-organisms and the high incidence of polymicrobic flora in this study give credence to the value of identifying one or more bacterial pathogens from wound cultures. The recognition of causative agents of wound infections can assist wound care practitioners in taking appropriate measures. Continuous dialogue between the microbiology department and wound care practitioners is strongly advised in order to improve treatment outcomes and slow the development of antibiotic-resistant bacteria.