Molecular characterization of non-aureus staphylococci and Mammaliicoccus from Hipposideros bats in Southwest Nigeria.

Bats are not only ecologically valuable mammals but also reservoirs of zoonotic pathogens. Their vast population, ability to fly, and inhabit diverse ecological niches could play some role in the spread of antibiotic resistance. This study investigated non-aureus staphylococci and Mammaliicoccus colonization in the Hipposideros bats at Obafemi Awolowo University, Ile-Ife, Nigeria. Pharyngeal samples (n = 23) of the insectivorous bats were analyzed, and the presumptive non-aureus staphylococcal and Mammaliicoccus isolates were confirmed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The isolates were characterized based on antibiotic susceptibility testing and whole-genome sequencing (WGS). Six bacterial genomes were assembled, and three species were identified, including Mammaliicoccus sciuri (n = 4), Staphylococcus gallinarum (n = 1), and Staphylococcus nepalensis (n = 1). All the isolates were resistant to clindamycin, while the M. sciuri and S. gallinarum isolates were also resistant to fusidic acid. WGS analysis revealed that the M. sciuri and S. gallinarum isolates were mecA-positive. In addition, the M. sciuri isolates possessed some virulence (icaA, icaB, icaC, and sspA) genes. Multi-locus sequence typing identified two new M. sciuri sequence types (STs) 233 and ST234. The identification of these new STs in a migratory mammal deserves close monitoring because previously known ST57, ST60, and ST65 sharing ack (8), ftsZ (13), glpK (14), gmk (6), and tpiA (10) alleles with ST233 and ST234 have been linked to mastitis in animals. Moreover, the broad host range of M. sciuri could facilitate the dispersal of antibiotic resistance genes. This study provides evidence of the importance of including migratory animals in monitoring the development and spread of antibiotic resistance.

Changing Ecotypes of Dengue Virus 2 Serotype in Nigeria and the Emergence of Cosmopolitan and Asian I Lineages, 1966-2019.

Dengue virus (DENV) is a leading mosquito-borne virus with a wide geographical spread and a major public health concern. DENV serotype 1 (DENV-1) and serotype 2 (DENV-2) were first reported in Africa in 1964 in Ibadan, Nigeria. Although the burden of dengue is unknown in many African countries, DENV-2 is responsible for major epidemics. In this study, we investigated the activities of DENV-2 to determine the circulating strains and to appraise the changing dynamics in the epidemiology of the virus in Nigeria. Nineteen DENV-2 sequences from 1966-2019 in Nigeria were retrieved from the GenBank of the National Center of Biotechnology Information (NCBI). A DENV genotyping tool was used to identify the specific genotypes. The evolutionary history procedure was performed on 54 DENV-2 sequences using MEGA 7. There is a deviation from Sylvatic DENV-2 to other genotypes in Nigeria. In 2019, the Asian I genotype of DENV-2 was predominant in southern Edo State, located in the tropical rainforest region, with the first report of the DENV-2 Cosmopolitan strain. We confirmed the circulation of other non-assigned genotypes of DENV-2 in Nigeria. Collectively, this shows that DENV-2 dynamics have changed from Sylvatic transmission reported in the 1960s with the identification of the Cosmopolitan strain and Asian lineages. Sustained surveillance, including vectorial studies, is required to fully establish the trend and determine the role of these vectors.

Japanese Encephalitis Enzootic and Epidemic Risks across Australia.

Japanese encephalitis virus (JEV) is an arboviral, encephalitogenic, zoonotic flavivirus characterized by its complex epidemiology whose transmission cycle involves reservoir and amplifying hosts, competent vector species and optimal environmental conditions. Although typically endemic in Asia and parts of the Pacific Islands, unprecedented outbreaks in both humans and domestic pigs in southeastern Australia emphasize the virus' expanding geographical range. To estimate areas at highest risk of JEV transmission in Australia, ecological niche models of vectors and waterbirds, a sample of piggery coordinates and feral pig population density models were combined using mathematical and geospatial mapping techniques. These results highlight that both coastal and inland regions across the continent are estimated to have varying risks of enzootic and/or epidemic JEV transmission. We recommend increased surveillance of waterbirds, feral pigs and mosquito populations in areas where domestic pigs and human populations are present.

Seroprevalence of Influenza A Virus in Dromedaries in North-Western Nigeria.

Although influenza A virus is endemic in wild waterfowl, domestic poultry, swine, humans, bats, cetaceans, dogs, and horses, there is a paucity of data on the potential role of camels in zoonotic transmission of the virus. To estimate the seroprevalence of the influenza A virus in camel populations, four local government areas of Nigeria that share an international border with the Niger Republic were selected. Blood samples from 184 one-hump camels (dromedaries) were collected and tested for influenza IgG antigen by ELISA. Each camel's demographic variable, such as age, gender, location, production system, and usage, was recorded. The overall seroprevalence rate of influenza virus IgG in this study was 10.33% (95%CI: 6.33-15.66%). In the bivariate model, there was no significant difference in gender, age, site location and production system, except for usage. There was a significantly lower seroprevalence rate among camels used for labour (odds ratio (OR) = 0.34, 95% CI: 0.10-0.97) than those used for meat consumption; however, not after adjusting for other variables in the model. Increase surveillance through early detection, prediction, and risk assessment of pathogens in animal reservoirs and environmental contamination as One Health strategies to reduce potential human spillover is recommended. Molecular epidemiology studies could better elucidate the role of camels in the dynamics of disease transmission pathways.

Detection of Crimean-Congo Hemorrhagic Fever Virus Antibodies in Cattle in Plateau State, Nigeria.

Crimean-Congo hemorrhagic fever (CCHF) is a vector-borne viral hemorrhagic disease with global clinical significance. Certain species of ticks are vectors of CCHF, which can be transmitted from animals to humans and humans to humans by direct exposure to blood or other body fluids. The zoonotic transmission at the human-animal interface from viremic animal hosts to humans is a public health concern with a paucity of data in Nigeria. Samples from 184 pastoral cattle from three local government areas (LGAs) of Plateau state, Nigeria, were screened for CCHF virus using a commercial enzyme-linked immunosorbent assay (ID Screen® CCHF Double Antigen for Multi-Species). Overall seropositivity of 30.4% (n = 56) (95% CI: 23.88%, 37.63%) was recorded from the study areas in Plateau State, while 48/126 (38.1%, 95% CI: 29.59%, 47.17%) sampled cows tested positive for CCHFV antibodies. Seropositivity was significantly higher (p < 0.001) among older cattle greater than two years, 54.69% (95% CI: 2.88%, 11.24%) compared to cattle younger than two years, 17.5% (95% CI: 11.17%, 25.50%). The location of farms played a significant role in the seropositivity of CCHF with the least risk observed in Wase LGA. CCHF is an important zoonotic disease in different parts of the globe with a high risk of transmission to pastoralists, livestock keepers/slaughterhouse workers, and veterinarians who handle animals. There is a need for a collaborative one-health approach with various stakeholders to unravel the dynamics of CCHFV epidemiology in Nigeria.

Concurrent Resistance to Carbapenem and Colistin Among Enterobacteriaceae Recovered From Human and Animal Sources in Nigeria Is Associated With Multiple Genetic Mechanisms.

Resistance to last resort drugs such as carbapenem and colistin is a serious global health threat. This study investigated carbapenem and colistin resistance in 583 non-duplicate Enterobacteriaceae isolates utilizing phenotypic methods and whole genome sequencing (WGS). Of the 583 isolates recovered from humans, animals and the environment in Nigeria, 18.9% (110/583) were resistant to at least one carbapenem (meropenem, ertapenem, and imipenem) and 9.1% (53/583) exhibited concurrent carbapenem-colistin resistance. The minimum inhibitory concentrations of carbapenem and colistin were 2-32 µg/mL and 8 to >64 µg/mL, respectively. No carbapenem resistant isolates produced carbapenemase nor harbored any known carbapenemase producing genes. WGS supported that concurrent carbapenem-colistin resistance was mediated by novel and previously described alterations in chromosomal efflux regulatory genes, particularly mgrB (M1V) ompC (M1_V24del) ompK37 (I70M, I128M) ramR (M1V), and marR (M1V). In addition, alterations/mutations were detected in the etpA, arnT, ccrB, pmrB in colistin resistant bacteria and ompK36 in carbapenem resistant bacteria. The bacterial isolates were distributed into 37 sequence types and characterized by the presence of internationally recognized high-risk clones. The results indicate that humans and animals in Nigeria may serve as reservoirs and vehicles for the global spread of the isolates. Further studies on antimicrobial resistance in African countries are warranted.

Molecular and Serological Detection of Piroplasms in Horses from Nigeria.

Equine piroplasmosis, an economically important disease of equids caused by the hemoprotozoan parasites Theileria equi, T. haneyi, and Babesia caballi, has a worldwide distribution. These parasites are transmitted by ixodid ticks. To improve the detection of horses in Nigeria exposed to piroplasm parasites, 72 horses with variable clinical signs of piroplasmosis were sampled from Northwest and Northcentral Nigeria and tested by nPCR and cELISA. Blood and serum samples were collected from each horse via jugular venesection. Individually, nPCR or cELISA failed to identify all horses exposed to piroplasms. A combination of species-specific nPCR and the OIE-approved T. equi and B. caballi cELISAs enhanced the detection of horses exposed to parasites. The results also demonstrated horses showing abnormal hematology were positive for only T. equi, except for one sample that was coinfected with T. equi and T. haneyi. We also identified ticks collected from some of the horses, with Rhipicephalus evertsi evertsi being the most prevalent. This study shows that a larger proportion of horses in the sample set were exposed to T. equi than B. caballi or T. haneyi. Additionally, ticks that have been previously reported as potential vectors for these parasites were found to have infested sampled horses. Further studies are needed to investigate which tick species are competent vectors for Theileria spp. and Babesia caballi in Nigeria.

Identification of mobile colistin resistance genes (mcr-1.1, mcr-5 and mcr-8.1) in Enterobacteriaceae and Alcaligenes faecalis of human and animal origin, Nigeria.

Colistin is a last-resort drug used to treat infections caused by multidrug-resistant Gram-negative bacteria that have developed carbapenem resistance. Emergence and rapid dissemination of the nine plasmid-mediated mobile colistin resistance genes (mcr-1 to mcr-9) has led to fear of pandrug-resistant infections worldwide. To date, there is only limited information on colistin resistance in African countries where the drug is widely used in agriculture. In this Nigerian study, 583 non-duplicate bacterial strains were isolated from 1119 samples from humans, camels, cattle, dogs, pigs and poultry using colistin-supplemented MacConkey agar, among which 17.0% (99/583) were colistin-resistant. PCR (mcr-1 to mcr-9) and whole-genome sequencing (WGS) identified mcr in 21.2% (21/99) of colistin-resistant isolates: mcr-1.1 (n = 13), mcr-8.1 (n = 5), mcr-1.1 and mcr-8.1 (n = 2), and mcr-1.1 and mcr-5 (n = 1). Of the 21 mcr-positive strains, 9 were isolated from human samples, with 8 being Klebsiella pneumoniae, and 6 of these human K. pneumoniae had a high colistin MIC (>64 µg/mL). In contrast, 9 of the 12 mcr-positive animal isolates were Escherichia coli, of which only 2 had a colistin MIC of >64 µg/mL. This study is the first to report mcr-1 in Alcaligenes faecalis and the emergence of mcr-5 and mcr-8 in Nigeria. WGS determined that mcr-1 was localised on an IncX4 plasmid and that 95.2% of mcr-1 harbouring isolates (20/21) transferred colistin resistance successfully by conjugation. These findings highlight the global spread of colistin resistance and emphasise the urgent need for co-ordinated global action to combat resistant bacteria.

Characterization of high level ampicillin- and aminoglycoside-resistant enterococci isolated from non-hospital sources.

PURPOSE: High level ampicillin- and aminoglycoside-resistant enterococci are being increasingly reported from non-hospital sources. This study was carried out to characterize these strains from non-hospital sources in Nigeria. METHODOLOGY: A collection of Enterococcus faecium isolated from vegetables, soil, farm animals and manure and observed to be resistant to ampicillin (n=63) and gentamicin (n=37) discs, were screened for resistance to high levels of ampicillin and aminoglycoside using E-test strips. Putative high level ampicillin- and aminoglycoside-resistant strains were screened for pbp5 and aminoglycoside modifying enzyme genes, respectively, by PCR. The C-terminal region of the amplified pbp5 gene was also sequenced. RESULTS: Five (5/63) and thirty-five (35/37) of the ampicillin- and aminoglycoside-resistant strains were identified as high level ampicillin- and aminoglycoside-resistant E. faecium strains, respectively, based on the MIC results. The amplified pbp5 gene from the high level ampicillin-resistant isolates displayed 96-99 % nucleotide sequence similarity with the reference strains and three novel insertions (500Glu→Leu, 502Asp→Arg and 614Ile→Phe) in the amino acid sequence. Aminoglycoside modifying enzyme genes aac(6')-Ie-aph(2″) (100 %), aph(2')-Ic (88.8 %), aph(3')-IIIa (90 %) and ant(4')-Ia (40 %) were detected among the high level aminoglycoside-resistant isolates. CONCLUSION: This is the first report on the characterization of high level ampicillin- and aminoglycoside-resistant Enterococcus faecium among animals and vegetables in Nigeria. The results show that non-hospital sources can constitute a reservoir for potential dissemination of these strains and genes to humans via the food chain or by direct contact.