Anthrax toxins-producing Bacillus spp. isolated from handwashing stations during COVID-19 pandemic in Lagos, Nigeria.

INTRODUCTION: The virulence binding factor, protective antigen (pag) and poly-D-γ-glutamate capsular (cap) genes, peculiar to Bacillus anthracis are located in the pXO1 and pXO2 plasmids which are transferable horizontally to related species called "cereus group". The cereus group are usually isolated from the environmental/food samples and have been implicated in debilitating human and animal anthrax-like diseases. This study was designed to investigate the presence of the anthrax virulence genes in different Bacillus spp. isolated from handwashing facilities during COVID-19 pandemic in Lagos, Nigeria. METHODOLOGY: The Bacillus anthracis (OK316847), B. thuringiensis (OK316855), B. amyloliquefaciens (OK316857), B. cereus (OK316858) and B. thuringiensis (OK316859) previously isolated from rinsates and bowl water in two local government areas (LGAs) of Lagos state were further investigated by the polymerase chain reaction (PCR) amplification of the pag and cap genes using specific primers. RESULTS: Bacillus anthracis and B. cereus co-harboured the two 578 bp cap and 364 bp pag genes while B. thuringiensis only harboured the cap gene. Similarly, the non-cereus B. amyloliquefaciens was found to habour the pag gene. CONCLUSIONS: The two anthrax toxin genes were amplified in the Bacillus spp isolated from rinsates and bowl water used in hand washing in the two study LGAs. Given that these virulence genes have a global consequence and are a potential threat to life, this study calls for an extensive surveillance, and reassessment of gene regulators and plasmid distribution among these strains in our environment.

Prevalence and molecular characterization of Mycobacterium tuberculosis complex in cattle and humans, Maiduguri, Borno state, Nigeria: a cross-sectional study.

INTRODUCTION: Globally, the highest burden of bovine and human tuberculosis resides in Africa and Asia. Tuberculosis (TB) is the second leading single infectious killer after severe acute respiratory syndrome corona virus-2 (SARSCOV-2). Bovine TB remains a treat to wild and domesticated animals, humans and hinders international trade in endemic countries like Nigeria. We aimed at determining the prevalence of bovine and human tuberculosis, and the spoligotypes of Mycobacterium tuberculosis complex in cattle and humans in Maiduguri. METHODS: We conducted a cross sectional study on bovine and human tuberculosis in Maiduguri, Borno state. We calculated sample size using the method of Thrusfield. Lesions suggestive of TB from 160 slaughtered cattle were obtained from Maiduguri Central Abattoir. Sputum samples from humans; 82 abattoir workers and 147 suspected TB patients from hospitals/clinics were obtained. Lesions and sputum samples were cultured for the isolation of Mycobacterium spp. Positive cultures were subjected genus typing, deletion analysis and selected isolates were spoligotyped. Data was analysed using SPSS VERSION 16.0. RESULTS: Prevalence of 32.5% (52/160) was obtained in cattle. Damboa local government area (LGA), where majority of the infected animals were obtained from had 35.5% bTB prevalence. All categories analysed (breed, age, sex, body conformation and score) had P-values that were not significant (P > 0.05). Sputum culture revealed a prevalence of 3.7% (3/82) from abattoir workers and 12.2% from hospitals/clinics. A significant P-value (0.03) was obtained when positive culture from abattoir and that of hospitals/clinics were compared. Out of the 52 culture positive isolates obtained from cattle, 26 (50%) belonged to M. tuberculosis complex (MTC) and 17/26 (65.4%) were characterized as M. bovis. In humans, 7/12 (58.3%) MTC obtained were characterized as M. tuberculosis. Spoligotyping revealed SB0944 and SB1025 in cattle, while SIT838, SIT61 of LAM10_CAM and SIT1054, SIT46 of Haarlem (H) families were obtained from humans. CONCLUSIONS: Cattle in Damboa LGA need to be screened for bTB as majority of the infected animals were brought from there. Our findings revealed the presence of SB0944 and SB1025 spoligotypes from cattle in Borno state. We isolated M. tuberculosis strain of the H family mainly domiciled in Europe from humans.

Identification of Mycoplasma mycoides subspecies mycoides from slaughtered cattle in two transboundary states of North­eastern Nigeria.

This study aimed to perform molecular typing of Mycoplasma mycoides subsp. mycoides from slaughtered cattle in Adamawa and Taraba States, north­eastern Nigeria. A total of four hundred and eighty (480) samples of lung tissues, nasal swabs, ear swabs and pleural fluids were collected from cattle at slaughter and processed according to standard laboratory protocols. Identification and confirmation were achieved with specific PCR and PCR­RFLP. An overall M. mycoides subsp. mycoides isolation rate of 6.87% (33/480) was obtained. In Adamawa State, 12 (10.91%) isolates of M. mycoides subsp. mycoides came from both, lung tissues and pleural fluids. While in Taraba State, 5 (7.14%) and 4 (5.71%) isolates of M. mycoides subsp. mycoides came from lung tissues and pleural fluids, respectively. The samples from nasal and ear swabs from the study states were negative for M. mycoides subsp. mycoides. Thirty­three out of the 37 culture positive isolates were confirmed to be Mycoplasma mycoides subspecies mycoides with the production of a band equivalent to 574­bp. Molecular typing with restriction endonuclease Vsp1 results in the two bands of 180­bp and 380­bp. In conclusion, the study has established an isolation rate of 6.87% for M. mycoides subsp. mycoides. Measures to strengthen movement control in order to minimise the spread of this dreaded disease of cattle were recommended.

Antimicrobial and Genomic Characterization of Salmonella Nigeria from Pigs and Poultry in Ilorin, North-central, Nigeria.

INTRODUCTION: Non-typhoidal Salmonella are major foodborne pathogens causing serious challenges to public health and food safety worldwide. This study aimed to determine the resistance, virulence genes, sequence type, using multi-locus sequence typing, plasmids and Single Nucleotide Polymorphisms (SNPs) of Salmonella enterica subsp. enterica serovar Nigeria (S. Nigeria) from livestock in Ilorin, North central Nigeria. METHODOLOGY: A total of 1,500 samples from pig (feces; n = 600) and poultry (feces, postmortem samples; n = 900) were collected and analyzed between 2014 to 2017. Presumptive Salmonella isolates were characterized by Whole Genome Sequencing (WGS). RESULTS: We recovered nine S. Nigeria serovars. All the isolates harbored a single point mutation parC(T57S) in addition to qnrB19 and the tetA gene. Furthermore, two plasmids, Col(pHAD28) and IncQ1 predicted to encode qnrB19 and tetA genes, respectively, were detected in all the strains. All the isolates belonged to a single sequence type (ST) 4911, the SNP-based phylogeny showed all the isolates to be highly related, in addition two clinical isolates from the United Kingdom (UK) and Canada, collected outside of this study, also fell into this cluster. Twenty virulence genes were identified from Salmonella Pathogenicity Islands (SPI), chromosomal and fimbriae loci. CONCLUSIONS: This study highlights the roles of pig and poultry in the emergence and spread of S. Nigeria serovar in Nigeria, sub-Sahara Africa. It also highlighted the importance of WGS in clinical and epidemiological surveillance. There is the need for collaborative research studies to investigate the public health importance of Salmonella enterica serovar Nigeria.

Virulent gene profile and antibiotic susceptibility pattern of Shiga toxin-producing Escherichia coli (STEC) from cattle and camels in Maiduguri, North-Eastern Nigeria.

Prevalence and distribution of Shiga toxin-producing Escherichia coli (STEC) serogroups from the faecal samples of cattle and camels slaughter in Maiduguri abattoir and their antibiotic resistance profile of the isolates were determined. The highest prevalence (24%) was recorded in the month of September and more STEC isolates came from cattle than the camels. There was significant (P < 0.05) seasonal trend in the prevalence of STEC among cattle and camel with more cases recorded during the wet season. Although, the study did not demonstrate significant influence of sex from the various sources. The serogroups recorded in this study were O157, O26, O91, O103 and O111. There was no significant difference (P < 0.05) between the detection rates of serogroups. The serogroup O26 was significantly (P < 0.05) the most observed serogroup in both camels and cattle. None of the STEC isolates tested positive for the O45 serogroup. PCR assays shows that 50 (63.3%) of the 86 STEC isolates carried the stx2 gene, 34 (43%) possessed the stx1 gene, and 14 (16.3%) carried both stx1 and stx2 genes. Other genes detected include eae and ehlyA. The antimicrobial resistance among the STEC O157 and non-O157 isolates from cattle and camels in Maiduguri abattoir were very high and the STEC isolates were resistant to at least one or more of the antimicrobial agents tested. There was also multidrug resistance with the most frequent occurring patterns been ampicillin/nalidixic acid and tetracycline/trimethoprim. However, all the 79 isolates were sensitive to chloramphenicol, ceftazidime and ceftriaxone; therefore, these drugs could be drugs of choice in the treatment of STEC infections.

Investigating Salmonella Eko from Various Sources in Nigeria by Whole Genome Sequencing to Identify the Source of Human Infections.

Twenty-six Salmonella enterica serovar Eko isolated from various sources in Nigeria were investigated by whole genome sequencing to identify the source of human infections. Diversity among the isolates was observed and camel and cattle were identified as the primary reservoirs and the most likely source of the human infections.

Serotypes, antimicrobial profiles, and public health significance of Salmonella from camels slaughtered in Maiduguri central abattoir, Nigeria.

AIM: This study aimed at determining the serotypes, antimicrobial profiles, and public health importance of Salmonella strains from camels slaughtered at Maiduguri central abattoir, Nigeria. MATERIALS AND METHODS: Two hundred samples were obtained from camel comprising of intestines, feces, liver, and spleen (n=50 each). Non-lactose fermenting dark center Salmonella colonies were identified using standard biochemical techniques, serotyped and subjected to antimicrobial susceptibility test using minimum inhibition concentration method. RESULTS: Out of the 200 samples collected, 17 were Salmonella positive (spleen=7, intestine=6, feces=3, and liver=1) with a prevalence of 8.5%. Five serotypes comprising Salmonella Eko, 7 (3.5%), Salmonella Uganda, 4 (2.0%), Salmonella Amager, 2 (1.0%), Salmonella Westhampton, 2 (1.0%), and Salmonella Give, 2 (1.0%) were incriminated. Majority of the serotypes were sensitive to the antimicrobials, but one Salmonella Amager exhibited resistance to streptomycin, and one each of Salmonella Uganda and Salmonella Eko were resistant to sulfamethoxazole. CONCLUSION: This study revealed the prevalence and the antibiotic resistance profile of newly emerging Salmonella from camels in the northeast of Nigeria, which can serve as a means for the transmission of Salmonella to human. Therefore, there is a need for the establishment of national Salmonella surveillance and control programs.

Piloting laboratory quality system management in six health facilities in Nigeria.

BACKGROUND: Achieving accreditation in laboratories is a challenge in Nigeria like in most African countries. Nigeria adopted the World Health Organization Regional Office for Africa Stepwise Laboratory (Quality) Improvement Process Towards Accreditation (WHO/AFRO- SLIPTA) in 2010. We report on FHI360 effort and progress in piloting WHO-AFRO recognition and accreditation preparedness in six health facility laboratories in five different states of Nigeria. METHOD: Laboratory assessments were conducted at baseline, follow up and exit using the WHO/AFRO- SLIPTA checklist. From the total percentage score obtained, the quality status of laboratories were classified using a zero to five star rating, based on the WHO/AFRO quality improvement stepwise approach. Major interventions include advocacy, capacity building, mentorship and quality improvement projects. RESULTS: At baseline audit, two of the laboratories attained 1- star while the remaining four were at 0- star. At follow up audit one lab was at 1- star, two at 3-star and three at 4-star. At exit audit, four labs were at 4- star, one at 3-star and one at 2-star rating. One laboratory dropped a 'star' at exit audit, while others consistently improved. The two weakest elements at baseline; internal audit (4%) and occurrence/incidence management (15%) improved significantly, with an exit score of 76% and 81% respectively. The elements facility and safety was the major strength across board throughout the audit exercise. CONCLUSION: This effort resulted in measurable and positive impact on the laboratories. We recommend further improvement towards a formal international accreditation status and scale up of WHO/AFRO- SLIPTA implementation in Nigeria.

Persistence of fluoroquinolone-resistant Salmonella enterica serovar Kentucky from poultry and poultry sources in Nigeria.

INTRODUCTION: This study investigated the antimicrobial resistance and clonality of Salmonella enterica serotype Kentucky in poultry and poultry sources in Nigeria, and compared the isolates with the clone of S. Kentucky STI98-X1 CIPR using (PFGE) and (MIC). METHODOLOGY: Fecal samples from chickens and poultry sources (litter, water, rodent and lizard fecal samples) were collected from  fourteen (14) poultry farms in 2007, 2010 and 2011 and were analyzed for S. Kentucky. RESULTS AND CONCLUSIONS: Six percent of the samples were positive for S. Kentucky - all resistant to nalidixic acid and ciprofloxacin. The isolates are grouped within the PFGE cluster X1 of S. Kentucky STI98 CIPR, indicating the association to the emerging and widely spread CIPR S. Kentucky clone with poultry and poultry sources.