Hepatitis B virus precore/core region mutations and genotypes among hepatitis B virus chronic carriers in South-Eastern, Nigeria.

OBJECTIVES: The study aimed at detecting the prevailing hepatitis B virus (HBV) genotypes and the presence of clinically relevant mutations in the precore/core gene of the HBV DNA, among patients with chronic infection in South-eastern, Nigeria. METHODS: A total of 72 participants with chronic HBV infection were enrolled into the study. Plasma samples from those with detectable HBV DNA were subjected to nested Polymerase Chain Reaction amplification using the precore/core specific primers. This resulted to the successful amplification and sequencing of the HBV precore/core region DNA from 16 participants. Mutation analysis on the precore/core region detected the presence of certain HBV precore/core gene mutations. Genotyping was carried out by phylogenetic analysis. RESULTS: The precore region mutation at nucleotide position 1896, which is a G to A change resulting to a nonsense mutation, was detected in 6.25% of the participants. Other HBV precore region mutations that were detected include: G1899A, T1846A, G1862C, G1888A, T1821C, C1826T, A1827C, A1850T, C1858T, precore start codon Kozak sequence mutations and some novel core region mutations such as G/A1951T and G1957A. Genotyping revealed the existence of HBV genotype/subgenotype A1 (87.5%) and D (12.5%) among the participants. There was no significant difference in the occurrence of specific precore/core mutations among the HBV/hepatitis C virus dually infected and HBV mono-infected participants. CONCLUSION: The data suggest the likelihood of a more severe outcome of hepatitis caused by HBV in South-eastern Nigeria due to the occurrence of a variety of precore/core mutation, which resulted to HBeAg-negative chronic HBV infection among the participants.

Pharmaceutical Quality of Selected Metronidazole and Ciprofloxacin Infusions Marketed in South Eastern Nigeria.

BACKGROUND: Pharmaceutical products need to be of good quality and it is even more critical when it comes to life saving medicaments like infusions. OBJECTIVE: This research surveyed the quality fitness of some ciprofloxacin and metronidazole infusion samples marketed in South-eastern of Nigeria. METHODS: Using Official Compendial methods, microbiological quality, active pharmaceutical ingredients quantitation, pH and particle count tests were evaluated on eighty infusion bottles (from eight pharmaceutical companies) of each of the two drugs. RESULTS: Out of the sixteen brands tested, 2 metronidazole brands and 1 ciprofloxacin brand (representing 18.75% of the total 16 brands/makes) were contaminated while the remaining 13 brands (81.25%) were found sterile. The active pharmaceutical ingredients quantitative assay showed that all the brands of ciprofloxacin infusion were between the 95% and 105% limit of label claim while one metronidazole brand has <95-110% limit label claim. Six brands each of the two drugs evaluated fall below the acceptable pH range [ciprofloxacin (3.5-4.6) and metronidazole (4.8-5.2)], while the other two brands of both drugs passed the test. In the antibacterial study, Pseudomonas aeruginosa and Escherichia coli were susceptible to the ciprofloxacin (5 µg). However, Salmonella typhi recorded inhibition zone diameters within resistant and intermediate range. Peptostrepococcus spp was susceptible (at minimum inhibitory concentrations of 100 µg/mL) to all the brands of metronidazole, while none of the brands were effective on Lactobacillus spp. All the brands passed the test for particulate contamination. The particles size range was <10µm. CONCLUSION: About eighty-one percent (81.25%) of the infusions have acceptable good microbiological quality. However, 18.75% that failed the tests is a concern knowing that these are lifesaving products.

Combinations of Alchornea cordifolia, Cassytha filiformis and Pterocarpus santalinoides in diarrhoegenic bacterial infections.

OBJECTIVES: This study examines the rationale, if any, behind combining the extracts from the fruits of Alchornea cordifolia and Pterocarpus santalinoides and aerial parts of Cassytha filiformis in the traditional treatment of diarrhoegenic bacterial infections. RESULTS: Four diarrhoegenic bacterial isolates: Salmonella typhi, Shigellae dysenteriae, Escherichia coli and Staphylococcus aureus were used and their antibiotic susceptibility screening showed that they were multi-antibiotic resistant. The extracts exhibited activity against all the test isolates with minimum inhibitory concentration values ranging from 3.125 to 12.5 mg/mL. From the checkerboard assay, the fractional inhibitory concentration indices showed that C. filiformis has antagonistic and indifference activities in combination with either P. santalinoides or A. cordifolia. This showed that the combination of extracts from the fruits of A. cordifolia and P. santalinoides and aerial parts of C. filiformis is counterproductive and invalidates any claim for positive results in the management of diarrhoegenic bacterial infections.

Whole-Genome Sequencing of an Uncommon Cryptococcus neoformans MLST43 Genotype Isolated in Nigeria.

Cryptococcosis is a human infection caused mainly by two species of the Cryptococcus genus, Cryptococcus neoformans and Cryptococcus gattii, whose populations contain several phylogenetically related haploid (VN/VG-types) and hybrid genotypes. Here, we report the whole-genome sequencing of a Nigerian C. neoformans VNII, Mat-α, strain with a rare multilocus-sequence-type (MLST) genotype (ST43).

Predominance of cervicitis agents with minimal testing rate within the student population in Benin city, Nigeria.

Here, we aimed to investigate the predominance of cervicitis agents with minimal testing rates among asymptomatic students in a Tertiary Institution and its associated risk factors. Endocervical swabs were collected from randomly selected 133 female students (15-34 years of age) in Benin City, Nigeria and screened for six genital pathogens (Chlamydia trachomatis, Mycoplasma hominis, M. genitalium, Ureaplasma urealyticum, U. parvum and Trichomonas vaginalis) using multiplex PCR. Out of the 133 subjects, 80 were positive for cervicitis with 46.3% of its agents. Five different species of the pathogens were identified with highest and lowest prevalence as: T. vaginalis (71.0%) and U. urealyticum (6.45%). Co-infection had predominance of 19.35 and 9.67% for three-organism and two-organism colonisation respectively. Strong associations were observed between the presence of cervicitis and co-infection with other genital pathogens, abortion, inconsistent condom use (p < .05). The high occurrence of cervicitis agents in our study combined with asymptomatic outcome among the subjects justifies screening for these pathogens in this population. Impact statement What is already known on this subject? Cervicitis is a genital condition among reproductive age women. It is characterised by inflammation of the uterine cervix which subsequently promotes the acquisition of certain genital conditions such as infertility and sexually transmitted infections. In various studies, there have been reports on 30-40% of cervicitis cases associated with known pathogens but left undetected within the general population. Studies have shown that majority of the cervicitis cases presents in an asymptomatic state. What the results of this study add? There may be some risk associated factors promoting the acquisition of cervicitis agents within the student population since the prevalence of these agents in this population which is underrated was relatively high. What the implication are of these findings for clinical practice and/or further research? Further investigation is needed to define the prospective influence of microbial load in colonisation of the organism and the association of new and higher sexual partners as their prevalence are not fully understood. Furthermore, our finding recommend inclusion of screening exercise for cervicitis agents within the student population which will control the infection, thereby improving female reproductive health, consequently limiting spread and serious sequelae.

Infection of Chick Chorioallantoic Membrane (CAM) as a Model for the Pathogenesis of Cryptococcus gattii.

The use of embryonated egg as an alternative in the study of the pathogenesis of fungi is evolving. Although murine models are the "gold standard," embryonated egg models are also used to screen determinants of virulence among fungi species. This study was aimed at determining the virulence potential of Cryptococcus gattii strains R265, R272, and EJB18, and Malassezia sympodialis using chorioallantoic membrane (CAM) of embryonated egg. At a concentration of 107 cfu/ml, C. gattii R272 was more virulent than R265 in the egg model, while EJB18 had low virulence. The CAM model supported the growth of Malassezia sympodialis strain and induced the formation of hyphae. The formation of lesions by the organism and its re-isolation from CAM suggest that the model can be used for evaluating the virulence of C. gattii. Histopathology of CAM from both strains also revealed massive disruption of CAM. This study suggests that embryonated egg is a useful alternative tool to pre-screen Cryptococcus gattii strains to select strains for subsequent testing in murine models and could also be a potential medium for studying the hyphal growth in Malassezia species.

Candida colonisation in asymptomatic HIV patients attending a tertiary hospital in Benin City, Nigeria.

BACKGROUND: Candidiasis is the commonest opportunistic fungal infection in patients infected with human immunodeficiency virus (HIV). CD4+ lymphocyte counts have been found to be a marker of HIV disease progression. AIM: This study focused on determining the spectrum of Candida isolates in urine, stool, and oral specimens among HIV patients in a tertiary hospital. METHODS: A total of 300 subjects comprising of 200 HIV patients and 100 non-HIV subjects were used for this study. Three samples (urine, stool, and oral swab) were collected from each subject. Each specimen was processed using standard microbiological techniques and emergent Candida isolates were identified with CHROMagar™ Candida and sugar fermentation tests. RESULTS: The overall prevalence of Candida colonisation among HIV patients was 52.5%. HAART-naive patients had a significantly higher prevalence (OR = 3.65; 95% CI = 2.03-6.56; p<0.0001) than their counterpart on highly active antiretroviral therapy (HAART) (OR = 1.99; 95% CI = 1.13-3.50; p=0.0232). Female gender was a significant risk factor for acquiring Candida infection (OR = 3.40; 95% CI = 1.14-10.13; p=0.0289). The effect of age on prevalence of candidiasis was observed among HIV patients on HAART (p=0.0161). A CD4+ count <200 cells/µl was a significant risk factor for acquiring candidal infection only among HAART-naive patients (OR = 4.37; 95% CI = 1.60-11.95; p=0.0042). The five species of Candida recovered from this study were C. albicans, C. krusei, C. parapsilosis, C. tropicalis, and C. glabrata. CONCLUSION: There is a significant relationship between antiretroviral therapy, CD4+ counts, and the prevalence of candidiasis.

Isolation and molecular characterization of Candida africana from Jos, Nigeria.

During a survey of the prevalence of Candida spp. in Jos, Plateau State, Nigeria, two atypical C. albicans isolates were recovered. These two yeasts were germ tube positive, chlamydospore-negative and gave a green color on CHROMagar Candida. Molecular analysis performed by amplification of the hwp1 gene showed that these two isolates belonged to C. africana, a newly proposed Candida species closely related to C. albicans. Based on the presence or absence of an intron in DNA sequences encoding rRNA, the two C. africana, including all C. albicans isolates examined, were found to belong to genotype A and no other genotypes or species such as C. dubliniensis were found. To our knowledge, this is the first isolation of C. africana in Nigeria.