Detection of Multidrug Resistant Tuberculosis (MDR-TB) among Rifampicin-resistant TB patients using Line Probe Assay (LPA) in Kano, Nigeria

The Emergence of drug-resistant Mycobacterium tuberculosis strains especially multidrug resistant-TB (MDR-TB) and indeed extensively drug resistant TB (XDR-TB) is considered a real threat to achieving TB control. Thus, the WHO identified the need for accelerated access to rapid testing for rifampicin resistance to improve case detection in all patients with suspected MDR and XDR-TB. This study aimed at detecting MDR status of rifampicin-resistant TB patients in north western part of Nigeria, as such information was found to be rare in this part of Nigeria. It was a descriptive cross-sectional study conducted at Aminu  Kano Teaching Hospital, North-West TB reference Laboratory and purposive sampling technique was used in selecting  patients for the study. Data generated was presented using descriptive statistics. Sputum samples were collected and  processed according to standard mycobacteriological procedures. The presence of M. tuberculosis complex (MTBC) was  confirmed using SD BIOLINE TB Ag MPT64 Rapid test. Genotypic assays were carried out on these samples by the  GenoType® MTBDRplus molecular line prove assay (LPA). The results of the study revealed that of the 88 sputum samples  collected from rifampicin resistant (confirmed by Genxpert from referring centres) TB patients, 74 (84.09%) were males and  14 (15.9%) were females. Most of them were aged 21-30 years and 31-40 years. Also, 86 of the sputum samples were  confirmed as MTBC. Genotypic assays revealed that 48/88 (54.5%) of the samples were identified as MDR-TB, 29 (32.9%) were rifampicin mono-resistant cases while only 1(1.36%) was resistant to isoniazid and 8 samples were found to be  susceptible to both isoniazid and rifampicin. The pattern of genetic mutations revealed that rifampicin resistance due to the missing rhoB WT gene and the presence of rhoB MUT gene was seen in 77/78 (98.7%) and 51/78 (61.4%) of the rifampicin-resistance samples respectively and isoniazid resistance due to katG missing WT gene and katG MUT gene region was seen in 35/78 (44.9%) of the samples. Among the MDR-TB cases rifampicin resistance was associated with rpoB WT gene and rpoB MUT gene in 100% and 62% of the isolates, and INH resistance was associated with katG WT gene and katG MUT gene in 70.8% and 72.9% of the isolates respectively. INH resistance in the MDR-TB cases was further associated with InhA WT and InhA MUT in 16.67% and 4.17% of the isolates respectively. The study concludes by identifying that the reported MDR-TB rate of 54.5% among rifampicin-resistant TB patients in the study was rather alarming and also identifies the need for expertize in the conduct and diagnosis of rifampicin-resistance status of TB patients using the GeneXpert techniques lack of which may be detrimental to the health of the patient.Thus, the study solicits for a much more committed TB control programmesincluding provision and training of manpower by the relevant agencies as a well as putting more efforts towards rapid and accurate diagnosis of TB patients, and proper treatment and management of the infected cases to minimize the spread and evolution of MDR-TB and indeed XDR-TB whose treatment is presented with more challenges.Keywords: Rifampicin resistance, MDR-TB, LPA, North-Western zone, Nigeria

Bacterial meningitis in pregnancy, neonate and adolescence: A report of five cases

Background: Bacterial meningitis is an infection of the meninges, the thin covering of the brain and spinal cord.Objective: This case study identifies the aetiology and the antimicrobial susceptibility of bacterial agents in suspected samples of acute meningitis.Methods: The bacterial agents were detected by culture, Bactec 9050, Directgen and Gram Techniques from five patients of Makoda and Tudun Wada Primary Health care (PHC) facilities as well as Hasiya Bayero Paediatric Hospital (HBPH) that were presented to Microbiology Laboratory unit of Aminu Kano Teaching Hospital (AKTH) – a tertiary hospital located within the meningitis belt of Nigeria. Among the five cases, three werepregnant women all in their second trimesters marked patients 1, 2 and 3 according to the increasing pattern of their ages, an adolescent marked patient 4 and a neonate marked patient 5.Results: The CSF sample of the respective patients was assayed for pus cell, protein, and RBC. The Directgen detected Haemophillus influenzae from patients 1 and same organism was isolated from patient 2. Streptococcus pneumoniae isolated from patient 3 and Neisseria meningitidis from patients 4 and 5 respectively. Antimicrobial susceptibility testing reveals sensitivity to penicillin and gentamicin by H. influenzae, S. pneumoniae and N. meningitidis. N. Meningitidis and S. pneumoniae were sensitive to ceftriazone, ciprofloxacin and ofloxacin. All the three organisms displayed intermediate sensitivity to chloramphenicol, erythromycin and tetracycline according to Clinical and Laboratory Standards Institute (CLSI) breakpoints.Conclusion: This study demonstrates the relevance of these agents as potential pathogens in the predisposed patients. The antimicrobial sensitivity depicted high sensitivity to the various classes of antibiotics

Assessing the in vitro antibacterial activity of honey against wound and respiratory tract infecting bacteria

Background: Honey is a natural product rich in compounds with antimicrobial properties. The antimicrobial properties are being harnessed in the treatment of respiratory tractinfections and wound care. Honey is locally available in Nigeria and is used in thetreatment of wounds as well as respiratory tract infections. However, the antimicrobialpotentials differ across honey sources and locality.AIM: We sought to obtain laboratory based-evidence by assessing the antibacterialactivities of three honey samples collected from Nsukka in Enugu State.Methods: Neat and diluted honey was tested on six (Escherichia coli, Pseudomonas aeruginosa, Salmonella specie, Staphylococcus aureus, Streptococcus faecium, and Streptococcus pneumoniae) bacterial isolates from infected wound and sputum specimen using in vitro agar diffusion assay.Results: The result obtained showed that the different honey samples had antibacterial activity on all the strains of bacteria tested except for Pseudomonas aeruginosa with minimum inhibitory concentration ranging from 10mg/ml to 27 mg/ml.Conclusion: This suggests the usefulness of honey as an antibacterial for wound and respiratory tract infecting bacteria. However, the need for isolation and susceptibility testing of infecting bacterial organisms to honey in clinical practice is recommended

Second line drug sensitivity test of multidrug resistant tuberculosis (MDR-TB) isolates using proportional method

Drug resistant Tuberculosis (DR-TB) continue to present challenges to the TB control programmes with Multidrug resistant TB (MDR-TB) becoming a major public health threat predisposing individuals to develop Extensively drug resistant TB (XDR-TB), and Totally dug resistant TB (XXDR-TB). The World Health Organization reemphasizes the need for drug Susceptibility testing (DST) for appropriate treatment of individual cases in order to minimize rapid transmission of tuberculosis especially . DR-TB. The study aimed at performing second line drug sensitivity test of MDR-TB isolates to some of the major second line anti-TB drugs. Samples for the study comprised of 22 purposely selected MDR-TB isolates obtained from sputum samples of new AFB smear positive (MDR-TB) patients that were brought to the North West Zonal Tuberculosis Reference Laboratory located at Aminu Kano Teaching Hospital, Kano. The isolates were re-confirmed as MDR-TB using Line prove assay (LPA), and subjected to second line drug sensitivity testing using Proportional methods to some of the second line anti-TB drugs (Ofloxacin, Capreomycin, Ethionamide, & Kanamycin). The results of the study revealed that all the 22 isolates were re-confirmed as MDR-TB using the LPA. The results further shows that all the 22 isolates were sensitive to Ofloxacin (100%) and Capreomycin (100%), followed by Kanamycin (90.9%) and Aminkacin (86.36%) and none of them was identified as XDR-TB. Most of the isolates 16 (72.72%) were however resistant to Ethionamide with only 6 (27.27%) of them being . sensitive. The study revealed absence of XDR-TB among the studied isolates as all of them were sensitive to Ofloxacin and Capreomycin, however majority of them were resistant to Ethionamide and very few to Amikasin and Kanamycin. The study identifies the need for conducting drug susceptibility tests for isolates from patients identified as new AFB smear positive for appropriate treatment so as to reduce transmission of DR-TB.KEYWORDS: Second Line Drug Sensitivity Test, Proportional Method, MDR-T

Enzymatic Hydrolysis and Bioethanol Production from Neem Tree Leaves (AzadirachtaIindica)

Bio-ethanol was produced from Neems tree leaves (Azadirachta indica) through enzymatic hydrolysis using Bacillus firmus as fermenting agent. Dried powdered of Neem tree leaves (Azadirachta indica) was hydrolyzed using 1.0cm3, 1.5cm3 and 2.0cm3 of Bacillus suspension. After that the hydrolyzed samples were fermented using 3cm3 suspension of cultured bacteria (Bacillus firmus). After that, the fermented broths formed were distillated to obtained bioethanol. Acidified K2Cr2O7 was used to determine the concentration of the bioethanol produced. In which the absorbance of the bioethanol obtained using UV-Visible Spectrophotometer was extrapolated with the series concentration prepared with standard glucose solution. The FTIR Spectroscopy analysis of bio-ethanol produced confirms the presence of alcohol functional group present in the sample. The percentage yield of the bioethanol from the leaves using 1.0cm3, 1.5cm3 and 2.0cm3 of Bacillus suspension were 1.42%, 1.85%, and 1.72% yield respectively. The result showed that the Neem tree leaves that are from lignocelluloses materials can produce some appreciable percentage of bio-ethanol. Among the samples 1.5cm3of Bacillus suspension has shown the highest yield of 1.85%.        &nbsp