Characterization of Mycobacterium Species Circulating among Tuberculosis Patients in Bayelsa State, Nigeria

Introduction: Tuberculosis is caused by infection with Mycobacterium tuberculosis. Looking at the evolution of the bacterium gene due to mutation is crucial to identify species circulating among patients in an area. WHO speculated that tuberculosis is caused by M. tuberculosis (MTB), but identification of the strains of MTB circulating in a particular area is important for the management of MTB and to identify pulmonary infections caused by non-tuberculosis mycobacterium. Contact tracing of drug resistant MTB in circulation in an area is also an important procedure of MTB therapeutic choice. Aim: This study aimed to isolate and identify Mycobacterium species circulating in Bayelsa State, Nigeria. Materials and Methods: A total of 102 sputum samples collected from MTB patients were cultured in Lowenstein Jensen (LJ) solid media. Isolates on LJ media were confirmed using Zeihl Nelseen staining method for AFB and Standard Diagnosis Bioline TB Ag MPT64 Rapid test kit. The 16s rRNA gene amplification, agarose gel electrophoresis, and gene sequencing were conducted. Phylogenic analysis and evolutional distances of the strains are computed using the Juke-cantor method. Result: Out of 102 sputum samples examined 15 (14.7%) had growth of Mycobacterium species (AFB positive). The extracted DNA of MTB amplified on agarose gel electrophoresis aligned horizontally at lanes 1 - 15 showing 16S gene band (1500 bp). Two 2 (2.0%) are non-tuberculosis Mycobacteria species, while 13 (12.7%) were M. tuberculosis. The non-tuberculosis Mycobacterium species isolated are Mycobacteriode abscesses and Mycobacterium kansassii strain FDAARGOS 1534. The tuberculosis strains are Mycobacterium tuberculosis MG003 and R2092 but the predominant strain was MG003. The degree of the genetic evolution of the non-MTB Mycobacterium kansassii strain FDAARGOS 1534 was 75.4%. Conclusion: The two major strains of Mycobacterium tuberculosis (MTB) circulating in Bayelsa State are MTB MG003 and MTB R2092;MTB MG003 was predominant. The non-tuberculosis species are Mycobacteriode abscesses and Mycobacterium kansasii.

Comparative Analyses of the Efficacy of Dry and Fresh <i>Mangifera indica</i>Ethanolic Extracts on <i>E. coli</i>and <i>S. aureus</i>

Background: The need to identify and characterize new antimicrobial agents is important due to the increasing development of resistance by microorganisms to the existing antimicrobial agents. Aim: This study examined the efficacies of Mangifera indica on Escherichia coli and Staphylococcus aureus. Method: Three parts (leaf [L], root [R], and bark [B]) of the plant were analyzed. The extraction of the samples was performed by aseptically grinding the samples, dissolving in absolute ethanol, and filtering through whatman filter paper. The efficacy of the extracts bothsingle and combined was determined using agar well diffusion assay with gentamycin [10 μl] (E. coli) and vancomycin [30 μl] (S. aureus) as control antibiotics. Results: The higher concentration (C2 = 3.0 g/ml) showed more antibacterial effectiveness than the lower concentration (C1 = 1.5 g/ml) against both bacterial isolates with significant differences (P < 0.05) in all extracts except for single extracts (E. coli dry leaf extract;fresh bark extract), double extracts (S. aureus: dry and fresh leaf extracts) and triple extract (E. coli and S. aureus dry extracts). For the single extracts the bacteria has the following significant results: E. coli L (dry 6.3 ± 2.5 mm, fresh 14.7 ± 0.6 mm, P = 0.0050), R (dry 11.3 ± 1.5 mm, fresh 7.3 ± 1.5 mm, P = 0.0327);for S. aureus L (dry 7.0 ± 1.7 mm, fresh 11.0 ± 1.0 mm, P = 0.0257), R (dry 7.0 ± 2.0 mm, fresh 11.7 ± 1.5 mm, P = 0.0325), and B (dry 5.0 ± 1.0 mm, fresh 16.0 ± 1.0 mm, P = 0.0002). For the double extracts the bacteria has the following significant results: E. coli L + R (dry 15.7 ± 2.3 mm, fresh 1.7 ± 1.5 mm, P = 0.0070), R + B (dry 18.7 ± 1.5 mm, fresh 9.7 ± 1.5 mm, P = 0.0020), and L + B (dry 9.7 ± 1.5 mm, fresh 6.3 ± 0.6 mm, P = 0.0241);S. aureus L + R (dry 14.7 ± 1.5 mm, fresh 7.0 ± 1.0 mm, P = 0.0019), R + B (dry 15.3 ± 1.5 mm, fresh 11.7 ± 1.5 mm, P = 0.0424). For the triple extracts, the fresh leaves showed significantly higher levels of efficacy than the dry for both E. coli L + R + B (P = 0.0101) and S. aureus (P = 0.0307). The fresh extracts showed higher levels of efficacy than dry extracts against both bacteria for all the single and three combined conditions. Conclusions: Fresh extracts show better efficacies against E. coli while dry extracts show greater efficacies against S. aureus for both single and triple combined extracts. The reverse is true for double combined extracts.

<i>Candida</i>Co-Infection with <i>Mycobacterium tuberculosis</i>in Tuberculosis Patients and Antifungal Susceptibility of the Isolates

It had been observed that tuberculosis (TB) subjects can be co-infected with Candida sp. which was previously assumed as normal flora of the oral cavity. Candida sp. might become an opportunistic pathogen in immune compromised individuals. Candida co-infection with Mycobacterium tuberculosis in TB patients might complicate underlying disease process in the lungs. Materials and Methods: A total of 400 sputum samples were collected from TB patients and examined using Ziehl-Neelsen staining technique and MDR/RIF Genexpert system for TB. Samples positive for Mycobacterium tuberculosis were cultured on Sabouraud Dextrose Agar with gentamicin and examined for the presence of budding yeast cells and pseudohyphae on Gram’s stain. Candida sp. isolated from TB positive sputa were cultured on CHROMager Candida for identification to species level and subjected to antifungal susceptibility testing. Results: Out of 400 sputum samples examined for TB 93 (23.3%) were positive and 32 (34.4%) out of 93 TB positive cases were co-infected with Candida sp. Candida albicans was the most predominant species with a prevalence of 23 (67.6%), C. tropicals 4 (11.8%), C. krusei 4 (11.8%) and C. parapsilosis 3 (8.8%). One sample had dual infection. Female subjects had high prevalence (19.4%) than the male (15.7%). Age group 31 - 40 years had both high prevalence of TB 32.3% and Candida 25.0%. Antifungal susceptibility testing showed that isolated Candida sp. were more susceptible to vericonazole and fluconazole compared to nystatin. Conclusions: Tuberculosis weakens the immune systems of infected persons especially when prophylactic administration proves abortive or there is non-adherence to treatment prescriptions. This may cause the development of multidrug resistance TB. Candida sp. may utilize these opportunities to establish alongside M. tuberculosis and worsen treatment and patient condition. For good treatment of TB, Candida co-infection should be screened concomitantly with TB in TB suspected individuals.

Evaluation of the Impact of Hydrocarbon-Generated Soot on Antibiotics Susceptibility of <i>Staphylococcus aureus</i>and <i>Escherichia coli</i>Isolates

Background: The antibiotic susceptibility of bacterial interaction with soot, a by-product of incomplete combustion of fossil fuel, has not been established. Aim: The study aimed to establish the antibiotics susceptibility of Staphylococcus aureus and Escherichia coli exposed to soot. Method: The bacteria were exposed to 12.5%, 25%, and 50% concentrations of soot at different time intervals. Control bacterial cultures without exposure to soot were also carried out. These cultures were incubated for 24 hrs. The numbers of surviving bacteria were determined by analyzing 10 μL of the incubated cultures at 6 hrs and 24 hrs on tryptone soy agar. Again, the bacteria were inoculated on Mueller Hinton agar and subjected to antibiotics susceptibility testing using the disk diffusion method. Results: After 6 hrs of exposure, the number of E. coli in the absence of soot was 102.50 ± 3.54 × 103 CFU/mL while at 12.5%, 25%, and 50% of soot, the surviving E. coli were 26.00 ± 1.41 (p = 0.0012), 21.00 ± 1.41 (p = 0.0011) and 5.50 ± 2.12 (p = 0.0009) × 103 CFU/mL respectively. Similarly, the population of S. aureus without soot was 122.5 ± 3.53 × 104 CFU/mL while at 12.5%, 25.0% and 50.0% of soot, the surviving S. aureus 46.00 ± 2.83 (p = 0.0017), 23.00 ± 1.41 (p = 0.0007) and 11.50 ± 2.12 (p = 0.0007) × 104 CFU/mL respectively. Similar results were obtained after 24 hrs of exposure. The soot shows some level of potency in reducing the number of E. coli and S. aureus significantly (p < 0.05). After 24 hrs, almost all treatment conditions (except for the Gentamicin for S. aureus), there was resistance to all the antibiotics while at 0 hr there was sensitivity to these drugs. Conclusion: These results suggest that while soot has some potency on E. coli and S. aureus, their exposure to soot could induce resistance.

Antimicrobial Properties and Phytochemical Composition of <i>Garcinia kola, Bryophyllum pinnatum</i>, and <i>Allium sativum</i>Juices on Some Clinical Pathogens

Background: Medicinal plants have been in use since the origin of man. Many important chemical substances with biological functions that could be used for treatment and prevention of attack from bacteria, fungi, herbivorous mammals and insects are produced by different plants. Such compounds with useful properties have been recorded in their numbers, about 12,000 accounting for about 10% of total plant species. Aim: The aim of the study was to determine the antimicrobial efficacies of herbal extracts on some clinical pathogens. Methods: The antimicrobial activities of pressed juices of Allium sativum (garlic), Bryophyllum pinnatum and Garcinia kola neats and their dilutions were tested on pathogens such as Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Staphylococcus aureus and Candida albicans to determine their susceptibility to the juices and their combinations. Agar well diffusion method was employed on Muller-Hinton agar to determine their antimicrobial susceptibility pattern. The phytochemical analysis of the plants’ juices were also determined. Results: At 100% (neat) the juices of G. kola, B. pinnatum and Garcinia kola showed substantial zones of inhibition against the pathogens with a zone diameter of about 22.0 mm and above. At 75% concentrations, the juices inhibited the pathogens tested against them. A. sativum (garlic) inhibited K. pneumonia, P. mirabilis, and S. aureus even at 50% concentration. C. albicans isolates were 60% susceptible to G. kola juice, 40% at 100% concentration. At 75% concentration of the juice, C. albicans isolates were also 60% susceptible to the juices. At 50% - 100% concentrations, C. albicans isolates were 100% sensitive to A. sativum extract. Conclusions: The medicinal plant juices tested against the pathogens possess some potentials worth exploiting as potent antimicrobial agents on gram-positive, gram-negative bacteria and the fungus.

Bacteriological Quality of Swimming Pools Water in Port Harcourt Metropolis

The bacteriological quality of swimming pool water in Port Harcourt Metropolis was investigated. Ten (10) swimming pools were examined for microbial quality. Out of the 10 swimming pools, 4 (2, 4, 7 and 9) had bacterial isolates of 40 (100%). Swimming pools 2, 4, 7 and 9 had the bacterial isolates of 10 (25%), 8 (20%), 10 (25%) and 12 (30%), respectively. Of the forty (40) bacterial isolates identified, which represented 100 percent, 22 (55%) were identified as Staphylococcus epidermidis, 10 (25%) Bacillus cereus, 6 (16%) Micrococcus and 2 (5%) Staphylococcus aureus. Among the four swimming pools, 2 and 9 did not have the isolates of Staphylococcus aureus and Micrococcus, respectively. Based on the World Health Organisation (WHO) standard for recreational waters, the absence of coliform and fecal coliform bacteria (E. coli) revealed that the ten (10) swimming pools used for this study are considered to be within the acceptable limits for certifying microbiological water quality. However, there is a need for care and continuous maintenance of the swimming pools.