Open Access Original Research Article

Antibiotic Resistance and Production of Extended Spectrum β-Lactamases by Clinical Gram-negative Bacteria in Benin

Wassiyath Moussé, Haziz Sina, Ibrahima A. Mama-Sirou, Eugénie Anago, Durand Dah-Nouvlessounon, Christine N’Tcha, Farid Bade, Sévérin Anagonou, Adolphe Adjanohoun, Lamine Baba-Moussa

Journal of Advances in Microbiology, Page 1-13
DOI: 10.9734/jamb/2019/v18i230158

Aims: The aim of this work was to determine the resistance profile and to investigate the production of extended spectrum β-lactamases (ESBL) by clinically relevant Gram-negative Bacillus (GNB) strains.

Methodology: About 191 strains were isolated from 1823 samples collected at the HKM National Hospital and University Center of Cotonou (Benin). Species identification was done with the Api 20th gallery. Two methods were used to search for β-lactamase production: the liquid acidimetric test for penicillinases and double halo method for ESBL. The susceptibility to conventional antibiotic molecules was investigated by the disk diffusion method. Polymerase Chain Reaction (PCR) was used to identify blaTEM and blaSHV genes in the β-lactamases.

Results: A prevalence of 10.48% of GNB was recorded. Among the isolated strains, 51.31% came from samples collected from in-patients and 48.69% from out-patients’ samples. The most contaminated samples were urine (43.98%), pus (34.58%) and blood (9.42%). Majority of the isolated species included: Klebsiella pneumoniae (28.27%), Acinetobacter spp. (18.32%), Pseudomonas aeruginosa (15.72%), Escherichia coli (14.15%) and Enterobacter cloacae (12.04%). More than the half (57.07%) of the strains produced penicillinases; whereas 16.76% were ESBL-producers and these occurred only among Klebsiella pneumoniae, Enterobacter cloacae, Escherichia coli and Enterobacter agglomerans. The ESBL-producing strains were cross-resistant to beta-lactams. Imipenem is the most effective antibiotic on all isolated strains. ESBL-producing GNB strains possessed both the blaTEM gene and the blaSHV gene in a proportion of 25%; 37.5% of the strains had only the blaTEM gene and 12.5% of the strains had only the blaSHV gene.

Conclusion: ESBL-producing strains of K. pneumonia in the hospital environment were the major carriers of blaTEM and blaSHV. Given this situation, it is necessary to continue research to identify resistance genes.

Open Access Original Research Article

Prevalence of Helicobacter pylori Infection in Patients with Dyspeptic Symptoms in Damaturu Metropolitan

S. Gide, Y. Ibrahim, G. Anas, S. D. Alegbe

Journal of Advances in Microbiology, Page 1-8
DOI: 10.9734/jamb/2019/v18i230159

Aim: To study the prevalence of Helicobacter pylori (H. pylori) contamination among dyspeptic patients in Damaturu and report on the relationship between H. pylori predominance and different age groups in the population under study.

Methods: A sum of 229 volunteers with dyspeptic symptoms (140 females and 89 males; mean period of 43.6 ± 14.2 years) took an interest in the investigation. The status of infection caused by H. pylori was determined using serological test. Information was gathered by the utilization of self-managed survey while status of H. pylori was resolved dependent on the serological examination (fast symptomatic test). The one Step H. Pylori serum whole blood rapid test kit was used to detect the presence of IgG antibodies specific to H. pylori infections in the participants.

Results: The prevalence of H. pylori disease was 51.96%. There was no noteworthy factual relationship between age and contamination rate at p-value (0.1515). In young subjects (under 11 years of age), the H. pylori contamination rate was moderately higher (50.00%). The most extreme number of the positive patients was found in the age range of 50-59 years (69.23%) and the base prevalence was in the age range of 10-19 years (50.00%).

The overall prevalence of H. pylori contamination between the gender is out of 89 males, 55 were found to be positive to the infection (61.79%), while out of 140 females, 64 were also positive (45.71%). Despite these findings, there was no critical factual relationship between the genders and H. pylori infection (p=0.113). The statistical qualities of members at the examination section demonstrate that out of 129 patients having a place with the upper lower class, 85 subjects (66.40%) were found to be positive to H. pylori. The lower-middle-class demonstrates that out of 89 subjects 30 were positive (33.70%), while upper-middle outcome demonstrates that out of 12 subjects 4 were confirmed with 33.33%. This demonstrates that subjects living under overcrowded  conditions with dense population have a high contamination rate of H. pylori and therefore is a noteworthy factual relationship between socio-statistic factors and overall prevalence of H. Pylori infection (p = 0.001).

Conclusion: We came to realize the overall prevalence of H. pylori contamination in patients with dyspeptic symptoms in Damaturu. The H. pylori contamination might be a hazard factor for peptic ulcer and more grounded gastritis.

Open Access Original Research Article

ESBL Mediated Antimicrobial Nonsusceptibility of Uropathogenic Escherichia coli and Klebsiella pneumoniae Isolates from Pregnant Women in Nnewi, Nigeria

O. Peculiar-Onyekere, Chioma, C. Agbo, Martina, A. Eze, Emmanuel

Journal of Advances in Microbiology, Page 1-13
DOI: 10.9734/jamb/2019/v18i230166

Background and Objective: Extended Spectrum β-lactamase (ESBL) producing Urinary Tract Infection (UTI) is an important public health issue due to lack of therapeutic antibiotic options and the danger it portends to the pregnant woman. This study was carried out to determine the prevalence and response to antimicrobials of ESBL-producing uropathogenic E. coli and K. pneumoniae, among pregnant women on ante natal care.

Study Design/Materials and Methods: Two hundred and fifteen pregnant women across three different hospitals in Nnewi North L.G.A of Anambra State were screened for these uropathogens. Modified Double Disc Synergy test (MDDST) was carried out on the isolates to phenotypically determine the presence of ESBL. Plasmid profiling as well as plasmid curing studies were undertaken. Molecular characterization of the phenotypically confirmed ESBL positive isolate via Polymearse Chain Reaction (PCR) was carried out using three ESBL primers (bla-TEM, bla-SHV, bla-CTX-M).

Results: 192 isolates were obtained of which 75(39.1%) were E. coli and 117(60.9%) were K. pneumoniae. A total of 130 (67.7%) of the pregnant women had ESBL-mediated UTI, the highest rate reported in recent times in Nigeria. Molecular characterization of the ESBL types revealed a predominance of bla-TEM (91.9%), followed by bla-SHV (73.3%) and bla-CTX-M (56.8%).

Conclusion: The Majority of the isolates harbored multiple ESBL genes. Curing studies were largely ineffectual as most of the isolates retained their resistance determinants regardless of the concentration of the curing agent (acridine orange).

Open Access Original Research Article

Molecular Identification of Plasmodium Species in Malaria in Zimbabwe by 18S Ribosequencing

Willlard Mbiri, Daniel Mukandabvute, Nyasha Chin’ombe

Journal of Advances in Microbiology, Page 1-7
DOI: 10.9734/jamb/2019/v18i230167

Background: Malaria remains a major cause of mortality and morbidity in Zimbabwe due to transmission of Plasmodium parasite by the Anopheles mosquitoes. Globally, several species of Plasmodium parasite have been identified, but only P. falciparum, P. ovale, P. vivax, P. malariae and P. knowlesi are known to cause malaria in humans. No studies have previously been done in Zimbabwe to identify the Plasmodium species in malaria using molecular methods. The the aim of this study was to identify circulating Plasmodium species in malaria in Zimbabwe using 18S ribosequencing.

Methods: The study was a cross-sectional survey in Zimbabwe from January to May in 2016. Venous blood was collected from malaria-suspected patients at three referral hospitals and subjected to microscopy and/or serology. Total DNA was isolated and Plasmodium species identified by 18S amplification and ribosequencing.

Results: A total of 160 patient samples were used in this study. Out of these, 130 were malaria-positive by microscopy and/or serology and 30 were negative. Total genomic DNA was extracted from 100 samples of the patients (80 malaria-positive and 20 malaria-negative samples by microscopy and/or serology). Amplification of the 18S ribosomal RNA gene of Plasmodium was performed on 74 malaria-positive and 6 malaria-negative samples. All the 74 samples showed 18S RNA gene amplification and the 6 negative controls did not show amplification. Only 50 amplicons were selected for sequencing. Ribosequencing and bioinformatics analyses showed that all (100%) the sequences belonged to Plasmodium falciparum.

Conclusion: The study was the first to provide molecular evidence of the existence of only P. falciparum in Zimbabwe. However, further studies with bigger sample sizes need to be done to ascertain whether P falciparum is the dominant circulating species in malaria in Zimbabwe.

Open Access Original Research Article

Effects of Moringa oleifera lam. Leaf Powder on Bifidobacteria and Escherichia coli in the Gut of Albino Rats

A. O. Gbadebo, O. T. Okareh, A. A. Ogunjobi, A. O. Dada

Journal of Advances in Microbiology, Page 1-11
DOI: 10.9734/jamb/2019/v18i230168

Aim: This study was carried out to determine the effects of dried Moringa oleifera leaves on Bifidobacteria and Escherichia coli in the gut of albino rats.

Location: The rats were habituated under laboratory conditions at the animal house of the Department of Zoology, Faculty of Science, University of Ibadan, for two weeks in other to adapt to the environmental conditions during the experiment.

Duration of Study: The rats were exposed to the M. oleifera feed for four weeks.

Design of Study: There were five groups in all. The 5 to 6 weeks old rats were fed with M. oleifera powder supplement except for the control groups.

Methods: No supplement of M. oleifera feed was administered to group A while group B received streptomycin antibiotics.

Groups C, D and E received dried leaf supplement of M. oleifera (DMO) 1.25 g/kg body weight (2.5%), 2.5 g/kg body weight (5%) and 5.0 g/kg body weight (10%) respectively.

Results: E.coli counts increased from 2.3*104 to 2.6*104 colony-forming units per gram (cfu/g) in group E, from 2.2*104 to 3.0*10 cfu/g in group B; but reduced from 4.1*104 to 3.7*104 cfu/g in group D and from 5.4*104 to 3.9*104 cfu/g in group C between day 20 and day 28. As from day 8, the isolates from the non-control groups were resistant to the M. oleifera extract except E. coli isolates in both 5% and 10% M. oleifera groups on day 8 with 6 mm zone of inhibition each. The rate of Bifidobacteria viable counts increase in group E was expressed as P = 0.05 at the beginning of the experiment, unlike E.coli counts where there was a decrease.

Conclusion: The M. oleifera leaf alters the microbiota in the gut, a situation which sends impulses to the brain. Thus, the M. oleifera leaf powder is a potential prebiotic for probiotics like Bifidobacteria, and as well as induce changes in the gut-brain axis.