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This present study was undertaken for detection of extended spectrum β-lactamases (ESBLS) enzyme genes among clinical isolates of Pseudomonas aeruginosa using phenotypic and molecular techniques. Thirty-four P. aeruginosa isolates from different hospitals in Nsukka and University of Nigeria Teaching Hospital (UNTH), Enugu were screened for the presence of ESBL-encoding genes. Phenotypic screening for ESBL producers was carried out using double disk synergy test and combined disk test. Genomic DNA was extracted from the isolates by modified boiling method. Extracted DNA was amplified by polymerase chain reaction (PCR) using ESBL specific primers namely Bla GES, PER, OXA-50, SHV, CTX-M and TEM. The results revealed that a total of 15 isolates of P. aeruginosa were identified as ESBL producer by phenotypic approaches which exhibited varying degrees of resistance to an array of antibiotics tested. While, the PCR screening revealed that 53.33% (n=8) of the isolates that were phenotypically ESBL positive harboured bla OXA-50 gene. However, the genes that encode PER, GES, SHV, TEM and CTX-M were not found in any of the P. aeruginosa isolates. This study highlights the need to establish antimicrobial resistance surveillance network to determine the appropriate empirical treatment regimen for Pseudomonas infections.

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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).

Studies have suggested that modulation of gut microbiota is a viable therapeutic possibility for diabetes. This study evaluated the ability of an edible plant, Gongronema (G.) latifolium Benth (Asclepiadaceae), to modulate the gut microbiome and reduce blood glucose of alloxan-induced diabetic rats. Thirty (30) young, male, albino rats were divided into 6 groups of 5 rats each: Group 1 comprised normal rats; Groups 2 to 4, diabetic rats treated with 200, 400 and 800 mg/Kg body weight of hydro-alcoholic leaf extract, respectively; Group 5, diabetic rats treated with 0.2 mg/Kg glibenclamide (an anti-diabetic drug); and Group 6 comprised untreated diabetic rats. Following induction of diabetes with alloxan injections, the treatments were administered twice daily on a 12-hourly basis by orogastric intubation for 21 days. Thereafter, faecal samples were collected from the animals and subjected to metagenomic analysis, to ascertain the composition and relative abundance of the gut microbiota. There were five dominant bacterial phyla in the rat gut: Firmicutes, Bacteroidetes, Actinobacteria, Spirochaetea and Proteobacteria. Induction of diabetes resulted in observable dysbiosis in the rats. However, treatment of the diabetic rats with G. latifolium extract, ameliorated the state of dysbiosis and resulted in significant increase in species like Lactobacillus (L.) johnsonii, L. reuteri and Prevotella corpri, which are associated with improved glucose metabolism. The plant extract produced the best result at the dose of 400 mg/Kg. The results from this study show that G. latifolium may be used as a therapeutic option for restoration of the microbiome in diabetic patients.

This study aims to determine the malaria parasite clearance rate of crude methanol extract of Cryptolepis sanguinolenta in mice infected with chloroquine sensitive strain of Plasmodium berghei. P. berghei was injected in mice and left for 3 days for establishment. Blood sample collected and diluted with phosphate buffer saline was used for infection. Five (5) groups of animals (mice) were used in this study each containing 5 animals each. The body weights of the entire animal were recorded before and after treatment. Group 1 (normal control), Group 2 (positive control, untreated malaria-passaged mice), Group 3 (standard control, malaria -passaged mice treated with 25 mg/kg body weight of chloroquine), Group 4 (malaria-passaged mice treated with 200 mg/kg body weight of extract), and Group 5 (malariapassaged mice treated with 400 mg/kg body weight of extract). Hematological assessments were carried out before the experiment, 5 days after infection and after treatment. The percentage of parasite load in malaria passaged mice was found to be significantly (p < 0.05) lower in animals treated with mid and high doses of the extract when compared to control groups. Before treatment, no significant (p > 0.05) elevation was observed in the body weight of mice. On day 5 after infection, dose-dependent significant (p < 0.05) decrease was observed in the test groups. After treatment period, the body weights of the animals exhibited dose-dependent increase. The study thus revealed that Cryptolepis sanguinolenta root extracts possesses antimalarial activity in the in vivo mice model and has the ability of re-establishing the blood cells by boosting and stabilizing the blood parameters.

Objective: This study was aimed to determine the antibiotic resistance patterns of clinical Pseudomonas aeruginosa isolates and to detect the presence of PstS gene. Methods: One hundred and ninety-two clinical isolates of P. aeruginosa were characterized using polymerase chain reaction (PCR) and 16S rDNA sequencing. Antibiotic resistance patterns were determined using the disk diffusion method, while the minimum inhibitory concentrations (MICs) of selected antibiotics against resistant isolates were determined by macro broth dilution and E-test strip methods. The resistant isolates were screened for the presence of PstS gene using PCR. Results: Of 192 clinical isolates of P. aeruginosa, 136 (70.83%) were resistant to at least two antibiotics. Of these, 135 (99%) could be classified as multidrug-resistant P. aeruginosa (MDR-PA), 63 (46%) were extensively drug-resistant (XDR-PA), while 38 (28%) were pandrug-resistant (PDR-PA). The isolates exhibited high level of resistance to cefotaxime and ticarcillin, and low levels of resistance to meropenem and imipenem. The MIC values for meropenem against the resistant isolates were generally <32 mg/L, while the values for other antibiotics ranged from 32 to >128 mg/L. Multiple antibiotic resistance indexes of the MDR-PA ranged from 0.27 to 0.91 and the most prevalent pattern of resistance was PiperacillinR – TicarcillinR – Piperacillin/TazobactamR – CefotaximeR – CeftazidimeR – GentamicnR – TobramycinR– CiprofloxacinR. About 50% of the resistant isolates possessed the PstS gene. Conclusions: The results confirmed the presence of XDR, PDRPA, and PstS gene in P. aeruginosa strains. There is an urgent need for healthcare practitioners to address the problem of multidrug resistance, by implementing a more rational and appropriate use of antibiotics.