Browsing by Author "Okoh, Anthony Ifeanyi"

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    Antibacterial Activity of Metallic Nanoparticles against MultidrugResistant Pathogens Isolated from Environmental Samples: Nanoparticles/Antibiotic Combination Therapy and Cytotoxicity Study
    (American Chemical Society, 2022-09-26) Adeniji, Oluwaseun Ola ; Ojemaye, Mike Onyewelehi ; Okoh, Anthony Ifeanyi
    Multidrug-resistant organisms have increased the prevalence of infectious diseases and have become the leading source of death globally. The adverse effects associated with conventional antibiotics cannot be underestimated, and as a result, the quest for antibacterial agents has received great attention over the years. Therefore, the current research was designed to synthesize and examine the antibacterial properties of two metallic nanoparticles, silver nanoparticles (AgNPs) and zinc oxide nanoparticles (ZnONPs), as well as their antibiotic combination therapy against multidrug-resistant bacteria. AgNPs and ZnONPs were synthesized by the coprecipitation method and characterized. Thereafter, their antibacterial activity against multidrug-resistant bacteria was investigated using the microdilution technique. Subsequently, the interactions between the synthesized nanoparticles and antibiotics were evaluated by checkerboard assay. Time-kill assays were carried out to assess bacteriostatic or bactericidal effects, and the cytotoxicity study was carried out by MTT assay. The SEM analysis of AgNPs and ZnONPs were spherical with an average size of 21.03 and 43.37 nm, respectively. FTIR analysis showed the characteristics of the metal−oxygen vibrational band for both materials around 450 cm−1 , which indicated the successful synthesis of these antibacterial agents. The EDX characterization revealed Zn and O with 77.89% and 18.24% abundance in ZnONPs and Ag with 95.65% abundance in AgNPs. UV−vis absorption spectra of AgNPs was obtained around 400 nm. ZnONPs showed a moderate antibacterial effect against Enterococcus species with a MIC range of 2.5−5 mg/mL, while AgNPs demonstrated a strong antibacterial effect against the tested bacterial strains with a MIC range of 0.078−0.039 mg/mL. The ZnONPs were found to be cytotoxic against Vero cell lines at the tested concentrations, whereas AgNPs had no cytotoxic effect at lower concentrations. Their combination activities showed synergetic and additive effects. These findings revealed that these synthesized materials could serve as alternate antibacterial agents against multidrug-resistant Acinetobacter baumanni and Enterococcus species.
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    Antibiotic resistance and virulence genes profiling of Vibrio cholerae and Vibrio mimicus isolates from some seafood collected at the aquatic environment and wet markets in Eastern Cape Province, South Africa
    (Public Library of Science, 2023-08-24) Abioye, Oluwatayo E.; Nontongana, Nolonwabo; Osunla, Charles A.; Okoh, Anthony Ifeanyi; Venkitanarayanan, K.
    The current study determines the density of Vibrio spp. and isolates V. cholerae and Vibrio mimicus from fish-anatomical-sites, prawn, crab and mussel samples recovered from fish markets, freshwater and brackish water. Virulence and antibiotic resistance profiling of isolates were carried out using standard molecular and microbiology techniques. Vibrio spp. was detected in more than 90% of samples [134/144] and its density was significantly more in fish than in other samples. Vibrio. cholerae and V. mimicus were isolated in at least one sample of each sample type with higher isolation frequency in fish samples. All the V. cholerae isolates belong to non-O1/non-O139 serogroup. One or more V. cholerae isolates exhibited intermediate or resistance against each of the eighteen panels of antibiotics used but 100% of the V. mimicus were susceptible to amikacin, gentamycin and chloramphenicol. Vibrio cholerae exhibited relatively high resistance against polymyxin, ampicillin and amoxicillin/clavulanate while V. mimicus isolates exhibited relatively high resistance against nitrofurantoin, ampicillin and polymixin. The multiple-antibiotic-resistance-index [MARI] for isolates ranges between 0 and 0.67 and 48% of the isolates have MARI that is >0.2 while 55% of the isolates exhibit MultiDrug Resistance Phenotypes. The percentage detection of acc, ant, drf18, sul1, mcr-1, blasvh, blaoxa, blatem, blaoxa48, gyrA, gyrB and parC resistance-associated genes were 2%, 9%, 14%, 7%, 2%, 25%, 7%, 2%, 2%, 32%, 25% and 27% respectively while that for virulence-associated genes in increasing other was ace [2%], tcp [11%], vpi [16%], ompU [34%], toxR [43%], rtxC [70%], rtxA [73%] and hyla [77%]. The study confirmed the potential of environmental non-O1/non-O139 V. cholerae and V. mimicus to cause cholera-like infection and other vibriosis which could be difficult to manage with commonly recommended antibiotics. Thus, regular monitoring of the environment to create necessary awareness for this kind of pathogens is important in the interest of public health.
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    Occurrence of virulence determinants in vibrio cholerae, vibrio mimicus, vibrio alginolyticus, and vibrio parahaemolyticus isolates from important water resources of Eastern Cape, South Africa
    (BioMed Central, 2023-10-23) Abioye, Oluwatayo E.; Osunla, Charles A.; Nontongana, Nolonwabo; Okoh, Anthony Ifeanyi
    Background: Virulence determinants are crucial to the risk assessment of pathogens in an environment. This study investigated the presence of eleven key virulence-associated genes in Vibrio cholerae (n=111) and Vibrio mimicus (n=22) and eight virulence determinants in Vibrio alginolyticus (n=65) and Vibrio parahaemolyticus (n=17) isolated from six important water resources in Eastern Cape, South Africa, using PCR techniques. The multiple virulence gene indexes (MVGI) for sampling sites and isolates as well as hotspots for potential vibriosis outbreaks among sampling sites were determined statistically based on the comparison of MVGI. Result: The PCR assay showed that all the V. cholerae isolates belong to non-O1/non-O139 serogroups. Of the isolates, Vibrio Cholera (84%), V. mimicus (73%), V. alginolyticus (91%) and V. parahaemolyticus (100%) isolates harboured at least one of the virulence-associated genes investigated. The virulence gene combinations detected in isolates varied at sampling site and across sites. Typical virulence-associated determinants of V. cholerae were detected in V. mimicus while that of V. parahaemolyticus were detected in V. alginolyticus. The isolates with the highest MVGI were recovered from three estuaries (Sunday river, Swartkopps river, buffalo river) and a freshwater resource (Lashinton river). The cumulative MVGI for V. cholerae, V. mimicus, V. alginolyticus and V. parahaemolyticus isolates were 0.34, 0.20, 0.45, and 0.40 respectively. The targeted Vibrio spp. in increasing order of the public health risk posed in our study areas based on the MVGI is V. alginolyticus>V. parahaemolyticus>V. cholerae>V. mimicus. Five (sites SR, PA5, PA6, EL4 and EL6) out of the seventeen sampling sites were detected as the hotspots for potential cholera-like infection and vibriosis outbreaks. Conclusions: Our findings suggest that humans having contact with water resources in our study areas are exposed to potential public health risks owing to the detection of virulent determinants in human pathogenic Vibrio spp. recovered from the water resources. The study affirms the relevancy of environmental Vibrio species to the epidemiology of vibriosis, cholera and cholera-like infections. Hence we suggest a monitoring program for human pathogenic Vibrio spp. in the environment most especially surface water that humans have contact with regularly
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    Prevalence of Class 1 Integron and In Vitro Effect of Antibiotic Combinations of Multidrug-Resistant Enterococcus Species Recovered from the Aquatic Environment in the Eastern Cape Province, South Africa
    (MDPI, 2023-02-03) Adeniji, Oluwaseun Ola; Nontongana, Nolonwabo; Okoh, Anthony Ifeanyi; Sharma, K.
    Enterococci are regarded as a better indication of faecal pollution in freshwater and marine waters. Their levels in seawater are positively connected with swimming-related gastrointestinal disorders. This study used an Enterococcus-specific polymerase chain reaction (PCR) to characterize the isolates. Classes 1 and 2 integrons were examined for environmental Enterococcus isolates using a standard biological procedure. All strains were assessed against a panel of 12 antibiotics from various classes using disc diffusion methods. The microdilution method was used to work out the minimum inhibitory concentration (MIC) according to the CLSI guiding principles. The combination therapy of the resistant drugs was evaluated using a checkerboard assay and a time-dependent test for assessing their bactericidal or bacteriostatic activity. The gene diversity of the tested organisms was analyzed with the aid of Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR. In total, 57 Enterococcus spp. environmental samples were recovered, in which Enterococcus faecalis (33.33%) and Enterococcus faecium (59.65%) were the dominant species. Resistance to linezolid, ciprofloxacin, erythromycin, gentamicin, vancomycin, rifampicin, and tetracycline was prevalent. Fifty (50) strains tested positive for class 1 integron, more frequent in Enterococcus faecium and Enterococcus faecalis isolates, with no gene cassette array discovered. A combination of gentamicin (MIC 4 µg/mL) with vancomycin (MIC 256 µg/mL) antibiotics against Enterococcus faecalis showed antibacterial activity. In contrast, the combination of ciprofloxacin (1 µg/mL) with Ampicillin (16 µg/mL) antibiotics against Enterococcus faecalis showed a bacteriostatic effect. The ERIC-PCR analysis pointed out that most of the assessed isolates have close genetic similarities.
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    Seasonal variations and associated health risks of polychlorinated naphthalenes in Markman Canal, Eastern Cape Province, South Africa
    (Springer, 2022-07-01) Agunbiade, Idowu Victoria; Adeniji, Abiodun Olagoke; Okoh, Anthony Ifeanyi; Okoh, Omobola Oluranti
    This study focused on evaluation of the levels, seasonal variations and human health risks associated with polychlorinated naphthalenes (PCNs) in water and sediment samples of Markman Canal using solid phase and soxhlet extraction methods respectively, followed by clean-up and quantification with gas chromatograph coupled with microelectron capture detector. The sum of eight PCNs congener’s (∑8PCNs) in water and sediments varied from 0.035 to 0.699 μg/L and 0.260 to 6744.16 μg/kg dw, respectively. Highest PCNs concentrations were recorded in water during winter, while sediment samples collected during spring contained maximum levels. The estimated toxic equivalency (TEQ) for water and sediments was 1.19× ­10–7–1.47× ­10-4 µg/L and 4.43× ­10−5–4.19× ­10−1 µg/kg consecutively. The PCN levels and TEQ values revealed that this waterbody is polluted but constitutes no excess health risk. Efforts should be made to control all the activities contributing to pollution of this canal.
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    Virulence Signatures, Integrons, and Antibiotic Resistance Genes in Bacterial Strains Recovered from Selected Commercial Dairy Products and Fresh Raw Meat
    (Springer, 2023-06-24) Aiyegoro, Olayinka A.; Moyane, Jeremia N.; Adegoke, Anthony A.; Jideani, Afam I.O.; Reddy, Poovendhree; Okoh, Anthony Ifeanyi
    Bacterial species responsible for food infections and intoxication are sometimes carried through the food production and processing. Very few published literatures exist on integrons among antibiotic-resistant staphylococcal strains from foods of animal origin in Gauteng Province, South Africa, hence this study. A total of 720 samples (360 meat and 360 dairies) from a community abattoir of a research farm in South Africa, using conventional bacteriological and molecular methods. Nine (9) bacterial strains, including Bacillus subtilis AYO-123, Acinetobacter baumannii AYO-241, Staphylococcus lentus AYO-352, among others were identified and submitted to GenBank. More bacterial strains were recovered from raw meat (90.5%) than dairy products (9.5%). Resistance was shown (0–100%) to Imipenem, Meropenem, Norfloxacin, Clindamycin, and 22 other antibiotics, without any carbapenem-resistant Acinetobacter baumannii and methicillin/vancomycin-resistant Staphylococcus species (MRSS/VRSS). Virulence genes for fibronectin-binding protein A (FnbA) were predominant (56.24%) followed by the circulating nucleic acids (cna) gene (43.75%). Others were staphylococcal enterotoxin A (sea, 41%), staphylococcal enterotoxin B (seb, 23.5%). Co-presence of sea and seb genes occurred in 11.76% of the isolates, but no coa genes was amplified. Antibiotic resistance genes (ARGs), tetK (70.58%), linA (29.4%), and ermA (11.76%) were detected, but none of the mecA and vat genes was amplified. Class 2 integron (50%) was more predominantly detected than integron 1 (25%), but no Class 3 integron was detected. Bacteria with both the detected virulence and antibiotic resistance genes are of potentialrisks to human health.