Hospital wastewater (HWW) harbours diverse microbial species and a miscellany of genome that woul... more Hospital wastewater (HWW) harbours diverse microbial species and a miscellany of genome that would facilitate the emergence of novel pathogen upon genome integration that manifests novel traits in infectious pathogens. The study aimed to determine the antibiogram, and virulence signatures of Pseudomonas aeruginosa (P. aeruginosa) recovered from selected agrestic hospital effluents in Eastern Cape, South Africa. Thirty-six (36) wastewater samples were collected from selected hospital drains between February 2018 and April 2018, processed and analyzed by culture-dependent methods for the isolation of P. aeruginosa. The identity confirmation of isolates was achieved by amplification of oprl and oprL genes. Antibiogram was done using standard disk diffusion technique of Kirby–Bauer as approved by CLSI 2018 guidelines. Virulence signatures (lasA, lasB, toxA, popB) among isolates were analysed using polymerase chain reaction. A total of 54 P. aeruginosa isolates were confirmed by amplific...
Keratinous biomass valorization for value-added products presents a high prospect in ecological m... more Keratinous biomass valorization for value-added products presents a high prospect in ecological management and the advancement of the bio-economy. Consequently, soil samples from the poultry dumpsite were collected. The bacteria isolated on the basal salt medium were screened for keratinolytic activity. The potent chicken feathers degrading bacteria were identified through 16S rRNA gene sequencing and phylogenetic analysis. Fermentation process conditions were optimized, and the amino acid compositions of the feather hydrolysate were likewise quantified. Ten (10) proteolytic bacteria evaluated on skimmed milk agar showed intact chicken feather degradation ranging from 33% (WDS-03) to 88% (FPS-09). The extracellular keratinase activity ranged from 224.52 ± 42.46 U/mL (WDS-03) to 834.55 ± 66.86 U/mL (FPS-07). Based on 16S rRNA gene sequencing and phylogenetic analysis, the most potent keratinolytic isolates coded as FPS-07, FPS-09, FPS-01, and WDS-06 were identified as <i>Chryse...
Background: The HbA1c estimates long-term glycaemic control in individuals. However, scanty data ... more Background: The HbA1c estimates long-term glycaemic control in individuals. However, scanty data exist on the determination of Diabetes Mellitus (DM) in newly diagnosed HIV patients using the HbA1c screening tool in the South African context. Thus, this study examines the prevalence of diabetes mellitus in newly diagnosed HIV-positive patients in Buffalo City Municipality, East London, South Africa. Methodology: This was a cross-sectional study of 335 newly HIV-diagnosed patients between August 2016 and July 2017. Demographic (age, gender, residence, employment status and level of education) and behavioural variables (smoking and alcohol use (past 30 days)) were by self-reporting. Information on HbA1c and other clinical variables were obtained from the medical records of the patients. Diabetes mellitus was defined based on the Society for Endocrinology, Metabolism and Diabetes of South Africa (SEMDSA) 2017 guideline of HbA1c of above 6.5%. Weight and height were measured using stand...
Background: Transmitted drug resistance (TDR) remains a significant threat to Human immunodeficie... more Background: Transmitted drug resistance (TDR) remains a significant threat to Human immunodeficiency virus (HIV) infected patients that are not exposed to antiretroviral treatment. Although, combined antiretroviral therapy (cART) has reduced deaths among infected individuals, emergence of drug resistance is gradually on the rise. Objective: To determine the drug resistance mutations and subtypes of HIV-1 among pre-treatment patients in the Eastern Cape of South Africa. Methods: Viral RNA was extracted from blood samples of 70 pre-treatment HIV-1 patients while partial pol gene fragment amplification was achieved with specific primers by RT-PCR followed by nested PCR and positive amplicons were sequenced utilizing ABI Prism 316 genetic sequencer. Drug resistance mutations (DRMs) analysis was performed by submitting the generated sequences to Stanford HIV drug resistance database. Results: Viral DNA was successful for 66 (94.3%) samples of which 52 edited sequences were obtained from ...
Sub-Saharan region in Africa still holds the highest burden of HIV/AIDS globally. HIV-1 requires ... more Sub-Saharan region in Africa still holds the highest burden of HIV/AIDS globally. HIV-1 requires coreceptor to gain entry into permissive cells to initiate infection. Molecular analysis of the chemokine coreceptor usage is important clinically and in the effective management of AIDS virus. This study aims to determine the coreceptor usage among HIV-1 drug naïve patients residing in the Eastern Cape, South Africa. We collected blood samples from 55 HIV infected patients into anticoagulant vacutainer. RNA was extracted from seperated plasma and RT-PCR performed followed by nested PCR to amplify the partial env fragment spanning the C2-C3 region. Sanger sequencing was done on the amplicons using the BigDye Terminator V3.1 sequencing kit (Applied Biosystems, Foster City, CA, USA) while sequences were manually edited using BioEdit and Geneious 10.2.6 tools. The WebPSSM and Geno2pheno online tools were also utilised to predict the coreceptor tropism while the phylogenetic analysis of the isolates was determined using MEGA 7. Of the 55 blood samples collected for the study, 50 (91%) were successfully amplified and sequenced. The mean age of the patients was 32 (18-56) years while the ratio of men to women was 35% and 65% correspondingly. Phylogenetic analysis revealed that all 50 sequences clustered with HIV-1 subtype C reference strains. Viral tropism of the V3 loop revealed 47 sequences to be R5 strains while three sequences (T1E, T10E and T11E,) were classified as X4 strains based on the WebPSSM and the Geno2pheno algorithm. HIV-1 R5 tropic strains were the most dominant virus obtained from this study while HIV-1 subtype C still drives the epidemic in South Africa suggesting greater invivo and host pathogen fitness. Documented data on mapping out cellular tropism based on viral tropism is important as Maraviroc and other CCR5 antagonist could be introduced as part of treatment regimen in South Africa.
The study investigated the occurrence of antimicrobial resistance genes and virulence determinant... more The study investigated the occurrence of antimicrobial resistance genes and virulence determinants in Vibrio species recovered from different freshwater sheds in rustic milieu. A total of 118 Vibrio isolates comprising Vibrio fluvialis (n=41), Vibrio mimicus (n=40) and V. vulnificus (n=37) was identified by amplification of ToxR, vmh and hsp60 genes. The amplification of virulence genes indicated that V. mimicus (toxR, zot, ctx, VPI, and ompU) genes were detected in 12.5%, 32.5%, 45%, 37.5% and 10% respectively. V. fluvialis genes (stn, hupO and vfh) were harboured in 48.8%, 14.6% and 19.5% isolates congruently. The other virulence genes that include vcgC and vcgE were observed in 63.1% and 29% of isolates belonging to V. vulnificus. With the exceptions of imipenem, meropenem and ciprofloxacin, most isolates exhibited more than 50% resistance to antibiotics. The antimicrobial resistance was more prevalent for polymyxin B (100%), azithromycin (100%) and least in ciprofloxacin (16.1%)...
OBJECTIVES Decaying wood samples were collected, and actinomycetes were isolated and screened for... more OBJECTIVES Decaying wood samples were collected, and actinomycetes were isolated and screened for laccase production. The identity of the efficient laccase-producing isolate was confirmed by using a molecular approach. Fermentation conditions for laccase production were optimized, and laccase biochemical properties were studied. RESULTS Based on the 16S rRNA gene sequencing and phylogenetic analysis, the isolate coded as HWP3 was identified as Streptomyces sp. LAO. The time-course study showed that the isolate optimally produced laccase at 84 h with 40.58 ± 2.35 U/mL activity. The optimized physicochemical conditions consisted of pH 5.0, ferulic acid (0.04%; v/v), pine back (0.2 g/L), urea (1.0 g/L), and lactose (1 g/L). Streptomyces sp. LAO laccase was optimally active at pH and temperature of 8.0 and 90 °C, respectively, with remarkable pH and thermal stability. Furthermore, the enzyme had a sufficient tolerance for organic solvents after 16 h of preincubation, with laccase activity > 70%. Additionally, the laccase maintained considerable residual activity after pretreatment with 100 mM of chemical agents, including sodium dodecyl sulphate (69.93 ± 0.89%), ethylenediaminetetraacetic acid (93.1 ± 7.85%), NaN3 (96.28 ± 3.34%) and urea (106.03 ± 10.72%). CONCLUSION The laccase's pH and thermal stability; and robust catalytic efficiency in the presence of organic solvents suggest its industrial and biotechnological application potentials for the sustainable development of green chemistry.
Hospital wastewater (HWW) harbours diverse microbial species and a miscellany of genome that woul... more Hospital wastewater (HWW) harbours diverse microbial species and a miscellany of genome that would facilitate the emergence of novel pathogen upon genome integration that manifests novel traits in infectious pathogens. The study aimed to determine the antibiogram, and virulence signatures of Pseudomonas aeruginosa (P. aeruginosa) recovered from selected agrestic hospital effluents in Eastern Cape, South Africa. Thirty-six (36) wastewater samples were collected from selected hospital drains between February 2018 and April 2018, processed and analyzed by culture-dependent methods for the isolation of P. aeruginosa. The identity confirmation of isolates was achieved by amplification of oprl and oprL genes. Antibiogram was done using standard disk diffusion technique of Kirby–Bauer as approved by CLSI 2018 guidelines. Virulence signatures (lasA, lasB, toxA, popB) among isolates were analysed using polymerase chain reaction. A total of 54 P. aeruginosa isolates were confirmed by amplific...
Keratinous biomass valorization for value-added products presents a high prospect in ecological m... more Keratinous biomass valorization for value-added products presents a high prospect in ecological management and the advancement of the bio-economy. Consequently, soil samples from the poultry dumpsite were collected. The bacteria isolated on the basal salt medium were screened for keratinolytic activity. The potent chicken feathers degrading bacteria were identified through 16S rRNA gene sequencing and phylogenetic analysis. Fermentation process conditions were optimized, and the amino acid compositions of the feather hydrolysate were likewise quantified. Ten (10) proteolytic bacteria evaluated on skimmed milk agar showed intact chicken feather degradation ranging from 33% (WDS-03) to 88% (FPS-09). The extracellular keratinase activity ranged from 224.52 ± 42.46 U/mL (WDS-03) to 834.55 ± 66.86 U/mL (FPS-07). Based on 16S rRNA gene sequencing and phylogenetic analysis, the most potent keratinolytic isolates coded as FPS-07, FPS-09, FPS-01, and WDS-06 were identified as <i>Chryse...
Background: The HbA1c estimates long-term glycaemic control in individuals. However, scanty data ... more Background: The HbA1c estimates long-term glycaemic control in individuals. However, scanty data exist on the determination of Diabetes Mellitus (DM) in newly diagnosed HIV patients using the HbA1c screening tool in the South African context. Thus, this study examines the prevalence of diabetes mellitus in newly diagnosed HIV-positive patients in Buffalo City Municipality, East London, South Africa. Methodology: This was a cross-sectional study of 335 newly HIV-diagnosed patients between August 2016 and July 2017. Demographic (age, gender, residence, employment status and level of education) and behavioural variables (smoking and alcohol use (past 30 days)) were by self-reporting. Information on HbA1c and other clinical variables were obtained from the medical records of the patients. Diabetes mellitus was defined based on the Society for Endocrinology, Metabolism and Diabetes of South Africa (SEMDSA) 2017 guideline of HbA1c of above 6.5%. Weight and height were measured using stand...
Background: Transmitted drug resistance (TDR) remains a significant threat to Human immunodeficie... more Background: Transmitted drug resistance (TDR) remains a significant threat to Human immunodeficiency virus (HIV) infected patients that are not exposed to antiretroviral treatment. Although, combined antiretroviral therapy (cART) has reduced deaths among infected individuals, emergence of drug resistance is gradually on the rise. Objective: To determine the drug resistance mutations and subtypes of HIV-1 among pre-treatment patients in the Eastern Cape of South Africa. Methods: Viral RNA was extracted from blood samples of 70 pre-treatment HIV-1 patients while partial pol gene fragment amplification was achieved with specific primers by RT-PCR followed by nested PCR and positive amplicons were sequenced utilizing ABI Prism 316 genetic sequencer. Drug resistance mutations (DRMs) analysis was performed by submitting the generated sequences to Stanford HIV drug resistance database. Results: Viral DNA was successful for 66 (94.3%) samples of which 52 edited sequences were obtained from ...
Sub-Saharan region in Africa still holds the highest burden of HIV/AIDS globally. HIV-1 requires ... more Sub-Saharan region in Africa still holds the highest burden of HIV/AIDS globally. HIV-1 requires coreceptor to gain entry into permissive cells to initiate infection. Molecular analysis of the chemokine coreceptor usage is important clinically and in the effective management of AIDS virus. This study aims to determine the coreceptor usage among HIV-1 drug naïve patients residing in the Eastern Cape, South Africa. We collected blood samples from 55 HIV infected patients into anticoagulant vacutainer. RNA was extracted from seperated plasma and RT-PCR performed followed by nested PCR to amplify the partial env fragment spanning the C2-C3 region. Sanger sequencing was done on the amplicons using the BigDye Terminator V3.1 sequencing kit (Applied Biosystems, Foster City, CA, USA) while sequences were manually edited using BioEdit and Geneious 10.2.6 tools. The WebPSSM and Geno2pheno online tools were also utilised to predict the coreceptor tropism while the phylogenetic analysis of the isolates was determined using MEGA 7. Of the 55 blood samples collected for the study, 50 (91%) were successfully amplified and sequenced. The mean age of the patients was 32 (18-56) years while the ratio of men to women was 35% and 65% correspondingly. Phylogenetic analysis revealed that all 50 sequences clustered with HIV-1 subtype C reference strains. Viral tropism of the V3 loop revealed 47 sequences to be R5 strains while three sequences (T1E, T10E and T11E,) were classified as X4 strains based on the WebPSSM and the Geno2pheno algorithm. HIV-1 R5 tropic strains were the most dominant virus obtained from this study while HIV-1 subtype C still drives the epidemic in South Africa suggesting greater invivo and host pathogen fitness. Documented data on mapping out cellular tropism based on viral tropism is important as Maraviroc and other CCR5 antagonist could be introduced as part of treatment regimen in South Africa.
The study investigated the occurrence of antimicrobial resistance genes and virulence determinant... more The study investigated the occurrence of antimicrobial resistance genes and virulence determinants in Vibrio species recovered from different freshwater sheds in rustic milieu. A total of 118 Vibrio isolates comprising Vibrio fluvialis (n=41), Vibrio mimicus (n=40) and V. vulnificus (n=37) was identified by amplification of ToxR, vmh and hsp60 genes. The amplification of virulence genes indicated that V. mimicus (toxR, zot, ctx, VPI, and ompU) genes were detected in 12.5%, 32.5%, 45%, 37.5% and 10% respectively. V. fluvialis genes (stn, hupO and vfh) were harboured in 48.8%, 14.6% and 19.5% isolates congruently. The other virulence genes that include vcgC and vcgE were observed in 63.1% and 29% of isolates belonging to V. vulnificus. With the exceptions of imipenem, meropenem and ciprofloxacin, most isolates exhibited more than 50% resistance to antibiotics. The antimicrobial resistance was more prevalent for polymyxin B (100%), azithromycin (100%) and least in ciprofloxacin (16.1%)...
OBJECTIVES Decaying wood samples were collected, and actinomycetes were isolated and screened for... more OBJECTIVES Decaying wood samples were collected, and actinomycetes were isolated and screened for laccase production. The identity of the efficient laccase-producing isolate was confirmed by using a molecular approach. Fermentation conditions for laccase production were optimized, and laccase biochemical properties were studied. RESULTS Based on the 16S rRNA gene sequencing and phylogenetic analysis, the isolate coded as HWP3 was identified as Streptomyces sp. LAO. The time-course study showed that the isolate optimally produced laccase at 84 h with 40.58 ± 2.35 U/mL activity. The optimized physicochemical conditions consisted of pH 5.0, ferulic acid (0.04%; v/v), pine back (0.2 g/L), urea (1.0 g/L), and lactose (1 g/L). Streptomyces sp. LAO laccase was optimally active at pH and temperature of 8.0 and 90 °C, respectively, with remarkable pH and thermal stability. Furthermore, the enzyme had a sufficient tolerance for organic solvents after 16 h of preincubation, with laccase activity > 70%. Additionally, the laccase maintained considerable residual activity after pretreatment with 100 mM of chemical agents, including sodium dodecyl sulphate (69.93 ± 0.89%), ethylenediaminetetraacetic acid (93.1 ± 7.85%), NaN3 (96.28 ± 3.34%) and urea (106.03 ± 10.72%). CONCLUSION The laccase's pH and thermal stability; and robust catalytic efficiency in the presence of organic solvents suggest its industrial and biotechnological application potentials for the sustainable development of green chemistry.
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