Genetic characterization of non-O1/non-O139 <i>Vibrio cholerae</i> mobilome: a strategy for understanding and discriminating emerging environmental bacterial strains

Acute diarrhea and cholera (AWD/C) result in more than 21000 to 143000 global mortality annually and are associated with Vibrio cholerae. The pathogen has shown increasing evolutionary/emerging dynamics linked with mobilome or ubiquitous nature of mobile integrative genetic and conjugative elements (MIGCE), however, such dynamics are rarely reported amongst somatic-antigen non-agglutinating Type-1/-139 V. cholerae (SA-NAG-T-1/139Vc). The study reports the genetic detection of mobilome-associated indices in SA-NAG-T-1/139Vc as a potential strategy for differentiating/discriminating emerging environmental bacteria. Presumptive V. cholerae isolates were retrieved from five water sources, while strains were characterized/serogrouped and confirmed using simplex and comparative-genomic-multiplex Polymerase Chain Reaction (PCR). Genomic island (GI-12det, GI-14det, GI-15det); Phages (TLC-phagedet, Kappa-phagedet) and ICEs of the SXT/R391 family genes (SXT/R391-ICEs integrase, SXT-Hotspot-IV, ICEVchInd5Hotspot-IV, ICEVchMoz10Hotspot-IV) were detected. Other rare ICE members such as the ICEVcBan8att gene and Vibrio Seventh Pandemic island detection (VSP-II Integrase, Prototypical VSP-II) were also detected. Results revealed that the 8.22% (61/742) SA-NAG-T-1/139Vc serogroup observed harbors the Vibrio Seventh Pandemic island integrase (34/61; 55.7%) and other rare genetic traits including; attB/attP (29/61; 47.5%, 14/61; 23%), integrative genetic elements (4/61; 6.56%), phage types (TLC-phagedet: 2/61; 3.28% and Kappa-phagedet: 7/61; 11.48%) as well as the integrase genes (INT1, Sul1, Sul2) (29/61: 47.5%; 21/61: 34.4%; 25/61: 41%). Such genetic detection of mobilome determinants/MIGCE suggests potential discriminatory tendencies amongst SA-NAG-T-1/139Vcwhich may be applied in mobilome typing of evolving/emerging environmental bacteria. The need to encourage the application of such mobilome typing indices and continuous study of these strains is suggestive of interest in controlling future potential emerging environmental strains

Distribution of Vibrio species in Shellfish and Water Samples Collected from the Atlantic Coastline of South-East Nigeria

Crayfish, lobster, and sea-water samples collected from five fishing islands on the Atlantic coast\u2013Bight of Biafra (Bonny)\u2013belonging to Ibaka Local Government Area of Akwa-Ibom State of Nigeria were bacteriologically evaluated on thiosulphate citrate bile-salt sucrose (TCBS) agar for Vibrio load and pathotypes. Mean log10 Vibrio counts of 7.64\ub12.78 cfu/g (in crayfish), 5.07\ub13.21 cfu/g (in lobster), and 3.06\ub12.27 cfu/mL (in sea-water) were obtained in rainy season (June-July) while counts in the dry season (November-December) were 6.25\ub11.93 cfu/g, 5.99\ub11.54 cfu/g, and 3.84\ub11.78 cfu/mL respectively. The physicochemical measurements (temperature, pH, and total dissolved solutes) of the sea-water did not vary significantly in the two seasons across all five islands. Vibrio species isolated were Vibrio cholerae (both O1 and non-O1 serotypes), V. parahaemolyticus , V. vulnificus , V. mimicus , and V. fluvialis . Both Ogawa and Inaba subtypes of V. cholerae O1 serotype were found. In addition, the Hikojima subtype, which had not been previously reported in the region, was isolated in two samples. The results show that these Vibrio species are endemic in the area

Bioflocculant production by a consortium of Streptomyces and Cellulomonas species and media optimization via surface response model

AbstractSpecies of actinobacteria previously isolated from Tyume River in the Eastern Cape Province of South Africa and identified by 16S rDNA sequence as Cellulomonas and Streptomyces species were evaluated as a consortium for the production of bioflocculant. Sucrose, peptone and magnesium chloride were the nutritional sources which supported optimal production of bioflocculant resulting in flocculation activities of 91%, 82% and 78% respectively. Response surface design revealed sucrose, peptone and magnesium chloride as critical media components following Plackett–Burman design, while the central composite design showed optimum concentration of the critical nutritional source as 16.0g/L (sucrose), 1.5g/L (peptone) and 1.6g/L (magnesium chloride) yielding optimal flocculation activity of 98.9% and bioflocculant yield of 4.45g/L. FTIR spectrometry of the bioflocculant indicated the presence of carboxyl, hydroxyl and amino groups, typical for heteropolysaccharide, while SEM imaging revealed an interwoven clump-like structure. The molecular weight distribution of the constituents of the bioflocculants ranged 494.81–18,300.26Da thus, an indication of heterogeneity in composition. Additionally, the chemical analyses of the purified bioflocculant revealed the presence of polysaccharides and proteins with neutral sugar, amino sugar and uronic acids in the following concentration: 5.7mg, 9.3mg and 17.8mg per 100mg. The high flocculation activity of the bioflocculant suggests commercial potential

Health Implications of Occupational Exposure of Butchers to Emissions from Burning Tyres

Background: Flames from burning scrap tyres are used in de-furring animals for human consumption in most parts of Nigeria. Emissions from tyres are known to contain a myriad of toxic mixtures especially particulate matter (PM), volatile organic compounds, hazardous air pollutants, and inspirable metals, some of which are known human carcinogens. This cross-sectional study investigated the deleterious health effects of these emissions in occupationally-exposed workers at the Dei-Dei Abattoir, Abuja, Nigeria. Methods: A total of 156 respondents were divided into two groups. Group 1 (124 butchers) and group 2 [32 administrative staff (AS)]. Data from digital spirometry were used to determine the association between chronic exposure to tyre emissions and lung function. Urinary 1-Hydroxypyrene concentration, phenolic compounds levels and heavy metal concentrations were determined. Also ambient PM and polycyclic aromatic hydrocarbons (PAHs) concentrations at 3 delineated points in the abattoir were measured. Findings: Spirometry results showed significant deterioration of lung function in the butchers. The concentration of 1-Hydroxypyrene (μg/molCret) in the post-shift urine samples of the butchers was significantly higher (P < 0.05) in butchers relative to the AS (0.52 ± 0.13 Vs 0.20 ± 0.07, respectively). Similarly the concentrations of zinc and nickel (mg/l) were significantly higher in the butchers compared to the AS (zinc: 0.91 ± 0.19 Vs 0.31 ± 0.28, respectively; nickel: 0.11 ± 0.06 Vs 0.06 ± 0.02, respectively). Anthracene, fluoranthene, pyrene, benzo-a- pyrene, and PM concentrations were significantly higher at the de-furring point when compared to the wash bay and the administrative building, especially between 8.00 and 8.30 am. Conclusion: Occupational exposure to scrap tyre emissions resulted in significant adverse health effects. The existing laws banning the use of burning tyres in meat processing should be enforced while the use of personal protective equipment should be encouraged in abattoirs

Studies on Bioflocculant Production by Arthrobacter sp. Raats, a Freshwater Bacteria Isolated from Tyume River, South Africa

A bioflocculant-producing bacteria was isolated from Tyume River in the Eastern Cape Province, South Africa and identified by 16S rRNA gene nucleotide sequence to have 91% similarity to Arthrobacter sp. 5J12A, and the nucleotide sequence was deposited in GenBank as Arthrobacter sp. Raats (accession number HQ875723). The bacteria produced an extracellular bioflocculant when grown aerobically in a production medium containing glucose as sole carbon source and had an initial pH of 7.0. Influences of carbon, nitrogen and metal ions sources, as well as initial pH on flocculating activity were investigated. The bacteria optimally produced the bioflocullant when lactose and urea were used as sole sources of carbon and nitrogen respectively with flocculating activities of 75.4% and 83.4% respectively. Also, the bacteria produced the bioflocculant optimally when initial pH of the medium was 7.0 (flocculating activity 84%), and when Mg2+ was used as cation (flocculating activity 77%). Composition analyses indicated the bioflocculant to be principally a glycoprotein made up of about 56% protein and 25% total carbohydrate

Decolourization of synthetic dyes by laccase produced from Bacillus sp. NU2

AbstractAdvanced industrialization has caused an increase in the continuous discharge of hazardous effluents in the environment. This study evaluated the potential of the laccase synthesized by Bacillus sp. NU2 to degrade five synthetic dyes. Sawdust, wheat bran and peels of banana and tangerine were utilized as carbon sources for bacterial growth and laccase production. The produced crude enzyme was purified to homogeneity to determine its molecular weight. The kinetic activity of the purified laccase was determined using 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). The toxicity of the laccase-treated dye solution was assessed on Bacillus sp. NU2 growth. The result showed optimum laccase yield from the tangerine peel medium. The purified laccase gave a specific activity of 349.94 U mg−1 and a molecular weight of 55 kDa, respectively. The purified laccase displayed a strong affinity for ABTS substrate with an enzyme activity of 31.21 U mg−1. It was optimum at 60 °C and pH 8, with catalytic efficiency (Kcat/Km) of 23.93 mmol L−1. The decolourization effects on Congo Red, Methyl Orange, Remazol Brilliant Blue R, Reactive Blue 4 and Malachite Green were 87%, 70%, 65%, 63% and 51%, respectively. The toxicity assay of laccase degraded dyes on Bacillus sp. NU2 showed a growth reduction of 36.75% (Malachite Green), 12.57% (Congo Red), 17.19% (Methyl Orange), 38.41% (Remazol Brilliant Blue R) and 28.14% (Reactive Blue 4). The laccase produced by Bacillus sp. NU2 holds a high catalytic potential for the detoxification of dye effluents in an environmental system

Effects of Fractionation and Combinatorial Evaluation of Tamarindus indica Fractions for Antibacterial Activity

Six fractions, named TiA – TiF, were obtained by fractionating the crude ethanol extract of the stem bark of Tamarindus indica using column chromatographic techniques. On TLC, fraction TiB showed five bands, TiC three bands, while TiD and TiE showed two bands each. TiC, TiD and TiE were re-eluted with different solvent systems to yield two fractions each, while TiB yielded four. These subfractions were designated B1-B4; C1-C2; D1-D2 and E1-E2, respectively. Tannins, flavonoids and alkaloids, among other components, were detected, albeit in different proportions with respect to fractions and subfractions and were compartmentalized with respect to the solvent systems used. The in vitro antibacterial activity of fractions and subfractions was tested separately and in combinations using the agar well diffusion technique. The susceptibly of test strains (expressed as %) were: 83.3% (TiA and TiB), 75.0% (crude extract and TiC), 66.7% (TiD), 50.0% (TiE) and 16.7% (TiF) when used singly, whereas in combination, the corresponding susceptibilities were 100% (CE), 83.3% (DE), 66.7% (AB, AF, BC, BD, DE and EF), 50% (AC and CD), 33.3% (BE and BF) and 16.7% (AD) against Gram negative bacteria strains and 100% (EF), 80% (DE), 60% (AB, BC and CE), 40% (AC, BD, BF, CF and DF) and 20% (AE, AF, BE and CD) against Gram positive strains. Percentage susceptibility with combinatorial use of re-fractions ranged from 85.7–57.1% and 60–40% against Gram negative and positive strains (TiB subfractions), respectively, 100–85.7% and 40–0% against Gram negative and positive strains (TiC, TiD and TiE sub-fractions)

Mixed Culture Fermentation and Media Optimization by Response Surface Model: Streptomyces and Brachybacterium Species in Bioflocculant Production

The biofloculant production potential of a consortium of Streptomyces and Brachybacterium species were evaluated. Optimum bioflocculant yields (g/L) and flocculation activities (%) were observed for the following preferred nutritional sources: glucose (56%; 2.78 ± 0.15 g/L), (NH4)2NO3 (53%; 2.81 ± 0.37 g/L) and CaSO4·H2O (47%; 2.19 ± 0.13 g/L). A Plackett-Burman design revealed the critical fermentation media components. The concentrations of these components were optimized [glucose; 16.0, (NH4)2NO3; 0.5 and CaSO4·H2O; 1.2 (g/L)] through a central composite design with optimum bioflocculant yield of 3.02 g/L and flocculation activity of 63.7%. The regression coefficient (R2 = 0.6569) indicates a weak estimation of the model’s adequacy and a high lack-of-fit value (34.1%). Lack of synergy in the consortium may have been responsible for the model inadequacy observed. FTIR spectrometry showed the bioflocculant to be a heteropolysaccharide, while SEM imaging revealed an amorphous loosely arranged fluffy structure with interstial spacing of less than 1 µm

Detection of Multidrug-Resistant RND Efflux Pumps and Regulatory Proteins in Antibiotic-Resistant <i>P. aeruginosa</i> Recovered from Hospital Wastewater Effluent in the Eastern Cape Province of South Africa

P. aeruginosa (P. aeruginosa) is a problematic hospital agent that is a global challenge due to the ineffectiveness of some conventional antimicrobial therapies. Multidrug-resistant (MDR) P. aeruginosa has distinct action modes, including beta-lactamase production, porin gene repression, and efflux pump overexpression. This current research work focuses on efflux pumps (MexAB-OprM, MexCD-OprJ, MexXY-OprN) and their regulatory proteins (NfxB, MexR, MexZ, NalC, NalD) in MDR P. aeruginosa isolated from hospital wastewater effluent. The sequence analysis of the main transporter MexB was also performed. Following antibiotic resistance profiling and polymerase chain reaction (PCR) amplification of the efflux pump genes, the association of the efflux pump proteins with antibiotic resistance was investigated and analysed statistically. Fifty-seven (57) multidrug-resistant isolates were obtained from 81 PCR-confirmed P. aeruginosa isolates. Of the MDR P. aeruginosa isolates, the following rates were recorded: MexA (96.5%), MexB (100%), OprM (96.5%), MexC (100%), MexD (74.1%), OprJ (63.7%), MexX (89.6%), and OprN (51.7%). Additionally, the regulatory proteins had the following rates: NfxB (31.6%), NalC (15.8%), NalD (12.2%), MexZ (3.5%), and MexR (3.5%). The efflux pumps and regulatory proteins are strongly associated with antibiotic resistance, implying that P. aeruginosa antibiotic resistance is heavily influenced by these efflux pumps and regulatory genes. The MexB DNA sequences had numerous substitutions and poor alignment with divergent regions, and hence they have a possible role in increased antibiotic resistance. The absence of regulatory genes in most MDR P. aeruginosa isolates in the current research may have permitted transcription of the efflux pump operons, thus also increasing the antibiotic resistance burden