Demographic dynamics of waterborne disease and perceived associated WASH factors in Bushenyi and Sheema districts of South-Western Uganda.

Water remains a significant player in spreading pathogens, including those associated with neglected tropical diseases. The implications of socio-demographic delineations of water quality, sanitation, and hygiene ("WASH") interventions are on the downswing. This study assessed waterborne diseases and perceived associated WASH factors in the Bushenyi and Sheema districts of South-Western Uganda. This study examines the linear relationship between WASH and identifies the association of specific demographic factors as well as their contributions/correlations to waterborne disease in the study area. A structured qualitative and quantitative data collection approach was adopted in face-to-face questionnaire-guided interviews of 200 respondents on eight surface water usage. Most participants, 65.5%, were females and had a higher score of knowledge of WASH (71%), 68% score on the improper practice of WASH, and 64% score on unsafe water quality. Low score for basic economic status was (57%), report of common diarrhoea was (47%), and a low incidence of waterborne disease outbreaks (27%). The principal component analysis (PCA) depicts the knowledge and practice of WASH to have a strong positive correlation (r = 0.84, p < 0.001; r = 0.82, p < 0.001); also economic status positively correlated with grade of water source, knowledge, and practice of WASH (correlation coefficient = 0.72; 0.99; 0.76 and p-values = 0.001; < 0.001; < 0.001 respectively). Occupation (p = 0.0001, OR = 6.798) was significantly associated with knowledge and practice of WASH, while age (r = -0.21, p < 0.001) was negatively associated with knowledge and practice of WASH. The basic economic status explains why "low economic population groups" in the remote villages may not effectively implement WASH, and diarrhoea was common among the population. Diarrhoea associated with unsafe water quality and improper practice of WASH is common among the study population, and there is a low incidence of waterborne disease outbreaks. Therefore, government, stakeholders, and non-governmental organisations should work together to promote proper practice of WASH conditions to limit the occurrence of diarrhoea and prevent potential waterborne disease outbreaks.

Non-serogroup O1/O139 agglutinable Vibrio cholerae: a phylogenetically and genealogically neglected yet emerging potential pathogen of clinical relevance.

Somatic antigen agglutinable type-1/139 Vibrio cholerae (SAAT-1/139-Vc) members or O1/O139 V. cholerae have been described by various investigators as pathogenic due to their increasing virulence potential and production of choleragen. Reported cholera outbreak cases around the world have been associated with these choleragenic V. cholerae with high case fatality affecting various human and animals. These virulent Vibrio members have shown genealogical and phylogenetic relationship with the avirulent somatic antigen non-agglutinable strains of 1/139 V. cholerae (SANAS-1/139- Vc) or O1/O139 non-agglutinating V. cholerae (O1/O139-NAG-Vc). Reports on implication of O1/O139-NAGVc members in most sporadic cholera/cholera-like cases of diarrhea, production of cholera toxin and transmission via consumption and/or contact with contaminated water/seafood are currently on the rise. Some reported sporadic cases of cholera outbreaks and observed change in nature has also been tracable to these non-agglutinable Vibrio members (O1/O139-NAGVc) yet there is a sustained paucity of research interest on the non-agglutinable V. cholerae members. The emergence of fulminating extraintestinal and systemic vibriosis is another aspect of SANAS-1/139- Vc implication which has received low attention in terms of research driven interest. This review addresses the need to appraise and continually expand research based studies on the somatic antigen non-serogroup agglutinable type-1/139 V. cholerae members which are currently prevalent in studies of water bodies, fruits/vegetables, foods and terrestrial environment. Our opinion is amassed from interest in integrated surveillance studies, management/control of cholera outbreaks as well as diarrhea and other disease-related cases both in the rural, suburban and urban metropolis.

Molecular Characterization and Antibiotic Susceptibility Profile of Acinetobacter baumannii Recovered from Hospital Wastewater Effluents.

Acinetobacter baumannii (A. baumannii) plays a significant part in nosocomial infections world over and is re-emerging as a formidable pathogen due to the wide range of antibiotic resistance factors it acquires and environmental resilience. The high attendance of patients (outpatients and inpatients) into the health care facilities formed the basis for the selection of the hospitals. Consequently, this study profiled the antibiogram and antibiotic resistance genes of A. baumannii isolated from selected hospital wastewater effluents. A total of twenty-four (24) wastewater samples from three selected hospital drainages were collected and analysed presumptively by culture-dependent methods for A. baumannii. The identity confirmation of A. baumannii was done by the amplification of recA and blaoxa-51 genes. Virulence and antibiotic resistance markers were assessed using polymerase chain reaction. A total of 53 A. baumannii isolates were confirmed and the highest antibiotic resistance profile was 93% (piperacillin). Multiple antibiotic resistance index (MARI) showed a range of 0.23 and 0.46. FimH virulence gene was detected in 29 (55%) of the isolates. Tetracycline and beta-lactam resistance markers were found; 70% and 92% of the isolates possessed tetA and ampC genes. The isolates showed high level of resistance to antibiotics. The multiple antibiotic resistance index (MARI) of ≥ 0.2 indicates that some of the isolates harbour virulence and resistance traits emerging from high-risk source thereby projecting a threat to public health.

Marine sediment derived bacteria Enterobacter asburiae ES1 and Enterobacter sp. Kamsi produce laccase with high dephenolisation potentials.

Purified laccases from bacterial species isolated from marine sediment were applied to degrade Bisphenol A (BPA). The Bacterial species were isolated from marine water sediments sampled from Cove Rock and Bonza Bay beach of the Eastern Cape Province, South Africa was tested for laccase activity on varied phenolic plates. The two most promising strains, Enterobacter asburiae ES1 and Enterobacter sp. Kamsi was subjected to extracellular laccase production and were identified using molecular methods. Both extracted bacterial laccases showed an affinity for ABTS and PFC substrates and were purified to homogeneity by ammonium sulfate precipitation, anion exchange, and size exclusion chromatography. A specific laccase activity of 231.67 and 218.15 U/mg of protein and a molecular weight of 50 and 55 kDa was obtained from the purified ES1 and Kamsi laccases. Laccase activity was optimum at pH8 and 5 and at 80 °C and 60 °C for ES1 and Kamsi laccases, and they manifested 71.7% and 65.8% BPA decolorizing effects. The optimized treatment condition applied showed maximum BPA removal effects of 85% and 86% at pH7 and 6, while 78% and 79% was degraded at 70 °C and 80 °C while at 250 µL enzyme volume, BPA was actively degraded to 85%, and 75% removal effect showed by ES1 and Kamsi laccases. The molecular identification of the pure colonies using 16S rRNA showed the isolate belonged to the class of gammaproteobacterial. Their nucleotide sequence has been deposited in NCBI with the accession number MN686602 and MN686603. Conclusively, marine habitat serves as a reservoir for active bacterial laccase producers suitable for bioprocess application.

Characterization of a thermostable and solvent-tolerant laccase produced by Streptomyces sp. LAO.

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.

Bacillus sp. CSK2 produced thermostable alkaline keratinase using agro-wastes: keratinolytic enzyme characterization.

BACKGROUND: Chicken feathers are the most abundant agro-wastes emanating from the poultry processing farms and present major concerns to environmentalists. Bioutilization of intractable feather wastes for the production of critical proteolytic enzymes is highly attractive from both ecological and biotechnological perspectives. Consequently, physicochemical conditions influencing keratinase production by Bacillus sp. CSK2 on chicken feathers formulation was optimized, and the keratinase was characterized. RESULTS: The highest enzyme activity of 1539.09 ± 68.14 U/mL was obtained after 48 h of incubation with optimized conditions consisting of chicken feathers (7.5 g/L), maltose (2.0 g/L), initial fermentation pH (5.0), incubation temperature (30 °C), and agitation speed (200 rpm). The keratinase showed optimal catalytic efficiency at pH 8.0 and a temperature range of 60 °C - 80 °C. The keratinase thermostability was remarkable with a half-life of above 120 min at 70 °C. Keratinase catalytic efficiency was halted by ethylenediaminetetraacetic acid and 1,10-phenanthroline. However, keratinase activity was enhanced by 2-mercaptoethanol, dimethyl sulfoxide, tween-80, but was strongly inhibited by Al3+ and Fe3+. Upon treatment with laundry detergents, the following keratinase residual activities were achieved: 85.19 ± 1.33% (Sunlight), 90.33 ± 5.95% (Surf), 80.16 ± 2.99% (Omo), 99.49 ± 3.11% (Ariel), and 87.19 ± 0.26% (Maq). CONCLUSION: The remarkable stability of the keratinase with an admixture of organic solvents or laundry detergents portends the industrial and biotechnological significance of the biocatalyst.

Bacillus sp. FPF-1 Produced Keratinase with High Potential for Chicken Feather Degradation.

Chicken feathers are predominantly composed of keratin; hence, valorizing the wastes becomes an imperative. In view of this, we isolated keratinase-producing bacteria and identified them through the 16S rDNA sequence. The process condition for keratinase activity was optimized, and electron micrography of the degradation timelines was determined. Keratinolytic bacteria were isolated and identified as Bacillus sp. FPF-1, Chryseobacterium sp. FPF-8, Brevibacillus sp. Nnolim-K2, Brevibacillus sp. FPF-12 and Brevibacillus sp. FSS-1; and their respective nucleotide sequences were deposited in GenBank, with the accession numbers MG214993, MG214994, MG214995, MG214996 and MG214999. The degree of feather degradation and keratinase concentration among the isolates ranged from 62.5 ± 2.12 to 86.0 ± 1.41(%) and 214.55 ± 5.14 to 440.01 ± 20.57 (U/mL), respectively. In the same vein, 0.1% (w/v) xylose, 0.5% (w/v) chicken feather, an initial fermentation pH of 5.0, fermentation temperature of 25 °C and an agitation speed of 150 rpm, respectively, served as the optimal physicochemical conditions for keratinase activity by Bacillus sp. FPF-1. The time course showed that Bacillus sp. FPF-1 yielded a keratinase concentration of 1698.18 ± 53.99(U/mL) at 120 h. The electron microscopic imaging showed completely structural dismemberment of intact chicken feather. Bacillus sp. FPF-1 holds great potential in the valorization of recalcitrant keratinous biomass from the agro sector into useful products.

Prevalence and capsular type distribution of Streptococcus agalactiae isolated from pregnant women in Namibia and South Africa.

BACKGROUND: Streptococcus agalactiae or Group B Streptococcus (GBS) is the leading cause of neonatal morbidity and mortality resulting in septicaemia, bacteraemia and meningitis. Long term problems in children range from loss of hearing to mental retardation. While Intrapartum Antibiotic Prophylaxis (IAP) has reduced the incidence of S. agalactiae infection, it still remains the leading cause of disease in neonates. GBS has ten capsular types whose distribution varies across the world. Therefore, this study sought to determine the prevalence of GBS in Namibia and South Africa amongst pregnant women between 35 and 37 weeks gestation and elucidate the capsular types. METHODS: Lower vaginal and rectal swabs were collected from pregnant women between 35 and 37 weeks gestation. Five hundred and thirty pregnant women were recruited into the study in Windhoek, Namibia while one hundred pregnant women were recruited in the Eastern Cape, South Africa. The swabs were cultured on 5% sheep blood agar (Biomerieux, New Jersey, USA) for isolation of GBS. Presumptive isolates were confirmed using both the Vitek (2) and molecular techniques targeting the scpB gene. Capsular typing was performed in a multiplex PCR with capsular specific primer pairs. RESULTS: The prevalence of GBS in Namibia was 13.6 and 37% in South Africa respectively. In both countries most women were dually colonised with GBS. Capsular types II, III and V were the most prevalent. CONCLUSIONS: The prevalence of GBS in Namibia was lower than in South Africa in this study. The prevalence in both countries was not different from those reported in other African countries and around the world. The predominant capsular types in this study are the ones commonly associated with adverse maternal outcomes.

Agrowastes utilization by Raoultella ornithinolytica for optimal extracellular peroxidase activity.

The industrial applications and prospects of microbial peroxidase are on the upwards trend, thus necessitating the search for sources with high turnaround time. Actinobacterial species have been a major source of peroxidase for the obvious reasons of having robust metabolite expression capabilities. However, other bacteria species have been underexplored for peroxidase production, hence the motivation for the investigation into the peroxidase production potential of Raoultella ornithinolytica OKOH-1 (KX640917). The bacteria expressed optimum specific peroxidase activity of 16.48 ± 0.89 U mg-1 , which is higher than those previously reported. The optimal fermentation conditions were pH 5 (3.44 ± 0.64 U mL-1 ), incubation temperature of 35 °C (5.25 ± 0.00 U mL-1 ), and agitation speed of 150 rpm (9.45 ± 2.57 U mL-1 ), with guaiacol and ammonium chloride as the best inducer and nitrogen supplement, respectively. On valorization of agrowastes as a sole carbon source for the secretion of peroxidase, sawdust gave the best peroxidase yield (15.21 ± 2.48 U mg-1 ) under solid-state fermentation. Also, a nonperoxide-dependent enzyme activity, which suggests probable laccase activity, was observed. The ability of the bacteria to utilize agrowastes is highly economical and as well a suitable waste management strategy. Consequently, R. ornithinolytica OKOH-1 is a promising industrial strain with dexterity for enhanced peroxidase production.

The activity of commercial antimicrobials, and essential oils and ethanolic extracts of Olea europaea on Streptococcus agalactiae isolated from pregnant women.

BACKGROUND: Streptococcus agalactiae also known as Group B Streptococcus (GBS) is a major cause of disease in pregnant women and new born babies where it causes early and late onset disease characterised by sepsis, pneumonia and meningitis. Ten to 37 % of pregnant women in the world are colonised with GBS while intrapartum antibiotic prophylaxis has led to significant reduction in early onset disease. The increase in drug resistant microorganisms has become a major threat. Development of vaccines is still in progress so there is need for new and safer alternatives to treatment. METHODS: Benzyl penicillin, Ampicillin, Cefotaxime, Ceftriaxone, Levofloxacin, Erythromycin, Clindamycin, Linezolid, Vancomycin, Tetracycline and Cotrimoxazole, Olea europaea leaf extracts and essential oil were tested against GBS isolates from South Africa and Namibia. RESULTS: The isolates showed 100% sensitivity to benzyl penicillin, ampicillin, ceftriaxone, levofloxacin, linezolid, vancomycin, O. europaea leaf extracts and essential oils. Only one isolate (0.6%) was resistant to cefotaxime and 23.4 and 10.4% were resistant to clindamycin and erythromycin respectively. CONCLUSION: GBS isolates showed sensitivity to O. europaea extracts at low minimum inhibitory concentrations. Β lactams are still the drugs of choice for treatment of GBS disease but O. europaea extracts potent as an alternative source of antimicrobials.

Utilization of agroindustrial wastes for the production of laccase by Achromobacter xylosoxidans HWN16 and Bordetella bronchiseptica HSO16.

Agroindustrial residual lignocellulosic biomaterial provides an economical and renewable natural bioresource for the large-scale, gainful biofuel production, as well as the production of fine bulk chemicals, which may include industrial biocatalysts. To this end, the laccase-inducing aptitude of some agroindustrial, lignocellulosic residues were appraised in submerged fermentation batch culture of two woodland betaproteobacteria (Hb9c; Achromobacter xylosoxidans HWN16, Hb16c; Bordetella bronchiseptica HSO16). Significant fermentation factors for laccase production were identified following a one-variable-at-a-time: OVAT screening method, levels of significant factors were optimized using response surface methodology: RSM. Mandarin peelings: MP and wheat bran: WB were suitable substrates for laccase production in Hb9c; 29.4 U/mL and Hb16c; 28.2 U/mL, respectively. However, the numerical optimization of significant factors for laccase production in both isolates presented an overall maximum laccase output encountered throughout the study (Hb9c; 169.39 U/mL, Hb16c; 45.22 U/mL), albeit the simulated conditions of the statistical model were outside the design space of the algorithm such as pH 5, 0.5 g MP, 100 rpm, 0.25 g NaNO3 for Hb9c and pH 3, 2.5 g WB, 50 rpm, 0.05 g yeast extract for Hb16c. Furthermore, a record 17.5 and 15.54 fold increase in laccase turnover depicts the astuteness of the statistical method in the valorization of these lignocellulosic residues for enhanced laccase production, hence, the incorporation of these outcomes at industrial scales might yield tremendous outputs.

Aptitude of Oxidative Enzymes for Treatment of Wastewater Pollutants: A Laccase Perspective.

Natural water sources are very often contaminated by municipal wastewater discharges which contain either of xenobiotic pollutants and their sometimes more toxic degradation products, or both, which frustrates the universal millenium development goal of provision of the relatively scarce pristine freshwater to water-scarce and -stressed communities, in order to augment their socioeconomic well-being. Seeing that both regulatory measures, as regards the discharge limits of wastewater, and the query for efficient treatment methods remain unanswered, partially, the prospects of enzymatic treatment of wastewater is advisable. Therefore, a reconsideration was assigned to the possible capacity of oxidative enzymes and the respective challenges encountered during their applications in wastewater treatment, and ultimately, the prospects of laccase, a polyphenol oxidase that oxidizes aromatic and inorganic substrates with electron-donating groups in treatment aromatic contaminants of wastewater, in real wastewater situations, since it is assumed to be a vehicle for a greener community. Furthermore, the importance of laccase-driven catalysis toward maintaining mass-energy balance, hence minimizing environmental waste, was comprehensibly elucidated, as well the strategic positioning of laccase in a model wastewater treatment facility for effective treatment of wastewater contaminants.

Ethanol extract and chromatographic fractions of Tamarindus indica stem bark inhibits Newcastle disease virus replication.

CONTEXT: The plethora of ethnomedicinal applications of Tamarindus indica Linn. (Leguminosae), tamarind, includes treatment of human and livestock ailments; preparations are recognized antipyretics in fevers, laxatives and carminatives. African folklore has various applications of tamarind. However, in Nyasaland, domestic fowl are fed with preparations for prophylactic properties. OBJECTIVES: The objective of this study is to evaluate the antiviral properties of T. indica extract. MATERIALS AND METHODS: Tamarindus indica stem bark was extracted through ethanol maceration over 24 h, and the crude extract was fractionated by gravity-propelled column chromatography. Newcastle disease virus (NDV) inhibitory activity of extract and fractions were evaluated in vivo using 10-d-old embryonated chicken egg (ECE) as the medium for virus cultivation and antivirus assay. About 240 ECE were grouped into eight (three controls and five experimental) and, 200 µL of the extract and fractions respectively inoculated into NDV pre-infected eggs and incubated at 37 °C. Allantoic fluid was harvested 5 d post-virus infection and assayed for haemagglutination (HA). RESULTS: Anti-NDV assessment showed 62.5 mg/mL of crude extract and fractions: TiA, TiC and TiD to yield a HA titre of 1:128 each, while TiB showed 1:64 HA titre. At 125 mg/mL, a titre of 1:16 was recorded against TiB and TiD and, 1:8 against TiA. Similarly, crude extract and TiC, each recorded 1:4 HA titre. However, the minimum concentrations of extract and fraction for virus inactivation were 0.24 mg/mL and 0.49 mg/mL, respectively. CONCLUSION: The antiviral activity shown by T. indica portends novel antiviral drugs and, perhaps, as scaffold for new drugs.

Bactericidal and antioxidant properties of essential oils from the fruits Dennettia tripetala G. Baker.

BACKGROUND: The antibacterial and antioxidant properties of the essential oils (EOs) of unripe and ripe fruits of Dennettia tripetala and their potential for the management of infectious and oxidative-stress diseases were investigated in-vitro in this study. METHOD: Essential oil obtained from the fruit in Clevenger modified apparatus, was characterized by high resolution GC-MS, while antioxidant and antibacterial properties were tested by spectrophotometric and agar diffusion methods respectively. RESULTS: The EO demonstrated strong antibacterial properties when subjected to multi -drug resistant bacterial strains: Enterococcus faecium (ATCC19434), Escherichia coli (ATCC 700728), Staphylococcus aureus (NCINB 50080), Listeria ivanovii (ATCC 19119), Enterobacter cloacae (ATCC13047) and four previously confirmed multi resistant bacterial isolates from our laboratory stock culture. The unripe fruit oil (UFO) demonstrated greater activity than the ripe fruit oil (RFO) against most of the tested bacteria with minimum inhibition concentrations (MIC) ranging between 0.05-0.20 mg/mL while that of the ripe fruit oil (RFO) ranged from 0.10-0.20 mg/mL. The IC50 for RFO (0.62 ± 0.12 mg/mL) showed that it has higher antioxidant strength than UFO and vitamin C (0.87 ± 0.23 and 3.39 ± 0.12 mg/mL) but a lower activity compared to ß-carotene (0.32 ± 0.22 mg/mL) in scavenging 2, 2-diphenyl-1-picrylhydrazyl radicals (DPPH•). The EOs also demonstrated strong ability in scavenging three other different radicals (ABTS, lipid peroxide and nitric oxide radicals) in concentration dependant -manner. CONCLUSION: Findings from this study suggest that apart from the local uses of the plant extracts, the EO has strong bioactive compounds, noteworthy antibacterial, antiradical properties and may be good candidates in the search for lead constituents for the synthesis of novel potent antibiotics.

Multiple antibiotic resistances among Shiga toxin producing Escherichia coli O157 in feces of dairy cattle farms in Eastern Cape of South Africa.

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) O157:H7 is a well-recognized cause of bloody diarrhea and hemolytic-uremic syndrome (HUS). The ability of STEC strains to cause human disease is due to the production of Shiga toxins. The objectives of this study were to determinate the prevalence, serotypes, antibiotic susceptibility patterns and the genetic capability for Shiga toxin production in Escherichia coli (STEC) strains isolated from dairy cattle farms in two rural communities in the Eastern Cape Province of South Africa. METHODS: Fecal samples were collected between March and May 2014, from individual cattle (n=400) in two commercial dairy farms having 800 and 120 cattle each. Three hundred presumptive isolates obtained were subjected to polymerase chain reactions (PCR) for identification of O157 serogroup and Shiga toxin producing genes (stx1, stx2) on genomic DNA extracted by boiling method. Susceptibility of the isolates to 17 antibiotics was carried out in vitro by the standardized agar disc-diffusion method. RESULTS: Based on direct PCR detection, 95 (31.7%) isolates were identified as O157 serogroup. The genetic repertoire for Shiga toxin production was present in 84 (88.42%) isolates distributed as stx1 (37), stx2 (38) and stx1/2 (9) respectively while 11 of the isolates did not harbor Shiga toxin producing genes. Multiple antibiotic resistances were observed among the isolates and genetic profiling of resistance genes identified bla ampC 90%, blaCMY 70%, blaCTX-M 65%, blaTEM 27% and tetA 70% and strA 80% genes among the antimicrobial resistance determinants examined. CONCLUSION: We conclude that dairy cattle farms in the Eastern Cape Province are potential reservoirs of antibiotic resistance determinants in the province.

Characterization and Flocculating Properties of a Biopolymer Produced by Halomonas sp. Okoh.

The flocculating efficiency and physiochemical properties of purified bioflocculant produced by Halomonas sp. Okoh were investigated. Approximately 1.213 g/L of bioflocculant was recovered after fermentation under predetermined ambient conditions. Jar test experimentation revealed optimum bioflocculant concentration as 0.2 mg/mL with flocculation activity of 66.1%. The thermostable bioflocculant retained high flocculation activity after heat treatment at 100 °C for 30 minutes; flocculation activity of 74% was achieved. Chemical analysis showed that the bioflocculant was composed of sugar (26.5%), protein (2.64%), and uronic acid (13.3%). The Fourier infrared spectroscopy spectrum of the purified bioflocculant revealed the presence of hydroxyl and carboxylic functional groups. Thermogravimetric analyses showed a varied decomposition step, thus, an indication of varied composition. Scanning electron micrograph revealed the amorphous structure of the bioflocculant. These results suggest potential applicability of the bioflocculant produced by Halomonas sp. Okoh industrially.

Characterization of a Bioflocculant (MBF-UFH) Produced by Bacillus sp. AEMREG7.

A bioflocculant named MBF-UFH produced by a Bacillus species isolated from sediment samples of Algoa Bay of the Eastern Cape Province of South Africa was characterized. The bacterial identification was through 16S rDNA sequencing; nucleotide sequences were deposited in GenBank as Bacillus sp. AEMREG7 with Accession Number KP659187. The production of the bioflocculant was observed to be closely associated with cell growth. The bioflocculant had the highest flocculating activity of 83.2% after 72 h of cultivation, and approximately 1.6 g of purified MBF-UFH was recovered from 1 L of fermentation broth. Its chemical analyses indicated that it is a glycoprotein composed of polysaccharide (76%) and protein (14%). Fourier transform infrared spectroscopy (FTIR) revealed that it consisted of hydroxyl, amide, carboxyl and methoxyl as the functional moieties. Scanning electron microscopy (SEM) revealed the amorphous structure of MBF-UFH and flocculated kaolin clay particles. The maximum flocculating activity of 92.6% against kaolin clay suspension was achieved at 0.3 mg/mL over pH ranges of 3-11 with the peak flocculating rate at pH 8 in the presence of MgCl2. The bioflocculant retained high flocculating activity of 90% after heating at 100 °C for 1 h. MBF-UFH appears to have immense potential as an alternative to conventional chemical flocculants.

Bacillus toyonensis strain AEMREG6, a bacterium isolated from South African marine environment sediment samples produces a glycoprotein bioflocculant.

A bioflocculant-producing bacteria, isolated from sediment samples of a marine environment in the Eastern Cape Province of South Africa demonstrated a flocculating activity above 60% for kaolin clay suspension. Analysis of the 16S ribosomal deoxyribonucleic acid (rDNA) nucleotide sequence of the isolate in the GenBank database showed 99% similarity to Bacillus toyonensis strain BCT-7112 and it was deposited in the GenBank as Bacillus toyonensis strain AEMREG6 with accession number KP406731. The bacteria produced a bioflocculant (REG-6) optimally in the presence of glucose and NH4NO3 as the sole carbon and nitrogen source, respectively, initial medium pH of 5 and Ca2+ as the cation of choice. Chemical analysis showed that purified REG-6 was a glycoprotein mainly composed of polysaccharide (77.8%) and protein (11.5%). It was thermally stable and had strong flocculating activity against kaolin suspension over a wide range of pH values (3-11) with a relatively low dosage requirement of 0.1 mg/mL in the presence of Mn2+. Fourier transform infrared spectroscopy (FTIR) revealed the presence of hydroxyl, carboxyl and amide groups preferred for flocculation. Scanning electron microscopy (SEM) revealed that bridging was the main flocculation mechanism of REG-6. The outstanding flocculating performance of REG-6 holds great potential to replace the hazardous chemical flocculants currently used in water treatment.

Molecular detection and characterization of resistant genes in Mycobacterium tuberculosis complex from DNA isolated from tuberculosis patients in the Eastern Cape province South Africa.

BACKGROUND: Tuberculosis (TB) in both animals and humans is caused by Mycobacterium tuberculosis complex (MTBC) primarily transmitted by inhalation of aerosolized droplets containing the organism. Multi-drug resistance (MDR) and extensive drug resistance (XDR) are evolutionary features of Mycobacterium tuberculosis to subvert the antibiotic regimes in place. The heavy burden of TB worsened by HIV endemic in South Africa motivated for the investigation of MTBC prevalence among TB patients in Port Elizabeth and the amplification and sequencing of the DNA amplicons known to confer resistance to TB drugs. METHODS: Three thousand eight hundred and ten (3810) sputum specimens were processed and DNA was isolated from sputum specimens collected from different hospitals and health care places in the Eastern Cape Province, South Africa. DNA was amplified using the Seeplex® MTB Nested ACE detection assay. The agar-dilution proportion method was used to perform drug-sensitivity testing using 7H10 Middlebrook medium. Target genes known to confer resistance to first and second-line drugs were amplified and the amplicons sequenced. RESULTS: One hundred and ninety (5%) DNA samples tested positive for MTBC and from the resistant profiles of the 190 positive samples, we noted that multidrug-resistant TB was identified in 189 (99.5%) with 190 (100%) patients infected with MTB resistant to isoniazid and 189 (99.5%) having MTB resistant to rifampicin. Other percentages of drug resistance observed including 40% pre-XDR and 60% of XDR. CONCLUSION: This study provides valuable data on the different kinds of mutations occurring at various target loci in resistant MTBC strains isolated from samples obtained from the Eastern Cape Province. The results obtained reveal a high incidence of MDR amongst the positive samples from Eastern Cape Province, South Africa.

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 (R(2) = 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.