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CHARACTERISATION, PATHOGENICITY AND ANTIBIOTIC RESISTANCE OF MYCOPLASMAS INVOLVED IN VULVOVAGINITIS IN GOATS.
Published 2013-04Call Number: Loading…
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FERMENTATION OF COCOA (Theobroma cacao L.) POD HUSK AND ITS HYDROLYSATE FOR ETHANOL PRODUCTION USING IMPROVED STARTER CULTURES
Published 2012-07Call Number: Loading…
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Page will reload when a filter is selected or excluded.- Antibiotic sensitivity 1 results 1
- Cocoa pod husk 1 results 1
- Ethanol production 1 results 1
- Fossil fuel, a main but dwindling energy source for automobiles, causes emission of environment unfriendly oxides of carbon. These contribute substantially to greenhouse gases which bring about climate change. There is therefore the need for sustainable source of energy like ethanol an environmental friendly bioenergy. Hence this study was aimed at the fermentation of cocoa pod husk for ethanol production. Isolates of yeast were obtained from sun-dried Cocoa Pod Husk (CPH), subjected to spontaneous submerged fermentation for 7 days. Five strains of Saccharomyces sp. (MX1, MX2, MX3, MX4 and MX5) with high frequency of occurrence were selected for further studies. The MX1 and MX2 were used for genetic modifications. Dried CPH was subjected to chemical analysis and pretreatment using particle size reduction and high pressure liquid hot water at 130oC for 30 minutes. Acid and enzymatic hydrolysis of the pretreated CPH was carried out using standard method. Products of the hydrolysis were analysed with high performance liquid chromatography. Two genes XL1 (xylose reductase) and XL2 (xylitol dehydrogenase) encoding pentose utilization were obtained from genomic DNA of Pichia stipitis (CBS 6054) using basic local alignment search tool. Primers of these genes were designed with Saccharomyces genome database, amplified with Polymerase Chain Reaction (PCR) and purified. The amplicon (genes) were ligated into plasmid vectors (pGAPZA and pVT100-U). Strains MX1 and MX2 were transformed with these construct using lithium acetate method. Physiological characterization of the selected unmodified yeast strains and the two genetically-modified strains was done under different environmental conditions including temperatures, pH and varied concentrations of acetic acid. The CPH hydrolysates were fermented for 120 hours using the unmodified and genetically-modified yeast strains respectively and the ethanol yield determined. Data were analysed using ANOVA. Twenty yeast isolates identified as Saccharomyces cerevisiae (80%) and Saccharomyces uvarum (20%) were obtained. Chemical composition of CPH included hemicellulose (13.9%) cellulose (18.6%) and lignin content (14.2%). Acid hydrolysis yielded 50.1% glucose, 11.97% xylose, 11.2% mannose while enzymatic hydrolysis gave 31.7% glucose, 4.8% mannose and 16.8% galactose. The inserted gene XL1 had 318 amino acids polypeptides while XL2 had 363 amino acid polypeptides. Restriction enzyme analysis and colony PCR confirmed the transformational integration of these constructs into Saccharomyces cerevisiae MX1 and MX2. The five isolates had optimal growth at 30 – 40oC and pH of 4.0 – 5.5. However the genetically-modified yeast strains were able to utilize xylose and arabinose carbon sources better than the unmodified types and also tolerated low concentration of acetic acid than the unmodified types. Ethanol production was highly significant (p0.05) in the modified starters (29.7g/L) than the unmodified strains (14.0g/L). Genetically-modified organisms performed better in ethanol production than the non-modified organisms. The application of genetic modification of microorganisms will aid the potential use of waste biomass like cocoa pod husk for bioenergy production and this will contribute significantly to reducing greenhouse gases associated with climate change. 1 results 1
- Genetic modification 1 results 1
- Mycoplasma species, 1 results 1
- Saccharomyces cerevisiae 1 results 1
- Vulvovaginitis is an inflammation of the vulva and vagina caused by bacteria, fungi, viruses, parasites and allergy. The condition results in reduction in mating ability, infertility, abortion and death of the affected animals with resultant economic loss to the livestock industry. Although mycoplasmas have been isolated from cases of vulvovaginitis, their role as sole causative agents and in the pathogenicity of the disease have not been investigated in goats in Nigeria. The aim of this study was to characterise mycoplasma isolates from cases of vulvovaginitis and also and determine the pathogenicity of mycoplasma-induced vulvovaginitis in goats. Two hundred and twenty-one vaginal swabs were obtained from identified cases of vulvovaginitis in goats aged 8- 11 months from markets in and around Lagos metropolis. Samples were analyzed bacteriologically and mycoplasmotologically. Mycoplasmas were identified biochemically using standard procedures and conventional polymerase chain reaction with specific primers. Antisera raised in rabbits using 0.5ml of 4.0 x107 CFU/ml of selected Mycoplasma species including M. bovis, M. capri, M. capricolum and M. arginini were used to group the mycoplasmas serologically by growth inhibition method. Sensitivity of the isolated Mycoplasmas and other bacteria to aminoglycosides, fluoroquinolones, cephalosporins and nitrofurans was carried out. Pathogenicity of the isolated M. bovis, M. capri, M. capricolum and M. arginini was evaluated using 2ml inoculum containing 4.0 x 10 7 CFU/ml of each isolate per vulva to reproduce vulvovaginitis over a six week experimental period. Four goats were used for each experimental group and the control group. Animals were observed for symptoms of vulvovaginitis such as hyperemia, vulva swelling and vaginal mucus discharges. Post-mortem gross and histopathological examination was carried out and findings reported. Other data were analysed using descriptive statistics. Two hundred and fifty-seven bacterial isolates were recovered from the 221 field samples as follows: Mycoplasma capricolum (1.6%), Mycoplasma arginini (1.6%), Mycoplasma capri (1.2%), Mycoplasma bovis (0.78%), Ureaplasma species (1.2%), Acholeoplasma species (0.8%). Other bacteria were Escherichia coli (35.4%), Streptococcus species (29.2%) and Staphylococcus species (28.4%). Mycoplasma isolates were confirmed with the production of specific 280bp bands. Isolated Mycoplasma species and other bacteria were sensitive to ciprofloxacin and nitrofurantoin. Six different combinations of antibiotic resistant patterns were observed with amoxicillin, norfloxacin and ampicillin having the highest level of resistance (100.0%) and nitrofurantoin the least (33.0%). Clinical symptoms, which included hyperemia, swollen vulva, vaginal mucus discharges with pyrexia (38.9 – 39.60 C), were first observed in the M. bovis- infected group on day three, ten and thirteen. Mortality was recorded on days 30, 33, 38 and 41 post- inoculation in the M. bovis, M. capricolum, M. arginini and M. capri- infected groups, respectively. M. bovis produced the most severe lesions marked by lymphoid necrosis of the vulva tissue, diffuse hyperaemia in the lung alveolar septa and massive alveolar infiltration with neutrophils while the mildest lesions were observed in the M. arginine-infected group. Vulvovaginitis was reproduced in goats with all the Mycoplasma species as the major infective agent and Mycoplasma bovis as the most pathogenic. Ciprofloxacin and nitrofurantoin were the most effective antibiotics against Mycoplasmas and other bacteria isolated. 1 results 1
- Vulvovaginitis, 1 results 1
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