Characterisation of antibacterial compounds produced by Pseudomonas spp. isolated from Hogsback wetlands, Eastern Cape, South Africa
- Authors: Obi, Akudo Odochi
- Date: 2017
- Subjects: Bacteriology -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/15688 , vital:40507
- Description: With microbial pathogens developing resistance against currently existing antimicrobials, there is an earnest need to discover and develop new therapeutic compounds with unique and/or precise modes of action. Natural products have provided the inspiration for most of the active ingredients in medicines and thus are regarded as a potential screening resource lead for discovery of new drugs. Pseudomonads are ubiquitous, Gram-negative, motile bacteria that flourish and inhabit a diversity of environments. Literature has vastly revealed that some members of the genus Pseudomonas are capable of producing a plethora of biologically active metabolites and thus are regarded as “attractive” bacteria for testing as potential sources for new therapeutic compounds. In this regard, this study was undertaken with an aim to isolate and characterize antibacterial compounds produced by Pseudomonas species isolated from aquatic milieu of the Hogsback wetlands, Eastern Cape, South Africa. A total of 12 water samples were aseptically collected from Hogsback wetlands. To ascertain antimicrobial potential of the isolates, 9 test bacterial (sensitive) strains, namely Escherichia coli ATCC 3695, Vibrio fluvialis, Salmonella typhimurium, Salmonella enteritidis, Pseudomonas aeruginosa ATCC 27853, Escherichia coli (2), Listeria ivanovii, Listeria monocytogenes and Enterococcus faecalis ATCC 19433 were used for antimicrobial screening. A total of 500 presumptive Pseudomonas spp. were isolated and screened for the production of antibacterial compound through the Agar-well diffusion method. About 8percent of the presumptive Pseudomonas spp. produced compound with activity against the test bacteria and based on the levels of activity shown, three isolates coded as A53, Y55 and Y138 were selected and identified by 16S rDNA sequence analysis and The Basic Local Alignment Search Tool analysis revealed that the three isolates (A53, Y55 and Y138) belonged to the Pseudomonas genus. Using MEGA6 software, a molecular phylogenetic tree was constructed by Maximum Likelihood Method. The phylogenetic analysis by this method showed that all three isolate codes (A53, Y55 and Y138) formed the same clade with Pseudomonas viridiflava strain P3 (Accession number: AY972186) and Pseudomonas plecoglossicida strain P9 (Accession number: AY972231) at a high bootstrap value of 100percent. Isolate code A53 and Y55 also grouped together at a bootstrap value of 96percent. Factors affecting antibacterial compound production by the three isolates were elucidated. The effect of pH on the three isolates revealed that isolate code A53 and Y55 could produce antibacterial compounds over all pH ranges (pH 4-9), however, antibacterial production was produced optimally at acidic conditions (pH 4) and a less acidic-neutral condition (pH 6) respectively. Isolate Y138 showed optimal production at pH 5. The effect of temperature on all three isolates was studied at different temperatures ranging between 25°C-40°C. Study on effect of temperature variations on antibacterial production revealed that all three isolates are capable of growing and producing antibacterial compounds over all the different temperature ranges, however, maximum/optimum production was observed at temperature of 30°C. In order to establish the role of plasmids in antibacterial production, two curing agents; Sodium dodecyl sulfate (SDS) and Ethidium bromide under different sub-lethal concentrations (1 mg/ml -11mg/ml and 2.5 μg/ml- 125 μg/ml respectively) were used to cure the selected isolates. Plasmid DNA samples of A53, Y55 and Y138, were separated by electrophoresis on a 0.7percent (w/v) agarose. Agarose gel electrophoresis revealed that not all the concentrations used were effective in curing the plasmid. A Quick Load 1 kb Extend DNA ladder marker was used for determining the size of the plasmids. Isolate code A53 and Y138 had a plasmid size of 48.5 kb while isolate Y55 had a plasmid size of approximately 20 kb. Antibacterial activity by agar-well diffusion method after the curing process revealed that antibacterial production was not lost or prevented, but rather increased. Thus, it is suggested that the genes responsible for antibiotic production in this study were chromosomal-encoded and not plasmid-encoded. This study revealed that Hogsback wetlands may represent a previously unexplored environment/source of discovering novel and potent antibacterial compounds.
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- Date Issued: 2017
The impact of oil prices on the trade balance of South Africa
- Authors: Mili, Akhona
- Date: 2017
- Subjects: Petroleum products -- Prices Balance of trade -- South Africa Petroleum industry and trade -- Economic aspects
- Language: English
- Type: Thesis , Masters , MCom
- Identifier: http://hdl.handle.net/10353/8630 , vital:33228
- Description: The Autoregressive Distributed Lag model was used to estimate and analyse the relationship between crude oil prices and the trade balance of South Africa. This study estimated the trade balance model and analysed the relationship between crude oil prices and the trade balance of South Africa from the period 1990 to 2015. The bounds cointegration test confirmed the existence of a long-run relationship between the trade balance and real oil prices, real effective exchange rate, domestic income, real interest rate and the inflation rate. The results confirm that changes in oil prices have a negative impact on the trade balance whereas, currency devaluation, real interest, inflation rate and domestic income had a positive effect on the trade balance. The negative sign and significance of the Error Correction Term and diagnostic tests confirmed the reliability of the results.
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- Date Issued: 2017
The use of behaviour, saliva cortisol and faecal glucocorticoid levels as stress indicators in beef cows
- Authors: Dzviti, Melody
- Date: 2017
- Subjects: Veterinary physiology Beef cattle -- Behavior Glucocorticoids
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/8165 , vital:31634
- Description: The study evaluated the use of saliva as a minimally invasive technique for cortisol level determination. This was done by assessing the relationship between serum and saliva cortisol based on short sampling intervals. In addition, the study also determined the effect of successive handling on faecal glucocorticoid metabolites (FGM), rectal temperature (RT), avoidance distance (AD), chute and crush scores as well as urination, defaecation and vocalization in multiparous cows. For trial one six non-pregnant, clinically healthy Nguni cows were used. They were administered with standardised doses of adrenocorticotropic hormone on treatment days and saline was used as a placebo on alternative days. Simultaneously, blood and saliva samples were collected to quantify cortisol levels. No relationship (r = 0.0131, P>0.05) was observed between serum and saliva cortisol. However, results showed that there was adrenal stimulation in saliva coupled by significant (P<0.05) effects of sampling intervals. For the second study 21 non-pregnant and clinically healthy Nguni cows of one herd were used for the trial. They were divided into parity groups and their weights were used as a covariate in data analysis. For five consecutive weeks, fresh faecal samples were extracted and rectal temperatures measured. In addition, fear of approaching humans was assessed by AD scores and behaviour scores in the crush chute, as well as vocalisation, urination and defaecation were observed. The effect of successive handling significantly (P<0.05) affected the physiological parameters, namely FGM and RT. Contrariwise, Chi-square tests revealed that parities had a significant (χ2 < 0.05) influence on AD and crush scores as well as on the occurrence of excretory behaviour. It was concluded that saliva cortisol can be used as a stress indicator in beef cows. However, plasma cortisol can be significantly volatile thereby becoming less sensitive to changes across time. This can cause a lack of relationship between plasma and saliva cortisol. On the other hand, repetitive handling of beef cows has an influence on faecal stress levels. In addition, beef cows of differing parities behave differently to handling stress.
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- Date Issued: 2017