Antibiogram profiling of Escherichia coli pathotypes isolated from Kat River and Fort Beaufort abstraction water
- Authors: Nontongana, Nolonwabo
- Date: 2014
- Subjects: Escherichia coli infections -- South Africa -- Eastern Cape , Water -- Pollution -- Environmental aspects -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11289 , http://hdl.handle.net/10353/d1019820 , Escherichia coli infections -- South Africa -- Eastern Cape , Water -- Pollution -- Environmental aspects -- South Africa -- Eastern Cape
- Description: Escherichia coli (E. coli) is a widespread species that includes a broad variety of strains, ranging from highly pathogenic strains causing worldwide outbreaks of severe disease to virulent isolates which are part of the normal intestinal flora or which are well characterized and safe laboratory strains. The pathogenicity of a given E. coli strain is mainly determined by specific virulence factors which include adhesins, invasins, toxins and capsule. The aim of this study was to evaluate the prevalence and antibiogram profiles of E. coli pathotypes previously isolated from Kat River and Fort Beaufort abstraction water. A total of 171 E. coli isolates showed at least one pathogenic determinant among the isolated 278 E. coli. The other 107 isolates were negative for the tested virulence genes. All 278 presumptive isolates tested positive for the UidA gene, and were therefore classified as non-categorized pathogenic E. coli. The 171 pathogenic isolates had at least one characteristic gene of pathogenic E. coli and were identified and classified as enteropathogenic E. coli (6%), enterotoxigenicE. coli (131), uropathogenic E. coli (6), neonatal meningitis E. coli (14), diffusely adherent E. coli (1) and enterohaemrrhagic E. coli (1). Interestingly, no virulence genes were detected for the enteroinvasive E. coli and the enteroaggregative E. coli. The antibiotic resistance profiles for all isolates that were identified as E. coli showed 100% resistance to penicillin G, 98% resistance to ampicillin, 38% resistance to trimethoprim-sulphamethoxazole and 8% resistance to streptomycin. Multiple antibacterial resistance (MAR) was also observed, where 44% of the isolates were resistant to three antibiotics and 8% resistant to four antibiotics. The results of this study showed the Kat River and Fort Beaufort abstraction water are reservoirs of pathogenic strains of E. coli which harbour antibiotic resistance determinants that can cause serious health risks to the people in the surrounding communities.
- Full Text:
- Date Issued: 2014
- Authors: Nontongana, Nolonwabo
- Date: 2014
- Subjects: Escherichia coli infections -- South Africa -- Eastern Cape , Water -- Pollution -- Environmental aspects -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11289 , http://hdl.handle.net/10353/d1019820 , Escherichia coli infections -- South Africa -- Eastern Cape , Water -- Pollution -- Environmental aspects -- South Africa -- Eastern Cape
- Description: Escherichia coli (E. coli) is a widespread species that includes a broad variety of strains, ranging from highly pathogenic strains causing worldwide outbreaks of severe disease to virulent isolates which are part of the normal intestinal flora or which are well characterized and safe laboratory strains. The pathogenicity of a given E. coli strain is mainly determined by specific virulence factors which include adhesins, invasins, toxins and capsule. The aim of this study was to evaluate the prevalence and antibiogram profiles of E. coli pathotypes previously isolated from Kat River and Fort Beaufort abstraction water. A total of 171 E. coli isolates showed at least one pathogenic determinant among the isolated 278 E. coli. The other 107 isolates were negative for the tested virulence genes. All 278 presumptive isolates tested positive for the UidA gene, and were therefore classified as non-categorized pathogenic E. coli. The 171 pathogenic isolates had at least one characteristic gene of pathogenic E. coli and were identified and classified as enteropathogenic E. coli (6%), enterotoxigenicE. coli (131), uropathogenic E. coli (6), neonatal meningitis E. coli (14), diffusely adherent E. coli (1) and enterohaemrrhagic E. coli (1). Interestingly, no virulence genes were detected for the enteroinvasive E. coli and the enteroaggregative E. coli. The antibiotic resistance profiles for all isolates that were identified as E. coli showed 100% resistance to penicillin G, 98% resistance to ampicillin, 38% resistance to trimethoprim-sulphamethoxazole and 8% resistance to streptomycin. Multiple antibacterial resistance (MAR) was also observed, where 44% of the isolates were resistant to three antibiotics and 8% resistant to four antibiotics. The results of this study showed the Kat River and Fort Beaufort abstraction water are reservoirs of pathogenic strains of E. coli which harbour antibiotic resistance determinants that can cause serious health risks to the people in the surrounding communities.
- Full Text:
- Date Issued: 2014
Parasite prevalence, nutritionally-related blood metabolites and pre-slaughter stress response in Nguni, Bonsmara and Angus steers raised on veld
- Authors: Ndlovu, Thulile
- Date: 2008
- Subjects: Parasites , Nguni cattle , Bonsmara cattle , Metabolites , Slaughtering and slaughter-houses , Aberdeen -- Angus cattle
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11258 , http://hdl.handle.net/10353/73 , Parasites , Nguni cattle , Bonsmara cattle , Metabolites , Slaughtering and slaughter-houses , Aberdeen -- Angus cattle
- Description: The effects of month on body weight, body condition scores, internal parasite prevalence and on nutritionally related blood metabolites were studied in Angus, Bonsmara and Nguni steers raised on sweet veld. Pre-slaughter stress was also determined using catecholamines, cortisol, dopamine, packed cell volume and serum creatinine levels. The blood chemical constituents evaluated included glucose, cholesterol, total protein, creatinine, urea, globulin, albumin, calcium, phosphorus, magnesium, aspartate amino transferase (AST), alkaline phosphatase (ALP) and creatinine kinase (CK). The Nguni steers maintained their body condition throughout the study period whereas Angus steers had the least body condition scores. Parasite levels were high during the rainy season and low during the dry season. The predominant internal parasites were Haemonchus (39.3 percent), Trichostrongylus (37.8 percent), Cooperia pectinita (25.5 percent), Fasciola gigantica (16.3 percent) and Ostertagia ostertagi (11.2 percent). The Nguni had the least parasite infestation levels and had high PCV levels. Nguni had higher levels of cholesterol and glucose (2.86 and 4mmol/l, respectively) than the other two breeds. Nguni and Bonsmara steers had higher (P<0.05) mineral levels. There were significant breed and month differences for glucose, cholesterol, creatinine, calcium, albumin, phosphorus, albumin-globulin ratio and ALP levels. Bonsmara was more susceptible to transport and pre-slaughter stress as it had the highest (P<0.05) levels of adrenalin (10.8nmol/mol), noradrenalin (9.7nmol/mol) and dopamine (14.8nmol/mol) levels, whereas the Nguni had the least levels of adrenalin (6.5nmol/mol), noradrenalin (4.6nmol/mol) and dopamine (4nmol/mol) levels. In conclusion, Nguni steers were better adapted to the local environmental conditions
- Full Text:
- Date Issued: 2008
- Authors: Ndlovu, Thulile
- Date: 2008
- Subjects: Parasites , Nguni cattle , Bonsmara cattle , Metabolites , Slaughtering and slaughter-houses , Aberdeen -- Angus cattle
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11258 , http://hdl.handle.net/10353/73 , Parasites , Nguni cattle , Bonsmara cattle , Metabolites , Slaughtering and slaughter-houses , Aberdeen -- Angus cattle
- Description: The effects of month on body weight, body condition scores, internal parasite prevalence and on nutritionally related blood metabolites were studied in Angus, Bonsmara and Nguni steers raised on sweet veld. Pre-slaughter stress was also determined using catecholamines, cortisol, dopamine, packed cell volume and serum creatinine levels. The blood chemical constituents evaluated included glucose, cholesterol, total protein, creatinine, urea, globulin, albumin, calcium, phosphorus, magnesium, aspartate amino transferase (AST), alkaline phosphatase (ALP) and creatinine kinase (CK). The Nguni steers maintained their body condition throughout the study period whereas Angus steers had the least body condition scores. Parasite levels were high during the rainy season and low during the dry season. The predominant internal parasites were Haemonchus (39.3 percent), Trichostrongylus (37.8 percent), Cooperia pectinita (25.5 percent), Fasciola gigantica (16.3 percent) and Ostertagia ostertagi (11.2 percent). The Nguni had the least parasite infestation levels and had high PCV levels. Nguni had higher levels of cholesterol and glucose (2.86 and 4mmol/l, respectively) than the other two breeds. Nguni and Bonsmara steers had higher (P<0.05) mineral levels. There were significant breed and month differences for glucose, cholesterol, creatinine, calcium, albumin, phosphorus, albumin-globulin ratio and ALP levels. Bonsmara was more susceptible to transport and pre-slaughter stress as it had the highest (P<0.05) levels of adrenalin (10.8nmol/mol), noradrenalin (9.7nmol/mol) and dopamine (14.8nmol/mol) levels, whereas the Nguni had the least levels of adrenalin (6.5nmol/mol), noradrenalin (4.6nmol/mol) and dopamine (4nmol/mol) levels. In conclusion, Nguni steers were better adapted to the local environmental conditions
- Full Text:
- Date Issued: 2008
Antibacterial properties of the methanol extract of helichrysum pedunculatum
- Authors: Ncube, Nqobile S
- Date: 2008
- Subjects: Medicinal plants , Methanol , Helichrysum
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11241 , http://hdl.handle.net/10353/461 , Medicinal plants , Methanol , Helichrysum
- Description: The methanol extract of Helichrisum pedunculatum was screened for antimicrobial activity up to a concentration of 5 mg/ml using the agar dilution technique. A number of test bacterial isolates, comprising both Gram negative and Gram positive organisms were susceptible to the crude extract of the plant. The minimum inhibitory concentrations (MICs) of the extract ranged between 1 and 5 mg/ml for the susceptible organisms. The MICs of the selected antibiotics, chloramphenicol and penicillin, ranged between 2 and 4 mg/L, and 2 and 32 mg/L respectively against Bacillus cereus, Proteus vulgaris and Staphylococcus aureus OKOH1. Bactericidal activity was determined by the time kill assay. The methanol extract of the plant was not bactericidal at 1 × MIC for B. cereus, P. vulgaris and Staph. aureus OKOH1. At 2 × MIC the extract was bacteriostatic against B. cereus but bactericidal against P. vulgaris and Staph. aureus OKOH1. Combination studies were done at 1/2 × MIC, 1 × MIC and 2 × MIC of the plant extract with 1 × MIC of the antibiotics. Combinations of the plant extract and chloramphenicol resulted in mostly indifferent interactions against P. vulgaris and Staph. aureus OKOH1 but synergistic interactions at higher concentration of the plant extract for B. cereus. Penicillin combinations gave synergistic interactions at lower concentrations of the plant for P.vulgaris and Staph. aureus OKOH1 but was mostly indifferent for B. cereus.
- Full Text:
- Date Issued: 2008
- Authors: Ncube, Nqobile S
- Date: 2008
- Subjects: Medicinal plants , Methanol , Helichrysum
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11241 , http://hdl.handle.net/10353/461 , Medicinal plants , Methanol , Helichrysum
- Description: The methanol extract of Helichrisum pedunculatum was screened for antimicrobial activity up to a concentration of 5 mg/ml using the agar dilution technique. A number of test bacterial isolates, comprising both Gram negative and Gram positive organisms were susceptible to the crude extract of the plant. The minimum inhibitory concentrations (MICs) of the extract ranged between 1 and 5 mg/ml for the susceptible organisms. The MICs of the selected antibiotics, chloramphenicol and penicillin, ranged between 2 and 4 mg/L, and 2 and 32 mg/L respectively against Bacillus cereus, Proteus vulgaris and Staphylococcus aureus OKOH1. Bactericidal activity was determined by the time kill assay. The methanol extract of the plant was not bactericidal at 1 × MIC for B. cereus, P. vulgaris and Staph. aureus OKOH1. At 2 × MIC the extract was bacteriostatic against B. cereus but bactericidal against P. vulgaris and Staph. aureus OKOH1. Combination studies were done at 1/2 × MIC, 1 × MIC and 2 × MIC of the plant extract with 1 × MIC of the antibiotics. Combinations of the plant extract and chloramphenicol resulted in mostly indifferent interactions against P. vulgaris and Staph. aureus OKOH1 but synergistic interactions at higher concentration of the plant extract for B. cereus. Penicillin combinations gave synergistic interactions at lower concentrations of the plant for P.vulgaris and Staph. aureus OKOH1 but was mostly indifferent for B. cereus.
- Full Text:
- Date Issued: 2008
In vitro bioactivity of crude extracts of Lippia javanica on clinical isolates of Helicobacter pylori: preliminary phytochemical screening
- Authors: Nkomo, Lindelwa Precious
- Date: 2010
- Subjects: Extracts , Helicobacter pylori , Antibiotics , Drug resistance in microorganisms , Materia medica, Vegetable
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11257 , http://hdl.handle.net/10353/508 , Extracts , Helicobacter pylori , Antibiotics , Drug resistance in microorganisms , Materia medica, Vegetable
- Description: Helicobacter pylori classified as a class 1 carcinogen is a common human pathogen implicated in certain gastrointestinal diseases. Helicobacter pylori infection is acquired mainly in childhood, especially in developing countries. H. pylori infection causes peptic ulcer, duodenitis, gastritis and cancer. The growing resistance of H. pylori to antibiotics used in its treatment as well as other innate limitations of the triple therapy has necessitated a search for alternative treatment from natural sources which could be readily available, less cost effective. The antimicrobial activity of solvents (acetone, ethanol, methanol, chloroform and water) crude extracts of Lippia javanica were investigated against 31 H. pylori strains by the agar well diffusion technique. The minimum inhibitory concentration (MIC) was determined by spectrophotometric analysis at 620 nm using the broth micro dilution method and the rate of kill by broth dilution method. Phytochemical analysis was also performed. H. pylori standard strain NCTC 11638 was included as a positive control. Metronidazole and amoxicillin were used as positive control antibiotics. The ANOVA test was used to analyze the results using SPSS version 17.0. The strains were inhibited by all the extracts with inhibition zones of diameter ranging from 0-36 mm and 0-35 mm for the control antibiotic, clarithromycin. The MIC90 ranged from 0.039- 0.625 mg/mL for acetone; 0.039-1.25mg/mL for methanol, 0.00195-0.313 mg/mL for ethanol; 0.01975-2.5 mg/mL for metronidazole and 0.0048-2.5 mg/mL for amoxicillin. Acetone extract completely inhibited strain PE369C at MIC (0.1 mg/mL) and 2× MIC (0.2 mg/mL) in 18h and at ½× MIC (0.05 mg/mL) in 36h. Strain PE466C was completely inhibited at 4× MIC in 72h. Phytochemical analysis revealed the presence of flavonoids, saponins, tannins, steroids and alkaloids. The results indicate that the extracts of the leaves of L. javanica may contain compounds with anti-H. pylori activity and merits further study to identify the compounds.
- Full Text:
- Date Issued: 2010
- Authors: Nkomo, Lindelwa Precious
- Date: 2010
- Subjects: Extracts , Helicobacter pylori , Antibiotics , Drug resistance in microorganisms , Materia medica, Vegetable
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11257 , http://hdl.handle.net/10353/508 , Extracts , Helicobacter pylori , Antibiotics , Drug resistance in microorganisms , Materia medica, Vegetable
- Description: Helicobacter pylori classified as a class 1 carcinogen is a common human pathogen implicated in certain gastrointestinal diseases. Helicobacter pylori infection is acquired mainly in childhood, especially in developing countries. H. pylori infection causes peptic ulcer, duodenitis, gastritis and cancer. The growing resistance of H. pylori to antibiotics used in its treatment as well as other innate limitations of the triple therapy has necessitated a search for alternative treatment from natural sources which could be readily available, less cost effective. The antimicrobial activity of solvents (acetone, ethanol, methanol, chloroform and water) crude extracts of Lippia javanica were investigated against 31 H. pylori strains by the agar well diffusion technique. The minimum inhibitory concentration (MIC) was determined by spectrophotometric analysis at 620 nm using the broth micro dilution method and the rate of kill by broth dilution method. Phytochemical analysis was also performed. H. pylori standard strain NCTC 11638 was included as a positive control. Metronidazole and amoxicillin were used as positive control antibiotics. The ANOVA test was used to analyze the results using SPSS version 17.0. The strains were inhibited by all the extracts with inhibition zones of diameter ranging from 0-36 mm and 0-35 mm for the control antibiotic, clarithromycin. The MIC90 ranged from 0.039- 0.625 mg/mL for acetone; 0.039-1.25mg/mL for methanol, 0.00195-0.313 mg/mL for ethanol; 0.01975-2.5 mg/mL for metronidazole and 0.0048-2.5 mg/mL for amoxicillin. Acetone extract completely inhibited strain PE369C at MIC (0.1 mg/mL) and 2× MIC (0.2 mg/mL) in 18h and at ½× MIC (0.05 mg/mL) in 36h. Strain PE466C was completely inhibited at 4× MIC in 72h. Phytochemical analysis revealed the presence of flavonoids, saponins, tannins, steroids and alkaloids. The results indicate that the extracts of the leaves of L. javanica may contain compounds with anti-H. pylori activity and merits further study to identify the compounds.
- Full Text:
- Date Issued: 2010
Distribution of virulence determinants in some members of the ESKAPE pathogens isolated from selected hospital wastewater effluents in the Eastern Cape, South Africa
- Authors: Mapipa, Qaqamba
- Date: 2019
- Subjects: Effluent quality Water -- Purification
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/14704 , vital:40061
- Description: Members of ESKAPE pathogens are grouped under the acronymn made up of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. ESKAPE pathogens, isolated from hospitals and communities, have just been characterized for new pathogenesis and transmission patterns and overcoming the usefulness of available drugs, this leads to infections that tend to increase the rate of disease in a population and death rate and cost our society massively. The distribution of virulence determinants and the antibiotic susceptibility patterns in some members of the ESKAPE pathogens isolated from hospital wastewater facilities that were selected within the Eastern Cape Province, South Africa was evaluated. Water samples from the hospital wastewater facilities were collected fortnightly over a period of three (3) months (February to April). Spread plate technique was employed for the enumeration and isolation of the members of ESKAPE pathogens. A total of 725 presumptive ESKAPE isolates were collected, screened and Polymerase Chain Reaction (PCR) amplification of target determinants was performed. Results showed that Klebsiella pneumoniae tested positive for ninety two (92=35.9%) samples, Enterobacter cloacae fifty seven (57=22.2%) samples tested positive, Pseudomonas aeruginosa fifty four (54=21.09%) isolates tested positive and fifty three (53=20.7%) isolates tested positive for Acinetobacter baumannii making a total of 256 isolates which tested positive for some members of ESKAPE pathogens. The detection of some members of the ESKAPE pathogens in these Hospitals is a cause of concern to public health. A total of 256 ESKAPE isolates that were obtained from the hospital wastewater facilities; were subjected to antibiotic susceptibility test in order to determine their antibiotic-resistant profiles. ESKAPE isolates recovered from the three hospital effluents showing resistance to antimicrobial agents in this study were further evaluated for resistance determinants namely, sulfonamides resistance determinants (sulI and sulII), β-lactams (ampC, blaTEM and blaZ), tetracyclines (tetA, tetB, tetC and tetM), 10 chloramphenicol’s (catI and catII) and aminoglycoside resistance determinants (aacC2, apHAI and apHAII). The resistant determinants targeted have been evaluated through their prevalence and the following distributions were obtained: [β-Lactams; ampC (80%), blaTEM (77%)]; [Tetracyclines; tetA (87%)]; [Sulfonamides; sulI (79%)]; [Aminoglycosides; aacC2 (12%), aphA2 (52%) and aadA (18%)]. With the worldwide increase in these resistant occurrences, the execution of strict infection control policies and treatment with different antimicrobials needs hasty detection. Comprehensive data are therefore needed so that the history of infection, identification of genes present in organisms and monitoring of changes in infection patterns can be traced correctly.
- Full Text:
- Date Issued: 2019
- Authors: Mapipa, Qaqamba
- Date: 2019
- Subjects: Effluent quality Water -- Purification
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/14704 , vital:40061
- Description: Members of ESKAPE pathogens are grouped under the acronymn made up of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. ESKAPE pathogens, isolated from hospitals and communities, have just been characterized for new pathogenesis and transmission patterns and overcoming the usefulness of available drugs, this leads to infections that tend to increase the rate of disease in a population and death rate and cost our society massively. The distribution of virulence determinants and the antibiotic susceptibility patterns in some members of the ESKAPE pathogens isolated from hospital wastewater facilities that were selected within the Eastern Cape Province, South Africa was evaluated. Water samples from the hospital wastewater facilities were collected fortnightly over a period of three (3) months (February to April). Spread plate technique was employed for the enumeration and isolation of the members of ESKAPE pathogens. A total of 725 presumptive ESKAPE isolates were collected, screened and Polymerase Chain Reaction (PCR) amplification of target determinants was performed. Results showed that Klebsiella pneumoniae tested positive for ninety two (92=35.9%) samples, Enterobacter cloacae fifty seven (57=22.2%) samples tested positive, Pseudomonas aeruginosa fifty four (54=21.09%) isolates tested positive and fifty three (53=20.7%) isolates tested positive for Acinetobacter baumannii making a total of 256 isolates which tested positive for some members of ESKAPE pathogens. The detection of some members of the ESKAPE pathogens in these Hospitals is a cause of concern to public health. A total of 256 ESKAPE isolates that were obtained from the hospital wastewater facilities; were subjected to antibiotic susceptibility test in order to determine their antibiotic-resistant profiles. ESKAPE isolates recovered from the three hospital effluents showing resistance to antimicrobial agents in this study were further evaluated for resistance determinants namely, sulfonamides resistance determinants (sulI and sulII), β-lactams (ampC, blaTEM and blaZ), tetracyclines (tetA, tetB, tetC and tetM), 10 chloramphenicol’s (catI and catII) and aminoglycoside resistance determinants (aacC2, apHAI and apHAII). The resistant determinants targeted have been evaluated through their prevalence and the following distributions were obtained: [β-Lactams; ampC (80%), blaTEM (77%)]; [Tetracyclines; tetA (87%)]; [Sulfonamides; sulI (79%)]; [Aminoglycosides; aacC2 (12%), aphA2 (52%) and aadA (18%)]. With the worldwide increase in these resistant occurrences, the execution of strict infection control policies and treatment with different antimicrobials needs hasty detection. Comprehensive data are therefore needed so that the history of infection, identification of genes present in organisms and monitoring of changes in infection patterns can be traced correctly.
- Full Text:
- Date Issued: 2019
Evaluation of some wastewater treatment facilities in Chris Hani and Amathole district municipalities as potential sources of Escherichia coli in the environment
- Authors: Mazwi, Sinazo Nomathamsanqa
- Date: 2014
- Subjects: Escherichia coli -- South Africa -- Eastern Cape , Water -- Purification
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11285 , http://hdl.handle.net/10353/d1019804 , Escherichia coli -- South Africa -- Eastern Cape , Water -- Purification
- Description: Access to clean and safe water is essential for the survival of human beings. Pollution of freshwater sources constitutes a major problem hindering access to safe water for drinking and other domestic uses. Wastewater effluent discharges often impact the microbiological qualities of surface waters with its attendant health and environmental problems. This study evaluated the microbiological qualities of the discharged effluents of four selected wastewater treatment plants in Amathole and Chris Hani District Municipalities of the Eastern Cape Province over a twelve-month sampling period. Microbiological analysis (faecal coliform, Escherichia coli and Escherichia coli O157:H7) was done using standard methods and polymerase chain reaction method was used to confirm identities ofbacterial isolates. Presumptive bacteria counts ranged as follows: faecal coliforms 0 to 1.6 × 103 CFU/100 ml, E. coli 0 to 1.4 × 103 CFU/100 ml and E. coli O157:H7 0 to 9.6 × 102 CFU/100 ml. Forty eight percent (305/626) of the presumptive E. coli isolates were confirmed using species-specific uidA gene which code for β-glucuronidase enzyme in E. coli. Antibiotic susceptibility profile of the isolate using a panel of 10 antibiotics shows 100% (150/150) resistance to antibiotics rifampicin and penicillin G while 49.3% (74/150) of the isolates and 46.7% (70/150) were susceptible to streptomycin and cefotaxime respectively. Multiple antibiotic resistance phenotypes (MARP) of the isolates showed resistance to two or more test antibiotics while the calculated multiple antibiotic resistance index (MARI) for the tested isolated is 0.49. The detection of potentially pathogenic E. coli in the final effluents suggestspotential danger to the receiving water bodies where the effluents are discharge. The high MARI valued obtained in this study indicates that the isolates are form environment where the tested antibiotics are being used and may further lead to the spread of multiple antibiotics resistance among other pathogens that may be present in the same environment.
- Full Text:
- Date Issued: 2014
- Authors: Mazwi, Sinazo Nomathamsanqa
- Date: 2014
- Subjects: Escherichia coli -- South Africa -- Eastern Cape , Water -- Purification
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11285 , http://hdl.handle.net/10353/d1019804 , Escherichia coli -- South Africa -- Eastern Cape , Water -- Purification
- Description: Access to clean and safe water is essential for the survival of human beings. Pollution of freshwater sources constitutes a major problem hindering access to safe water for drinking and other domestic uses. Wastewater effluent discharges often impact the microbiological qualities of surface waters with its attendant health and environmental problems. This study evaluated the microbiological qualities of the discharged effluents of four selected wastewater treatment plants in Amathole and Chris Hani District Municipalities of the Eastern Cape Province over a twelve-month sampling period. Microbiological analysis (faecal coliform, Escherichia coli and Escherichia coli O157:H7) was done using standard methods and polymerase chain reaction method was used to confirm identities ofbacterial isolates. Presumptive bacteria counts ranged as follows: faecal coliforms 0 to 1.6 × 103 CFU/100 ml, E. coli 0 to 1.4 × 103 CFU/100 ml and E. coli O157:H7 0 to 9.6 × 102 CFU/100 ml. Forty eight percent (305/626) of the presumptive E. coli isolates were confirmed using species-specific uidA gene which code for β-glucuronidase enzyme in E. coli. Antibiotic susceptibility profile of the isolate using a panel of 10 antibiotics shows 100% (150/150) resistance to antibiotics rifampicin and penicillin G while 49.3% (74/150) of the isolates and 46.7% (70/150) were susceptible to streptomycin and cefotaxime respectively. Multiple antibiotic resistance phenotypes (MARP) of the isolates showed resistance to two or more test antibiotics while the calculated multiple antibiotic resistance index (MARI) for the tested isolated is 0.49. The detection of potentially pathogenic E. coli in the final effluents suggestspotential danger to the receiving water bodies where the effluents are discharge. The high MARI valued obtained in this study indicates that the isolates are form environment where the tested antibiotics are being used and may further lead to the spread of multiple antibiotics resistance among other pathogens that may be present in the same environment.
- Full Text:
- Date Issued: 2014
Productions of high quality wastewater final effluents remain a challenge in the Eastern Cape Province of South Africa
- Authors: Gusha, Siyabulela Stability
- Date: 2012
- Subjects: Water-supply, Rural -- Health aspects -- South Africa , Pathogenic microorganisms -- South Africa -- Eastern Cape , Water-supply, Rural -- South Africa -- Eastern Cape , Effluent quality -- Testing , Sewage disposal plants -- South Africa -- Eastern Cape , Escherichia coli
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11265 , http://hdl.handle.net/10353/489 , Water-supply, Rural -- Health aspects -- South Africa , Pathogenic microorganisms -- South Africa -- Eastern Cape , Water-supply, Rural -- South Africa -- Eastern Cape , Effluent quality -- Testing , Sewage disposal plants -- South Africa -- Eastern Cape , Escherichia coli
- Description: Water is an indispensible and yet a difficult resource to be renewed, thus water scarcity has become one of the major challenges faced worldwide, with the Southern regions of Africa being the most impacted and affected, especially the Eastern Cape Province of South Africa where rural communities depend on receiving waterbodies that are often negatively impacted by wastewater final effluents. This present study was conducted between August and December 2010 to assess the physicochemical and microbial qualities of the final effluents of peri-urban and rural communities based wastewater treatment plants in the Eastern Cape Province. The physicochemical parameters were determined on site and in the laboratory, while bacteriological qualities were determined using culture based techniques. The virological qualities were determined by molecular methods using reverse transcriptase polymerase chain reaction for the target RNA virus and the conventional polymerase chain reaction for the target DNA virus. For both wastewater treatment plants, the physicochemical parameters ranged as follows: chemical oxygen demand (5.95-45 mg/L); total dissolved solids (114.5-187.0 mg/L); salinity (0.12-0.20 psu); temperature (14.2-25.7oC); pH (6.0-7.6); nitrate and nitrites (1.55-6.7 mg/L and 0.023-1.15 mg/L respectively); biological oxygen demand (3.5-7.8 mg/L); turbidity (1.49-6.98 NTU); and chlorine residual (0-2.97 mg/L). Feacal indicator bacteria counts ranged as follows: feacal coliforms (0-1.25×104 cfu/100 ml); total coliforms (0-3.95×104 cfu/100 ml); and enterococci (0-5.0×103 cfu/100 ml). xviii Seventy five percent of the rural community based plant and 80 percent of the peri-urban community based plant were positive for coxsackie A virus, while hepatitis A virus was detected in all the rural community based plant 80 percent of the peri-urban community based plant. This study suggests the need for intervention by appropriate regulatory agencies to ensure regular monitoring of the qualities of final effluents of wastewater treatment facilities in the Eastern Cape Province and ensure compliance to established guidelines.
- Full Text:
- Date Issued: 2012
- Authors: Gusha, Siyabulela Stability
- Date: 2012
- Subjects: Water-supply, Rural -- Health aspects -- South Africa , Pathogenic microorganisms -- South Africa -- Eastern Cape , Water-supply, Rural -- South Africa -- Eastern Cape , Effluent quality -- Testing , Sewage disposal plants -- South Africa -- Eastern Cape , Escherichia coli
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11265 , http://hdl.handle.net/10353/489 , Water-supply, Rural -- Health aspects -- South Africa , Pathogenic microorganisms -- South Africa -- Eastern Cape , Water-supply, Rural -- South Africa -- Eastern Cape , Effluent quality -- Testing , Sewage disposal plants -- South Africa -- Eastern Cape , Escherichia coli
- Description: Water is an indispensible and yet a difficult resource to be renewed, thus water scarcity has become one of the major challenges faced worldwide, with the Southern regions of Africa being the most impacted and affected, especially the Eastern Cape Province of South Africa where rural communities depend on receiving waterbodies that are often negatively impacted by wastewater final effluents. This present study was conducted between August and December 2010 to assess the physicochemical and microbial qualities of the final effluents of peri-urban and rural communities based wastewater treatment plants in the Eastern Cape Province. The physicochemical parameters were determined on site and in the laboratory, while bacteriological qualities were determined using culture based techniques. The virological qualities were determined by molecular methods using reverse transcriptase polymerase chain reaction for the target RNA virus and the conventional polymerase chain reaction for the target DNA virus. For both wastewater treatment plants, the physicochemical parameters ranged as follows: chemical oxygen demand (5.95-45 mg/L); total dissolved solids (114.5-187.0 mg/L); salinity (0.12-0.20 psu); temperature (14.2-25.7oC); pH (6.0-7.6); nitrate and nitrites (1.55-6.7 mg/L and 0.023-1.15 mg/L respectively); biological oxygen demand (3.5-7.8 mg/L); turbidity (1.49-6.98 NTU); and chlorine residual (0-2.97 mg/L). Feacal indicator bacteria counts ranged as follows: feacal coliforms (0-1.25×104 cfu/100 ml); total coliforms (0-3.95×104 cfu/100 ml); and enterococci (0-5.0×103 cfu/100 ml). xviii Seventy five percent of the rural community based plant and 80 percent of the peri-urban community based plant were positive for coxsackie A virus, while hepatitis A virus was detected in all the rural community based plant 80 percent of the peri-urban community based plant. This study suggests the need for intervention by appropriate regulatory agencies to ensure regular monitoring of the qualities of final effluents of wastewater treatment facilities in the Eastern Cape Province and ensure compliance to established guidelines.
- Full Text:
- Date Issued: 2012
Phytochemical analysis and bioactivity of Garcinia Kola (Heckel) seeds on selected bacterial pathogens
- Seanego, Christinah Tshephisho
- Authors: Seanego, Christinah Tshephisho
- Date: 2012
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11259 , http://hdl.handle.net/10353/420 , Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Description: Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.
- Full Text:
- Date Issued: 2012
- Authors: Seanego, Christinah Tshephisho
- Date: 2012
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11259 , http://hdl.handle.net/10353/420 , Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants , Microbial sensitivity tests , Streptococcal infections , Streptococcus , Staphylococcus aureus infections , Salmonella typhimurium , Traditional medicine
- Description: Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.
- Full Text:
- Date Issued: 2012
Assessment of the antibacterial properties of n-Hexane extract of Cocos Nucifera and its interactions with some conventional antibiotics
- Authors: Akinyele, Taiwo Adesola
- Date: 2011
- Subjects: Coconut palm , Microbial sensitivity tests , Gram-negative bacterial infections , Vibrio infections , Antibiotics , Hexane , Extracts
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11245 , http://hdl.handle.net/10353/416 , Coconut palm , Microbial sensitivity tests , Gram-negative bacterial infections , Vibrio infections , Antibiotics , Hexane , Extracts
- Description: Cocos nucifera belong to the family Aracaceae (palm Family). The English name is coconut and it is used extensively as medicinal remedies against infections such as urinary tract infections, gastro intestinal infections, skin and wound infections. The in vitro antibacterial (including anti-listerial and anti-vibrio) properties as well as the evaluation of the combination potentials of the plant extract with six front-line antibiotics were evaluated in this study using standard procedures. The in vitro anti-listerial properties of the crude aqueous and n-Hexane extract of the husk of Cocos nucifera were carried out against 37 Listeria isolates. Twenty-nine of the test organisms were susceptible to the aqueous extract while thirty were susceptible to the n-Hexane extract both at the screening concentration of 25 mg/ml. Minimum Inhibitory Concentration (MIC) values for all the susceptible bacteria ranged between 0.6 - 5.0 mg/ml. For the aqueous extract, average log reduction in viable cell count ranged between 0.32 Log10 and 4.8 Log10 CFU/ml after 8 hours interaction in 1 × MIC and 2 × MIC. For the n-Hexane extract, the log reduction ranged between 2.4 Log10 and 6.2 Log10 CFU/ml after 8 hours interaction in 1 × MIC and 2 × MIC. The time-kill characteristics of the two extracts suggest that at higher concentration (2 × MIC) and longer duration of interaction (8 hr), more bacteria were killed. In vitro anti-vibrio and antibacterial properties experiment revealed that of all the 45 vibrio and 25 bacteria strains that was tested, 37 were susceptible to the aqueous extract and 38 to the n-Hexane extract, while 17 were susceptible to the aqueous extract and 21 to the n-Hexane extract. Minimum Inhibitory Concentration (MIC) values for all the susceptible bacteria ranged between 0.3 - 5.0 mg/ml. viii The time kill studies revealed that for the aqueous extract, average log reduction in viable cell count in time kill assay ranged between 0.12 Log10 and 4.2 Log10 CFU/ml after 8 hr interaction at 1 × MIC and 2 × MIC. For the n-Hexane extract, the log reduction ranged between 0.56 Log10 and 6.4 Log10 CFU/ml after 8 hr interaction in 1 × MIC and 2 × MIC. In the test for the combination interactions, the checkerboard method revealed synergy of 67% and indifferent of 33%, while the time kill assay detected synergy in 72% and indifferent in 28% of the combinations tested. The synergy detected was not specific to any of the antibiotics or the Gram reaction of the bacteria, and no antagonism was detected. We conclude that the aqueous and n-Hexane extract of the husk of C. nucifera contains potential broad spectrum antibiotics resistance modulating compounds that could be relevant in the treatment of infections caused by these pathogens. In addition, the husk which is being discarded as agro waste will opens up a vista of opportunities for utilization for therapeutic purposes
- Full Text:
- Date Issued: 2011
- Authors: Akinyele, Taiwo Adesola
- Date: 2011
- Subjects: Coconut palm , Microbial sensitivity tests , Gram-negative bacterial infections , Vibrio infections , Antibiotics , Hexane , Extracts
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11245 , http://hdl.handle.net/10353/416 , Coconut palm , Microbial sensitivity tests , Gram-negative bacterial infections , Vibrio infections , Antibiotics , Hexane , Extracts
- Description: Cocos nucifera belong to the family Aracaceae (palm Family). The English name is coconut and it is used extensively as medicinal remedies against infections such as urinary tract infections, gastro intestinal infections, skin and wound infections. The in vitro antibacterial (including anti-listerial and anti-vibrio) properties as well as the evaluation of the combination potentials of the plant extract with six front-line antibiotics were evaluated in this study using standard procedures. The in vitro anti-listerial properties of the crude aqueous and n-Hexane extract of the husk of Cocos nucifera were carried out against 37 Listeria isolates. Twenty-nine of the test organisms were susceptible to the aqueous extract while thirty were susceptible to the n-Hexane extract both at the screening concentration of 25 mg/ml. Minimum Inhibitory Concentration (MIC) values for all the susceptible bacteria ranged between 0.6 - 5.0 mg/ml. For the aqueous extract, average log reduction in viable cell count ranged between 0.32 Log10 and 4.8 Log10 CFU/ml after 8 hours interaction in 1 × MIC and 2 × MIC. For the n-Hexane extract, the log reduction ranged between 2.4 Log10 and 6.2 Log10 CFU/ml after 8 hours interaction in 1 × MIC and 2 × MIC. The time-kill characteristics of the two extracts suggest that at higher concentration (2 × MIC) and longer duration of interaction (8 hr), more bacteria were killed. In vitro anti-vibrio and antibacterial properties experiment revealed that of all the 45 vibrio and 25 bacteria strains that was tested, 37 were susceptible to the aqueous extract and 38 to the n-Hexane extract, while 17 were susceptible to the aqueous extract and 21 to the n-Hexane extract. Minimum Inhibitory Concentration (MIC) values for all the susceptible bacteria ranged between 0.3 - 5.0 mg/ml. viii The time kill studies revealed that for the aqueous extract, average log reduction in viable cell count in time kill assay ranged between 0.12 Log10 and 4.2 Log10 CFU/ml after 8 hr interaction at 1 × MIC and 2 × MIC. For the n-Hexane extract, the log reduction ranged between 0.56 Log10 and 6.4 Log10 CFU/ml after 8 hr interaction in 1 × MIC and 2 × MIC. In the test for the combination interactions, the checkerboard method revealed synergy of 67% and indifferent of 33%, while the time kill assay detected synergy in 72% and indifferent in 28% of the combinations tested. The synergy detected was not specific to any of the antibiotics or the Gram reaction of the bacteria, and no antagonism was detected. We conclude that the aqueous and n-Hexane extract of the husk of C. nucifera contains potential broad spectrum antibiotics resistance modulating compounds that could be relevant in the treatment of infections caused by these pathogens. In addition, the husk which is being discarded as agro waste will opens up a vista of opportunities for utilization for therapeutic purposes
- Full Text:
- Date Issued: 2011
Enterococcus pathotypes as reservoirs of antibiotic resistance determinants in the Kat River and Fort Beaufort abstraction waters
- Authors: Ntloko, Phindiwe
- Date: 2014
- Subjects: Enterococcus , Drug resistance
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11290 , http://hdl.handle.net/10353/d1019821 , Enterococcus , Drug resistance
- Description: In this study, 400 presumptive Enterococcus isolates previously recovered from Kat River and Fort Beaufort Abstraction water dam were subjected to molecular confirmation and pathotyping. Two hundred and seventy-four (68%) of these isolates were confirmed to be enterococci species. Confirmations studies were polymerase chain reaction (PCR) based, using enterococci specific primers targeting the tuf gene. The confirmed enterococci isolates were further differentiated into their pathotypes, the targets of which were: E. faecalis, E. avium, E. hirae, E. casseliflavarus and E. gallinarum using well documented species specific primer sequences. E. faecalis accounted for 20% of the isolates, followed by E. avium (16%), E. hirae (13%), E. casseliflavarus (5%) and E. gallinarum (3%). Furthermore, all the confirmed isolates were analysed for antibiotic susceptibilities using a panel of nine different antibiotics, namely vancomycin, linezolid, ciprofloxacin, ampicillin, gentamicin, chloramphenicol, tetracycline, erythromycin, penicillin, and those that were resistant were assayed for the presence of relevant antibiotic resistance genes. All the 274 isolates were found to harbour vanA resistance gene confirming their phenotypic resistance to the vancomycin. Similarly, 60% (109/180) of the isolates showed phenotypic resistance to erythromycin which was further confirmed by the presence of ermA genes in these isolates. The presence of antibiotic resistant bacteria in surface waters poses a risk to public health.
- Full Text:
- Date Issued: 2014
- Authors: Ntloko, Phindiwe
- Date: 2014
- Subjects: Enterococcus , Drug resistance
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11290 , http://hdl.handle.net/10353/d1019821 , Enterococcus , Drug resistance
- Description: In this study, 400 presumptive Enterococcus isolates previously recovered from Kat River and Fort Beaufort Abstraction water dam were subjected to molecular confirmation and pathotyping. Two hundred and seventy-four (68%) of these isolates were confirmed to be enterococci species. Confirmations studies were polymerase chain reaction (PCR) based, using enterococci specific primers targeting the tuf gene. The confirmed enterococci isolates were further differentiated into their pathotypes, the targets of which were: E. faecalis, E. avium, E. hirae, E. casseliflavarus and E. gallinarum using well documented species specific primer sequences. E. faecalis accounted for 20% of the isolates, followed by E. avium (16%), E. hirae (13%), E. casseliflavarus (5%) and E. gallinarum (3%). Furthermore, all the confirmed isolates were analysed for antibiotic susceptibilities using a panel of nine different antibiotics, namely vancomycin, linezolid, ciprofloxacin, ampicillin, gentamicin, chloramphenicol, tetracycline, erythromycin, penicillin, and those that were resistant were assayed for the presence of relevant antibiotic resistance genes. All the 274 isolates were found to harbour vanA resistance gene confirming their phenotypic resistance to the vancomycin. Similarly, 60% (109/180) of the isolates showed phenotypic resistance to erythromycin which was further confirmed by the presence of ermA genes in these isolates. The presence of antibiotic resistant bacteria in surface waters poses a risk to public health.
- Full Text:
- Date Issued: 2014
Studies on the antimicrobial, antioxidant and antiproliferative potential of the ethyl acetate extract and compounds of Peltophorum africanum
- Authors: Okeleye, Benjamin Ifeoluwa
- Date: 2014
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11282 , http://hdl.handle.net/10353/d1016165
- Description: Cells are constantly exposed to a variety of oxidizing agents, some of which are necessary for life. Oxidants produced in excess can cause an imbalance, leading to oxidative stress, especially in chronic bacterial, viral, and parasitic infections. This can result to damage of biomolecules such as lipids, proteins, and DNA, hence, an increased risk for cancer. Plants have a long history of use in the treatment of cancer. Plant secondary metabolites have proved to be an excellent reservoir of new medical compounds. Fruits, vegetables, and whole grains contain a wide variety of antioxidant phytochemicals, such as phenolics and carotenoids, and may help protect cellular systems from oxidative damage and also may lower the risk of chronic diseases. Peltophorum africanum, a member of the family Fabaceae (Sond) is also known as the African weeping wattle and is used in traditional medicine in South Africa. This study investigated the antimicrobial, antioxidant and antiproliferative potential of the ethyl acetate extract and compounds of Peltophorum africanum in order to validate its pharmacological use. The study assessed the in vitro antimicrobial activity of ethyl acetate extract (EAE) of Peltophorum africanum stem bark and its fractions by the agar well and macrodilution methods. The toxicity on a normal human liver cell (Chang liver cell) and antiproliferation of human breast (MCF-7), colon (HT-29) and cervical (HeLa) cancer cell lines were determined using the CellTiter-Blue cell viability assay and the mechanism of action delineated using the Nucleic Acid and Protein Purification Nucleospin® Tissue Kit, Scanning Electron Microscopy (SEM), Propidium iodide (PI) and Acridine orange (AO) double-staining techniques, the Cleaved Caspase 3 (Asp 175) Alexa Fluor® 488 Antibody and the Coulter® DNA PrepTM Reagents Kit. Purification and identification of the compounds from EAE and fractions as well as the morphological alteration of bacteria, yeast and cancer cells were determined using thin layer chromatography, infrared spectra fingerprint and GC-MS analysis, micro-dilution and scanning electron microscopy with energy-dispersive X-ray analysis. In vitro antioxidant activity of EAE was determined by means of radical scavenging and ferric reducing power analysis using 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 2`-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) kit, hydrogen peroxide (H2O2), iron (iii) chloride (Fe3+) and nitric oxide (NO). To assess the likely effects of secondary metabolites on the activities observed; total proanthocyanidins, phenolics, flavonols, and flavonoids were determined using standard phytochemical methods. Data were analyzed by one way analysis of variance (ANOVA; SPSS Version 17.0, 2011), regression analysis (MINITAB, version 12 for windows), probit analysis test (software NCSS, 2007) and GraphPad Prism4 software package. The p-values < 0.05 were considered significant. Marked activity of the extract was observed against Plesiomonas shigelloides ATCC 51903, with MIC and MLC values of 0.15625 and 0.3125mg/mL, respectively. The extract was both bactericidal (MICindex ≤ 2) and bacteriostatic/fungistatic (MICindex > 2) in activity. Lethal dose at 50 (LD50) showed 82.64 ± 1.40 degree of toxicity at 24 hrs, and 95 percentile of cell death dose activity ranged from log 3.12 ± 0.01 to 4.59 ± 0.03. The activity of the eight fractions tested ranged from 1.0 ± 0.5 to 3.7 ± 1.6 mg/mL (IC50) and from 2.1 ± 0.8 to 6.25 ± 0 mg/mL (IC90) (Chapter 3). Due to the effect of compounds present in the crude extract and fractions, the P. aeruginosa treated with EAE had a reduction of sodium from 5.55 % (untreated) - 1.50 %. For C. albicans, pottasium was reduced from 4.16 % (untreated) - 0.76 % (T1). Remarkable morphological alterations were observed including deformation of the germ tubes and perforation of the cell wall (Chapter 4). Extract scavenging activity of 88.73± 6.69 % (25 μg mL-1), 53.93±1.09 % (25 μg mL-1) were recorded for H2O2 and NO respectively with proanthocyanidins (92.18±4.68 mg/g) occurring more (p < 0.05) in the extract compared to all other phenolics compounds (Chapter 5). Significant reduction in cell viability of the cells was noted as the MCF-7 cells were reduced from 100 - 54.33±1.84 % after 72 hrs of treatment with 5 μg/mL of EAE (P. value < 0.05). TEt10 was cytotoxic against human normal cells (chang liver cell) at EC50 of 37 μg/mL and 74 μg/mL after 24 and 48 h of treatment respectively. Marked antiproliferative activity of 13.2 μg/mL (EC50) was observed when HeLa cells were treated for 48 h. Internucleosomal DNA of MCF-7, HT-29 and HeLa cells randomly fragmented into an uninterrupted spectrum of sizes, complemented by the intercalation of nucleic acid-specific fluorochromes by PI and AO spotting two phases of apoptosis; early (EA) and late (LA) apoptosis. Distinctive ultramorphological changes observed include; cell shrinkage, membrane blebbing, and typical cell induced death. The study also recorded 705.102 ± 28.56 % TEt10 caspase-3 activity compared to curcumin 592.857 ± 165.76 % (positive control) and untreated (negative control; 100 ± 15.81 %) cells. Percentage HeLa cell with Sub-G1 DNA phase increased from 0.13 ± 0.06 % (negative control) to 13.8 ± 3.04 % compared to curcumin (8.17 ± 2.20 %) after treatment with TEt10. The compounds identified in the fractions including Colchicine, N-(trifluoroacetyl)methyl-N-deacetyl-, Lupeol and .gamma.-Sitosterol may be responsible for the induction of apoptosis observed and could be further studied in vivo as a potential template for new anticancer treatment (Chapter 6 & 7).
- Full Text:
- Date Issued: 2014
- Authors: Okeleye, Benjamin Ifeoluwa
- Date: 2014
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11282 , http://hdl.handle.net/10353/d1016165
- Description: Cells are constantly exposed to a variety of oxidizing agents, some of which are necessary for life. Oxidants produced in excess can cause an imbalance, leading to oxidative stress, especially in chronic bacterial, viral, and parasitic infections. This can result to damage of biomolecules such as lipids, proteins, and DNA, hence, an increased risk for cancer. Plants have a long history of use in the treatment of cancer. Plant secondary metabolites have proved to be an excellent reservoir of new medical compounds. Fruits, vegetables, and whole grains contain a wide variety of antioxidant phytochemicals, such as phenolics and carotenoids, and may help protect cellular systems from oxidative damage and also may lower the risk of chronic diseases. Peltophorum africanum, a member of the family Fabaceae (Sond) is also known as the African weeping wattle and is used in traditional medicine in South Africa. This study investigated the antimicrobial, antioxidant and antiproliferative potential of the ethyl acetate extract and compounds of Peltophorum africanum in order to validate its pharmacological use. The study assessed the in vitro antimicrobial activity of ethyl acetate extract (EAE) of Peltophorum africanum stem bark and its fractions by the agar well and macrodilution methods. The toxicity on a normal human liver cell (Chang liver cell) and antiproliferation of human breast (MCF-7), colon (HT-29) and cervical (HeLa) cancer cell lines were determined using the CellTiter-Blue cell viability assay and the mechanism of action delineated using the Nucleic Acid and Protein Purification Nucleospin® Tissue Kit, Scanning Electron Microscopy (SEM), Propidium iodide (PI) and Acridine orange (AO) double-staining techniques, the Cleaved Caspase 3 (Asp 175) Alexa Fluor® 488 Antibody and the Coulter® DNA PrepTM Reagents Kit. Purification and identification of the compounds from EAE and fractions as well as the morphological alteration of bacteria, yeast and cancer cells were determined using thin layer chromatography, infrared spectra fingerprint and GC-MS analysis, micro-dilution and scanning electron microscopy with energy-dispersive X-ray analysis. In vitro antioxidant activity of EAE was determined by means of radical scavenging and ferric reducing power analysis using 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 2`-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) kit, hydrogen peroxide (H2O2), iron (iii) chloride (Fe3+) and nitric oxide (NO). To assess the likely effects of secondary metabolites on the activities observed; total proanthocyanidins, phenolics, flavonols, and flavonoids were determined using standard phytochemical methods. Data were analyzed by one way analysis of variance (ANOVA; SPSS Version 17.0, 2011), regression analysis (MINITAB, version 12 for windows), probit analysis test (software NCSS, 2007) and GraphPad Prism4 software package. The p-values < 0.05 were considered significant. Marked activity of the extract was observed against Plesiomonas shigelloides ATCC 51903, with MIC and MLC values of 0.15625 and 0.3125mg/mL, respectively. The extract was both bactericidal (MICindex ≤ 2) and bacteriostatic/fungistatic (MICindex > 2) in activity. Lethal dose at 50 (LD50) showed 82.64 ± 1.40 degree of toxicity at 24 hrs, and 95 percentile of cell death dose activity ranged from log 3.12 ± 0.01 to 4.59 ± 0.03. The activity of the eight fractions tested ranged from 1.0 ± 0.5 to 3.7 ± 1.6 mg/mL (IC50) and from 2.1 ± 0.8 to 6.25 ± 0 mg/mL (IC90) (Chapter 3). Due to the effect of compounds present in the crude extract and fractions, the P. aeruginosa treated with EAE had a reduction of sodium from 5.55 % (untreated) - 1.50 %. For C. albicans, pottasium was reduced from 4.16 % (untreated) - 0.76 % (T1). Remarkable morphological alterations were observed including deformation of the germ tubes and perforation of the cell wall (Chapter 4). Extract scavenging activity of 88.73± 6.69 % (25 μg mL-1), 53.93±1.09 % (25 μg mL-1) were recorded for H2O2 and NO respectively with proanthocyanidins (92.18±4.68 mg/g) occurring more (p < 0.05) in the extract compared to all other phenolics compounds (Chapter 5). Significant reduction in cell viability of the cells was noted as the MCF-7 cells were reduced from 100 - 54.33±1.84 % after 72 hrs of treatment with 5 μg/mL of EAE (P. value < 0.05). TEt10 was cytotoxic against human normal cells (chang liver cell) at EC50 of 37 μg/mL and 74 μg/mL after 24 and 48 h of treatment respectively. Marked antiproliferative activity of 13.2 μg/mL (EC50) was observed when HeLa cells were treated for 48 h. Internucleosomal DNA of MCF-7, HT-29 and HeLa cells randomly fragmented into an uninterrupted spectrum of sizes, complemented by the intercalation of nucleic acid-specific fluorochromes by PI and AO spotting two phases of apoptosis; early (EA) and late (LA) apoptosis. Distinctive ultramorphological changes observed include; cell shrinkage, membrane blebbing, and typical cell induced death. The study also recorded 705.102 ± 28.56 % TEt10 caspase-3 activity compared to curcumin 592.857 ± 165.76 % (positive control) and untreated (negative control; 100 ± 15.81 %) cells. Percentage HeLa cell with Sub-G1 DNA phase increased from 0.13 ± 0.06 % (negative control) to 13.8 ± 3.04 % compared to curcumin (8.17 ± 2.20 %) after treatment with TEt10. The compounds identified in the fractions including Colchicine, N-(trifluoroacetyl)methyl-N-deacetyl-, Lupeol and .gamma.-Sitosterol may be responsible for the induction of apoptosis observed and could be further studied in vivo as a potential template for new anticancer treatment (Chapter 6 & 7).
- Full Text:
- Date Issued: 2014
Evaluation of the final effluents of some wastewater treatment plants in Amathole and Chris Hani District Municipality of the Eastern Cape Province as sources of vibrio pathogens in the aquatic environment
- Authors: Nongogo, Vuyokazi
- Date: 2014
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11287 , http://hdl.handle.net/10353/d1019813
- Description: Certain areas in the world still depend on the receiving water bodies as sources of domestic water and for recreational purposes. The discharge of poor quality effluents from wastewater treatment plants can impact negatively on these water bodies, as they can act as vehicles for pathogens to the environment, posing a threat to humans if such water is used without precaution. Vibrio species are amongst those pathogens that can survive wastewater treatment processes, ending up in the environment, hence the aim of this study was to evaluate the final effluents of some wastewater treatment plants as sources of vibrio pathogens. Five wastewater treatment plants (WWTP) located in Amathole and Chris Hani District Municipalities in the Eastern Cape were used in this study. Samples were collected monthly from September 2012 – August 2013 and analysed using the standard membrane filtration technique. Yellow and green colonies on TCBS agar were enumerated as presumptive Vibrio species and expressed as CFU/100ml for each plant. Colonies were later picked based on their phenotypic characteristics, sub-cultured on fresh TCBS agar to ascertain purity. These presumptive isolates were then subjected to Gram staining and Oxidase test. Gram negative and Oxidase positive isolates were selected for further confirmation using Polymerised Chain Reaction (PCR). PCR was also employed for characterisation of Vibrio into three species viz V. parahaemolyticus, V. fluvialis and V. vulnificus. Antibiogram profile of the characterised species was then determined together with the presence of relevant antibiotic resistance genes Vibrio densities for the twelve month period ranged between 0 - 1.48×104 CFU/100ml with two of the plants located in East bank and Queenstown characterized by extremely high counts and one plant( Reeston) with very low counts.Three hundred presumptive Vibrio isolates were screened for identity confirmation. Of these, the dominating species found was V. fluvialis (28.6%) followed by V. vulnificus (28%) and the least was found to be V. parahaemolyticus (11.6%). The remaining unidentified 31.6% were suspected to belong to other Vibrio species not covered within the scope of this study. All the confirmed isolates i.e., V. parahaemolyticus, V. vulnificus and V. fluvialis were susceptible to imipenem, gentamicin and meropenem and resistant to only tetracycline. Between 60-100% of the V. parahaemolyticus isolates, 7.1% to 100 % V. vulnificus isolates and 2.5 to 100 % V. fluvialis showed resistances to polymixin B, sulfamethazole, erythromycin, penicillin G, chloramphenicol, trimethroprim and trimethroprim & sulfamethazole. Antibiotic Resistance Genes that were assessed included dfRA, SXT, floR and Sul2 varying in proportion with each species showing diversity in the Vibrio community. The dfR A gene was detected in all the V. parahaemolyticus isolates while floR gene was not detected in any of the isolates belonging to the three species. The distribution of sul2 gene cut across the species being 1% (1) in V. fluvialis, 3% (1) in V. parahaemolyticus and 4% (3) in V. vulnificus. The SXT gene was only determined in V. parahaemolyticus. It is clear that the final effluents of the selected plants are reservoirs for Vibrio pathogens as well as antibiotic resistance genes in the environment. The isolation of Vibrio from WWTP shows that this pathogen is in circulation in some pockets of the population. Therefore, wastewater treatment plants need to be properly monitored to ensure that they comply with set guidelines.
- Full Text:
- Date Issued: 2014
- Authors: Nongogo, Vuyokazi
- Date: 2014
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11287 , http://hdl.handle.net/10353/d1019813
- Description: Certain areas in the world still depend on the receiving water bodies as sources of domestic water and for recreational purposes. The discharge of poor quality effluents from wastewater treatment plants can impact negatively on these water bodies, as they can act as vehicles for pathogens to the environment, posing a threat to humans if such water is used without precaution. Vibrio species are amongst those pathogens that can survive wastewater treatment processes, ending up in the environment, hence the aim of this study was to evaluate the final effluents of some wastewater treatment plants as sources of vibrio pathogens. Five wastewater treatment plants (WWTP) located in Amathole and Chris Hani District Municipalities in the Eastern Cape were used in this study. Samples were collected monthly from September 2012 – August 2013 and analysed using the standard membrane filtration technique. Yellow and green colonies on TCBS agar were enumerated as presumptive Vibrio species and expressed as CFU/100ml for each plant. Colonies were later picked based on their phenotypic characteristics, sub-cultured on fresh TCBS agar to ascertain purity. These presumptive isolates were then subjected to Gram staining and Oxidase test. Gram negative and Oxidase positive isolates were selected for further confirmation using Polymerised Chain Reaction (PCR). PCR was also employed for characterisation of Vibrio into three species viz V. parahaemolyticus, V. fluvialis and V. vulnificus. Antibiogram profile of the characterised species was then determined together with the presence of relevant antibiotic resistance genes Vibrio densities for the twelve month period ranged between 0 - 1.48×104 CFU/100ml with two of the plants located in East bank and Queenstown characterized by extremely high counts and one plant( Reeston) with very low counts.Three hundred presumptive Vibrio isolates were screened for identity confirmation. Of these, the dominating species found was V. fluvialis (28.6%) followed by V. vulnificus (28%) and the least was found to be V. parahaemolyticus (11.6%). The remaining unidentified 31.6% were suspected to belong to other Vibrio species not covered within the scope of this study. All the confirmed isolates i.e., V. parahaemolyticus, V. vulnificus and V. fluvialis were susceptible to imipenem, gentamicin and meropenem and resistant to only tetracycline. Between 60-100% of the V. parahaemolyticus isolates, 7.1% to 100 % V. vulnificus isolates and 2.5 to 100 % V. fluvialis showed resistances to polymixin B, sulfamethazole, erythromycin, penicillin G, chloramphenicol, trimethroprim and trimethroprim & sulfamethazole. Antibiotic Resistance Genes that were assessed included dfRA, SXT, floR and Sul2 varying in proportion with each species showing diversity in the Vibrio community. The dfR A gene was detected in all the V. parahaemolyticus isolates while floR gene was not detected in any of the isolates belonging to the three species. The distribution of sul2 gene cut across the species being 1% (1) in V. fluvialis, 3% (1) in V. parahaemolyticus and 4% (3) in V. vulnificus. The SXT gene was only determined in V. parahaemolyticus. It is clear that the final effluents of the selected plants are reservoirs for Vibrio pathogens as well as antibiotic resistance genes in the environment. The isolation of Vibrio from WWTP shows that this pathogen is in circulation in some pockets of the population. Therefore, wastewater treatment plants need to be properly monitored to ensure that they comply with set guidelines.
- Full Text:
- Date Issued: 2014
Assessment of some bacteria species isolated from woodlands of Raymond Mhlaba Local Municipality for high activity laccase production
- Authors: Gogotya, Asemahle
- Date: 2019
- Subjects: Laccase
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/19098 , vital:39879
- Description: The function of enzymes in keeping the earth clean is enormous; being executed in the biodegradation of different natural pollutants and biocatalysis of different responses by substituting the ecologically risky and harmful concoction impetuses offering a situation inviting option, laccases is an example of an enzyme described as it doesn’t produce harmful byproducts. Laccases are employed in several industrial processes that play a key responsibility in transformation of life and making the environment a better place. Bacteria have been described as best producers of laccases with a potential in the industrial scale processes. Laccase was produced from different bacterial isolates identified and named as Bacillus sp. strain GFN1 isolated from soil sample, Bacillus sp. strain GLN and Streptomyces sp. strain LAO both isolated from decaying wood samples in Raymond Mhlaba local municipality with accession numbers MK290988 to MK290990 respectively, as identified by partial sequencing, these were the best producers some of which were positive for Napthol and guaiacol; even upon quantitative screening they were better laccase producers. For quantitative laccase screening, 2,2'-azino-bis(3-ethylbenzothiazoline-6- sulphonic acid) was utilised as the substrate for laccase assays. These laccase producing bacteria were subjected to optimization of growth conditions using submerged fermentation which increased the activity of the produced laccase in great amounts. In optimizing basal medium growth conditions where laccase was harvested after every 72 hours where optimal activity was 16 obtained, studying several factors such as pH which turned out to be pH 5 for all isolates, effect of supplemented carbon and nitrogen sources with the be best being lactose and urea respectively with their effective concentrations using lignin as the main carbon and nitrogen source. Copper sulfate was used as the main inducer and the species preferred guaiacol and ferullic acid and the Fe2+ asthe best supplemented metal ion. The time course was done investigating parameters such as cell growth which was determined by observing the optical density, laccase activity, protein concentration and pH and the presented results suggested that when there was an increase in cell growth, enzyme activity decreased pH had no much effect on the enzyme production as it was almost stable all the time with protein concentration exhibiting no direct effect on enzyme activity also. Characterization of the crude enzyme was done to check the stability of the enzyme produced in various parameters. The enzymes produced by the different strains were thermophilic as they were able to withstand elevated temperatures between 90 and 100 C, with pH stability within an extensive variety of alkaline pH, typical of most bacterial laccases. Various metal ions affected the stability of the enzyme with CuSO4 increasing the stability of two of the bacterial enzyme and appeared to slightly decrease the stability of one enzyme. The studied inhibitors only decreased the stability on the enzyme and couldn’t completely inhibit the enzyme, and the enzymes showed specificity towards varying substrates. The studied bacterial laccases exhibit tremendous characteristics which are of great significance in the industries and will add to the novelty of bacterial laccases and their stability amongst the most studied fungal laccases.
- Full Text:
- Date Issued: 2019
- Authors: Gogotya, Asemahle
- Date: 2019
- Subjects: Laccase
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/19098 , vital:39879
- Description: The function of enzymes in keeping the earth clean is enormous; being executed in the biodegradation of different natural pollutants and biocatalysis of different responses by substituting the ecologically risky and harmful concoction impetuses offering a situation inviting option, laccases is an example of an enzyme described as it doesn’t produce harmful byproducts. Laccases are employed in several industrial processes that play a key responsibility in transformation of life and making the environment a better place. Bacteria have been described as best producers of laccases with a potential in the industrial scale processes. Laccase was produced from different bacterial isolates identified and named as Bacillus sp. strain GFN1 isolated from soil sample, Bacillus sp. strain GLN and Streptomyces sp. strain LAO both isolated from decaying wood samples in Raymond Mhlaba local municipality with accession numbers MK290988 to MK290990 respectively, as identified by partial sequencing, these were the best producers some of which were positive for Napthol and guaiacol; even upon quantitative screening they were better laccase producers. For quantitative laccase screening, 2,2'-azino-bis(3-ethylbenzothiazoline-6- sulphonic acid) was utilised as the substrate for laccase assays. These laccase producing bacteria were subjected to optimization of growth conditions using submerged fermentation which increased the activity of the produced laccase in great amounts. In optimizing basal medium growth conditions where laccase was harvested after every 72 hours where optimal activity was 16 obtained, studying several factors such as pH which turned out to be pH 5 for all isolates, effect of supplemented carbon and nitrogen sources with the be best being lactose and urea respectively with their effective concentrations using lignin as the main carbon and nitrogen source. Copper sulfate was used as the main inducer and the species preferred guaiacol and ferullic acid and the Fe2+ asthe best supplemented metal ion. The time course was done investigating parameters such as cell growth which was determined by observing the optical density, laccase activity, protein concentration and pH and the presented results suggested that when there was an increase in cell growth, enzyme activity decreased pH had no much effect on the enzyme production as it was almost stable all the time with protein concentration exhibiting no direct effect on enzyme activity also. Characterization of the crude enzyme was done to check the stability of the enzyme produced in various parameters. The enzymes produced by the different strains were thermophilic as they were able to withstand elevated temperatures between 90 and 100 C, with pH stability within an extensive variety of alkaline pH, typical of most bacterial laccases. Various metal ions affected the stability of the enzyme with CuSO4 increasing the stability of two of the bacterial enzyme and appeared to slightly decrease the stability of one enzyme. The studied inhibitors only decreased the stability on the enzyme and couldn’t completely inhibit the enzyme, and the enzymes showed specificity towards varying substrates. The studied bacterial laccases exhibit tremendous characteristics which are of great significance in the industries and will add to the novelty of bacterial laccases and their stability amongst the most studied fungal laccases.
- Full Text:
- Date Issued: 2019
Characterization of some virulence and antibiotic resistance genes of Staphylococcus aureus isolated from cases of Bovine Mastitis in Nkonkobe Municipality, Eastern Cape Province, RSA
- Authors: Pekana, Abongile
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11293 , http://hdl.handle.net/10353/d1021133
- Description: Staphylococcus aureus is one of the predominant causative agents of mastitis disease in dairy herds. Mastitis disease has a negative impact in the economic losses in the dairy sector across the globe. The aim of this study is to detect some of the virulence genes in the S. aureus isolated from 400 milk samples of subclinical and clinical mastitis dairy cows in Fort Hare dairy farm and Middle Drift dairy farm in Alice in the Eastern Cape province of South Africa. In addition antibiotic resistance pattern and antibiotic resistance genes were investigated. Gram-staining, oxidase test, catalase test and API Staph kit were preliminary biochemical tests used for the identification of S. aureus isolates. The MALDI-TOF-MS was also used for further identification. Polymerase chain reaction was performed of genes encoding antibiotic resistance as well as clumping (clfA), coagulase (coa) gene, toxic shock syndrome (tsst), exfoliative toxin A and B (eta and etb), and the gene segment encoding the immunoglobulin G binding region and X region of protein gene spa. A total of 20 (5%) S. aureus strains obtained from 400 milk samples from the two farms were subjected to 16 antibiotics for antibiotic susceptibility testing. In Middle Drift dairy farm 11 (5.5%) isolates were obtained from 200 samples and 9 (4.5%) isolates were obtained in Fort Hare dairy farm from 200 samples. A large percent of the isolates were resistant to penicillin G (60%), followed by trimethoprim (60%) and tetracycline (60%), trimethoprim-sulfamethaxazole (55%), telithroprim (55%) and doxycycline (45%). Most of the isolates were sensitive to several (50-85%) antibiotics. Of the twenty isolates tested 12 samples contained the penicillin antibiotic resistance gene (blaZ gene), 8 samples contained at least one aminoglycoside-modifying enzyme gene (AME gene); the (aac(6’)/aph(2’’) gene and no amplification occurred for aph(3’)-IIIa and ant(4’)-Ia) genes. In the case of the tetracycline antibiotic resistance gene (tetK and tetM), 2 samples contained tetM and a single sample contained tetK gene. No amplification was observed for the erythromycin antibiotic resistance genes (ermA, ermB, ermC, Mef and msrA). All the samples tested were negative for the expression of toxic syndrome gene (tsst), etb, and Immunoglobulin G binding region. However, amplification of the clumping factor was observed in 7 (35%) isolates of S. aureus, exfoliative toxin (eta) expressed 4(20%) isolates; coagulase gene (coa) yielded six DNA bands of six differences sizes from 16 (80%) isolates. A total of four different bands size were expressed for the spa X region from 12 (60%) isolates. The data obtained in this study suggests that poor hygienic practices and inadequate management practices are responsible for the increase in Staphylococcus aureus isolation. The high resistance of S. aureus to antibiotics and the distribution of virulence genes contribute in bovine mastitis in these farms may cause health problems in the community consuming raw milk purchased from these farms.
- Full Text:
- Date Issued: 2015
- Authors: Pekana, Abongile
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11293 , http://hdl.handle.net/10353/d1021133
- Description: Staphylococcus aureus is one of the predominant causative agents of mastitis disease in dairy herds. Mastitis disease has a negative impact in the economic losses in the dairy sector across the globe. The aim of this study is to detect some of the virulence genes in the S. aureus isolated from 400 milk samples of subclinical and clinical mastitis dairy cows in Fort Hare dairy farm and Middle Drift dairy farm in Alice in the Eastern Cape province of South Africa. In addition antibiotic resistance pattern and antibiotic resistance genes were investigated. Gram-staining, oxidase test, catalase test and API Staph kit were preliminary biochemical tests used for the identification of S. aureus isolates. The MALDI-TOF-MS was also used for further identification. Polymerase chain reaction was performed of genes encoding antibiotic resistance as well as clumping (clfA), coagulase (coa) gene, toxic shock syndrome (tsst), exfoliative toxin A and B (eta and etb), and the gene segment encoding the immunoglobulin G binding region and X region of protein gene spa. A total of 20 (5%) S. aureus strains obtained from 400 milk samples from the two farms were subjected to 16 antibiotics for antibiotic susceptibility testing. In Middle Drift dairy farm 11 (5.5%) isolates were obtained from 200 samples and 9 (4.5%) isolates were obtained in Fort Hare dairy farm from 200 samples. A large percent of the isolates were resistant to penicillin G (60%), followed by trimethoprim (60%) and tetracycline (60%), trimethoprim-sulfamethaxazole (55%), telithroprim (55%) and doxycycline (45%). Most of the isolates were sensitive to several (50-85%) antibiotics. Of the twenty isolates tested 12 samples contained the penicillin antibiotic resistance gene (blaZ gene), 8 samples contained at least one aminoglycoside-modifying enzyme gene (AME gene); the (aac(6’)/aph(2’’) gene and no amplification occurred for aph(3’)-IIIa and ant(4’)-Ia) genes. In the case of the tetracycline antibiotic resistance gene (tetK and tetM), 2 samples contained tetM and a single sample contained tetK gene. No amplification was observed for the erythromycin antibiotic resistance genes (ermA, ermB, ermC, Mef and msrA). All the samples tested were negative for the expression of toxic syndrome gene (tsst), etb, and Immunoglobulin G binding region. However, amplification of the clumping factor was observed in 7 (35%) isolates of S. aureus, exfoliative toxin (eta) expressed 4(20%) isolates; coagulase gene (coa) yielded six DNA bands of six differences sizes from 16 (80%) isolates. A total of four different bands size were expressed for the spa X region from 12 (60%) isolates. The data obtained in this study suggests that poor hygienic practices and inadequate management practices are responsible for the increase in Staphylococcus aureus isolation. The high resistance of S. aureus to antibiotics and the distribution of virulence genes contribute in bovine mastitis in these farms may cause health problems in the community consuming raw milk purchased from these farms.
- Full Text:
- Date Issued: 2015
Prevalence and antibiogram of some swine associated Shiga toxin producing Escherichia coli Serogroups and Salmonella species in Nkonkobe Municipality, Eastern Cape Province, South Africa
- Authors: Iwu, Chinwe Juliana
- Date: 2015
- Subjects: Escherichia coli -- South Africa -- Eastern Cape , Salmonella infections -- South Africa -- Eastern Cape , Water -- Pollution -- Environmental aspects -- South Africa -- Eastern Cape , Escherichia coli infections -- South Africa -- Eastern Cape , Water quality management -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11294 , http://hdl.handle.net/10353/d1021273 , Escherichia coli -- South Africa -- Eastern Cape , Salmonella infections -- South Africa -- Eastern Cape , Water -- Pollution -- Environmental aspects -- South Africa -- Eastern Cape , Escherichia coli infections -- South Africa -- Eastern Cape , Water quality management -- South Africa -- Eastern Cape
- Description: Gastrointestinal illnesses have continually become a global public health issue. Exposure to zoonotic food borne pathogens such as Salmonella and diarrhoegenic E. coli either by direct or indirect contact through the consumption of food producing animals is likely an important mode of infection to humans. More so, the use of antibiotics in farm animals similar to those used in humans can select for resistance in bacteria frequently harboured by them. These resistant strains can be passed on to humans through contaminated meat products and water leading to resistant infections with consequences such as prolonged illnesses, treatment failures, and increased morbidity and mortality. In animals, these can lead to reduced productivity. Monitoring the level of resistance among bacteria from animal isolates will help in generating data that could be used to create awareness of their presence in the environment and aid in preventing a potential epidemic in the community. In this study, we investigated the prevalence and antimicrobial resistance profile of Escherichia coli serogroups and Salmonella species in faecal samples collected from pigs in Nkonkobe Municipality in the Eastern Cape Province, South Africa between April – July, 2014. A total of 310 presumptive Shiga toxin producing Escherichia coli (STEC) were confirmed as E. coli spp using polymerase chain reaction (PCR) technique by amplification of the uidA gene, out of which 179 (58%) were confirmed positive. Approximately, serogrougs O157:H7, O145 and O26 made up 24% (n=43), 8% (n=14) and 20% (n=35) of the E. coli population respectively. Only E. coli O26 was positive for stx2 gene in 31% of the isolates harbouring the gene, while the other serogroups were non-pathogenic. Susceptibility of the isolates to 18 antibiotics was carried out in vitro by the standardized agar disc-diffusion method. All the isolates were susceptible to imipenem. Similarly, a relatively high susceptibility was observed in norfloxacin (83-100%), ciprofloxacin (63-100%), gentamycin (77-100%), and chloramphenicol (77-100%). However, all the isolates were resistant to tetracycline and its long acting counterpart oxytetracycline. Resistances observed against other antimicrobials are as follows: ampicillin (84-91%), streptomycin (14-100%), erythromycin (91-100%), ceftazidime (35%). Multiple antimicrobial resistance patterns and indices ranged from 3 to 12 and 0.2 to 0.7 to respectively. Genes encoding resistances to ampicillin (ampC), streptomycin (strA) and tetracycline (tetA) were frequently detected in 50-100%, 22-29% and 40-86% of the resistant isolates respectively. In the other arm of the dissertation, two hundred and fifty eight presumptive isolates of Salmonella were recovered from the faecal samples of pigs. Specific primers targeting serogroups A, B, C1, C2, and D were used to delineate the isolates into different serogroups using PCR. Only serogroup A (n=48) was detected. These isolates were examined for antimicrobial susceptibility by disc diffusion method using 18 antibiotics. The results showed that a large proportion of the isolates were resistant to tetracycline (100%), oxytetracycline (100%), ampicillin (75%), sulphamethoxazole/trimethoprim (75%) and streptomycin (75%). Majority of the isolates exhibited multidrug resistances with the predominant multiple antibiotic resistance (MAR) phenotype being against eleven antibiotics. A high multiple antibiotic resistance (MAR) index in a range of 0.3- 0.6 was observed. The incidence of genes encoding resistance against tetracycline (tetA), streptomycin (stra), and ampicillin (ampC) were 54%, 44% and 61% respectively. These findings reveal that pigs within the Nkonkobe Municipality in the Eastern Cape Province could harbour Shiga toxins and multidrug resistant serogroups of E. coli as well as resistant Salmonella which could be transmitted to humans through the food chain. To ensure public health safety, continuous monitoring and sufficient sanitation in swine industries must be ensured.
- Full Text:
- Date Issued: 2015
- Authors: Iwu, Chinwe Juliana
- Date: 2015
- Subjects: Escherichia coli -- South Africa -- Eastern Cape , Salmonella infections -- South Africa -- Eastern Cape , Water -- Pollution -- Environmental aspects -- South Africa -- Eastern Cape , Escherichia coli infections -- South Africa -- Eastern Cape , Water quality management -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11294 , http://hdl.handle.net/10353/d1021273 , Escherichia coli -- South Africa -- Eastern Cape , Salmonella infections -- South Africa -- Eastern Cape , Water -- Pollution -- Environmental aspects -- South Africa -- Eastern Cape , Escherichia coli infections -- South Africa -- Eastern Cape , Water quality management -- South Africa -- Eastern Cape
- Description: Gastrointestinal illnesses have continually become a global public health issue. Exposure to zoonotic food borne pathogens such as Salmonella and diarrhoegenic E. coli either by direct or indirect contact through the consumption of food producing animals is likely an important mode of infection to humans. More so, the use of antibiotics in farm animals similar to those used in humans can select for resistance in bacteria frequently harboured by them. These resistant strains can be passed on to humans through contaminated meat products and water leading to resistant infections with consequences such as prolonged illnesses, treatment failures, and increased morbidity and mortality. In animals, these can lead to reduced productivity. Monitoring the level of resistance among bacteria from animal isolates will help in generating data that could be used to create awareness of their presence in the environment and aid in preventing a potential epidemic in the community. In this study, we investigated the prevalence and antimicrobial resistance profile of Escherichia coli serogroups and Salmonella species in faecal samples collected from pigs in Nkonkobe Municipality in the Eastern Cape Province, South Africa between April – July, 2014. A total of 310 presumptive Shiga toxin producing Escherichia coli (STEC) were confirmed as E. coli spp using polymerase chain reaction (PCR) technique by amplification of the uidA gene, out of which 179 (58%) were confirmed positive. Approximately, serogrougs O157:H7, O145 and O26 made up 24% (n=43), 8% (n=14) and 20% (n=35) of the E. coli population respectively. Only E. coli O26 was positive for stx2 gene in 31% of the isolates harbouring the gene, while the other serogroups were non-pathogenic. Susceptibility of the isolates to 18 antibiotics was carried out in vitro by the standardized agar disc-diffusion method. All the isolates were susceptible to imipenem. Similarly, a relatively high susceptibility was observed in norfloxacin (83-100%), ciprofloxacin (63-100%), gentamycin (77-100%), and chloramphenicol (77-100%). However, all the isolates were resistant to tetracycline and its long acting counterpart oxytetracycline. Resistances observed against other antimicrobials are as follows: ampicillin (84-91%), streptomycin (14-100%), erythromycin (91-100%), ceftazidime (35%). Multiple antimicrobial resistance patterns and indices ranged from 3 to 12 and 0.2 to 0.7 to respectively. Genes encoding resistances to ampicillin (ampC), streptomycin (strA) and tetracycline (tetA) were frequently detected in 50-100%, 22-29% and 40-86% of the resistant isolates respectively. In the other arm of the dissertation, two hundred and fifty eight presumptive isolates of Salmonella were recovered from the faecal samples of pigs. Specific primers targeting serogroups A, B, C1, C2, and D were used to delineate the isolates into different serogroups using PCR. Only serogroup A (n=48) was detected. These isolates were examined for antimicrobial susceptibility by disc diffusion method using 18 antibiotics. The results showed that a large proportion of the isolates were resistant to tetracycline (100%), oxytetracycline (100%), ampicillin (75%), sulphamethoxazole/trimethoprim (75%) and streptomycin (75%). Majority of the isolates exhibited multidrug resistances with the predominant multiple antibiotic resistance (MAR) phenotype being against eleven antibiotics. A high multiple antibiotic resistance (MAR) index in a range of 0.3- 0.6 was observed. The incidence of genes encoding resistance against tetracycline (tetA), streptomycin (stra), and ampicillin (ampC) were 54%, 44% and 61% respectively. These findings reveal that pigs within the Nkonkobe Municipality in the Eastern Cape Province could harbour Shiga toxins and multidrug resistant serogroups of E. coli as well as resistant Salmonella which could be transmitted to humans through the food chain. To ensure public health safety, continuous monitoring and sufficient sanitation in swine industries must be ensured.
- Full Text:
- Date Issued: 2015
Molecular study of mycobacterium tuberculosis complex (MTBC) DNA from Port Elizabeth
- Authors: Londiwe, Bhembe Nolwazi
- Date: 2014
- Subjects: Mycobacterium tuberculosis
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11281 , http://hdl.handle.net/10353/d1016163 , Mycobacterium tuberculosis
- Description: Mycobacterium tuberculosis complex (MTBC) is a causative agent of tuberculosis (TB) in humans and animals. The burden of tuberculosis in South Africa is worsened by the concurrent epidemic of HIV. The dynamic of TB epidemics has been investigated and yet little data has been given about the Eastern Cape, particularly Port Elizabeth. The study aimed to investigate the prevalence of drug resistant MTBC and to determine the mutations causing resistance in Port Elizabeth. One hundred and ninety (190) DNA samples isolated from sputum specimen in humans suspected of having TB were amplified using the Seeplex® MTB Nested ACE detection assay. To differentiate Mycobacterium tuberculosis complex (MTBC) members for surveillance purposes a multiplex polymerase chain reaction (PCR) method was done based on genomic regions of differences such as RD1, RD1mic, RD2seal, RD4, RD9 and RD12. Target genes known to confer resistance to first and second-line drugs were amplified and the amplicons sequenced using Big Dye Terminator DNA sequencing kit v3.1 (Applied Biosystems, UK). The patient’s demographic profiles were obtained from the National Health Laboratory Service (NHLS). All hundred and ninety DNA samples tested positive for MTBC using the Seeplex® MTB Nested ACE assay. Results show a high prevalence of extensive drug resistant TB in Port Elizabeth, Eastern Cape Province. One hundred and eighty four (184) DNA isolates were used in the identification of different MTBC species. We ended up working with 184 DNA isolates because we ran out of DNA, and we could not go back to isolate DNA from the affected individuals due to the fact that some patients died, while some have been released to go to their homes. From the 184 DNA isolates 45 (24.5%) isolates were identified to be M. tuberculosis, 94 isolates (51.1%) to be M. bovis BCG and 3 isolates (1.6%) to be M. cannetti. Sequencing results show the position of mutation in each DNA isolate; however in the study we got resistance to MDR to be 100% and 42% pre-XDR while 58% was XDR. These results raise an alarm for the prevalence MDR in MTBC from Port Elizabeth. This is a serious health concern which calls for a need to strategise on the identification of extensive drug resistant TB patients from multi-drug resistant TB patients and ensure monitoring of their treatment.
- Full Text:
- Date Issued: 2014
- Authors: Londiwe, Bhembe Nolwazi
- Date: 2014
- Subjects: Mycobacterium tuberculosis
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11281 , http://hdl.handle.net/10353/d1016163 , Mycobacterium tuberculosis
- Description: Mycobacterium tuberculosis complex (MTBC) is a causative agent of tuberculosis (TB) in humans and animals. The burden of tuberculosis in South Africa is worsened by the concurrent epidemic of HIV. The dynamic of TB epidemics has been investigated and yet little data has been given about the Eastern Cape, particularly Port Elizabeth. The study aimed to investigate the prevalence of drug resistant MTBC and to determine the mutations causing resistance in Port Elizabeth. One hundred and ninety (190) DNA samples isolated from sputum specimen in humans suspected of having TB were amplified using the Seeplex® MTB Nested ACE detection assay. To differentiate Mycobacterium tuberculosis complex (MTBC) members for surveillance purposes a multiplex polymerase chain reaction (PCR) method was done based on genomic regions of differences such as RD1, RD1mic, RD2seal, RD4, RD9 and RD12. Target genes known to confer resistance to first and second-line drugs were amplified and the amplicons sequenced using Big Dye Terminator DNA sequencing kit v3.1 (Applied Biosystems, UK). The patient’s demographic profiles were obtained from the National Health Laboratory Service (NHLS). All hundred and ninety DNA samples tested positive for MTBC using the Seeplex® MTB Nested ACE assay. Results show a high prevalence of extensive drug resistant TB in Port Elizabeth, Eastern Cape Province. One hundred and eighty four (184) DNA isolates were used in the identification of different MTBC species. We ended up working with 184 DNA isolates because we ran out of DNA, and we could not go back to isolate DNA from the affected individuals due to the fact that some patients died, while some have been released to go to their homes. From the 184 DNA isolates 45 (24.5%) isolates were identified to be M. tuberculosis, 94 isolates (51.1%) to be M. bovis BCG and 3 isolates (1.6%) to be M. cannetti. Sequencing results show the position of mutation in each DNA isolate; however in the study we got resistance to MDR to be 100% and 42% pre-XDR while 58% was XDR. These results raise an alarm for the prevalence MDR in MTBC from Port Elizabeth. This is a serious health concern which calls for a need to strategise on the identification of extensive drug resistant TB patients from multi-drug resistant TB patients and ensure monitoring of their treatment.
- Full Text:
- Date Issued: 2014
Quality indices of the final effluents of two sub-urban-based wastewater treatment plants in Amathole District Municipality in the Eastern Cape Province of South Africa
- Authors: Gcilitshana, Onele
- Date: 2014
- Subjects: Whole effluent toxicity testing -- South Africa -- Eastern Cape , Sewage disposal plants -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape , Effluent quality -- Testing , Viruses -- South Africa -- Eastern Cape , Water reuse -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11288 , http://hdl.handle.net/10353/d1019816 , Whole effluent toxicity testing -- South Africa -- Eastern Cape , Sewage disposal plants -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape , Effluent quality -- Testing , Viruses -- South Africa -- Eastern Cape , Water reuse -- South Africa -- Eastern Cape
- Description: Worldwide, water reuse is promoted as an alternative for water scarcity, however, wastewater effluents have been reported as possible contaminants to surface water. The failure of some wastewater treatment processes to completely remove organic matter and some pathogenic microorganisms allows them to initiate infections. This manifests more in communities where surface water is used directly for drinking. To assess water quality, bacteria alone cannot be used as it may be absent in virus-contaminated water. This study was carried out to assess the quality of two wastewater treatment plant effluents from the Eastern Cape Province of South Africa. Physicochemical parameters and microbiological parameters like faecal coliforms, adenovirus, rotavirus, hepatitis A virus, norovirus and enterovirus were evaluated over a projected period of one year. Physicochemical parameters were measured on site using multiparameters, faecal coliforms enumerated using culture-based methods and viruses are detected using both conventional and real-time PCR. Physicochemical parameters like electrical conductivity, turbidity, free chlorine and phosphates were incompliant with the standards set by the Department of Water affairs for effluents to be discharged. Faecal coliform counts were nil for one plant (WWTP-R) where they correlated inversely (P < 0.01) with the high free chlorine. For WWTP-K, faecal coliforms were detected in 27% of samples in the range of 9.9 × 101 to 6.4× 104 CFU/100ml. From the five viruses assessed, three viruses were detected with Rotavirus being the most abundant (0-2034176 genome copies/L) followed by Adenovirus (0–275 genome copies/L) then Hepatitis A virus (0–71 genome copies/L) in the WWTP-K while none of the viruses was detected in WWTP-R. Species B, species C and Adv41 serotypes were detected from the May 2013 and June 2013 samples where almost all parameters were incompliant in the plant. The detection of these viruses in supposedly treated effluents is suggestive of these being the sources of contamination to surface water and therefore renders surface waters unsafe for direct use and to aquatic life. Although real-time PCR is more sensitive and reliable in detection of viruses, use of cell-culture techniques in this study would have been more efficient in confirming the infectivity of the viruses detected, hence the recommendation of these techniques in future projects of this nature.
- Full Text:
- Date Issued: 2014
- Authors: Gcilitshana, Onele
- Date: 2014
- Subjects: Whole effluent toxicity testing -- South Africa -- Eastern Cape , Sewage disposal plants -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape , Effluent quality -- Testing , Viruses -- South Africa -- Eastern Cape , Water reuse -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11288 , http://hdl.handle.net/10353/d1019816 , Whole effluent toxicity testing -- South Africa -- Eastern Cape , Sewage disposal plants -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape , Effluent quality -- Testing , Viruses -- South Africa -- Eastern Cape , Water reuse -- South Africa -- Eastern Cape
- Description: Worldwide, water reuse is promoted as an alternative for water scarcity, however, wastewater effluents have been reported as possible contaminants to surface water. The failure of some wastewater treatment processes to completely remove organic matter and some pathogenic microorganisms allows them to initiate infections. This manifests more in communities where surface water is used directly for drinking. To assess water quality, bacteria alone cannot be used as it may be absent in virus-contaminated water. This study was carried out to assess the quality of two wastewater treatment plant effluents from the Eastern Cape Province of South Africa. Physicochemical parameters and microbiological parameters like faecal coliforms, adenovirus, rotavirus, hepatitis A virus, norovirus and enterovirus were evaluated over a projected period of one year. Physicochemical parameters were measured on site using multiparameters, faecal coliforms enumerated using culture-based methods and viruses are detected using both conventional and real-time PCR. Physicochemical parameters like electrical conductivity, turbidity, free chlorine and phosphates were incompliant with the standards set by the Department of Water affairs for effluents to be discharged. Faecal coliform counts were nil for one plant (WWTP-R) where they correlated inversely (P < 0.01) with the high free chlorine. For WWTP-K, faecal coliforms were detected in 27% of samples in the range of 9.9 × 101 to 6.4× 104 CFU/100ml. From the five viruses assessed, three viruses were detected with Rotavirus being the most abundant (0-2034176 genome copies/L) followed by Adenovirus (0–275 genome copies/L) then Hepatitis A virus (0–71 genome copies/L) in the WWTP-K while none of the viruses was detected in WWTP-R. Species B, species C and Adv41 serotypes were detected from the May 2013 and June 2013 samples where almost all parameters were incompliant in the plant. The detection of these viruses in supposedly treated effluents is suggestive of these being the sources of contamination to surface water and therefore renders surface waters unsafe for direct use and to aquatic life. Although real-time PCR is more sensitive and reliable in detection of viruses, use of cell-culture techniques in this study would have been more efficient in confirming the infectivity of the viruses detected, hence the recommendation of these techniques in future projects of this nature.
- Full Text:
- Date Issued: 2014
Bioactivity and phytochemical analysis of Hydnora Africana on some selected bacterial pathogens
- Authors: Nethathe, Bono Bianca
- Date: 2011
- Subjects: Helicobacter pylori , Medicinal plants -- South Africa -- Eastern Cape , Microbial sensitivity tests , Herbs -- Therapeutic use -- South Africa -- Eastern Cape , Plants -- Analysis , Staphylococcus aureus , Aeromonas hydrophila , Drug resistance in microorganisms , Plant-pathogen relationships
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11247 , http://hdl.handle.net/10353/d1001063 , Helicobacter pylori , Medicinal plants -- South Africa -- Eastern Cape , Microbial sensitivity tests , Herbs -- Therapeutic use -- South Africa -- Eastern Cape , Plants -- Analysis , Staphylococcus aureus , Aeromonas hydrophila , Drug resistance in microorganisms , Plant-pathogen relationships
- Description: Abstract Medicinal plants have been for long remedies for human diseases because they contain components of therapeutic value. The growing problem of antibiotic resistance by organisms demands the search for novel compounds from plant based sources. The present study was aimed at evaluating the bioactivity and phytochemical analysis of Hydnora africana on clinical and standard strains of Helicobacter pylori (PE 252C and ATCC 43526), Aeromonas hydrophila ATCC 35654, and Staphylococcus aureus NCT 6571 in an effort to identify potential sources of cheap starting materials for the synthesis of new drugs against these strains. Ethyl acetate, acetone, ethanol, methanol, and water crude extracts of H. africana were screened for activity against the test organisms using the agar well diffusion assay. The Minimum Inhibitory Concentration (MIC50) and Minimum Bactericidal Concentration (MBC) of the most potent extracts were determined by the microdilution method, followed by qualitative phytochemical analysis. Results were analyzed statistically by ANOVA one - way test. Different concentrations (200,100, 50mg/mL) of the methanol, acetone, ethanol and ethyl acetate extracts showed activity against S. aureus and A. hydrophila while for H. pylori, only methanol and ethyl acetate extracts were active; water showed no activity for all studied bacterial pathogens. Mean zone diameter of inhibition which ranged from 0-22mm were observed for all test bacterial pathogens and 14-17mm for ciprofloxacin. The activity of methanol and ethyl acetate extracts were statistically significant (P< 0.05) compared to all the other extracts. MIC50 and MBC ranged from 0.078 – 2.5mg/mL, 0.78-25mg/mL respectively for all tested bacterial pathogens. For ciprofloxacin, the MIC50 and MBC ranged from 0.00976 – 0.078mg/mL and 0.098– 0.78mg/mL respectively. There was no statistically significant difference between extracts (methanol, acetone, ethanol, ethyl acetate) and the control antibiotic (ciprofloxacin) (P> 0.05). Qualitative phytochemical analysis confirmed the presence of alkaloids, saponins, steroids, tannins and flavonoids in the methanol, acetone,ethanol and ethyl acetate extracts. The results demonstrate that H. africana may contain compounds with therapeutic potentials which can be lead molecules for semi-synthesis of new drugs.
- Full Text:
- Date Issued: 2011
- Authors: Nethathe, Bono Bianca
- Date: 2011
- Subjects: Helicobacter pylori , Medicinal plants -- South Africa -- Eastern Cape , Microbial sensitivity tests , Herbs -- Therapeutic use -- South Africa -- Eastern Cape , Plants -- Analysis , Staphylococcus aureus , Aeromonas hydrophila , Drug resistance in microorganisms , Plant-pathogen relationships
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11247 , http://hdl.handle.net/10353/d1001063 , Helicobacter pylori , Medicinal plants -- South Africa -- Eastern Cape , Microbial sensitivity tests , Herbs -- Therapeutic use -- South Africa -- Eastern Cape , Plants -- Analysis , Staphylococcus aureus , Aeromonas hydrophila , Drug resistance in microorganisms , Plant-pathogen relationships
- Description: Abstract Medicinal plants have been for long remedies for human diseases because they contain components of therapeutic value. The growing problem of antibiotic resistance by organisms demands the search for novel compounds from plant based sources. The present study was aimed at evaluating the bioactivity and phytochemical analysis of Hydnora africana on clinical and standard strains of Helicobacter pylori (PE 252C and ATCC 43526), Aeromonas hydrophila ATCC 35654, and Staphylococcus aureus NCT 6571 in an effort to identify potential sources of cheap starting materials for the synthesis of new drugs against these strains. Ethyl acetate, acetone, ethanol, methanol, and water crude extracts of H. africana were screened for activity against the test organisms using the agar well diffusion assay. The Minimum Inhibitory Concentration (MIC50) and Minimum Bactericidal Concentration (MBC) of the most potent extracts were determined by the microdilution method, followed by qualitative phytochemical analysis. Results were analyzed statistically by ANOVA one - way test. Different concentrations (200,100, 50mg/mL) of the methanol, acetone, ethanol and ethyl acetate extracts showed activity against S. aureus and A. hydrophila while for H. pylori, only methanol and ethyl acetate extracts were active; water showed no activity for all studied bacterial pathogens. Mean zone diameter of inhibition which ranged from 0-22mm were observed for all test bacterial pathogens and 14-17mm for ciprofloxacin. The activity of methanol and ethyl acetate extracts were statistically significant (P< 0.05) compared to all the other extracts. MIC50 and MBC ranged from 0.078 – 2.5mg/mL, 0.78-25mg/mL respectively for all tested bacterial pathogens. For ciprofloxacin, the MIC50 and MBC ranged from 0.00976 – 0.078mg/mL and 0.098– 0.78mg/mL respectively. There was no statistically significant difference between extracts (methanol, acetone, ethanol, ethyl acetate) and the control antibiotic (ciprofloxacin) (P> 0.05). Qualitative phytochemical analysis confirmed the presence of alkaloids, saponins, steroids, tannins and flavonoids in the methanol, acetone,ethanol and ethyl acetate extracts. The results demonstrate that H. africana may contain compounds with therapeutic potentials which can be lead molecules for semi-synthesis of new drugs.
- Full Text:
- Date Issued: 2011
Prevalence of pathogenic Escherichia coli strains in the final effluents of four wastewater treatment plants in the Eastern Cape Province of South Africa
- Authors: Seti, Nozuko Zukiswa
- Date: 2014
- Subjects: Escherichia coli -- South Africa -- Eastern Cape , Bacterial diseases -- Pathogenesis , Sewage -- Purification -- South Africa -- Eastern Cape , Sewage disposal plants -- South Africa -- Eastern Cape , Whole effluent toxicity testing -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11286 , http://hdl.handle.net/10353/d1019808 , Escherichia coli -- South Africa -- Eastern Cape , Bacterial diseases -- Pathogenesis , Sewage -- Purification -- South Africa -- Eastern Cape , Sewage disposal plants -- South Africa -- Eastern Cape , Whole effluent toxicity testing -- South Africa -- Eastern Cape
- Description: Water is an essential need that stimulates health and well being. Increase in population size and urbanization negatively affect water resources due to high demands of effluent outputs. Wastewater is an important reservoir for Escherichia coli and can present significant acute toxicity if released into receiving water sources without being adequately treated. E. coli is used as indicator organism for the detection of faecal contamination. These strains have been considered to be one of the primary causes of diarrhoeal infections worldwide. The present study was conducted between September 2012 and June 2013 to assess the prevalence of pathogenic E. coli strains in the final effluents of four wastewater treatment plants in Chris Hani and Buffalo City Municipalities in the Eastern Cape Province of South Africa. Standard membrane filtration technique was used for bacteriological analysis and molecular based technique was used for identification of E. coli pathotypes. The results were recorded in colony forming units/100 ml. Faecal coliforms ranged between 0-9.6×10³ CFU/100 ml for the wwtp-Q and E. coli densities ranged between 0-8.4×10³ CFU/100ml. Faecal coliforms ranged between 4×10²-9.7×10³ CFU/100 ml for wwtp-M and E. coli densities ranged between 1.2×10¹-8.4×10³ CFU/100 ml. The wwtp-E showed to have bacterial counts of faecal coliforms ranging between 4.0×10³-8.2×10³ CFU/100 ml and E. coli densities ranging between 3.5×10¹-7.1×10³ CFU/100 ml. The WWTP-K in this study was only assessed for the presence of E. coli. Faecal coliforms were assessed by the other members of the group. This plant showed to have E. coli densities ranging between 0-7.5×10²CFU/100 ml. A total of 200 presumptive E. coli isolates were subjected to screening by conventional PCR in which (29%) of the wwtp-M isolates were positively identified as E. coli, (16%) of the wwtp-K, (22%) of the wwtp-Q and (34%) of the wwtp-E isolates were positively confirmed as E. coli. A total of 100 randomly selected E. coli isolates were characterised into different pathotypes. (16%) of positive isolates were detected as EPEC and 11% were detected as UPEC strains. There was no detection for the ETEC strains. Antibiotic susceptibility patterns of E. coli strains showed high levels of resistance to Penicillin G, Erythromycin, Tetracycline and Sulfamethoxazole. High levels of Susceptibility were observed in antibiotics such as Chloramphenicol, Amoxicillin and Tetracycline. The results of this study reveal that the plants were above the recommended Standard limit of zero CFU/100 ml for effluents meant to be discharge into receiving water sources. This study reveals inadequacy of the plants studied to produce effluents of acceptable quality.
- Full Text:
- Date Issued: 2014
- Authors: Seti, Nozuko Zukiswa
- Date: 2014
- Subjects: Escherichia coli -- South Africa -- Eastern Cape , Bacterial diseases -- Pathogenesis , Sewage -- Purification -- South Africa -- Eastern Cape , Sewage disposal plants -- South Africa -- Eastern Cape , Whole effluent toxicity testing -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11286 , http://hdl.handle.net/10353/d1019808 , Escherichia coli -- South Africa -- Eastern Cape , Bacterial diseases -- Pathogenesis , Sewage -- Purification -- South Africa -- Eastern Cape , Sewage disposal plants -- South Africa -- Eastern Cape , Whole effluent toxicity testing -- South Africa -- Eastern Cape
- Description: Water is an essential need that stimulates health and well being. Increase in population size and urbanization negatively affect water resources due to high demands of effluent outputs. Wastewater is an important reservoir for Escherichia coli and can present significant acute toxicity if released into receiving water sources without being adequately treated. E. coli is used as indicator organism for the detection of faecal contamination. These strains have been considered to be one of the primary causes of diarrhoeal infections worldwide. The present study was conducted between September 2012 and June 2013 to assess the prevalence of pathogenic E. coli strains in the final effluents of four wastewater treatment plants in Chris Hani and Buffalo City Municipalities in the Eastern Cape Province of South Africa. Standard membrane filtration technique was used for bacteriological analysis and molecular based technique was used for identification of E. coli pathotypes. The results were recorded in colony forming units/100 ml. Faecal coliforms ranged between 0-9.6×10³ CFU/100 ml for the wwtp-Q and E. coli densities ranged between 0-8.4×10³ CFU/100ml. Faecal coliforms ranged between 4×10²-9.7×10³ CFU/100 ml for wwtp-M and E. coli densities ranged between 1.2×10¹-8.4×10³ CFU/100 ml. The wwtp-E showed to have bacterial counts of faecal coliforms ranging between 4.0×10³-8.2×10³ CFU/100 ml and E. coli densities ranging between 3.5×10¹-7.1×10³ CFU/100 ml. The WWTP-K in this study was only assessed for the presence of E. coli. Faecal coliforms were assessed by the other members of the group. This plant showed to have E. coli densities ranging between 0-7.5×10²CFU/100 ml. A total of 200 presumptive E. coli isolates were subjected to screening by conventional PCR in which (29%) of the wwtp-M isolates were positively identified as E. coli, (16%) of the wwtp-K, (22%) of the wwtp-Q and (34%) of the wwtp-E isolates were positively confirmed as E. coli. A total of 100 randomly selected E. coli isolates were characterised into different pathotypes. (16%) of positive isolates were detected as EPEC and 11% were detected as UPEC strains. There was no detection for the ETEC strains. Antibiotic susceptibility patterns of E. coli strains showed high levels of resistance to Penicillin G, Erythromycin, Tetracycline and Sulfamethoxazole. High levels of Susceptibility were observed in antibiotics such as Chloramphenicol, Amoxicillin and Tetracycline. The results of this study reveal that the plants were above the recommended Standard limit of zero CFU/100 ml for effluents meant to be discharge into receiving water sources. This study reveals inadequacy of the plants studied to produce effluents of acceptable quality.
- Full Text:
- Date Issued: 2014
Isolation and molecular characterization of Bacillus cereus from cow’s raw milk
- Authors: Lukanji, Zinathi , Ndip, R N
- Date: 2015
- Subjects: Milk contamination -- South Africa -- Eastern Cape , Bacillus (Bacteria) -- South Africa -- Eastern Cape , Foodborne diseases -- Molecular diagnosis -- South Africa -- Eastern Cape , Dairy products -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11296 , http://hdl.handle.net/10353/d1021284 , Milk contamination -- South Africa -- Eastern Cape , Bacillus (Bacteria) -- South Africa -- Eastern Cape , Foodborne diseases -- Molecular diagnosis -- South Africa -- Eastern Cape , Dairy products -- South Africa -- Eastern Cape
- Description: Bacillus cereus is a group of ubiquitous facultative anaerobic spore forming Gram-positive rods commonly found in soil. It has been detected and implicated in several contaminated food products and raw milk in dairy farms and it causes foodborne gastroenteritis by producing several toxins. This study is aimed at characterizing virulence determinants of B. cereus from cow‟s raw milk. A total of 400 raw milk samples were collected in Fort Hare Dairy Trust and Middledrift Dairy Farm; and cultured on Polymyxin pyruvate Egg-Yolk Mannitol Bromothymol Agar (PEMBA) for 48 hours at 37°C. DNA was isolated from the isolates and 16S rDNA was amplified and sent to Central Analytical Laboratory for sequencing. The gyrB gene of B. cereus was also used to confirm the identity of the isolates. Antibiotic susceptibility profiles of the isolates together with virulence genes were investigated. Multilocus Sequence typing was used to investigate the genetic relatedness of some selected isolates. Furthermore, spores of the isolates were produced, harvested and their concentrations determined. All (100%) of the isolates were identified as having a 96-99% similarity to B. cereus, B. thuringiensis and B. anthracis using sequencing; while gyrB gene was observed in all (100%) of the isolates. Three virulence genes nheA, nheB, nheC encoding for non haemolysin enterotoxin were amplified in all (100%) the isolates. All (100%) of the isolates were susceptible to doxycycline, gentamycin, tetracycline, ciprofloxacin, chloramphenicol and streptomycin. Resistance to rifampicin and penicillin G was predominant with equal rate of 100%, while susceptibility to erythromycin, clindamycin and doxycycline ranged from 60% to 100%. The selected isolates were related and are descendants of the same ancestor. All (100%) the isolates produced spores. The B. cereus isolates contain virulence genes, has multiple antibiotic drug resistance and produce spores, which poses a health risk to the public and cannot be used as probiotics.
- Full Text:
- Date Issued: 2015
- Authors: Lukanji, Zinathi , Ndip, R N
- Date: 2015
- Subjects: Milk contamination -- South Africa -- Eastern Cape , Bacillus (Bacteria) -- South Africa -- Eastern Cape , Foodborne diseases -- Molecular diagnosis -- South Africa -- Eastern Cape , Dairy products -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11296 , http://hdl.handle.net/10353/d1021284 , Milk contamination -- South Africa -- Eastern Cape , Bacillus (Bacteria) -- South Africa -- Eastern Cape , Foodborne diseases -- Molecular diagnosis -- South Africa -- Eastern Cape , Dairy products -- South Africa -- Eastern Cape
- Description: Bacillus cereus is a group of ubiquitous facultative anaerobic spore forming Gram-positive rods commonly found in soil. It has been detected and implicated in several contaminated food products and raw milk in dairy farms and it causes foodborne gastroenteritis by producing several toxins. This study is aimed at characterizing virulence determinants of B. cereus from cow‟s raw milk. A total of 400 raw milk samples were collected in Fort Hare Dairy Trust and Middledrift Dairy Farm; and cultured on Polymyxin pyruvate Egg-Yolk Mannitol Bromothymol Agar (PEMBA) for 48 hours at 37°C. DNA was isolated from the isolates and 16S rDNA was amplified and sent to Central Analytical Laboratory for sequencing. The gyrB gene of B. cereus was also used to confirm the identity of the isolates. Antibiotic susceptibility profiles of the isolates together with virulence genes were investigated. Multilocus Sequence typing was used to investigate the genetic relatedness of some selected isolates. Furthermore, spores of the isolates were produced, harvested and their concentrations determined. All (100%) of the isolates were identified as having a 96-99% similarity to B. cereus, B. thuringiensis and B. anthracis using sequencing; while gyrB gene was observed in all (100%) of the isolates. Three virulence genes nheA, nheB, nheC encoding for non haemolysin enterotoxin were amplified in all (100%) the isolates. All (100%) of the isolates were susceptible to doxycycline, gentamycin, tetracycline, ciprofloxacin, chloramphenicol and streptomycin. Resistance to rifampicin and penicillin G was predominant with equal rate of 100%, while susceptibility to erythromycin, clindamycin and doxycycline ranged from 60% to 100%. The selected isolates were related and are descendants of the same ancestor. All (100%) the isolates produced spores. The B. cereus isolates contain virulence genes, has multiple antibiotic drug resistance and produce spores, which poses a health risk to the public and cannot be used as probiotics.
- Full Text:
- Date Issued: 2015