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
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
Assessment of antibacterial potentials of Garcinia Kola seed extracts and their interactions with antibiotics
- Authors: Sibanda, Thulani
- Date: 2007
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11242 , http://hdl.handle.net/10353/71 , Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants
- Description: The antibacterial potency of the extracts of the seed of Garcinia kola (bitter kola) was investigated in this study against a panel of referenced, environmental and clinical bacterial strains. The killing rates of the active extract as well as their potential for combination antibacterial therapy with standard antibiotics were also elucidated using standard procedures. The aqueous and acetone extracts of the seed were screened for activity against 27 bacterial isolates. The aqueous extract exhibited activity mainly against Gram positive organisms with Minimum inhibitory concentration (MIC) values ranging from 5 mgml-1 – 20 mgml-1, while the acetone extract showed activity against both Gram negative and Gram positive organisms with MIC values ranging from 10 mgml-1 - 0.156 mgml-1. The acetone extract also showed rapid bactericidal activity against Staphylococcus aureus ATCC 6538 with a 3.097 Log10 reduction in counts within 4 hours at 0.3125 mgml-1 and a 1.582 Log10 reduction against Proteus vulgaris CSIR 0030 at 5 mgml-1 after 1 hour. In addition, the aqueous, methanol and acetone extracts of the seeds also exhibited activity against four clinical strains of Staphylococcus isolated from wound sepsis specimens. The MIC values for the aqueous extract were 10 mgml-1 for all the isolates while the acetone and methanol extracts had lower values ranging from 0.3125 - 0.625 mgml-1. The acetone extract was strongly bactericidal against Staphylococcus aureus OKOH3 resulting in a 2.70 Log10 reduction in counts at 1.25 mgml-1 within 4 hours of exposure and a complete elimination of the organism after 8 hours. The bactericidal vi activity of the same extract against Staphylococcus aureus OKOH1 was weak, achieving only a 2.92 Log10 reduction in counts at 1.25 mgml-1 (4× MIC) in 24 hours. In the test for interactions between the acetone extract of the seeds and antibiotics, synergistic interactions were observed largely against Gram positive organisms using the FIC indices, (indices of 0.52 - 0.875) with combinations against Gram negatives yielding largely antagonistic interactions (indices of 2.0 to 5.0). Synergy (≥ 1000 times or ≥ 3 Log10 potentiation of the bactericidal activity) against both Gram negative and Gram positive organisms was detected by time kill assays mainly involving the antibiotics tetracycline, chloramphenicol, amoxycillin and penicillin G. Combinations involving erythromycin and ciprofloxacin consistently gave antagonistic or indifferent interactions. We conclude that the acetone extract of Garcinia kola seeds possess strong bactericidal activities against both Gram positive and Gram negative organisms and can be therapeutically useful in the treatment of bacterial infections including the problematic staphylococcal wound infections. In addition, the acetone extract can be a potential source of broad spectrum resistance modifying compounds that can potentially improve the performance of antibiotics in the treatment of drug resistant infections.
- Full Text:
- Date Issued: 2007
- Authors: Sibanda, Thulani
- Date: 2007
- Subjects: Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11242 , http://hdl.handle.net/10353/71 , Drug resistance in microorganisms , Garcinia , Antibiotics , Medicinal plants
- Description: The antibacterial potency of the extracts of the seed of Garcinia kola (bitter kola) was investigated in this study against a panel of referenced, environmental and clinical bacterial strains. The killing rates of the active extract as well as their potential for combination antibacterial therapy with standard antibiotics were also elucidated using standard procedures. The aqueous and acetone extracts of the seed were screened for activity against 27 bacterial isolates. The aqueous extract exhibited activity mainly against Gram positive organisms with Minimum inhibitory concentration (MIC) values ranging from 5 mgml-1 – 20 mgml-1, while the acetone extract showed activity against both Gram negative and Gram positive organisms with MIC values ranging from 10 mgml-1 - 0.156 mgml-1. The acetone extract also showed rapid bactericidal activity against Staphylococcus aureus ATCC 6538 with a 3.097 Log10 reduction in counts within 4 hours at 0.3125 mgml-1 and a 1.582 Log10 reduction against Proteus vulgaris CSIR 0030 at 5 mgml-1 after 1 hour. In addition, the aqueous, methanol and acetone extracts of the seeds also exhibited activity against four clinical strains of Staphylococcus isolated from wound sepsis specimens. The MIC values for the aqueous extract were 10 mgml-1 for all the isolates while the acetone and methanol extracts had lower values ranging from 0.3125 - 0.625 mgml-1. The acetone extract was strongly bactericidal against Staphylococcus aureus OKOH3 resulting in a 2.70 Log10 reduction in counts at 1.25 mgml-1 within 4 hours of exposure and a complete elimination of the organism after 8 hours. The bactericidal vi activity of the same extract against Staphylococcus aureus OKOH1 was weak, achieving only a 2.92 Log10 reduction in counts at 1.25 mgml-1 (4× MIC) in 24 hours. In the test for interactions between the acetone extract of the seeds and antibiotics, synergistic interactions were observed largely against Gram positive organisms using the FIC indices, (indices of 0.52 - 0.875) with combinations against Gram negatives yielding largely antagonistic interactions (indices of 2.0 to 5.0). Synergy (≥ 1000 times or ≥ 3 Log10 potentiation of the bactericidal activity) against both Gram negative and Gram positive organisms was detected by time kill assays mainly involving the antibiotics tetracycline, chloramphenicol, amoxycillin and penicillin G. Combinations involving erythromycin and ciprofloxacin consistently gave antagonistic or indifferent interactions. We conclude that the acetone extract of Garcinia kola seeds possess strong bactericidal activities against both Gram positive and Gram negative organisms and can be therapeutically useful in the treatment of bacterial infections including the problematic staphylococcal wound infections. In addition, the acetone extract can be a potential source of broad spectrum resistance modifying compounds that can potentially improve the performance of antibiotics in the treatment of drug resistant infections.
- Full Text:
- Date Issued: 2007
Assessment of antibiotic production by some marine Streptomyces isolated from the Nahoon Beach
- Ogunmwonyi, Isoken Nekpen Henrietta
- Authors: Ogunmwonyi, Isoken Nekpen Henrietta
- Date: 2010
- Subjects: Streptomyces , Actinobacteria , Gram-positive bacteria , Antibiotics , Antibiotics -- Testing , Drug resistance in microorganisms
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11243 , http://hdl.handle.net/10353/264 , Streptomyces , Actinobacteria , Gram-positive bacteria , Antibiotics , Antibiotics -- Testing , Drug resistance in microorganisms
- Description: Rapidly emerging strains of bacteria resistant to most advanced antibiotics have become issues of very important public health concern. Research currently directed towards marine actinomycetes presents a vast potential for new compounds that could be able to safely and effectively target resistant species. In this regard, ten putative Streptomyces strains isolated from the Nahoon beach were selected and assessed for antibiotic production and activity against a wide range of bacteria including reference strains, environmental strain and clinical isolates. The ethyl acetate extracts of the putative Streptomyces isolates showed activities against at least 6 and up to 26 of the 32 test bacteria. Inhibition zones were found to range between 9-32 mm diameters at a concentration of 10 mg/ml. The minimum inhibitory concentrations (MICs) of the crude extracts ranged from 0.039 - 10 mg/ml and the least minimum bactericidal concentration (MBC) demonstrated was 0.625 mg/ml against a reference strain Staphylococcus aureus ATCC 6538. Time kill kinetics of all extracts revealed bacteristatic and bactericidal activities. Average Log reductions in viable cell counts for all the extracts ranged from 0.86 Log10 and 3.99 Log10 cfu/ml after 3 h interaction and 0.01 Log10 and 4.86 Log10 after 6 h interaction at MIC, 2 × MIC, 3 × MIC and 4 × MIC concentrations. Most of the extracts were speedily bactericidal at 3 × MIC and 4 × MIC resulting in over 50 % elimination of most of the test bacteria within 3 h and 6 h interaction. The partial characterization of the crude extracts by IR spectral analysis revealed possibility of terpenoid, long chain fatty acids and secondary amine derivatives compounds in the extracts. It is therefore recommended that further investigation should address the relationship between the structure of the active component of the extracts and the broad spectrum activity, as well as a rapid method for large scale production and purification and whether this group of antibiotics has any application in managing human infectious disease.
- Full Text:
- Date Issued: 2010
- Authors: Ogunmwonyi, Isoken Nekpen Henrietta
- Date: 2010
- Subjects: Streptomyces , Actinobacteria , Gram-positive bacteria , Antibiotics , Antibiotics -- Testing , Drug resistance in microorganisms
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11243 , http://hdl.handle.net/10353/264 , Streptomyces , Actinobacteria , Gram-positive bacteria , Antibiotics , Antibiotics -- Testing , Drug resistance in microorganisms
- Description: Rapidly emerging strains of bacteria resistant to most advanced antibiotics have become issues of very important public health concern. Research currently directed towards marine actinomycetes presents a vast potential for new compounds that could be able to safely and effectively target resistant species. In this regard, ten putative Streptomyces strains isolated from the Nahoon beach were selected and assessed for antibiotic production and activity against a wide range of bacteria including reference strains, environmental strain and clinical isolates. The ethyl acetate extracts of the putative Streptomyces isolates showed activities against at least 6 and up to 26 of the 32 test bacteria. Inhibition zones were found to range between 9-32 mm diameters at a concentration of 10 mg/ml. The minimum inhibitory concentrations (MICs) of the crude extracts ranged from 0.039 - 10 mg/ml and the least minimum bactericidal concentration (MBC) demonstrated was 0.625 mg/ml against a reference strain Staphylococcus aureus ATCC 6538. Time kill kinetics of all extracts revealed bacteristatic and bactericidal activities. Average Log reductions in viable cell counts for all the extracts ranged from 0.86 Log10 and 3.99 Log10 cfu/ml after 3 h interaction and 0.01 Log10 and 4.86 Log10 after 6 h interaction at MIC, 2 × MIC, 3 × MIC and 4 × MIC concentrations. Most of the extracts were speedily bactericidal at 3 × MIC and 4 × MIC resulting in over 50 % elimination of most of the test bacteria within 3 h and 6 h interaction. The partial characterization of the crude extracts by IR spectral analysis revealed possibility of terpenoid, long chain fatty acids and secondary amine derivatives compounds in the extracts. It is therefore recommended that further investigation should address the relationship between the structure of the active component of the extracts and the broad spectrum activity, as well as a rapid method for large scale production and purification and whether this group of antibiotics has any application in managing human infectious disease.
- Full Text:
- Date Issued: 2010
Assessment of bioflocculant production by some marine bacteria isolated from the bottom sediment of Algoa Bay
- Authors: Cosa, Sekelwa
- Date: 2010
- Subjects: Flocculants , Bacteria -- South Africa -- Algoa Bay
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11244 , http://hdl.handle.net/10353/404 , Flocculants , Bacteria -- South Africa -- Algoa Bay
- Description: Several problems concerning the use of conventional synthetic flocculants has necessitated the need for alternative cost effective, safe and efficient bioflocculants from microorganisms inhabiting many environments, particularly those from unusual environments. Hence, this study assessed bioflocculant production by three marine bacteria isolated from the bottom sediment of Algoa Bay in the Eastern Cape Province of South Africa. Analysis of the 16S rDNA sequences led to their identification as Halobacillus sp. Mvuyo, Virgibacillus sp. Rob and Oceanobacillus sp. Pinky. Several factors affecting the production and activity of the bioflocculant(s) were studied. Halobacillus sp. Mvuyo produced bioflocculant optimally with glucose (76%) and ammonium chloride (93%) as sole carbon and nitrogen sources, respectively and at neutral pH and in the presence of Ca2+. On the other hand, Virgibacillus sp. Rob preferred glucose (70.4 %) and iron sulphate (74%) as carbon and nitrogen source respectively; an alkaline pH (12.0) and Fe2+. Oceanobacillus sp. Pinky produced bioflocculant optimally when sucrose (80%) and peptone (72.4 %) were used as carbon and nitrogen source respectively, at neutral pH, and in the presence of Ca2+ cation. The chemical analyses of the partially purified bioflocculants revealed that the bioflocculants produced by Halobacillus sp. Mvuyo and Oceanobacillus sp. Pinky were glycoproteins, while that produced by Virgibacillus sp. Rob was a polysaccharide. We thus conclude that Halobacillus sp. Mvuyo, Virgibacillus sp. Rob and Oceanobacillus sp. Pinky hold promise as producers of new and efficient bioflocculant(s). We recommended development of process conditions for large scale production of the bioflocculants followed by their detailed characterization, as well as pilot scale assessment of the applicability of the purified bioflocculant in water/wastewater treatment and other industrial uses
- Full Text:
- Date Issued: 2010
- Authors: Cosa, Sekelwa
- Date: 2010
- Subjects: Flocculants , Bacteria -- South Africa -- Algoa Bay
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11244 , http://hdl.handle.net/10353/404 , Flocculants , Bacteria -- South Africa -- Algoa Bay
- Description: Several problems concerning the use of conventional synthetic flocculants has necessitated the need for alternative cost effective, safe and efficient bioflocculants from microorganisms inhabiting many environments, particularly those from unusual environments. Hence, this study assessed bioflocculant production by three marine bacteria isolated from the bottom sediment of Algoa Bay in the Eastern Cape Province of South Africa. Analysis of the 16S rDNA sequences led to their identification as Halobacillus sp. Mvuyo, Virgibacillus sp. Rob and Oceanobacillus sp. Pinky. Several factors affecting the production and activity of the bioflocculant(s) were studied. Halobacillus sp. Mvuyo produced bioflocculant optimally with glucose (76%) and ammonium chloride (93%) as sole carbon and nitrogen sources, respectively and at neutral pH and in the presence of Ca2+. On the other hand, Virgibacillus sp. Rob preferred glucose (70.4 %) and iron sulphate (74%) as carbon and nitrogen source respectively; an alkaline pH (12.0) and Fe2+. Oceanobacillus sp. Pinky produced bioflocculant optimally when sucrose (80%) and peptone (72.4 %) were used as carbon and nitrogen source respectively, at neutral pH, and in the presence of Ca2+ cation. The chemical analyses of the partially purified bioflocculants revealed that the bioflocculants produced by Halobacillus sp. Mvuyo and Oceanobacillus sp. Pinky were glycoproteins, while that produced by Virgibacillus sp. Rob was a polysaccharide. We thus conclude that Halobacillus sp. Mvuyo, Virgibacillus sp. Rob and Oceanobacillus sp. Pinky hold promise as producers of new and efficient bioflocculant(s). We recommended development of process conditions for large scale production of the bioflocculants followed by their detailed characterization, as well as pilot scale assessment of the applicability of the purified bioflocculant in water/wastewater treatment and other industrial uses
- Full Text:
- Date Issued: 2010
Assessment of bioflocculant production by two marine bacteria isolated from the bottom sediment of marine Algoa Bay
- Authors: Ntozonke, Ncedo
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11298 , http://hdl.handle.net/10353/d1021290
- Description: Bioflocculants are polymers, mostly, of microbial origin which floc out suspended particles from liquid medium. The ability of these biopolymers to remove suspended particles from solutions is termed bioflocculation, and the efficiency of flocculation activities depends on the characteristics of the flocculants. In comparison with conventionally used flocculants, bioflocculants have the advantage of being safe (no toxic effects known), biodegradable and harmlessness to the environment. The study assessed production of bioflocculant by two marine bacteria from the bottom sediment of marine environment. The 16S rDNA was used for identification, and the two bacteria species were identified as Enterococcus hirae and Bacillus thuringiensis. Factors affecting the production and activity of the bioflocculants produced by these two organisms were studied. The bacteria optimally produced bioflocculant with fructose (91.7%) and urea (91%) as sole carbon and nitrogen sources respectively. Mg2+ (87%) and Ca2+ (86%), likewise, served as best cation sources on the production of the bioflocculant at pH 5(93%). Additionally, the flocculating activity of the bioflocculant increased with the addition of Mg2+ (81%) and Na+ (81%), and the highest flocculating activity was at pH 5 of the kaolin clay. The Fourier transform infrared spectroscopy (FTIR) shows that the bioflocculant is a glycoprotein. The second bacterium (Bacillus thuringiensis) produced bioflocculant optimally when the media had mixed nitrogen sources (Urea, ammonium chloride and tryptone (67%)) and glucose (85.65%) as a sole carbon source, also Ca2+ (74.6%) was the best cation that induced the production of bioflocculant. After purification, the bioflocculant flocculated optimally in alkaline pH 12 (81%) in the presence of Mn2+ (73%) and Ca2+ (72.8%). Chemical analysis of the bioflocculant revealed it to be a polysaccharide. Both bioflocculants flocculate efficiently and can be used to replace synthetic flocculants in water treatment, wastewater, in downstream processing, and processing of food and chemicals and other industrial uses of flocculants. Challenges though (i) are to develop conditions for large scale production of the bioflocculant, (ii) to do further characterization of the both bioflocculants (iii) to assess the bioflocculants for treatments of water/wastewater, and to apply it in various industrial processes.
- Full Text:
- Date Issued: 2015
- Authors: Ntozonke, Ncedo
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11298 , http://hdl.handle.net/10353/d1021290
- Description: Bioflocculants are polymers, mostly, of microbial origin which floc out suspended particles from liquid medium. The ability of these biopolymers to remove suspended particles from solutions is termed bioflocculation, and the efficiency of flocculation activities depends on the characteristics of the flocculants. In comparison with conventionally used flocculants, bioflocculants have the advantage of being safe (no toxic effects known), biodegradable and harmlessness to the environment. The study assessed production of bioflocculant by two marine bacteria from the bottom sediment of marine environment. The 16S rDNA was used for identification, and the two bacteria species were identified as Enterococcus hirae and Bacillus thuringiensis. Factors affecting the production and activity of the bioflocculants produced by these two organisms were studied. The bacteria optimally produced bioflocculant with fructose (91.7%) and urea (91%) as sole carbon and nitrogen sources respectively. Mg2+ (87%) and Ca2+ (86%), likewise, served as best cation sources on the production of the bioflocculant at pH 5(93%). Additionally, the flocculating activity of the bioflocculant increased with the addition of Mg2+ (81%) and Na+ (81%), and the highest flocculating activity was at pH 5 of the kaolin clay. The Fourier transform infrared spectroscopy (FTIR) shows that the bioflocculant is a glycoprotein. The second bacterium (Bacillus thuringiensis) produced bioflocculant optimally when the media had mixed nitrogen sources (Urea, ammonium chloride and tryptone (67%)) and glucose (85.65%) as a sole carbon source, also Ca2+ (74.6%) was the best cation that induced the production of bioflocculant. After purification, the bioflocculant flocculated optimally in alkaline pH 12 (81%) in the presence of Mn2+ (73%) and Ca2+ (72.8%). Chemical analysis of the bioflocculant revealed it to be a polysaccharide. Both bioflocculants flocculate efficiently and can be used to replace synthetic flocculants in water treatment, wastewater, in downstream processing, and processing of food and chemicals and other industrial uses of flocculants. Challenges though (i) are to develop conditions for large scale production of the bioflocculant, (ii) to do further characterization of the both bioflocculants (iii) to assess the bioflocculants for treatments of water/wastewater, and to apply it in various industrial processes.
- Full Text:
- Date Issued: 2015
Assessment of phytochemical constituents, antioxidant properties and cytotoxicity potentials of Leonutis leonurus and Boophone disticha extracts used in the management of Alzheimer’s disease in Hogsback, Eastern Cape, South Africa
- Authors: Tonisi, Sipho
- Date: 2019
- Subjects: Medicinal plants Alzheimer's disease
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/14726 , vital:40067
- Description: In this present study, the phytochemical compounds, antioxidant activities, and cytotoxicity potentials of solvent extracts (polar solvents) of Leonotis leonurus (L. leonurus) and Boophone disitcha (B. disticha) commonly used in the management of Alzheimer’s diseases in Hogsback, in the Eastern Cape, South Africa were investigated. The phytochemical compounds were determined by standard protocols of Folin–Ciocalteu method, aluminium chloride colorimetric method as well as Mayer’s and Wagner’s method. The antioxidant capabilities were assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and nitric oxide radical scavenging assays. The majority of extracts from both plant species exhibited free radical scavenging potential for oxidants they were tested against viz; DPPH, hydrogen peroxide, ABTS and nitric oxide. They further displayed good reducing potential against ferric iron. The results have shown a concentration-dependent dose of inhibition/scavenging activity (in DPPH, hydrogen peroxide, ABTS and nitric oxide assays) and reducing ability (in ferric reducing assay). However, leaf extracts of L. leonurus displayed ineffective scavenging activity against nitric oxide whereas extracts from L. leonurus flower and B. disticha bulb had higher scavenging potential against the production of nitric oxide radical. The GC-MS profiles of the plants extracts revealed the presence of numerous bioactive compounds that are known to possess excellent anti-inflammatory and most importantly antioxidant properties. Another interesting observation of GC-MS analysis data was the presence of compounds reported to inhibit Acetycholinesterase (AChE), which is a key enzyme targeted in the treatment of Alzheimers’ Disease (AD). The cytotoxicy tests for these extracts were carried out in order to assess the potentential detrimental effects on human health using Alamar Blue assay. It was noted that an increase in the extract concentration of both plant species resulted in a noticeable decline in cell population. These extracts were then declared xix to be quite toxic against HeLa (human cervix adenocarcinoma cells) cells as the cell viability in some extracts was quite low. Hence, toxicity testing of the studied plants’ extracts against normal cells is highly imperative in the future study. The results from this present study scientifically validated the use of the studied plants in folkoric medicine for the management of Alzheimer’s diseases.
- Full Text:
- Date Issued: 2019
- Authors: Tonisi, Sipho
- Date: 2019
- Subjects: Medicinal plants Alzheimer's disease
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/14726 , vital:40067
- Description: In this present study, the phytochemical compounds, antioxidant activities, and cytotoxicity potentials of solvent extracts (polar solvents) of Leonotis leonurus (L. leonurus) and Boophone disitcha (B. disticha) commonly used in the management of Alzheimer’s diseases in Hogsback, in the Eastern Cape, South Africa were investigated. The phytochemical compounds were determined by standard protocols of Folin–Ciocalteu method, aluminium chloride colorimetric method as well as Mayer’s and Wagner’s method. The antioxidant capabilities were assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and nitric oxide radical scavenging assays. The majority of extracts from both plant species exhibited free radical scavenging potential for oxidants they were tested against viz; DPPH, hydrogen peroxide, ABTS and nitric oxide. They further displayed good reducing potential against ferric iron. The results have shown a concentration-dependent dose of inhibition/scavenging activity (in DPPH, hydrogen peroxide, ABTS and nitric oxide assays) and reducing ability (in ferric reducing assay). However, leaf extracts of L. leonurus displayed ineffective scavenging activity against nitric oxide whereas extracts from L. leonurus flower and B. disticha bulb had higher scavenging potential against the production of nitric oxide radical. The GC-MS profiles of the plants extracts revealed the presence of numerous bioactive compounds that are known to possess excellent anti-inflammatory and most importantly antioxidant properties. Another interesting observation of GC-MS analysis data was the presence of compounds reported to inhibit Acetycholinesterase (AChE), which is a key enzyme targeted in the treatment of Alzheimers’ Disease (AD). The cytotoxicy tests for these extracts were carried out in order to assess the potentential detrimental effects on human health using Alamar Blue assay. It was noted that an increase in the extract concentration of both plant species resulted in a noticeable decline in cell population. These extracts were then declared xix to be quite toxic against HeLa (human cervix adenocarcinoma cells) cells as the cell viability in some extracts was quite low. Hence, toxicity testing of the studied plants’ extracts against normal cells is highly imperative in the future study. The results from this present study scientifically validated the use of the studied plants in folkoric medicine for the management of Alzheimer’s diseases.
- Full Text:
- Date Issued: 2019
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
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
Assessment of the prevalence of faecal coliforms and Escherichia coli o157:h7 in the final effluents of two wastewater treatment plants in Amahlathi Local Municipality of Eastern Cape Province, South Africa
- Authors: Ajibade, Adefisoye Martins
- Date: 2014
- Subjects: Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Escherichia coli O157:H7 , Escherichia coli , Effluent quality -- Testing , Whole effluent toxicity testing , Water -- Purification
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11283 , http://hdl.handle.net/10353/d1016166 , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Escherichia coli O157:H7 , Escherichia coli , Effluent quality -- Testing , Whole effluent toxicity testing , Water -- Purification
- Description: The production of final effluents that meet discharged requirements and guidelines remain a major challenge particularly in the developing world with the resultant problem of surface water pollution. This study assessed the physicochemical and microbiological qualities of two wastewater treatment works in the Eastern Cape Province of South Africa in terms of the prevalence of faecal coliforms and Escherichia coli O157:H7 over a five month period. All physicochemical and microbiological analyses were carried out using standard methods. Data were collected in triplicates and analysed statistically using IBM SPSS version 20.0. The ranges of some of the physicochemical parameters that complied with set guidelines include pH (6.7 – 7.6), TDS (107 – 171 mg/L), EC (168 – 266 μS/cm), Temperature (15 – 24oC), NO3- (0 – 8.2 mg/L), NO2- (0.14 – 0.71 mg/L) and PO4 (1.05 – 4.50 mg/L). Others including Turbidity (2.64 – 58.00 NTU), Free Cl (0.13 – 0.65 mg/L), DO (2.20 – 8.48 mg/L), BOD (0.13 – 6.85 mg/L) and COD (40 – 482 mg/L) did not comply with set guidelines. The microbiological parameters ranged 0 – 2.7 × 104 CFU/100 ml for FC and 0 – 9.3 × 103 for EHEC CFU/100 ml, an indication of non-compliance with set guidelines. Preliminary identification of 40 randomly selected presumptive enterohemorrhagic E. coli isolates by Gram’s staining and oxidase test shows 100% (all 40 selected isolates) to be Gram positive while 90% (36 randomly selected isolates) were oxidase negative. Statistical correlation between the physicochemical and the microbiological parameters were generally weak except in the case of free chlorine and DO where they showed inverse correlation with the microbiological parameters. The recovery of EHEC showed the inefficiency of the treatment processes to effectively inactivate the bacteria, and possibly other pathogenic bacteria that may be present in the treated wastewater. The assessment suggested the need for proper monitoring and a review of the treatment procedures used at these treatment works.
- Full Text:
- Date Issued: 2014
- Authors: Ajibade, Adefisoye Martins
- Date: 2014
- Subjects: Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Escherichia coli O157:H7 , Escherichia coli , Effluent quality -- Testing , Whole effluent toxicity testing , Water -- Purification
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11283 , http://hdl.handle.net/10353/d1016166 , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Escherichia coli O157:H7 , Escherichia coli , Effluent quality -- Testing , Whole effluent toxicity testing , Water -- Purification
- Description: The production of final effluents that meet discharged requirements and guidelines remain a major challenge particularly in the developing world with the resultant problem of surface water pollution. This study assessed the physicochemical and microbiological qualities of two wastewater treatment works in the Eastern Cape Province of South Africa in terms of the prevalence of faecal coliforms and Escherichia coli O157:H7 over a five month period. All physicochemical and microbiological analyses were carried out using standard methods. Data were collected in triplicates and analysed statistically using IBM SPSS version 20.0. The ranges of some of the physicochemical parameters that complied with set guidelines include pH (6.7 – 7.6), TDS (107 – 171 mg/L), EC (168 – 266 μS/cm), Temperature (15 – 24oC), NO3- (0 – 8.2 mg/L), NO2- (0.14 – 0.71 mg/L) and PO4 (1.05 – 4.50 mg/L). Others including Turbidity (2.64 – 58.00 NTU), Free Cl (0.13 – 0.65 mg/L), DO (2.20 – 8.48 mg/L), BOD (0.13 – 6.85 mg/L) and COD (40 – 482 mg/L) did not comply with set guidelines. The microbiological parameters ranged 0 – 2.7 × 104 CFU/100 ml for FC and 0 – 9.3 × 103 for EHEC CFU/100 ml, an indication of non-compliance with set guidelines. Preliminary identification of 40 randomly selected presumptive enterohemorrhagic E. coli isolates by Gram’s staining and oxidase test shows 100% (all 40 selected isolates) to be Gram positive while 90% (36 randomly selected isolates) were oxidase negative. Statistical correlation between the physicochemical and the microbiological parameters were generally weak except in the case of free chlorine and DO where they showed inverse correlation with the microbiological parameters. The recovery of EHEC showed the inefficiency of the treatment processes to effectively inactivate the bacteria, and possibly other pathogenic bacteria that may be present in the treated wastewater. The assessment suggested the need for proper monitoring and a review of the treatment procedures used at these treatment works.
- 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
Biodiversity of Salmonella strains isolated from selected water sources and wastewater discharge points in the Easern Cape Province of South Africa
- Authors: Mafu, Nwabisa Charity
- Date: 2008
- Subjects: Biodiversity conservation -- South Africa -- Eastern Cape , Biodiversity -- South Africa -- Eastern Cape , Salmonella typhimurium
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11248 , http://hdl.handle.net/10353/74 , Biodiversity conservation -- South Africa -- Eastern Cape , Biodiversity -- South Africa -- Eastern Cape , Salmonella typhimurium
- Description: In this study, the diversity of forty Salmonella isolates from selected drinking water and wastewater sources in the Eastern Cape Province of South Africa was assessed using parameters such as protein and lipopolysaccharide profile analysis, DNA fingerprinting and antibiotic susceptibility profile as test indices. Wastewater samples from Amalinda, Shornville and Fort Hare wastewater plants, and water samples from Gogogo and Tyume rivers were collected on ice and transported to the laboratory of the department of Microbiology and Biochemistry, University of Fort Hare for processing. The DNA dendograms of Salmonella and the applied UPGMA revealed 4 similarity groups of the strains. Most of the strains recovered from Amalinda, Shornville, Fort Hare wastewater plants, Gogogo and Tyume rivers show a high percentage of genetic similarity. On the other hand, protein dendograms of Salmonella isolates revealed 2 similarity groups which varied widely. Also, the lipopolysaccharide dendograms revealed three similarity groups with the first similarity groups showing a very high relatedness between strains from different water sources. The second similarity group included 16 strains which formed a rather homogenous group, and the third similarity group formed a distinct group. Of the seven antibiotics and sulfonamides tested against the Salmonella species, five namely, neomycin, chloramphenicol, kanamycin, streptomycin and cotrimoxazole were significantly inhibitory, while the bacteria showed considerable resistance to doxycycline and sulphamethoxazole. Our results based on restriction digestion, SDS/PAGE and dendogram construction show that there is a high similarity between the forty Salmonella strains studied, and that these methods are valuable tools for evaluating the relatedness ofSalmonella species. Our observations have proffered a veritable reference point on the diversity of Salmonella strains in the studied area.
- Full Text:
- Date Issued: 2008
- Authors: Mafu, Nwabisa Charity
- Date: 2008
- Subjects: Biodiversity conservation -- South Africa -- Eastern Cape , Biodiversity -- South Africa -- Eastern Cape , Salmonella typhimurium
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11248 , http://hdl.handle.net/10353/74 , Biodiversity conservation -- South Africa -- Eastern Cape , Biodiversity -- South Africa -- Eastern Cape , Salmonella typhimurium
- Description: In this study, the diversity of forty Salmonella isolates from selected drinking water and wastewater sources in the Eastern Cape Province of South Africa was assessed using parameters such as protein and lipopolysaccharide profile analysis, DNA fingerprinting and antibiotic susceptibility profile as test indices. Wastewater samples from Amalinda, Shornville and Fort Hare wastewater plants, and water samples from Gogogo and Tyume rivers were collected on ice and transported to the laboratory of the department of Microbiology and Biochemistry, University of Fort Hare for processing. The DNA dendograms of Salmonella and the applied UPGMA revealed 4 similarity groups of the strains. Most of the strains recovered from Amalinda, Shornville, Fort Hare wastewater plants, Gogogo and Tyume rivers show a high percentage of genetic similarity. On the other hand, protein dendograms of Salmonella isolates revealed 2 similarity groups which varied widely. Also, the lipopolysaccharide dendograms revealed three similarity groups with the first similarity groups showing a very high relatedness between strains from different water sources. The second similarity group included 16 strains which formed a rather homogenous group, and the third similarity group formed a distinct group. Of the seven antibiotics and sulfonamides tested against the Salmonella species, five namely, neomycin, chloramphenicol, kanamycin, streptomycin and cotrimoxazole were significantly inhibitory, while the bacteria showed considerable resistance to doxycycline and sulphamethoxazole. Our results based on restriction digestion, SDS/PAGE and dendogram construction show that there is a high similarity between the forty Salmonella strains studied, and that these methods are valuable tools for evaluating the relatedness ofSalmonella species. Our observations have proffered a veritable reference point on the diversity of Salmonella strains in the studied area.
- Full Text:
- Date Issued: 2008
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
Comparative in-vitro activities of trimethoprimsulfamethoxazole and the new fluoroquinolones against confirmed extended spectrum beta-lactamase producing Stenotrophomonas maltophilia in Nkonkobe Municipality, Eastern Cape environment
- Adeyemi, Oluwatosin Oluwakemi
- Authors: Adeyemi, Oluwatosin Oluwakemi
- Date: 2012
- Subjects: Antibiotics , Microbial sensitivity tests , Drug resistance in microorganisms , Pathogenic microorganisms , Gram-negative bacterial infections
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11275 , http://hdl.handle.net/10353/d1007576 , Antibiotics , Microbial sensitivity tests , Drug resistance in microorganisms , Pathogenic microorganisms , Gram-negative bacterial infections
- Description: Stenotrophomonas maltophilia is increasingly emerging as an opportunistic pathogen of global concern. Due to its inherent resistance to several classes of antibiotics including carbapenems and its ability to acquire mobile resistance elements, treatment of infections caused by S. maltophilia is a constant challenge for clinicians. Trimethoprim-sulphamethoxazole (TMP-SMX) is the generally accepted antibiotic of choice for the treatment of infections caused by this organism, but resistance to the drug is increasingly being reported; hence, the need for alternative therapeutic options. In this study, the antimicrobial susceptibility profile of 110 commensal S. maltophilia isolates obtained from Nkonkobe municipality, Eastern Cape Province, Republic of South Africa was investigated. Twenty-one antibiotics including TMP-SMX and the newer fluoroquinolones; levofloxacin, gatifloxacin and moxifloxacin were included in the antibiotic panel. About 63.4 percent of the isolates were susceptible to TMP-SMX with a resistance rate of 28.2 percent. The fluoroquinolones were more effective with susceptibilities ranging from 76 percent to 94.7 percent. Resistance to the fluoroquinolones ranged from 1.3 percent to 2.7 percent. Levofloxacin was the most effective fluoroquinolone tested. Phenotypic dectection of extended spectrum β-lactamases (ESBLs) showed double disc synergy test (DDST) positivity in 59.5 percent of the isolates. Cefepime was the most sensitive indicator cephalosporin in the DDST with 77.3 percent of suspected ESBL-producing isolates showing cefepime-clavulanic acid synergy. Isolates exhibited nine different ESBL phenotypes, however, PCR amplification of the bla genes revealed four isolates that possessed genes belonging to the CTX-M group (CTX-M-1 and CTX-M-8 groups). ESBL genes are usually carried on mobile elements such as plasmids and transposons which may also bear genes that mediate resistance to aminoglycosides, tetracyclines, TMP-SMX and fluoroquinolones. ESBL positive isolates appeared more susceptible to the fluoroquinolones compared to TMP-SMX but there was no significant relationship between ESBL production and susceptibility to these drugs (p > 0.05). The newer fluoroquinolones are a possible alternative treatment option for S. maltophilia infections in this environment but further studies and clinical investigations are needed to determine the in vivo efficacy of these drugs.
- Full Text:
- Date Issued: 2012
- Authors: Adeyemi, Oluwatosin Oluwakemi
- Date: 2012
- Subjects: Antibiotics , Microbial sensitivity tests , Drug resistance in microorganisms , Pathogenic microorganisms , Gram-negative bacterial infections
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11275 , http://hdl.handle.net/10353/d1007576 , Antibiotics , Microbial sensitivity tests , Drug resistance in microorganisms , Pathogenic microorganisms , Gram-negative bacterial infections
- Description: Stenotrophomonas maltophilia is increasingly emerging as an opportunistic pathogen of global concern. Due to its inherent resistance to several classes of antibiotics including carbapenems and its ability to acquire mobile resistance elements, treatment of infections caused by S. maltophilia is a constant challenge for clinicians. Trimethoprim-sulphamethoxazole (TMP-SMX) is the generally accepted antibiotic of choice for the treatment of infections caused by this organism, but resistance to the drug is increasingly being reported; hence, the need for alternative therapeutic options. In this study, the antimicrobial susceptibility profile of 110 commensal S. maltophilia isolates obtained from Nkonkobe municipality, Eastern Cape Province, Republic of South Africa was investigated. Twenty-one antibiotics including TMP-SMX and the newer fluoroquinolones; levofloxacin, gatifloxacin and moxifloxacin were included in the antibiotic panel. About 63.4 percent of the isolates were susceptible to TMP-SMX with a resistance rate of 28.2 percent. The fluoroquinolones were more effective with susceptibilities ranging from 76 percent to 94.7 percent. Resistance to the fluoroquinolones ranged from 1.3 percent to 2.7 percent. Levofloxacin was the most effective fluoroquinolone tested. Phenotypic dectection of extended spectrum β-lactamases (ESBLs) showed double disc synergy test (DDST) positivity in 59.5 percent of the isolates. Cefepime was the most sensitive indicator cephalosporin in the DDST with 77.3 percent of suspected ESBL-producing isolates showing cefepime-clavulanic acid synergy. Isolates exhibited nine different ESBL phenotypes, however, PCR amplification of the bla genes revealed four isolates that possessed genes belonging to the CTX-M group (CTX-M-1 and CTX-M-8 groups). ESBL genes are usually carried on mobile elements such as plasmids and transposons which may also bear genes that mediate resistance to aminoglycosides, tetracyclines, TMP-SMX and fluoroquinolones. ESBL positive isolates appeared more susceptible to the fluoroquinolones compared to TMP-SMX but there was no significant relationship between ESBL production and susceptibility to these drugs (p > 0.05). The newer fluoroquinolones are a possible alternative treatment option for S. maltophilia infections in this environment but further studies and clinical investigations are needed to determine the in vivo efficacy of these drugs.
- Full Text:
- Date Issued: 2012
Distribution of virulence determinants in Plesiomonas shigelloides and Vibrio species isolated from selected hospital wastewater effluents in the Eastern Cape Province, South Africa
- Authors: Mashudu, Mavhungu
- Date: 2018
- Subjects: Vibrio
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/14505 , vital:39996
- Description: Plesiomonas shigelloides and some Vibrio species are amongst the bacteria species classified into the emerging pathogenic bacteria grouping, and they portend increasing virulence and high resistance to some of the most potent antimicrobial agents used as most important and critical lines of defence against pathogenic infectious agents. Hospital wastewater has attracted significant attention as a pool for the regurgitation of pathogens into the environment which eventually becomes a problem to public health. It has been variously reported that hospital wastewater critically serves also as a puddle for microbial genetic interaction with outcomes including the development of resistance to antimicrobial agents among other pathogenic traits. And, in the same vein, exchanges and gene recombination may have been leading to the emergence of virulence determinants not previously known owing to the capacity of hospital wastewater to encompass a vast diversity of pathogenic bacteria and antimicrobial agents at the same material time. Hence, this research aimed at the evaluation of the prevalence of virulence and antibiotic resistance in Plesiomonas shigelloides and some human pathogenic Vibrio species isolated from selected hospital wastewater in the Eastern Cape Province of South Africa. Wastewater samples were collected at two weeks intervals throughout three months sampling duration from hospital wastewater facility in the Eastern Cape Province. The wastewater samples were from the secondary hospital and, tertiary hospital along with Limbede community wastewater facilities based in the Eastern Cape Province of South Africa. The wastewater samples were aseptically screened for the presence of presumptive Plesiomonas shigelloides and selected Vibrio species using the culture-based method. The identity of the presumptive Plesiomonas shigelloides and Vibrio species (Vibrio cholerae, V. fluvialis, V. parahaemolyticus, and V. vulnificus) isolates were confirmed using species specific primers. After that, the presence 14 of virulence determinants and the antimicrobial susceptibility profiles of the positive strains as well as the antimicrobial resistance factors were all determined. The distribution of the presumptive isolates was as follows; 322 Plesiomans shigelloides and 378 Vibrio species respectively. However, upon molecular confirmation studies, 85 out of 322; 26% of the total presumptive isolates, were confirmed as positive for Plesiomans shigelloides and, on the other hand, from the 378 presumptive Vibrio species, 71% (270) was as positive to Vibrio genus, and upon speciation of the positive isolates; 27% (73) was confirmed as Vibrio cholerae, 9% (24) as Vibrio parahaemolyticus, 4% (12) as Vibrio vulnificus and 3% (8) as Vibrio fluvialis. The remaining 57% (153) which was unaccounted for may belong to other Vibrio species not included in this studies. Various virulence determinants were shown to be present in the Vibrio species identified and the degree varied in line with the species. P. shigelloides showed high levels of resistance to impregnated antibiotics (18 different types). The antibiogram characteristics showed P. shigelloides to possess 100% resistance to Ciprofloxacin, Nalidixic acid, Trimethoprim, Ampicillin, Penicillin, Tetracycline, Doxycycline, Erythromycin, and Cephalothin. These findings are of crucial importance to public health and the science of infectology as it gives insight into the role of the hospital wastewater as a puddle for the recombination of antibiotic resistance determinants and virulence emergence in bacteria. On the strength of the finding of this research, it is essential to indicate that there is a need for an immediate review of hospital wastewater disposal process. Some form of treatment or pretreatment of the hospital wastewater is required before discharge into municipal wastewater treatment facilities. And, perhaps, continuous monitoring of hospital wastewater to reduce the risk posed to public health.
- Full Text:
- Date Issued: 2018
- Authors: Mashudu, Mavhungu
- Date: 2018
- Subjects: Vibrio
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/14505 , vital:39996
- Description: Plesiomonas shigelloides and some Vibrio species are amongst the bacteria species classified into the emerging pathogenic bacteria grouping, and they portend increasing virulence and high resistance to some of the most potent antimicrobial agents used as most important and critical lines of defence against pathogenic infectious agents. Hospital wastewater has attracted significant attention as a pool for the regurgitation of pathogens into the environment which eventually becomes a problem to public health. It has been variously reported that hospital wastewater critically serves also as a puddle for microbial genetic interaction with outcomes including the development of resistance to antimicrobial agents among other pathogenic traits. And, in the same vein, exchanges and gene recombination may have been leading to the emergence of virulence determinants not previously known owing to the capacity of hospital wastewater to encompass a vast diversity of pathogenic bacteria and antimicrobial agents at the same material time. Hence, this research aimed at the evaluation of the prevalence of virulence and antibiotic resistance in Plesiomonas shigelloides and some human pathogenic Vibrio species isolated from selected hospital wastewater in the Eastern Cape Province of South Africa. Wastewater samples were collected at two weeks intervals throughout three months sampling duration from hospital wastewater facility in the Eastern Cape Province. The wastewater samples were from the secondary hospital and, tertiary hospital along with Limbede community wastewater facilities based in the Eastern Cape Province of South Africa. The wastewater samples were aseptically screened for the presence of presumptive Plesiomonas shigelloides and selected Vibrio species using the culture-based method. The identity of the presumptive Plesiomonas shigelloides and Vibrio species (Vibrio cholerae, V. fluvialis, V. parahaemolyticus, and V. vulnificus) isolates were confirmed using species specific primers. After that, the presence 14 of virulence determinants and the antimicrobial susceptibility profiles of the positive strains as well as the antimicrobial resistance factors were all determined. The distribution of the presumptive isolates was as follows; 322 Plesiomans shigelloides and 378 Vibrio species respectively. However, upon molecular confirmation studies, 85 out of 322; 26% of the total presumptive isolates, were confirmed as positive for Plesiomans shigelloides and, on the other hand, from the 378 presumptive Vibrio species, 71% (270) was as positive to Vibrio genus, and upon speciation of the positive isolates; 27% (73) was confirmed as Vibrio cholerae, 9% (24) as Vibrio parahaemolyticus, 4% (12) as Vibrio vulnificus and 3% (8) as Vibrio fluvialis. The remaining 57% (153) which was unaccounted for may belong to other Vibrio species not included in this studies. Various virulence determinants were shown to be present in the Vibrio species identified and the degree varied in line with the species. P. shigelloides showed high levels of resistance to impregnated antibiotics (18 different types). The antibiogram characteristics showed P. shigelloides to possess 100% resistance to Ciprofloxacin, Nalidixic acid, Trimethoprim, Ampicillin, Penicillin, Tetracycline, Doxycycline, Erythromycin, and Cephalothin. These findings are of crucial importance to public health and the science of infectology as it gives insight into the role of the hospital wastewater as a puddle for the recombination of antibiotic resistance determinants and virulence emergence in bacteria. On the strength of the finding of this research, it is essential to indicate that there is a need for an immediate review of hospital wastewater disposal process. Some form of treatment or pretreatment of the hospital wastewater is required before discharge into municipal wastewater treatment facilities. And, perhaps, continuous monitoring of hospital wastewater to reduce the risk posed to public health.
- Full Text:
- Date Issued: 2018
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
Effects of genetically modified maize (MON810) and its residues on the functional diversity of microorganisms in two South African soils
- Authors: Puta, Usanda
- Date: 2011
- Subjects: Genetically modified foods -- South Africa , Transgenic plants -- South Africa , Crops -- Genetic engineering -- South Africa , Soil microbiology -- South Africa , Microorganisms , Microbial ecology , Rhizosphere -- Microbiology , Vesicular-arbuscular mycorrhizas , Corn -- South Africa
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11250 , http://hdl.handle.net/10353/419 , Genetically modified foods -- South Africa , Transgenic plants -- South Africa , Crops -- Genetic engineering -- South Africa , Soil microbiology -- South Africa , Microorganisms , Microbial ecology , Rhizosphere -- Microbiology , Vesicular-arbuscular mycorrhizas , Corn -- South Africa
- Description: Genetically modified (GM) crops are commercially cultivated worldwide but there are concerns on their possible negative impacts on soil biodiversity. A glasshouse study was conducted to determine effects of Bt maize residues on soil microbial diversity. Residues of Bt maize (PAN 6Q-308B) and non-Bt maize (PAN 6Q-121) were incorporated into the soil and corresponding maize seeds planted. The treatments were replicated three times. Fertilizer and water application were similar for both treatments. Rhizosphere and bulk soil was destructively sampled from each treatment and analyzed for microbial community level physiological profiles using Biolog plates with 31 different carbon substrates. Absorbance in the Biolog plates was recorded after 72 h of incubation at 20oC. Arbuscular mycorrhizal fungi spore counts were also determined. Field studies were conducted at the University of Free State and University of Fort Hare Research Farms to determine the effects of growing Bt maize on soil microbial diversity. One Bt maize cultivar (PAN6Q-308B) and non-Bt maize (PAN6Q-121) were grown in a paired experiment at University of Free State farm, while two Bt maize (DKC61-25B and PAN6Q-321B) and their near-isogenic non-Bt maize lines (DKC61-24 and PAN6777) were grown in a randomized complete block design with three replicates. Fertilization, weed control and water application, were similar for both Bt maize cultivars and their non-Bt maize counterparts. Rhizosphere soil samples were collected by uprooting whole plants and collecting the soil attached to the roots. The samples were analysed for microbial diversity and for arbuscular mycorrhizae fungal spore counts. Principal component analysis showed that soil microbial diversity was affected more by sampling time whereas genetic modification had minimal effects. Presence of residues also increased the diversity of microorganisms. Mycorrhizal fungal spores were not affected by the presence of Bt maize residues. Growing Bt maize had no effect on the soil microbial diversity in the rhizosphere.
- Full Text:
- Date Issued: 2011
- Authors: Puta, Usanda
- Date: 2011
- Subjects: Genetically modified foods -- South Africa , Transgenic plants -- South Africa , Crops -- Genetic engineering -- South Africa , Soil microbiology -- South Africa , Microorganisms , Microbial ecology , Rhizosphere -- Microbiology , Vesicular-arbuscular mycorrhizas , Corn -- South Africa
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11250 , http://hdl.handle.net/10353/419 , Genetically modified foods -- South Africa , Transgenic plants -- South Africa , Crops -- Genetic engineering -- South Africa , Soil microbiology -- South Africa , Microorganisms , Microbial ecology , Rhizosphere -- Microbiology , Vesicular-arbuscular mycorrhizas , Corn -- South Africa
- Description: Genetically modified (GM) crops are commercially cultivated worldwide but there are concerns on their possible negative impacts on soil biodiversity. A glasshouse study was conducted to determine effects of Bt maize residues on soil microbial diversity. Residues of Bt maize (PAN 6Q-308B) and non-Bt maize (PAN 6Q-121) were incorporated into the soil and corresponding maize seeds planted. The treatments were replicated three times. Fertilizer and water application were similar for both treatments. Rhizosphere and bulk soil was destructively sampled from each treatment and analyzed for microbial community level physiological profiles using Biolog plates with 31 different carbon substrates. Absorbance in the Biolog plates was recorded after 72 h of incubation at 20oC. Arbuscular mycorrhizal fungi spore counts were also determined. Field studies were conducted at the University of Free State and University of Fort Hare Research Farms to determine the effects of growing Bt maize on soil microbial diversity. One Bt maize cultivar (PAN6Q-308B) and non-Bt maize (PAN6Q-121) were grown in a paired experiment at University of Free State farm, while two Bt maize (DKC61-25B and PAN6Q-321B) and their near-isogenic non-Bt maize lines (DKC61-24 and PAN6777) were grown in a randomized complete block design with three replicates. Fertilization, weed control and water application, were similar for both Bt maize cultivars and their non-Bt maize counterparts. Rhizosphere soil samples were collected by uprooting whole plants and collecting the soil attached to the roots. The samples were analysed for microbial diversity and for arbuscular mycorrhizae fungal spore counts. Principal component analysis showed that soil microbial diversity was affected more by sampling time whereas genetic modification had minimal effects. Presence of residues also increased the diversity of microorganisms. Mycorrhizal fungal spores were not affected by the presence of Bt maize residues. Growing Bt maize had no effect on the soil microbial diversity in the rhizosphere.
- 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
Evaluation of incidence of Mycobacterium tuberculosis complex associated with soil, hayfeed and water in three Agricultural facilities in Amathole District Municipality in the Eastern Cape Province, South Africa
- Authors: Ntloko, Athini
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/756 , vital:26494
- Description: Mycobacterium bovis and other species of Mycobacterium tuberculosis complex (MTBC) can result to a zoonotic infection known as Bovine tuberculosis (bTB). MTBC has members that may contaminate an extensive range of hosts, including wildlife. Diverse wild species are known to cause disease in domestic livestock and are acknowledged as TB reservoirs. It has been a main study worldwide to deliberate on bTB risk factors as a result some studies focused on particular parts of risk factors such as wildlife and herd management. The objectives of this study were to design questionnaires from commercial farms and smallholding farms; isolate and identify MTBC from collected samples using culture and PCR assays recovered from Fort Hare, Middledrift and Seven star dairy farms; and assessing genotypic drug resistance through detection of mutations conferring resistance to INH and RMP associated with first line treatment for MTBC infection. Questionnaires were administered to thirty (30) smallholding farm owners in the two villages (kwaMasele and Qungqwala) and three (3) three commercial farms ( Fort Hare dairy farm, Middledrift dairy farm and Seven star dairy farm). Detection of M. tuberculosis complex was achieved by Polymerase Chain Reaction using primers for IS6110; whereas a genotypic drug resistance mutation was detected using Genotype MTBDRplus assays. Nine percent (9%) of respondents had more than 40 cows in their herd, while 60% reported between 10 and 20 cows in their herd. Relationship between farm size and vaccination for TB differed from forty one percent (41%) being the highest to the least five percent (5%). The highest number of respondents who knew about relationship between TB cases and cattle location was ninety one percent (91%). Approximately fifty one percent (51%) of respondents had knowledge about wild life access to the farms. Relationship between import of cattle and farm size ranged from nine percent (9%) to thirty five percent (35%). Cattle sickness in relation to farm size differed from forty three (43%) being the highest to the least three percent (3%); while thirty three percent (33%) of respondents had knowledge about health management. Respondents with knowledge about the occurrence of TB infections in farms were forty eight percent (48%). The frequency of DNA isolation from samples ranged from the highest forty five percent (45%) from water to the least twenty two percent (22%) from soil. Fort Hare dairy farm had the highest number of positive samples forty four percent (44%) from water samples; whereas Middledrift dairy farm had the lowest positive from water, seventeen percent (17%). Twelve (22%) out of 55 isolates showed resistance to INH and RMP that is, multi-drug resistance (MDR) and nine percent (9%) were sensitive to either INH or RMP. The mutations at rpoB gene differed from 58% being the highest to the least (23%). Fifty seven percent (57%) of samples showed a S315T1 mutation while only 14% possessed a S531L in the katG gene. The highest inhA mutations were detected in T8A (80%) eighty percent and the least was observed in A16G (17%). The results of this study reveals that risk factors for bTB in cattle and dairy farm workers is a serious issue abound in the Eastern Cape of South Africa; with the possibility of widespread dissemination of multidrug resistant determinants in MTBC from the environment.
- Full Text:
- Date Issued: 2015
- Authors: Ntloko, Athini
- Date: 2015
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: http://hdl.handle.net/10353/756 , vital:26494
- Description: Mycobacterium bovis and other species of Mycobacterium tuberculosis complex (MTBC) can result to a zoonotic infection known as Bovine tuberculosis (bTB). MTBC has members that may contaminate an extensive range of hosts, including wildlife. Diverse wild species are known to cause disease in domestic livestock and are acknowledged as TB reservoirs. It has been a main study worldwide to deliberate on bTB risk factors as a result some studies focused on particular parts of risk factors such as wildlife and herd management. The objectives of this study were to design questionnaires from commercial farms and smallholding farms; isolate and identify MTBC from collected samples using culture and PCR assays recovered from Fort Hare, Middledrift and Seven star dairy farms; and assessing genotypic drug resistance through detection of mutations conferring resistance to INH and RMP associated with first line treatment for MTBC infection. Questionnaires were administered to thirty (30) smallholding farm owners in the two villages (kwaMasele and Qungqwala) and three (3) three commercial farms ( Fort Hare dairy farm, Middledrift dairy farm and Seven star dairy farm). Detection of M. tuberculosis complex was achieved by Polymerase Chain Reaction using primers for IS6110; whereas a genotypic drug resistance mutation was detected using Genotype MTBDRplus assays. Nine percent (9%) of respondents had more than 40 cows in their herd, while 60% reported between 10 and 20 cows in their herd. Relationship between farm size and vaccination for TB differed from forty one percent (41%) being the highest to the least five percent (5%). The highest number of respondents who knew about relationship between TB cases and cattle location was ninety one percent (91%). Approximately fifty one percent (51%) of respondents had knowledge about wild life access to the farms. Relationship between import of cattle and farm size ranged from nine percent (9%) to thirty five percent (35%). Cattle sickness in relation to farm size differed from forty three (43%) being the highest to the least three percent (3%); while thirty three percent (33%) of respondents had knowledge about health management. Respondents with knowledge about the occurrence of TB infections in farms were forty eight percent (48%). The frequency of DNA isolation from samples ranged from the highest forty five percent (45%) from water to the least twenty two percent (22%) from soil. Fort Hare dairy farm had the highest number of positive samples forty four percent (44%) from water samples; whereas Middledrift dairy farm had the lowest positive from water, seventeen percent (17%). Twelve (22%) out of 55 isolates showed resistance to INH and RMP that is, multi-drug resistance (MDR) and nine percent (9%) were sensitive to either INH or RMP. The mutations at rpoB gene differed from 58% being the highest to the least (23%). Fifty seven percent (57%) of samples showed a S315T1 mutation while only 14% possessed a S531L in the katG gene. The highest inhA mutations were detected in T8A (80%) eighty percent and the least was observed in A16G (17%). The results of this study reveals that risk factors for bTB in cattle and dairy farm workers is a serious issue abound in the Eastern Cape of South Africa; with the possibility of widespread dissemination of multidrug resistant determinants in MTBC from the environment.
- Full Text:
- Date Issued: 2015
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