Development of a nystatin-loaded micellar system for oral mucoadhesion
- Authors: Sarpong, Nancy Owusu Akyere
- Date: 2019
- Subjects: Medical microbiology , HIV infections -- Treatment Clinical pharmacology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/43556 , vital:36918
- Description: Oropharyngeal Candidiasis (OPC) is an opportunistic fungal infection that affects mostly infants and immunocompromised patients. In recent years, the disease has been on the rise due to an increased life span, the HIV pandemic and the increased use of broad spectrum antibiotics. OPC may be treated using a nystatin suspension, which is not as effective as it could be due to the mechanism by which the suspension is applied to the affected tissue. This research aims to improve the effectiveness of nystatin, for the treatment of OPC, by proposing a nystatin loaded micellar system incorporated into a mucoadhesive system for drug delivery. This will ensure that nystatin is gradually released from the film, thus increasing retention time of nystatin at the affected area. In this study, a mixture design was developed, which was used to determine the most appropriate solvent system for nystatin solubilization. Optimisation of the micelle formulation was achieved by using the central composite rational design (CCRD). The two factors that were taken into consideration were the temperature of the hydration medium (water) and the length of time the micellar solution was exposed to the temperature environment of the rotary evaporator. The responses that were investigated were the mean particle size, mean polydispersity index (PDI), mean zeta potential, change in mean particle size and change in mean PDI. The optimal micelle formulation was characterised for size, stability, morphology and drug encapsulation efficiency. The micelles were found to be spherical and stable with an acceptable size range. However, their drug encapsulation efficiency was low. The mucoadhesive film was formulated and characterised for physical characteristics, pliability, percentage swelling index and drug release profile. The film was found to be highly pliable and evenly distributed with a smooth surface; no bumps or bubbles were visible. The film was able to swell to up to 550 %. In vitro studies showed that nystatin was gradually released from the film.
- Full Text:
- Date Issued: 2019
- Authors: Sarpong, Nancy Owusu Akyere
- Date: 2019
- Subjects: Medical microbiology , HIV infections -- Treatment Clinical pharmacology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/43556 , vital:36918
- Description: Oropharyngeal Candidiasis (OPC) is an opportunistic fungal infection that affects mostly infants and immunocompromised patients. In recent years, the disease has been on the rise due to an increased life span, the HIV pandemic and the increased use of broad spectrum antibiotics. OPC may be treated using a nystatin suspension, which is not as effective as it could be due to the mechanism by which the suspension is applied to the affected tissue. This research aims to improve the effectiveness of nystatin, for the treatment of OPC, by proposing a nystatin loaded micellar system incorporated into a mucoadhesive system for drug delivery. This will ensure that nystatin is gradually released from the film, thus increasing retention time of nystatin at the affected area. In this study, a mixture design was developed, which was used to determine the most appropriate solvent system for nystatin solubilization. Optimisation of the micelle formulation was achieved by using the central composite rational design (CCRD). The two factors that were taken into consideration were the temperature of the hydration medium (water) and the length of time the micellar solution was exposed to the temperature environment of the rotary evaporator. The responses that were investigated were the mean particle size, mean polydispersity index (PDI), mean zeta potential, change in mean particle size and change in mean PDI. The optimal micelle formulation was characterised for size, stability, morphology and drug encapsulation efficiency. The micelles were found to be spherical and stable with an acceptable size range. However, their drug encapsulation efficiency was low. The mucoadhesive film was formulated and characterised for physical characteristics, pliability, percentage swelling index and drug release profile. The film was found to be highly pliable and evenly distributed with a smooth surface; no bumps or bubbles were visible. The film was able to swell to up to 550 %. In vitro studies showed that nystatin was gradually released from the film.
- Full Text:
- Date Issued: 2019
Incidence of bacterial infections and colonisation in patients admitted to a tuberculosis hospital
- Authors: Annear, Dale John
- Date: 2018
- Subjects: Medical microbiology , Microbiology Bacteriology Tuberculosis -- Hospitals -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/21482 , vital:29526
- Description: Patients with drug resistant tuberculosis (TB) are treated with multiple antibiotics including moxifloxacin, linezolid, and meropenem, which puts them at greater risk for colonisation by multi-drug resistant (MDR) bacteria. The objectives of this study were to: (i) assess the antimicrobial prescribing patterns practiced within the hospital by retrospective patient file review; (ii) determine the spectrum of bacterial colonisation in TB patients upon admission and during hospitalisation; (iii) identify bacterial isolates and evaluate antimicrobial susceptibility profiles; (iv) detect antimicrobial resistance genes in the bacterial isolates by PCR and DNA sequencing; and (v) investigate genetic relatedness of Klebsiella pneumoniae isolates using Multi Locus Sequence Typing. Nasal, groin and rectal swabs [for the detection of extended spectrum beta lactamases (EBSLs), carbapenem-resistant Enterobacteriaceae (CRE), vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA)] were analysed from a cohort of patients (n=37) admitted either from the community (n = 28) or from other healthcare facilities (n=9) to a TB hospital. Swab samples were collected at admission and at four week intervals thereafter during hospitalization. Identification and antimicrobial susceptibility testing of bacterial isolates (n=62) were determined at the National Health Laboratory Services (NHLS) by the VITEK-MS and Vitek 2 systems respectively. Additional antimicrobial susceptibility testing was conducted by Sensititre Gram Negative Xtra (GNFX2) MIC plates. PCR and DNA sequencing were used for detection of resistance genes. Patients (n=13/37; 35%) were colonized by MDR bacteria (ESBLs [n=11], MRSA [n=2]) on admission. Colonization rates were lower in patients admitted from the community (9/28; 32%) compared to those transferred from other healthcare facilities (4/9; 44%). All admitted patients who did not exhibit colonization at baseline and who were resident within the hospital for longer than 4 weeks (17/37; 46% of total patients) became colonised by an ESBL-producing Enterobacteriaceae species. No patients acquired MRSA during hospitalisation. Among ESBL Enterobacteriaceae, Escherichia coli (41/62; 66%) and K. pneumoniae [14/62; 23%]) predominated. Nineteen percent (7/37) of patients demised during their hospitalization. Both the Vitek system and Sensititre Gram Negative Xtra (GNFX2) MIC plates susceptibilities were similar for most antimicrobials, however there were discrepancies for tigecycline susceptibility profiles. A high number of isolates exhibited resistance to aminoglycosides and fluoroquinolones. Genes encoding for ESBLs (CTX-M-14, CTX-M-15, SHV-28, OXA-1, and OXY-2-9) were detected among ESBL Enterobacteriaceae. Two Enterobacteriaceae isolates with reduced carbapenem susceptibility did not contain carbapenemase-encoding genes. MLST revealed unique sequence types and genetic diversity among the K. pneumoniae isolates from hospitalised patients. However, the source and colonization routes of these isolates could not be determined, which requires further investigation. This study provides insight into the spectrum of bacterial pathogen colonisation in hospitalised TB patients and suggests a review of infection control programs and practices at the TB hospital.
- Full Text:
- Date Issued: 2018
- Authors: Annear, Dale John
- Date: 2018
- Subjects: Medical microbiology , Microbiology Bacteriology Tuberculosis -- Hospitals -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10948/21482 , vital:29526
- Description: Patients with drug resistant tuberculosis (TB) are treated with multiple antibiotics including moxifloxacin, linezolid, and meropenem, which puts them at greater risk for colonisation by multi-drug resistant (MDR) bacteria. The objectives of this study were to: (i) assess the antimicrobial prescribing patterns practiced within the hospital by retrospective patient file review; (ii) determine the spectrum of bacterial colonisation in TB patients upon admission and during hospitalisation; (iii) identify bacterial isolates and evaluate antimicrobial susceptibility profiles; (iv) detect antimicrobial resistance genes in the bacterial isolates by PCR and DNA sequencing; and (v) investigate genetic relatedness of Klebsiella pneumoniae isolates using Multi Locus Sequence Typing. Nasal, groin and rectal swabs [for the detection of extended spectrum beta lactamases (EBSLs), carbapenem-resistant Enterobacteriaceae (CRE), vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA)] were analysed from a cohort of patients (n=37) admitted either from the community (n = 28) or from other healthcare facilities (n=9) to a TB hospital. Swab samples were collected at admission and at four week intervals thereafter during hospitalization. Identification and antimicrobial susceptibility testing of bacterial isolates (n=62) were determined at the National Health Laboratory Services (NHLS) by the VITEK-MS and Vitek 2 systems respectively. Additional antimicrobial susceptibility testing was conducted by Sensititre Gram Negative Xtra (GNFX2) MIC plates. PCR and DNA sequencing were used for detection of resistance genes. Patients (n=13/37; 35%) were colonized by MDR bacteria (ESBLs [n=11], MRSA [n=2]) on admission. Colonization rates were lower in patients admitted from the community (9/28; 32%) compared to those transferred from other healthcare facilities (4/9; 44%). All admitted patients who did not exhibit colonization at baseline and who were resident within the hospital for longer than 4 weeks (17/37; 46% of total patients) became colonised by an ESBL-producing Enterobacteriaceae species. No patients acquired MRSA during hospitalisation. Among ESBL Enterobacteriaceae, Escherichia coli (41/62; 66%) and K. pneumoniae [14/62; 23%]) predominated. Nineteen percent (7/37) of patients demised during their hospitalization. Both the Vitek system and Sensititre Gram Negative Xtra (GNFX2) MIC plates susceptibilities were similar for most antimicrobials, however there were discrepancies for tigecycline susceptibility profiles. A high number of isolates exhibited resistance to aminoglycosides and fluoroquinolones. Genes encoding for ESBLs (CTX-M-14, CTX-M-15, SHV-28, OXA-1, and OXY-2-9) were detected among ESBL Enterobacteriaceae. Two Enterobacteriaceae isolates with reduced carbapenem susceptibility did not contain carbapenemase-encoding genes. MLST revealed unique sequence types and genetic diversity among the K. pneumoniae isolates from hospitalised patients. However, the source and colonization routes of these isolates could not be determined, which requires further investigation. This study provides insight into the spectrum of bacterial pathogen colonisation in hospitalised TB patients and suggests a review of infection control programs and practices at the TB hospital.
- Full Text:
- Date Issued: 2018
Immunological and molecular characterization of Cryptosporidium species in HIV-Positive and HIV-Negative diarrhoea patients in the Nkonkobe Municipality of the Eastern Cape Province of South Africa: a pilot study
- Authors: Etinosa, Omoruyi Beauty
- Date: 2010
- Subjects: Protozoa, Pathogenic , Pathogenic microorganisms -- Detection , Medical microbiology , HIV-positive persons , Cryptosporidium , Diarrhea , HIV-infections
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11252 , http://hdl.handle.net/10353/392 , Protozoa, Pathogenic , Pathogenic microorganisms -- Detection , Medical microbiology , HIV-positive persons , Cryptosporidium , Diarrhea , HIV-infections
- Description: Cryptosporidiosis is an infection caused by Cryptosporidium; a protozoan parasite that infects the gastrointestinal tract. The infection is of major public health concern in both developed and developing countries. Faecal samples were collected from 160 in-patient adults, with complaint of diarrhoea, admitted at Victoria hospital in Alice, Nkonkobe Municipality. Twenty apparently healthy subjects were included as controls. All diarrhoea positive patients were interviewed to record socio-demographic information, water supply and animal contact. Initial screening was carried out by microscopy and ELISA to detect positive Cryptosporidium. Genomic DNA was extracted from microscopically positive samples and a PCR reaction was perform to amplify the (18S) SSUrRNA gene for further identification and epidemiology of Cryptosporidium. Data were analysed using Pearson‘s χ2 and Fisher‘s exact test to assess the univariate association between Cryptosporidium infection and the possible risk factors. Of the 180 subjects screened for cryptosporidial infection, Cryptosporidium antigen was detected in 122 giving an overall prevalence of 67.8 percent. In HIV-positive diarrhoea patients, prevalence increased with ages; between 31-43 (mean age 36.5 yr) and 70-82 (mean age 75.8 yr) had a higher prevalence (100 percent) of the antigen than 18-30 (mean age 23.2 yr) and 83-95 (mean age 88.8 yr) (50.0 percent) in HIV-positive diarrhoea patients (P > 0.05). In HIV-negative diarrhoea patients, prevalence was highest in the 18-30 (mean age 23.2 yr) (87.5 percent) and least (35.7 percent) in those aged 83-95 (mean age 88.8 yr) (P > 0.05). Cryptosporidium antigen was higher in females than in males. Of 115 females (mean age 46.7yr) who participated in the study, antigen was detected in 90 (78.2 percent) against 32 (71.1 percent) of 45 males (mean age 42.6yr). None of the 20 apparently healthy control subjects was found to be infected with Cryptosporidium. Cryptosporidium was detected in 27 HIV-positive and 97 HIV-negative diarrhoea patients by any one of the techniques. Antigen detection by ELISA 14 showed the highest positivity 96 (76.8 percent) in HIV- negative and 26 (74.3 percent) in HIV- positive diarrhoea patients. PCR detected eighty-nine (71.2 percent) cases in HIV-negative and 23 (65.7 percent) in HIV-positive patients with diarrhoea. Only 13 (37.1 percent) HIV-positive and 34 (27.2 percent) HIV-negative diarrhoea patients were found positive for Cryptosporidium by modified ZN. No significant difference was observed in sensitivity of antigen detection by ELISA and PCR (96.9 percent) in HIV-negative diarrhoea patients, respectively. Specificity of the staining technique was 88.9 percent in HIV-positive and 96.6 percent in HIV-negative diarrhoea patients. No significant difference was found in specificity of antigen detection by ELISA and PCR in HIV-positive and HIV-negative diarrhoea patients, respectively. Positive predictive value of ZN staining in both HIV-positive and HIV-negative diarrhoea patients (92.3 and 96.9 percent) was statistically higher than ELISA and PCR. No significant difference was observed in negative predictive value of ZN technique for detection of Cryptosporidium between HIV-positive and HIV- negative diarrhoea patients. Differences found in prevalence rates due to water source, suggest that the high infection rates of specific groups are associated with their exposure to the contaminated water supply. The results indicate that Cryptosporidium infection is highly prevalent in adult faecal specimens in the Nkonkobe Municipality, an indication of active infection that is likely to emerge as major human pathogen in this location due to socioeconomic changes which favour transmission. However, sequencing analysis is required to differentiate between Cryptosporidium genotypes in the various outbreaks
- Full Text:
- Date Issued: 2010
- Authors: Etinosa, Omoruyi Beauty
- Date: 2010
- Subjects: Protozoa, Pathogenic , Pathogenic microorganisms -- Detection , Medical microbiology , HIV-positive persons , Cryptosporidium , Diarrhea , HIV-infections
- Language: English
- Type: Thesis , Masters , MSc (Biochemistry)
- Identifier: vital:11252 , http://hdl.handle.net/10353/392 , Protozoa, Pathogenic , Pathogenic microorganisms -- Detection , Medical microbiology , HIV-positive persons , Cryptosporidium , Diarrhea , HIV-infections
- Description: Cryptosporidiosis is an infection caused by Cryptosporidium; a protozoan parasite that infects the gastrointestinal tract. The infection is of major public health concern in both developed and developing countries. Faecal samples were collected from 160 in-patient adults, with complaint of diarrhoea, admitted at Victoria hospital in Alice, Nkonkobe Municipality. Twenty apparently healthy subjects were included as controls. All diarrhoea positive patients were interviewed to record socio-demographic information, water supply and animal contact. Initial screening was carried out by microscopy and ELISA to detect positive Cryptosporidium. Genomic DNA was extracted from microscopically positive samples and a PCR reaction was perform to amplify the (18S) SSUrRNA gene for further identification and epidemiology of Cryptosporidium. Data were analysed using Pearson‘s χ2 and Fisher‘s exact test to assess the univariate association between Cryptosporidium infection and the possible risk factors. Of the 180 subjects screened for cryptosporidial infection, Cryptosporidium antigen was detected in 122 giving an overall prevalence of 67.8 percent. In HIV-positive diarrhoea patients, prevalence increased with ages; between 31-43 (mean age 36.5 yr) and 70-82 (mean age 75.8 yr) had a higher prevalence (100 percent) of the antigen than 18-30 (mean age 23.2 yr) and 83-95 (mean age 88.8 yr) (50.0 percent) in HIV-positive diarrhoea patients (P > 0.05). In HIV-negative diarrhoea patients, prevalence was highest in the 18-30 (mean age 23.2 yr) (87.5 percent) and least (35.7 percent) in those aged 83-95 (mean age 88.8 yr) (P > 0.05). Cryptosporidium antigen was higher in females than in males. Of 115 females (mean age 46.7yr) who participated in the study, antigen was detected in 90 (78.2 percent) against 32 (71.1 percent) of 45 males (mean age 42.6yr). None of the 20 apparently healthy control subjects was found to be infected with Cryptosporidium. Cryptosporidium was detected in 27 HIV-positive and 97 HIV-negative diarrhoea patients by any one of the techniques. Antigen detection by ELISA 14 showed the highest positivity 96 (76.8 percent) in HIV- negative and 26 (74.3 percent) in HIV- positive diarrhoea patients. PCR detected eighty-nine (71.2 percent) cases in HIV-negative and 23 (65.7 percent) in HIV-positive patients with diarrhoea. Only 13 (37.1 percent) HIV-positive and 34 (27.2 percent) HIV-negative diarrhoea patients were found positive for Cryptosporidium by modified ZN. No significant difference was observed in sensitivity of antigen detection by ELISA and PCR (96.9 percent) in HIV-negative diarrhoea patients, respectively. Specificity of the staining technique was 88.9 percent in HIV-positive and 96.6 percent in HIV-negative diarrhoea patients. No significant difference was found in specificity of antigen detection by ELISA and PCR in HIV-positive and HIV-negative diarrhoea patients, respectively. Positive predictive value of ZN staining in both HIV-positive and HIV-negative diarrhoea patients (92.3 and 96.9 percent) was statistically higher than ELISA and PCR. No significant difference was observed in negative predictive value of ZN technique for detection of Cryptosporidium between HIV-positive and HIV- negative diarrhoea patients. Differences found in prevalence rates due to water source, suggest that the high infection rates of specific groups are associated with their exposure to the contaminated water supply. The results indicate that Cryptosporidium infection is highly prevalent in adult faecal specimens in the Nkonkobe Municipality, an indication of active infection that is likely to emerge as major human pathogen in this location due to socioeconomic changes which favour transmission. However, sequencing analysis is required to differentiate between Cryptosporidium genotypes in the various outbreaks
- Full Text:
- Date Issued: 2010
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