Antibiotic resistance in bacteria is a growing threat to global health. Many of the genes responsible for resistance are carried on mobile genetic elements which can be transferred laterally between strains and species. The most important of these are conjugative and mobilisable elements including plasmids and integrating and conjugating elements, ICEs. Haemophi/us influenzae is an important human pathogen, which was first identified as carrying antibiotic resistance genes in the 1970s. Much of this resistance is encoded by ICEHin1056, which is present in H. influenzae strains worldwide. The aims of this study were to describe features of the biology of ICEHin1056, with particular reference to the genetic site and control mechanisms responsible for instigating conjugative transfer. The origin of transfer has been localised to a sequence on ICEHin1056 and an environmental stressor initiating conjugative transfer, oxidative stress, has been identified. In addition, detailed phylogenetic analysis has demonstrated ICEHin1056 to be part of a much larger family of mobile genetic elements, widely distributed in proteobacteria and carrying accessory genes responsible for survival in adverse environments, virulence and antibiotic resistance. The ICEs in the family have conserved homology of gene content and synteny of gene arrangement over deep evolutionary time, challenging the accepted paradigm of modular mosaicism of mobile genetic elements. A key event in increasing dissemination of the ICE, acquisition of a phage type integrase gene has also been identified. The findings presented provide significant insight into the behaviour of ICEs and may in future allow predictions about the spread of virulence factors and antibiotic resistance genes, with important implications for human and animal health.

Background: The use of antimicrobials has been presented as a significant risk factor for Clostridium difficile infection (CDI). Nevertheless, it remains unclear which antimicrobials are more likely to be associated with CDI. It is mandatory for acute trusts to report the numbers of diagnosed CDI cases to the Health Protection Agency (HPA). There is no national system to collect and analyse antimicrobial usage data from the trusts. The company IMS collects antimicrobial usage data from the trusts for creating marketing research statistics. Therefore, it was hypothesised that data collected from the HPA and from IMS could be used to undertake an ecological study about the association between CDI cases and antimicrobial use in English trusts. Methods: A trust-level Antimicrobial Usage Database provided by IMS and a database, including the numbers of CDI cases for patients aged 65 years old and above, provided by the HPA, were utilised in this work. These referred to 26 out of the 29 NW SHA trusts (that managed 64 hospitals) for the quarters between 2005 and 2008 inclusive. A sample of antimicrobial usage data collected directly from trusts was used to investigate potential limitations in using the Antimicrobial Usage Database for the purpose of this work. Multilevel models were used to study antimicrobial usage and the number of CDI cases over time. These models were also used to investigate the association between the CDI cases and antimicrobial usage in the trusts. The trends of trust antimicrobial usage over time were compared with DH recommendations for the prevention of CDI through antimicrobial prescribing published in 1994, 2005 and 2008. Results: Discrepancies between the antimicrobial usage recorded in the IMS database and in a sample of antimicrobial usage data collected from trusts were found for 31 out of 155 antimicrobial usage records; only 1 of these referred to an antimicrobial with high usage. Eight out of the 23 antimicrobial groups and 10 out of 63 antimicrobials were presented as having high usage. The antimicrobial usage over time increased significantly for 7 antimicrobial groups, decreased significantly for 2 groups and remained constant for 54 groups. The number of CDI cases reported for patients aged 65 years old and above decreased significantly over the time. Trust antimicrobial usage over time changed in the opposite direction compared to the DH recommendations published in 1994, 2004 and 2008, respectively, for 2 out of 11, 3 out of 12 and 3 out of 14 antimicrobial groups/antimicrobials. The increased usage of 5 antimicrobial groups was significantly associated with an increase in the number of CDI cases and an increased usage of 4 antimicrobial groups was significantly associated with a decreased number of CDI cases. Within the antimicrobial groups that were significantly associated with an increased number of CDI cases, the usage of 8 individual antimicrobials was significantly associated with the CDI cases. Discussion/Conclusion: Collecting antimicrobial usage over time for large groups of trusts is very time consuming and requires extensive data manipulation. The similarity of the results of this study with those of previously published studies suggest that HPA and IMS data may be used to investigate the association between CDI cases and antimicrobial usage in English trusts.

M. Tech. (Chemistry, Department of Chemistry), Vaal University of Technology / Infectious diseases, a group of illnesses caused by specific pathogens or its toxins is a leading cause of death globally. Treatment with antibiotics is a key intervention in the control and management of many infectious disease. However, the increasing incidence of antibiotics failure, due to the emergence of drug resistant pathogens, is rendering the use of antibiotics chemotherapy ineffective. A possible solution is to synthesize new compounds with broad spectrum characteristics and superior drug performances as alternative to conventional antibiotics. Schiff Bases are biologically active ligands. They form metal complexes with superior biological activities. This research aims to synthesize some Schiff Base metal complexes and investigate their biological effects on Staphylococcus aureus and Enterococcus faecalis. Metal acetylacetonates of Vanadium, Copper, Cobalt, Zinc, Magnesium, Manganese, Cadmium, Nickel and Iron were synthesized and characterized by Fourier transform infrared spectroscopy. Four Schiff bases, LI, L2, L3 and L4 were also synthesized by the condensation of 4- (diethylamino)-2-hydroxybenzaldehyde with 4-nitrobenzohydrazide and 4-methoxybenzohydrazide to form L1 and L2. 4-(dimethylamino) benzaldehyde was reacted with 4-nitrobenzohydrazide and 4-methoxybenzohydrazide to form L3 and L4 respectively. The Schiff base ligands were then reacted with synthesized Vanadium, manganese, cobalt and magnesium acetylacetonates to form Schiff base complexes (SBC 1A to 4D). Schiff bases ligands and complexes were characterized by FT-IR, 1H-NMR, 13C-NMR, TGA and DTA. Fourier Transform infrared spectroscopy (FTIR) of the acetylacetonates showed the formation of metal acetylacetonates as characterized by the absence of the carbonyl stretching n(C=O) vibration in metal acetylacetonate spectra as compared to pure acetylacetone. Metal acetylacetonates also showed the presence of metal oxygen vibration frequency, n(M-O-C), in the spectra obtained. Thermogravimetric analysis (TGA) and Derivative or Differential Thermogravimetric analysis (DTA) of the Schiff base ligands showed the presence of a single decomposition product in L1, L2, L3 and L4 indicating the formation of a single reaction product while those of Schiff base complexes showed the formation of several decomposition products. Proton and carbon thirteen Nuclear Magnetic Resonance (1H- and 13C-NMR) spectroscopy of the Schiff base ligands indicated the presence of hydrogen and carbon-13 in different environments. The chemical shifts of the hydrogens and carbon-13 provided evidence that Schiff base ligands were formed. The strongest evidence is the presence of the azomethine hydrogen and carbon in the spectra of the Schiff base ligands. The presence of aromatic hydrogens and carbon at chemical shift environments found in literature also confirmed the formation of Schiff base ligand. The NMR spectra of Schiff base complexes showed the presence of azomethine (HC=N) and aromatic hydrogens at expected chemical shifts. The synthesized Schiff bases and their corresponding metal complexes were screened for their invitro antibacterial activities against two Gram-positive (Staphylococcus aureues and Enterococcus feacalis) bacterial strains by the Agar-well diffusion methods.The ligands and complexes were tested against confirmed S. aureus and E. faecalis strains and only 4 exhibited antimicrobial activities. The ligands and complexes were effective against the S. aureusand E. faecalis isolates. / VUT

BACKGROUND AND OBJECTIVES: Streptococcus agalactiae (Group B Streptococcus, GBS) is the leading cause of neonatal infections and deaths in human. It can also cause infections in pregnant women and non-pregnant adults. Penicillin and ampicillin are antibiotics of choice for the treatment of GBS infections. Erythromycin and clindamycin are used as alternative therapy in penicillin allergic patients, however resistance to these agents has been increasingly observed. This present study was undertaken to determine the colonization rate of GBS, susceptibility profile and the mechanism of antibiotic resistance in pregnant women and their babies at Dr. George Mukhari Academic Hospital in Pretoria. METHODS: Rectal and vaginal swabs were collected from pregnant women; ear and umbilical swabs from newborns over an 11 month period. Samples were cultured on selective media (CNA agar and Todd-Hewitt broth) and GBS positively identified using morphological and biochemical tests including Gram staining, hemolytic activity, catalase test, bile esculin, CAMP test and Latex agglutination test. The susceptibility testing was done using the Kirby-Bauer and E-test methods. The D-test method was used to determine the inducible clindamycin resistance. Multiplex PCR with were used to detect different genes coding for resistance. RESULTS: Out of the 413 patients evaluated, 128 (30.9%) were positive with GBS. All isolates were sensitive to penicillin and ampicillin. Erythromycin and clindamycin resistance was 21.1% and 17.2% respectively; of which 69% harbouring constitutive MLBB, 17.4% inducible MLSB. The alteration of ribosomal target encoded by ermB genes was the commonest mechanism of resistance observed in 55% of isolates, 38% of isolates had both ermB and linB genes and efflux pump mediated by mefA genes was detected in one of isolates. Conclusion: This study reaffirms the appropriateness of penicillin as the antibiotic of choice for treating GBS infection. However it raises the challenges of resistance to the macrolides and lincosamides. More GBS treatment options for penicillin allergic patients need to be researched. / Health Studies / M.Sc. (Life Sciences (Microbiology))

BACKGROUND AND OBJECTIVES: Streptococcus agalactiae (Group B Streptococcus, GBS) is the leading cause of neonatal infections and deaths in human. It can also cause infections in pregnant women and non-pregnant adults. Penicillin and ampicillin are antibiotics of choice for the treatment of GBS infections. Erythromycin and clindamycin are used as alternative therapy in penicillin allergic patients, however resistance to these agents has been increasingly observed. This present study was undertaken to determine the colonization rate of GBS, susceptibility profile and the mechanism of antibiotic resistance in pregnant women and their babies at Dr. George Mukhari Academic Hospital in Pretoria. METHODS: Rectal and vaginal swabs were collected from pregnant women; ear and umbilical swabs from newborns over an 11 month period. Samples were cultured on selective media (CNA agar and Todd-Hewitt broth) and GBS positively identified using morphological and biochemical tests including Gram staining, hemolytic activity, catalase test, bile esculin, CAMP test and Latex agglutination test. The susceptibility testing was done using the Kirby-Bauer and E-test methods. The D-test method was used to determine the inducible clindamycin resistance. Multiplex PCR with were used to detect different genes coding for resistance. RESULTS: Out of the 413 patients evaluated, 128 (30.9%) were positive with GBS. All isolates were sensitive to penicillin and ampicillin. Erythromycin and clindamycin resistance was 21.1% and 17.2% respectively; of which 69% harbouring constitutive MLBB, 17.4% inducible MLSB. The alteration of ribosomal target encoded by ermB genes was the commonest mechanism of resistance observed in 55% of isolates, 38% of isolates had both ermB and linB genes and efflux pump mediated by mefA genes was detected in one of isolates. Conclusion: This study reaffirms the appropriateness of penicillin as the antibiotic of choice for treating GBS infection. However it raises the challenges of resistance to the macrolides and lincosamides. More GBS treatment options for penicillin allergic patients need to be researched. / Health Studies / M. Sc. (Life Sciences (Microbiology))