STUDIES ON ANTIMICROBIAL AND HAEMOLYTIC ACTIVITIES, PROTEIN PROFILE AND TRANSCRIPTOMES OF AGELENOPSIS NAEVIA WALCKENAER, 1842 (GRASS SPIDER)VENOM

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dc.contributor.authorAHMED, JAMILA
dc.date.accessioned2017-05-17T08:14:25Z
dc.date.available2017-05-17T08:14:25Z
dc.date.issued2016-08
dc.descriptionA DISSERTATION SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY, ZARIA. IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF A MASTER DEGREE IN BIOLOGY DEPARTMENT OF BIOLOGICAL SCIENCES, FACULTY OF SCIENCE AHMADU BELLO UNIVERSITY, ZARIAen_US
dc.description.abstractThe study was carried out to investigate the antimicrobial and haemolytic activities,profile protein component of Agelenopsis naevia venom as well as transcript coding for protein in A. naevia venom. Venom was collected from the spiders by micro-dissection after homogenization of the venom gland and the concentration of venom was determined in a nanodrop spectrophotometer. Antimicrobial activity of venom againstBacilus subtilis, Candida albicans, and Salmonella typhi was carried out by disc diffusion and well diffusion assay. Haemolytic activity was carried out using purified 1% human erythrocyte. Crude venom was subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) using a precasted 4-20% gel. Gel was stained with commassie blue and destained to reveal the protein bands. Two dimension gel electrophoresis was also carried out in 13-cmimmobilized pH gradient(IPG) strip with a linear range of ƿH 3 to 10. Electrofocusing was carried out and the gel was rehydrated and subjected to a second dimension electrophoresis using a 15% SDS gel. The spots were visualized using commasie blue and detected using dynamic spot detector software. The messenger RNAs (mRNA) were isolated from venom gland and the first and second complimentary DNA (cDNA) strand were synthesized using ThermoScientific kit following manufacturer’s instructions. The cDNA library was constructed and purified. Pair-end sequencing was carried out using illumina NextSeq 500. The transcriptomes derived were searched against public databases using BLASTx alogrithm. Multiple sequence alignment andphylogenetic analysis was achieved using ClustalX and MEGA softwares respectively. Antimicrobial activity data were subjected to ANOVA to compare means, where significant, Duncan multiple range test was used to separate means. Effective concentration of crude venom was calculated using probit analysis. The crude venom of A. naevia showed significant activity against Bacillus subtilis(23.5±0.5) when compared to the Controls and no activity against Candida albicans and Salmonella typhi on the disc diffusion assay. However, the venom did not show activity against the three micro-organisms using well diffusion assay. The crude venom also showed haemolytic activity on human erythrocytes with activity within the first 1hr (42.40%) for all the three concentrations (0.579mg/ml, 2.843mg/ml and 4.044mg/ml) used. Percentage haemolysis ranged between 42.40%-52.52% within 1-6hr using three different concentrations. The venom has an EC50 of 2.07mg/ml. Six bands were evident on the SDS-PAGE gel with molecular weights ranging from below 6-64kDa. The crude venom showed both protein (>10kDa) and peptide (<10kDa) resolutions. Over 300 spots were detected on the 2D gel with molecular weight ranging from below 14kDa to 94kDa. Twelve transcriptomes homologous to sequences in databases were identified. Transcript from Agelenopsis naeviavenom gland clustered with that of Stegodyphus mimosarum. It was concluded that A. naevia venom have both antimicrobial and heamolytic activities. Its venom is made up of both proteins and peptides that are both cationic and anionicwhich could be harnessed as a potential bioinsecticide and antimicrobial agenten_US
dc.identifier.urihttp://hdl.handle.net/123456789/8995
dc.language.isoenen_US
dc.subjectSTUDIES,en_US
dc.subjectANTIMICROBIAL,en_US
dc.subjectHAEMOLYTIC ACTIVITIES,en_US
dc.subjectPROTEIN PROFILE,en_US
dc.subjectTRANSCRIPTOMES,en_US
dc.subjectAGELENOPSIS NAEVIA WALCKENAER,en_US
dc.subject1842,en_US
dc.subject(GRASS SPIDER)VENOM,en_US
dc.titleSTUDIES ON ANTIMICROBIAL AND HAEMOLYTIC ACTIVITIES, PROTEIN PROFILE AND TRANSCRIPTOMES OF AGELENOPSIS NAEVIA WALCKENAER, 1842 (GRASS SPIDER)VENOMen_US
dc.typeThesisen_US
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