Browsing by Author "Sumari, Deborah"
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Item Genetic Characterization of Osmotolerant Fermentative Saccharomyces Yeasts from Tanzania Suitable for Industrial Very High Gravity Fermentation(2010) Sumari, Deborah; Hosea, Ken M.; Magingo, F. S. S.Naturally occurring yeasts were sought from diverse Tanzanian environments and screened for industrial application. The yeasts were isolated from environments such as traditional brews and wines from various parts of Tanzania. In this regard, a total of thirty yeast isolates were screened for their suitability in Industrial Very High Gravity Fermentation (VHGF). Five of these isolates were able to grow and ferment medium with 40% initial sucrose concentration. Whereby, three were able to grow and ferment medium with 700 g/Litre (70% w/w) initial sucrose concentration. One of the three isolates coded LB2 isolated from a traditional Makonde sorghum brew was able to ferment a medium with initial sucrose concentration of 1000 g/Litre (100% w/w) at 30°C. On the basis of PCR-RFLP of the internal transcribed spacer (ITS) of the ribosomal DNA (rDNA), all the three most osmotolerant isolates were identified to belong to the Saccharomyces sensu stricto clade. Phylogenetic analysis of the 650 bp D1D2 domain of the large subunit 26S rDNA of the isolate LB2 clustered this isolate away from the so far known typical osmotolerant yeasts. The fermentation by LB2 isolate of 100% gravity substrate observed in this work is higher than any other encountered in the literatures reviewed.Item Genetic Diversity of Plasmodium falciparum Strains in Children under Five Years of Age in Southeastern Tanzania(2010) Sumari, Deborah; Hosea, Ken M.; Mugasa, Joseph P.; Abdulla, SalimStrain diversity may play a role in delaying development of protective immunity in endemic areas. We evaluated genetic diversity of Plasmodium falciparum infected children before being treated with Sulphadoxine Pyrimethamine (SP) and Coartem™ in Southeastern Tanzania. Allelic diversity of P. falciparum strains were determined in order to further assist in correct estimation of recrudescent and new infections. P. falciparum isolated from 300 children aged 1-59 months was used in the study, where nested PCR followed by Restriction Fragment Length Polymorphism (RFLP) of highly polymorphic Merozoite surface protein 2 (msp2) was employed to understand the genetic diversity of the parasites population. Frequency of msp2 gene alleles was calculated and further associated with multiplicity of infection of children under five years of age. A total of 71 and 83 different msp2 alleles were found in Rufiji and Ulanga districts respectively. Children infected with either FC27 or 3D7 allelic type in Rufiji were 42% single, 55.3% double and 2.7% triple, while in Ulanga, 36.7% single, 62% double and 1.3% triple infections. Mean numbers of multiplicity of infections (MOI) in Rufiji and Ulanga were 1.6 and 1.3, respectively. These findings show a high genetic diversity of P. falciparum strains in study areas and low MOI could reflect production of susceptible parasites that immune response can accommodate or can be cleared by the drugs. Furthermore, 3D7 allelic type of msp2 gene was more prevalent than FC27 in Ulanga district, indicating association between msp2 allelic type and disease severity, hence predict possible vaccine candidate in the future.Item Malaria prevalence in asymptomatic and symptomatic children in Kiwangwa, Bagamoyo district, Tanzania(Springer Nature, 2017-05-25) Sumari, Deborah; Mwingira, Felista; Majige, Selemani; Mugittu, Kefas; Mugasa, Joseph; Gwakisa, PaulBackground Malaria prevalence continues to decline across sub-Saharan Africa as a result of various intervention strategies. However, the diseases still poses a public health concern in the region. While symptomatic malaria is recognized and treated, asymptomatic infections become increasingly important for interrupting transmission. A cross-sectional survey was conducted to assess malaria prevalence in symptomatic and asymptomatic children in Kiwangwa ward in Bagamoyo District in Tanzania. Methods Four hundred school-aged children in Kiwanga ward were recruited in the study; 200 from Kiwangwa dispensary and 200 from nearby schools. Primary health parameters were examined and blood samples collected and examined for Plasmodium falciparum prevalence using rapid diagnostic test (RDT), light microscopy (LM) and reverse transcription quantitative PCR (RT-qPCR) targeting transcripts of A-type 18s rRNA of P. falciparum. Gametocytes were detected by LM and RT-qPCR targeting transcripts of gametocyte specific marker, Pfs25. Results Overall P. falciparum prevalence was 73.3, 40.8 and 36.3% by RT-qPCR, RDT and LM in the study area, respectively (P < 0.001). As expected symptomatic children had a significantly higher prevalence of 89, 67.5 and 64.5% by qPCR, RDT and LM, compared to 57.5, 14 and 8% in the asymptomatic group, respectively. However, gametocyte prevalence in asymptomatic individuals was higher by both LM (2%) and qPCR (14%) than in symptomatic individuals LM (0.5%) and qPCR (3%). Conclusions A substantial difference in prevalence of symptomatic and asymptomatic infections observed in Kiwangwa ward underpins the use of molecular tools in malaria surveillance aiming at estimating prevalence and transmission. Notably, the higher gametocytaemia observed in asymptomatic children indicates the reservoir infections and points to the need for detection and treatment of both asymptomatic and symptomatic malaria.Item Plasmodium falciparum msp1 , msp2 and glurp allele frequency and diversity in sub-Saharan Africa(Malaria Journal, 2011) Mwingira, Felista; Nkwengulila, Gamba; Schoepflin, Sonja; Sumari, Deborah; Beck, Hans-Peter; Snounou, Georges; Felger, Ingrid; Olliaro, Piero; Mugittu, KefasBackground: The efficacy of anti-malarial drugs is assessed over a period of 28-63 days (depending on the drugs’ residence time) following initiation of treatment in order to capture late failures. However, prolonged follow-up increases the likelihood of new infections depending on transmission intensity. Therefore, molecular genotyping of highly polymorphic regions of Plasmodium falciparum msp1, msp2 and glurp loci is usually carried out to distinguish recrudescence (true failures) from new infections. This tool has now been adopted as an integral part of anti-malarial efficacy studies and clinical trials. However, there are concerns over its utility and reliability because conclusions drawn from molecular typing depend on the genetic profile of the respective parasite populations, but this profile is not systematically documented in most endemic areas. This study presents the genetic diversity of P. falciparum msp1, msp2 and glurp markers in selected sub-Saharan Africa countries with varying levels of endemicity namely Malawi, Tanzania, Uganda, Burkina Faso and São Tomé. Methods: A total 780 baseline (Day 0) blood samples from children less than seven years, recruited in a randomized controlled clinical trials done between 1996 and 2000 were genotyped. DNA was extracted; allelic frequency and diversity were investigated by PCR followed by capillary electrophoresis for msp2 and fragment sizing by a digitalized gel imager for msp1 and glurp. Results and Conclusion: Plasmodium falciparum msp1, msp2 and glurp markers were highly polymorphic with low allele frequencies. A total of 17 msp1 genotypes [eight MAD20-, one RO33- and eight K1-types]; 116 msp2 genotypes [83 3D7 and 33 FC27- types] and 14 glurp genotypes were recorded. All five sites recorded very high expected heterozygosity (HE) values (0.68 - 0.99). HE was highest in msp2 locus (HE = 0.99), and lowest for msp1 (HE = 0.68) (P < 0.0001). The genetic diversity and allelic frequency recorded were independent of transmission intensity (P = 0.84, P = 0.25 respectively. A few genotypes had particularly high frequencies; however the most abundant showed only a 4% probability that a new infection would share the same genotype as the baseline infection. This is unlikely to confound the distinction of recrudescence from new infection, particularly if more than one marker is used for genotyping. Hence, this study supports the use of msp1, msp2 and glurp in malaria clinical trials in sub-Saharan Africa to discriminate new from recrudescent infections.Item Plasmodium Falciparum Msp1, Msp2 And Glurp Allele Frequency and Diversity in Sub-Saharan Africa(BioMed Central, 2011) Mwingira, Felista; Nkwengulila, Gamba; Schoepflin, Sonja; Sumari, Deborah; Beck, Hans-Peter; Snounou, Georges; Felger, Ingrid; Olliaro, Piero; Mugittu, KefasThe efficacy of anti-malarial drugs is assessed over a period of 28-63 days (depending on the drugs’ residence time) following initiation of treatment in order to capture late failures. However, prolonged follow-up increases the likelihood of new infections depending on transmission intensity. Therefore, molecular genotyping of highly polymorphic regions of Plasmodium falciparum msp1, msp2 and glurp loci is usually carried out to distinguish recrudescence (true failures) from new infections. This tool has now been adopted as an integral part of anti-malarial efficacy studies and clinical trials. However, there are concerns over its utility and reliability because conclusions drawn from molecular typing depend on the genetic profile of the respective parasite populations, but this profile is not systematically documented in most endemic areas. This study presents the genetic diversity of P. falciparum msp1, msp2 and glurp markers in selected sub-Saharan Africa countries with varying levels of endemicity namely Malawi, Tanzania, Uganda, Burkina Faso and São Tomé. A total 780 baseline (Day 0) blood samples from children less than seven years, recruited in a randomized controlled clinical trials done between 1996 and 2000 were genotyped. DNA was extracted; allelic frequency and diversity were investigated by PCR followed by capillary electrophoresis for msp2 and fragment sizing by a digitalized gel imager for msp1 and glurp. Plasmodium falciparum msp1, msp2 and glurp markers were highly polymorphic with low allele frequencies. A total of 17 msp1 genotypes [eight MAD20-, one RO33- and eight K1-types]; 116 msp2 genotypes [83 3D7 and 33 FC27- types] and 14 glurp genotypes were recorded. All five sites recorded very high expected heterozygosity (HE) values (0.68 - 0.99). HE was highest in msp2 locus (HE = 0.99), and lowest for msp1 (HE = 0.68) (P < 0.0001). The genetic diversity and allelic frequency recorded were independent of transmission intensity (P = 0.84, P = 0.25 respectively. A few genotypes had particularly high frequencies; however the most abundant showed only a 4% probability that a new infection would share the same genotype as the baseline infection. This is unlikely to confound the distinction of recrudescence from new infection, particularly if more than one marker is used for genotyping. Hence, this study supports the use of msp1, msp2 and glurp in malaria clinical trials in sub-Saharan Africa to discriminate new from recrudescent infections.