Developing DNA barcoding (matK) primers for marama bean [Tylosema esculentum (Burchell) Schreiber]

DNA barcoding is based on the premise that a short standardized DNA barcoding sequence can distinguish individuals of a species because the genetic variation between species exceeds that within species. Information on genetic variation of breeding materials helps to maintain genetic diversity and sustains long term selection gain. This information is a prerequisite for the genetic improvement of any plant species for effective use of germplasm in breeding and for conservation. Marama bean [Tylosema esculentum (Burchell) Schreiber] is found in the arid, dry parts of Southern Africa and due to the high nutrient value of the seeds and tubers, richness in protein, oil and starch, it is a potential crop for arid areas where few conventional crops can survive. The effective conservation and use of marama bean genetic resources for domestication involves investigating the extent of genetic variation. The matK gene, formerly known as orfK, is emerging as a DNA barcoding gene with potential contribution to plant molecular systematics and evolution. The gene matK, approximately 1500 base pairs (bp), is believed to code for a maturase-related protein based on structural similarities to other such genes. This gene was investigated for potential contribution in genetic variation studies of marama bean and also establishing a barcode for T. esculentum. The matK gene was amplified in marama bean and we reported herein, the first record of sequences of this gene for the species that were found to be related to other legume matK sequences deposited in GenBank. The homology found with Tylosema fassoglensis (trnK gene) and Pisum sativum (matK gene) suggests that an identical region was amplified for Tylosema esculentum. A phylogenetic tree was constructed based on the matK sequences and the results suggest that the matK region can also be used in determining levels of genetic variation and for barcoding.Key words: Marama bean, DNA barcoding, genetic variation, maturase kinase

Infrastructure development in economic crisis and recovery : the rail cargo sector in Zimbabwe since 2000.

Thesis (M.Dev.Studies)-University of KwaZulu-Natal, Durban, 2014.The study sought to investigate infrastructure development in economic crisis and recovery, with specific regard to the rail cargo sector in Zimbabwe since 2000. The research was prompted by firstly, the need to assess the current situation of the National Railways of Zimbabwe (NRZ), secondly, to examine the root cause of the challenges being faced by the National Railways of Zimbabwe and lastly, to review the plans going forward to resolve the challenges. In the study, a sample of 17 respondents was selected. Questions were used to guide the interview schedule. The data was collected through semi-structured interviews and was presented and analysed using thematic analysis. The findings were then interpreted in relation to research objectives and questions. The research findings showed that most of the challenges that were being experienced by the National Railways of Zimbabwe were as a result of the economic meltdown of the Zimbabwean economy. It is evident from the findings that the rail cargo in Zimbabwe can be improved if the parastatal can be given autonomy to run the day to day operations without political interference from the government. The results further suggest that the National Railways of Zimbabwe can be in a position to breakeven or become financially viable if the government, who is the major shareholder, opens up lines or credit and helps secure strategic partners who will bring in capital. In light of the above findings, the study recommends that the government explore more private sector participation and private concession options as this will help the National Railways of Zimbabwe improve its current challenges. Secondly, the 1997 Railway Act has to be signed and officialised so as to enable the organisation to operate commercially. Thirdly, the staff composition and the election of the Board of Directors should be revisited so as to engage to stakeholders in promoting transparency, integrity and efficiency. Lastly, the National Railways of Zimbabwe needs to revive its Railway Motor Services Private Limited because it will help the organisation to cut down on competition from the road sector

The role of Anopheles arabiensis (Diptera: Culicidae) in malaria transmission and control in Gokwe and Binga districts, Zimbabwe..

Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1996.Opportunistic feeding behaviour and partial exophily make An. arabiensis much more difficult to control by indoor residual spraying than any other vector in the Afro-tropical region. The persistent malaria outbreaks in Zimbabwe despite decades of indoor house spraying prompted this investigation into the role of An. arabiensis in malaria transmission and assessment of the possible impact of this control measure. The study was conducted in the malaria endemic districts of Binga and Gokwe. An. gambiae complex mosquitoes were collected from artificial outdoor resting sites, and from human dwellings by i) daytime hut searches, ii) pyrethrum spray catches and iii) exit window traps. Mosquito components were processed to enable: i) the distinction of An. arabiensis from An. quadriannulatus and An. merus on the basis of the pale band at the junction of the hind leg 3/4 tarsomeres; ii) species identification and scoring of inversion polymorphism on the basis of the X chromosome and autosomes respectively; iii) the determination of blood meal sources using the Ouchterlony precipitin test; and iv) identification of An. gambiae s.l. using polymerase chain reaction (PCR) and enzyme electrophoresis techniques. Entomological assessment of residual spraying included determining: the vector resting densities indoors and outdoors, bioassay and insecticides susceptibility tests. Data were also collected on hut profiles, knowledge-attitudes-practices surveys, and household malaria prevalence . surveys. An. arabiensis and An. quadriannulatus were found in sympatry in Binga and Gokwe, and in addition, An. merus was found in Gokwe. Most species identifications were made using PCR; which was found to have 7.5% and 41.6% levels of error for An. arabiensis and An. quadriannulatus respectively, using the cytogenetic technique as benchmark. The pale band technique yielded > 80% correct identification for An. arabiensis but the extent of overlap in the pale band lengths between An. arabiensis and An. quadriannulatus renders the method unsuitable for distinguishing these two species. Inversions 2Rb and 3Ra were found floating in An. arabiensis, with 60% frequency in the former. The Wright's F statistic value of -0.0416 indicated an excess of heterozygotes, and a state of panmixis in the vector population. No significant differences were observed between 2Rb karyotypes in host choice. Human blood indices among indoor (0.82), exit trap (0.98) and outdoor resting (0.30) specimens suggested exophilic behaviour. This was corroborated by the high fed:gravid ratios of 6.8: 1 and 11.6: 1 in sprayed and non-sprayed dwellings respectively. This was worsened by a high feeder-survivor index (FSI) of 93 % among exit trap specimens. The susceptibility to deltamethrin coupled with residual efficacy nine weeks post-spray indicated the suitability of the insecticide. Rural dwellings were suitably built for spraying but had no mosquito proofing. Personal protective measures are hardly known; sleeping outdoors occurs in Siabuwa. While An. arabiensis bites humans indoors the partial exophily it exhibits is a threat to indoor residual insecticide spraying. An integrated malaria control approach is recommended

A review of geographical distribution of marama bean [Tylosema esculentum (Burchell) Schreiber] and genetic diversity in the Namibian germplasm

Marama bean [Tylosema esculentum (Burchell) Schreiber] occurs naturally in the drier areas of Southern Africa, including Botswana and Namibia. The implementation of molecular breeding is important to advance the process of securing the world’s food supply. The development of molecular markers is vital for mapping important traits that can then be followed in subsequent breeding programs. This study assessed the distribution of marama bean in Namibia and isolated microsatellite regions for microsatellite primer design for the purpose of determining genetic diversity and construction of molecular genetic maps for marama. This will greatly enhance the process ofdomesticating marama bean, currently a wild plant that is still picked wild and unsustainably. The geographical distribution was geo-referenced using Geographic Positioning System (GPS) points andmicrosatellites were isolated from the germplasm using a modified FIASCO technique. The study revealed widespread, but patchy distribution of marama bean in Namibia. Five Marama beanmicrosatellite enriched libraries were created. The initial results provided vital information for the ongoing marama bean conservation function and improvement program

Cholera: A comparison of the 2008-9 and 2010 Outbreaks in Kadoma City, Zimbabwe

Introduction: Kadoma City experienced cholera outbreaks in 2008-9, and 2010, affecting 6,393 and 123 people, respectively. A study wasconducted to compare epidemiology of the cholera outbreaks. Methods: a descriptive cross sectional study was conducted, analyzing line list data for the 2 outbreaks. Proportions, means were generated and compared using the Chi Square test at 5% level of significance.Results: cholera cases were similar by gender and age, with the 20-30 years group being most affected. Rimuka township contributed 80% and 100% of city cases in 2008-9 and 2010, respectively, p value=0.000. In 2008-9, 91% of cholera cases presented within 2 days compared to 98% in 2010. Delay seeking treatment increased from 58% to 73% (p  value=0.001), with gender, and place equally affected. The 2010 outbreak evolved faster, resulting in higher proportion being managed in CTU. CFR was 2% in 2008-9, and 3.3% in 2010 (p value =0.31). Conclusion: the 2008-9 and 2010 cholera outbreaks were similar by age and gender.  Rimuka Township was most affected by the outbreaks. There was  worsening of delay seeking treatment. The 2010 outbreak was more rapid, leading to early opening of CTC. CFR was consistently above 1%

Indoor household residual spraying program performance in Matabeleland South province, Zimbabwe: 2011 to 2012; a descriptive cross-sectional study

Introduction: Matabeleland South launched the malaria pre-elimination campaign in 2012 but provincial spraying coverage has failed to attain95% target, with some districts still encountering malaria outbreaks. A study was conducted to evaluate program performance against achievingmalaria pre-elimination. Methods: a descriptive cross sectional study was done in 5 districts carrying out IRS using the logical framework involving inputs, process, outputs and outcome evaluation. Health workers recruited into the study included direct program implementers, district and provincial program managers. An interviewer administered questionnaire, checklists, key informant  interviewer guide and desk review of records were used to collect data. Results: we enrolled 37 primary respondents and 5 key informants. Pre-elimination, Epidemic Preparedness and Response plans were absent in all districts. Shortages of inputs were reported by 97% of respondents, with districts receiving 80% of requested budget. Insecticides were procured centrally at national level. Spraying started late and districts failed to spray all targeted households by end of December. The province is using makeshift camps with inappropriate evaporation ponds where liquid DDT waste is not safely accounted for. The provincial IHRS coverage for 2011 was 84%. Challenges cited included; food shortages for spraymen, late delivery of inputs and poor state of IHRS equipment. Conclusion: the province has failed to achieve Malaria pre-elimination IRS coverage targets for 2011/12 season. Financial and logistical challenges led to delays in supply of program inputs, recruitment and training of sprayers. The Province should establish camping infrastructure with standard evaporation ponds to minimise contamination of the  environment

Delayed initiation of anti-retroviral therapy in TB/HIV co-infected patients, Sanyati District, Zimbabwe, 2011-2012

Introduction: tuberculosis (TB) remains a public health problem and is driven by HIV. Recent studies indicate that anti-retroviral therapy (ART) initiated during the first two months of anti-TB treatment (ATT) reduces risk of HIV morbidity and mortality. In Sanyati district, 14% of TB/HIV coinfected patients were initiated on ART during TB treatment, in 2010. The study was conducted to determine the magnitude and determinants of delay in ART initiation, in TB/HIV co-infected patients. Methods: an analytic cross sectional study was conducted at three study sites in Sanyati district. The outcome was delayed ART initiation, being failure to be initiated on ART during the first two months of ATT. Respondents were interviewed using pre-tested questionnaires. Epi-InfoTM was used to generate frequencies, means, odds ratios and 95% confidence intervals. Stratified and logistic regression analysis was done. Results: of the 186 respondents, 63% had delayed ART initiation. Median delay from initiation of ATT to ART was 48 days (Q1=20; Q3=82). Risk factors for delayed ART initiation were: being treated for TB first time, AOR=2.23 (p=0.03); initially registered for HIV care outside Sanyati, AOR=3.08 (p<0.01); staying more than 5km from a clinic, AOR=3.29 (p<0.01). Enabling factors for early ART initiation was having a family member on ART, AOR=0.23 (p<0.01). Conclusion: significant delay and barriers to ART initiation were identified. Decentralization of ART initiation should be expedited. ART initiation should be expedited in patients with identified risk factors fordelaying ART initiation. Peer support should be strengthened in families and community. Periodic evaluation of magnitude of delay and impact of early ART initiation in TB/HIV patients is recommended.Keywords: Tuberculosis, HIV, delay, initiation, anti-retroviral therapy, Sanyati, Zimbabw

Typhoid outbreak investigation in Dzivaresekwa, suburb of Harare City, Zimbabwe, 2011

Introduction: Typhoid fever is a systemic infection caused by a Gram negative bacterium, Salmonella  typhi. Harare City reported 1078 cases of suspected typhoid fever cases from October  011 to January 2012. We initiated an investigation to identify possible source of transmission so as to institute control measures. Methods: An unmatched 1:1 case-control study was conducted. A questionnaire was administered to study participants to identify risk factors for contracting typhoid. A case was a resident of Dzivaresekwa who presented with signs and symptoms of typhoid between October and December 2011. Water  samples were collected for microbiological analysis. Results: 115 cases and 115 controls were enrolled. Drinking water from a well (OR= 6.2 95% CI  (2.01-18.7)), attending a gathering (OR= 11.3 95% CI (4.3-29.95)), boiling drinking water (OR= 0.21 95% CI (0.06-0.76)) and burst sewer pipe at home (OR= 1.19 95% CI (0.67-2.14)) were factors  associated with contracting typhoid. Independent risk factors for contracting typhoid were drinking water from a well (AOR=5.8; 95% CI (1.90-17.78)), and burst sewer pipe at home (AOR=1.20; 95% CI (1.10-2.19)). Faecal coli forms and E. coli were isolated from 8/8 well water samples. Stool, urine and blood specimens were cultured and serotyped for Salmonella typhi and 24 cases were confirmed positive. Shigella, Giardia and E coli were also isolated. Ciprofloxacin, X-pen and Rocephin were used for case  management. No complications were reported. Conclusion: Contaminated water from unprotected water sources was the probable source of the  outbreak. Harare City Engineer must invest in repairing water and sewage reticulation systems in the city.Key words: Typhoid, outbreak investigation, contaminated water, Zimbabwe

Factors associated with contracting malaria in Ward 29 of Shamva District, Zimbabwe, 2014

Background. Malaria cases at Wadzanayi Clinic in Shamva District, Zimbabwe, increased drastically, surpassing the epidemic threshold, in week four of December 2013. This rise was sustained, which necessitated an investigation of the outbreak.Objectives. To identify risk factors and system weaknesses to improve epidemic preparedness and response.Methods. An unmatched 1:1 case-control study was conducted in Ward 29 of Shamva District in Zimbabwe. Epidemic preparedness and response were assessed using the Zimbabwean epidemic preparedness and response guidelines.Results. The sociodemographic characteristics of all participants were similar, except for gender. The risk factors for contracting malaria were performing early morning chores (odds ratio (OR) 2.75; 95% confidence interval (CI) 1.20 - 6.32), having a body of water near the home (OR 3.41; 95% CI 1.62 - 7.20) and having long grass near the home (OR 2.61; 95% CI 1.10 - 6.37). Protective factors were staying indoors at night (OR 0.13; 95% CI 0.06 - 0.28) and staying in a sprayed home (OR 0.36; 95% CI 0.21 - 0.92). All cases were diagnosed with a malaria rapid diagnostic test. All complicated cases were treated with quinine. Four out of 58 uncomplicated cases were treated with quinine. The rest were treated with co-artemether. There was no documentation of the outbreak response by the district health executive. Respraying (indoor residual spraying) was carried out, with a coverage of 78% of rooms sprayed. One nurse out of seven at Wadzanayi Clinic was trained in integrated disease surveillance and response, and malaria case management. District malaria thresholds were outdated. Malaria commodities such as drugs and sprays did not have reorder limits.Conclusion. This study re-emphasises the importance of environmental- and personal-level factors as determinants of malaria. Poor outbreak preparedness and response may have propagated the malaria outbreak in this setting. Health education and the use of mosquito repellants should be emphasised. Larvicide may reduce the malaria burden. Epidemic preparedness and response need to be strengthened. Outbreak investigation remains important. This study emphasises the need for malaria interventions to be tailored to locally prevailing determinants to avert outbreaks