Characterization of foot-and-mouth disease viruses in Zambia-implications for the epidemiology of the disease in southern Africa

The livestock industry supports livelihood and nutritional security of at least 42% of people in the Southern African Development Community region. However, presence of animal diseases such as foot-and-mouth disease poses a major threat to the development of this industry. Samples collected from FMD outbreaks in Zambia during 2015–2020, comprising epithelial tissues samples (n = 47) and sera (n = 120), were analysed. FMD virus was serotyped in 26 samples, while 92 sera samples tested positive on NSP-ELISA. Phylogenetic analysis revealed notable changes in the epidemiology of FMD in Zambia, which included: (i) introduction of a novel FMDV SAT-3 (topotype II) causing FMD cases in cattle in Western Province; (ii) emergence of FMDV serotype O (topotype O/EA-2) in Central, Southern, Copperbelt, Western, Lusaka Provinces; and (iii) new outbreaks due to SAT -2 (topotypes I) in Eastern Zambia. Together, these data describe eight different epizootics that occurred in Zambia, four of which were outside the known FMD high-risk areas. This study highlights the complex epidemiology of FMD in Zambia, where the country represents an interface between East Africa (Pool 4) and Southern Africa (Pool 6). These changing viral dynamics have direct impacts on FMD vaccine selection in the SADC region.The Government of Zambia through the Ministry of Fisheries and Livestock, Department of Veterinary; the UK Department of the Environment, Food and Rural Affairs (Defra SE2944) and the European Union.http://www.mdpi.com/journal/virusespm2022Veterinary Tropical Disease

Galactosemia : the South African study

Thesis (Ph.D. (Biochemistry))--North-West University, Potchefstroom Campus, 2007Classic galactosemia and congenital hypothyroidism are both inborn errors of metabolism. The biochemical and molecular detection of both disorders is already defined in scientific literature. Their management is also well documented, yet their detection especially in poorer communities remains uncoordinated moreover in developing countries. These disorders can both be detected early in childhood if neonatal screening is instituted. When diagnosed early in childhood, congenital hypothyroidism can be successfully treated. In the case of classic galactosemia, preliminary clinical outcome is satisfactory although long-term prognosis is disappointing. In South Africa, neonatal screening has not been instituted. The reasons are related to the economics of the country as well as the lack of sufficient data about the prevalence and incidence of the diseases. In this study, a pilot newborn screening program for classic galactosemia and congenital hypothyroidism that take into consideration the socio-economic status of the surveyed populations has been established. The total protocol includes the sampling, storage of sampled specimens, biochemical and molecular diagnostic techniques. Using this protocol, an incidence ratio of one in a thousand congenital hypothyroidism cases was established in the Nkangala region of the Mpumalanga province of South Africa. Appropriate management therapy was also instituted in affected individuals. Furthermore, to initiate a lead into the investigation of mechanistic imperatives that result in poor long-term prognosis of classic galactosemia, a novel approach towards application of RNA interference in disease mechanism study was introduced. In this approach, the design and development of a mammalian cell model with GALT gene knockdown using RNAi was utilized to create a cellular state reminiscent of a classic galactosemia rather than to elucidate the gene function as is conventionally applied. The model was successfully completed and was compatible with the enzymatic activity prerequisites when compared to the control sets. However, the mammalian cellular model still needs to be rigorously tested to confirm its application towards studying long-term biochemical outcomes and identification of galactosemia secondary biomarkers.Doctora

Molecular characterisation of suspected heterozygotes of trimethylaminuria

Thesis, MSc, Potchefstroom University for Christian Higher Education 2002.Trimethylaminuria (McKusick 602079) or Fish odour syndrome is inherited recessively as a defect in hepatic nitrogen-oxidation of dietary derived trimethylamine (TMA), which causes excess excretion of trimethylamine such that affected individuals have a body odour reminiscent of rotten fish (Zhang et al., 1995). Trimethylaminuria is a result of either partial or total incapacity to oxygenate trimethylarninuria to its oxide, trimethylamine oxide (TMAO), by an enzyme known as the flavin-containing monooxygenase 3 (FM03). Mutations in the gene of the major human liver enzyme isoform, FM03, are responsible for causing trimethylaminuria (Akerman et al., 1999a and Dolphin et al., 1997b ). Clinical symptoms of this disorder of metabolism include fish-like to garbage-like odour of urine (trimethylaminuria), sweat (ishthyhidrosis) and breath (halitosis) as well as psycho-clinical symptoms such as depression (Akerman et al., l999a). To establish the percentage of homo- and heterozygous trimethylaminuric individuals, a screening programme was introduced for the Potchefstroom first year students. Evaluation of the screening results through the liquid chromatograph-mass spectrometer, which is based on the accurate determination of the TMA:TMAO ratio, showed a 1.46% of mild trimethylaminuria individuals. In this study, clinical symptoms induced by the loading test prior to urine evaluation of the TMA:TMAO ratio is described. This was followed by isolation of the FM03 gene from the blood of suspected individuals and subsequent amplification using the PCR. Amplification was succeeded by restriction fragment length polymorphism analysis for the determination of known common mutations throughout the different exons of the FM03 gene. Single stranded conformation polymorphism and heteroduplex analysis were performed to validate their applicability towards screening the FM03 gene. Preliminary work was also done towards establishing the usage of the denaturing gel gradient gel electrophoresis to screen the FM03 gene for aberrant sequences. xvi Results obtained through restriction fragment length polymorphism indicated the possible presence of the A52T mutation in ex on 3 of both patients that showed symptoms of mild trimethylaminuria. The A52T mutation may be the most prevalent in the South African population although more research still have to be done to investigate this possibility. The main objective of this study was to establish the suitability of different methods towards mutation screening of the FM03 gene. The methods attempted so far include polymerase chain reaction, restriction fragment length polymorphism, single stranded conformation polymorphism, heteroduplex analysis as well as denaturing gradient gel electrophoresis. All methods were applicable, although to different extents and with different limitations and resolutions. This study was a preliminary evaluation for a bigger study, which will include family members of the suspected heterozygotes. In the subsequent study, all nucleotide sequence fragments suspected of having mutations will be sequenced to confirm the presence and the type of the mutation present. xviiMaster

Additional file 2 of Investigation of foot and mouth disease virus and other animal pathogens in cattle, buffaloes and goats at the interface with Akagera National Park 2017 – 2020

Additional file 2. FMD serological NSP results from cattle and goats in Eastern Rwanda

Internalisation of Salmonella spp. by Typha latifolia and Cyperus papyrus in vitro and implications for pathogen removal in constructed wetlands

Summarization: Freshwater contamination by enteric pathogens is implicated in the high frequency of diarrhoeal diseases in low to middle income countries, typically due to poor wastewater management. Constructed Wetlands are a cost-effective and sustainable alternative to conventional/mechanical treatment technologies, but the pathogen removal mechanisms in Constructed Wetlands are not fully understood. This study investigated for the first time the internalisation of Salmonella spp. by Typha latifolia and Cyperus papyrus in hydroponic microcosms. Presence of Salmonella spp. within roots, rhizomes and shoots was assayed using agar-based methods over a period of 12 days. Concentration of Salmonella spp. in growth media showed 2.7 and 4.8 log unit reduction with T. latifolia and C. papyrus, respectively, and 1.8 and 6.0 log unit in unplanted units. Salmonella spp. was recovered from root and rhizome tissues of T. latifolia (up to 4.4 logCFU/g) and C. papyrus (up to 3.4 logCFU/g), and the bacteria were highly concentrated in the epidermis and cortex. However, Salmonella spp. was not detected in the stems and leaves of the two plant species. The present study demonstrates for the first time that these macrophytes internalise cells of Salmonella spp., which could be one pathogen removal mechanism employed by wetland plants.Presented on: Environmental Technolog

Additional file 1 of Investigation of foot and mouth disease virus and other animal pathogens in cattle, buffaloes and goats at the interface with Akagera National Park 2017 – 2020

Additional file 1. Retrieved sequences used in this study