The boom of the internet economy and the smartphone explosion currently experienced in South Africa is making it increasingly more convenient for South African residents to purchase electronically supplied goods and services from suppliers located all across the globe with the mere click of a button. The increasing number of purchases online is making it almost impossible for the South African Revenue Service to collect the Value-Added Tax on specifically electronically supplied services purchased from foreign suppliers. It is further placing South African suppliers of similar goods and services in a disadvantaged position in respect of pricing when compared to foreign suppliers. In the 2013 budget speech, the Minister of Finance indicated that the Value-Added Tax implications of the supply of digitised products by foreign suppliers are an area of concern. By July 2013 proposals were contained in the Draft Taxation Laws Amendment Bill to address this very problem. The aim of this study is to critically evaluate the proposed methods to collect Value-Added Tax on electronically supplied services, as contained in the Draft Taxation Laws Amendment Bill through a detailed literature review and critical evaluation. The literature review focuses on sections 7(1)(c) and 14 of the Value-Added Tax Act, the sections of the Value-Added Tax Act which currently regulates the collection of Value-Added Tax on electronically supplied services, as well as the methods used in the European Union and New Zealand to collect Value-Added Tax and Goods and Services Tax on electronically supplied services. The critical evaluation of the proposals identifies shortcomings and provides recommendations to overcome the shortcomings to ensure that the proposed changes will address the concerns and align South African legislation with global practices. The study concludes that, although the proposed changes to the method to collect Value-Added Tax on electronically supplied services are a step in the right direction, there is still work to be done by legislators on the details of the proposals to ensure successful implementation in a South African environment. / Dissertation (MCom)--University of Pretoria, 2014. / am2014 / Taxation / unrestricted

Solenopsis invicta is an invasive ant that has caused detrimental impacts to ecosystems and economies in the Southeastern United States, recently including Virginia. In this study, we explored the invasion ecology of S. invicta through two main objectives. First, we established a comprehensive distribution map of S. invicta in Virginia through multiple survey techniques. We then compared our findings with published models quantifying the potential spread of S. invicta and created our own species distribution model. In 2020-2021, S. invicta occurrences were found in 7 counties beyond the current Quarantine and our data show that S. invicta has spread further than predicted. Our own species distribution model suggests that the distribution area for S. invicta is likely to increase under the projected climate change. This study provides insights into the range expansion of S. invicta at the border of its suitable habitat and allow for improvements to models of its spread under these conditions. Additionally, it provides useful information to inform county extension agents to know where they are to expect new infestations of S. invicta. Second, we investigated the impacts of pesticide residue on the behavior of S. invicta through neonicotinoid exposure. We found detectable levels of neonicotinoids in the soil of the ant mounds as well as in the ants themselves. In addition, we investigated the effects of dietary exposure to imidacloprid on foraging behavior in a laboratory setting. We found that unexposed colonies were able to locate the food source more quickly during the second trial while exposed ants were unable to improve their performance. We also found that more exposed ant workers were unable to successfully navigate the maze as compared to unexposed workers. Our results suggest impaired learning of maze tasks and impaired navigational skills in neonicotinoid-exposed ants. / Master of Science in Life Sciences / The red imported fire ant (RIFA) is an invasive ant species found throughout the Southeastern United States that has negatively impacted ecosystems and economies. In the past few decades, RIFA has invaded the Coastal Plain of Virginia, resulting in legislation that restricts the movements of soil, plant products, and some equipment in and out of several southeastern counties and independent cities. To develop better management practices, there is a critical need to understand the spread, establishment, and impacts of RIFA in greater detail. We aimed to do this by surveying the current distribution of RIFA in Virginia and investigate the impact of insecticide use on their spread and behavior. In 2020-2021, we found RIFA occurrences in 7 counties beyond the current Quarantine, which is further than previously predicted. We then built a model using climatic variables that predicts the distribution of RIFA and found their habitable range is likely to increase under the projected climate change. Additionally, we investigated the impacts of common agricultural pesticides on the behavior of RIFA. We found these chemicals present in both the soil of the ant mounds and in the ants themselves. We also found that dietary exposure to imidacloprid, a neonicotinoid pesticide, altered the foraging behavior of RIFA. This study provides useful information to advise county extension agents to know where they are to expect new infestations of RIFA. Our results also suggest that human activity alters the invasion ecology of recent arrivals such as the red imported fire ant.

Introdução - O pescado é amplamente utilizado para a produção de alimentos à base de matéria-prima de origem animal. A importação de pescado vem crescendo ano a ano, sendo o Porto de Santos/SP uma das principais rotas de entrada desse produto no país. Objetivos - Avaliar as condições higienicossanitárias de pescado importado no Porto de Santos/SP. Métodos - A pesquisa foi desenvolvida com base em duas abordagens: estudo dos requisitos obrigatórios para nacionalização do produto importado com base na legislacão vigente e trabalho desempenhado pelo Vigiagro/MAPA, assim como, estudo laboratorial. Foram coletadas 60 amostras de 30 diferentes produtos de pescado sendo cada amostra constituída por, no mínimo, 500 g do produto de uma mesma marca, lote, data de fabricação e validade. Uma parte das amostras, 30 embalagens de 500 g no mínimo, foi acondicionada em caixas isotérmicas com temperatura controlada e encaminhada para realização das análises microbiológicas. As demais 30 embalagens foram encaminhadas, sob as mesmas condições, para realização das análises físico-químicas. As análises físico-químicas e microbiológicas foram realizadas em Laboratórios credenciados no Mapa. Para produtos congelados foram realizadas as análises microbiológicas para pesquisa de Salmonella spp em 25 gramas e contagem de Staphylococcus coagulase positiva e realização das análises físico-químicas de N-BVT. Para produtos salgados refrigerados, foram realizadas análises microbiológicas para pesquisa de Salmonella spp em 25 gramas, contagem de Staphylococcus coagulase positiva e contagem de coliformes termotolerantes a 450 C, assim como, realização das análises físico-químicas de umidade e resíduo mineral fixo. Resultados - Nos produtos congelados, as análises físico-químicas para o N-BVT e as análises microbiológicas para pesquisa de Salmonella spp em 25 gramas e contagem de Staphylococcus coagulase positiva indicaram 100,0% de conformidade com a legislação vigente. Nos produtos salgados refrigerados, as análises microbiológicas para pesquisa de Salmonella spp em 25 gramas, contagem de Staphylococcus coagulase positiva e contagem de coliformes termotolerantes a 450 C também indicaram 100,0% de conformidade com a legislação vigente. Por outro lado, 100,0% das análises físico-química de umidade e 44,4% das análises físico-químicas para resíduo mineral fixo no pescado salgado resfriado estavam com valores acima da legislação vigente. Todos os produtos apresentaram documentação obrigatória e rótulo de acordo com a legislação vigente. Conclusão Embora os resultados das análises microbiológicas tenham demonstrado que os produtos estivessem próprios para o consumo, as análises físico-químicas, em específico os parâmetros referentes a umidade no pescado salgado refrigerado, não estavam de acordo com os valores determinados pela legislação vigente. A legislação sanitária vigente para importação de pescados, assim como a metodologia de inspeção e fiscalização desses produtos, se mostrou eficiente comparado aos resultados laboratoriais encontrados, com possibilidade de inclusão de novas ferramentas e análises laboratoriais visando garantir, ainda mais, as condições higiênicossanitárias e segurança alimentar. / Introduction - Seafood is widely used for the production of food based on raw materials of animal origin. The fish import has been growing year by year, and the Port of Santos / SP one of the main routes of entry of that product in the country. Objectives - To evaluate the hygienic-sanitary conditions of imported fish in the Port of Santos / SP. Methodology - The study was based up on two approaches: study of the mandatory requirements for nationalization of the imported product on the basis of current legislation and work performed by Vigiagro / MAPA, as well as laboratory study. It was collected 60 samples from 30 different products of fish each sample being composed by 500 g from the same brand, batch number, manufacturing and expiration date. A share of these samples, 30 packages of 500 g at least, was accommodated and transported in temperature-controlled isothermal box to the microbiological analysis. The remaining 30 packages were sent, under the same conditions, to physico-chemical analysis. The physico-chemical and microbiological analyzes were performed in laboratories accredited on MAPA. For frozen products were performed microbiological analyzes for Salmonella spp research in 25 g Staphylococcus and count coagulase positive and achievement of physical and chemical analysis of N-BVT. For salted products, microbiological analyzes were performed for detection of Salmonella spp in 25 g, Staphylococcus coagulase positive count and count of coliform to 450 C and physical-chemical analysis for humidity and fixed mineral residue. Results - In the frozen products, physico-chemical analysis for the N-BVT and microbiological analyzes for Salmonella spp research in 25 g and Staphylococcus coagulase positive counts indicated 100.0% of compliance with current regulation. In salted products, microbiological analyzes for Salmonella spp research in 25 g, Staphylococcus coagulase positive and coliform to 450 C counts also indicated 100.0% compliance with current legislation. On the other, 100.0% of the physico-chemical analysis of humidity and 44,4 % in the ashs physico-chemical results, in salted fish, were with higer limits established by Brazilian legislation. All products submitted mandatory documents required for import and labeling in accordance with current regulation. Conclusion - Although the results of the microbiological analyzes showed that the imported fish were apropriate for consumption, the physico-chemical analysis, in particularly the humidity parameters were not in accordance with values indicated in the current regulation. The current health regulation for the imported fish, as well as the inspection methodology and supervision of these products, was efficient compared to laboratory findings, with the possibility of adding new tools and laboratory tests to ensure even more the higienic-sanitary conditions and food safety.

Thelohania solenopsae are intracellular pathogens found in the red imported fire ant, Solenopsis invicta. These pathogens cause detrimental effects to their fire ant hosts. The present study revealed that the midgut and the meconium materials from pupating fourth instar larvae were possible vehicles for the horizontal transmission of the disease. The pathogen was further found to cause a reduction of humeral proteins. In SDS-PAGE stained with silver, several proteins were observed only in controls but not in infected fire ant queens. Different queens were found to have variable proteins reduced due to infection of this pathogen. Furthermore, vitellogenin titers were found to be significantly reduced in infected fire ant queens, although the infection rates of the fat body cells was found to be less than 20%. Finally, although the pathogens did not directly induce apoptosis in infected cells, there were more infected cells undergoing apoptosis than uninfected cells. There was no evidence to support the idea that infected fat body cells became more resistant to apoptosis inducers.

Solenopsis invicta, the red imported fire ant, has recently become associated with Antonina graminis, an invasive pest, and Neodusmetia sangwani, biological control agent, and maybe negatively affecting established biological control. A preliminary survey outlined the range of A. graminis and its parasitoids, and found N. sangwani was present at a reduced rate in South Texas and in the southeastern United States. A greenhouse experiment demonstrated that S. invicta decreased the rate of parasitism of A. graminis by N. sangwani, with S. invicta directly interfering with oviposition. Interactions between S. invicta and A. gaminis may be facilitating the spread and establishment of two invasive pests which has a negative impact on established classical biological control of A. graminis by N. sangwani.

Solenopsis invicta, the red imported fire ant, has recently become associated with Antonina graminis, an invasive pest, and Neodusmetia sangwani, biological control agent, and maybe negatively affecting established biological control. A preliminary survey outlined the range of A. graminis and its parasitoids, and found N. sangwani was present at a reduced rate in South Texas and in the southeastern United States. A greenhouse experiment demonstrated that S. invicta decreased the rate of parasitism of A. graminis by N. sangwani, with S. invicta directly interfering with oviposition. Interactions between S. invicta and A. gaminis may be facilitating the spread and establishment of two invasive pests which has a negative impact on established classical biological control of A. graminis by N. sangwani.

Ants are capable of building close associations with plants, insects, fungi and bacteria. Symbionts can provide essential nutrients to their insect host, however, the development of new molecular tools has allowed the discovery of new microorganisms that manipulate insect reproduction, development and even provide defense against parasitoids and pathogens. In this study we investigated the presence of bacteria inside the Red Imported Fire Ant midgut using molecular tools and transmission electron microscopy. The midgut bacteria were also characterized by their morphology, biochemical activity, and antibiotic resistance profile. After isolation, culture, and characterization of these bacteria, the molecular analysis revealed ten unique profiles which were identified to at least the genus level, Enterococcus sp./durans, Klebsiella ornithinolytica, Kluyvera cryocrescens, Lactococ-cus garvieae, Pseudomonas aeruginosa, Achromobacter xylosoxidans, Bacillus pumilus, Listeria innucua, Serratia marcescens, and an uncultured bacterium from the Entero-bacteriaceae. New SEM and TEM techniques revealed a possible functional association of endosymbiotic bacteria with the insect host, and it also showed the absence of bacteriocytes in the epithelial cells of the midgut. The PCR results, from the bacteria abundance and distribution studies, showed that Enterococcus sp., Kluyvera cryocres-cens and Lactococcus garvieae are the most abundant species, but they are not consistently found in all sites throughout the southeastern United States. Kluyvera cryocrescens, Serratia marcescens, and an uncultured bacterium (isolate #38: Enterobacteriaceae) were genetically modified with the plasmid vector pZeoDsRed and successfully reintroduced into fire ant colonies. Strong fluorescence of DsRed was detected up to seven days after introduction. The transformed bacteria can still be rescued after pupal emergence; however most were passed out in the meconium. We further demonstrated that nurses contributed to the spread of the transformed bacteria within the colony by feeding the meconium to naive larvae. Although the role of midgut bacteria in the fire ant is still unknown, we have no indication that they cause any pathology. Studies emphasizing the role of these bacteria in fire ant physiology are still ongoing. These results are the foundation for a fire ant biological control program using endosymbiotic bacteria as vectors to introduce foreign genes that express proteins with insecticidal properties.

Thelohania solenopsae are intracellular pathogens found in the red imported fire ant, Solenopsis invicta. These pathogens cause detrimental effects to their fire ant hosts. The present study revealed that the midgut and the meconium materials from pupating fourth instar larvae were possible vehicles for the horizontal transmission of the disease. The pathogen was further found to cause a reduction of humeral proteins. In SDS-PAGE stained with silver, several proteins were observed only in controls but not in infected fire ant queens. Different queens were found to have variable proteins reduced due to infection of this pathogen. Furthermore, vitellogenin titers were found to be significantly reduced in infected fire ant queens, although the infection rates of the fat body cells was found to be less than 20%. Finally, although the pathogens did not directly induce apoptosis in infected cells, there were more infected cells undergoing apoptosis than uninfected cells. There was no evidence to support the idea that infected fat body cells became more resistant to apoptosis inducers.

I examined the impacts of the invasive red imported fire ant (RIFA, Solenopsis invicta) on native ants (Monomorium minimum, Paratrechina sp., S. krockowi, Pheidole metallescens, Forelius pruinosus, and Camponotus americanus) and litter removal in a post oak savannah community in central Texas. The study site was divided into 3 adjacent areas, and ant-toxic bait was used, along with additional colonies of RIFA, to establish 3 different densities of RIFA (naturally occurring, low, and high). I surveyed the ants in the 3 density areas and calculated the catch per unit effort for each species. Litter baits were placed in the 3 density areas for 14 12-hour trials. The masses of the litter removed were measured, and means were calculated for each speciesdensity/ trial/date/period/bait combination. The average amounts of litter removed by RIFA and native ant were different in the 3 density areas (0.42 g, 0.0 g, and 0.75 g for RIFA in the natural RIFA density area, low RIFA density area, and high RIFA density area, respectively; 0.0 g, 0.16 g, and 0.15 g for native ants in the natural RIFA density area, low RIFA density area, and high RIFA density area, respectively), indicating that RIFA does have an effect on native ant habitat use.

This research evaluated the new red imported fire ant (RIFA) bait Advion™. Advion™ contains the active ingredient indoxacarb which, following ingestion, must be metabolized into an N-decarbomethoxyllated metabolite known as JT333 in order to become acutely toxic. Laboratory experimentation was conducted to determine the most effective chemical form and concentration of indoxacarb, as well as the most appropriate grit size, for use in Advion™. The results indicated that Advion™ containing indoxacarb was more effective than Advion™ containing JT333, that 0.10%, 0.06%, and 0.045% were the most effective concentrations of indoxacarb, and that standard sized grit (~2 mm) was more effective than small sized grit (<1 mm). Field experimentation was conducted to determine the most effective concentration and quantity of Advion™, as well as the most effective placement of the bait, for obtaining maximum control of RIFA colonies via individual mound treatments. The results indicated that 10 g (2 Tbsp) of 0.045% Advion™ placed around the mound in a circle with a radius of 0.5-3.0 m was the most effective manner in which to treat individual RIFA mounds. Field experimentation was also conducted to compare the effectiveness of label-rate broadcast treatment with Advion™ to the effectiveness of both label-rate broadcast treatment with Amdro® and pre-baiting broadcast treatment with Advion™. Both pre-baiting broadcast treatment and label-rate broadcast treatment with Advion™ ultimately resulted in 98-99% RIFA colony mortality, which was significantly greater than the 87% colony mortality resulting from broadcast treatment with Amdro®. The 6.2 d LT90 for label-rate broadcast treatment with Advion™ was nearly one half that of the LT90 for pre-baiting broadcast treatment. Laboratory experimentation was conducted to determine whether RIFA workers were capable of metabolizing indoxacarb into the toxic metabolite, and the results clearly indicated that they were. Finally, field experimentation was conducted to evaluate the effectiveness of label-rate broadcast treatment with Advion™ at controlling multiple ant species. The results indicated that label-rate broadcast treatment with Advion™ resulted in control of the RIFA and Pogonomyrmex barbatus for at least 7 wk, and Monomorium pharaonis and Dorymyrmex pyramicus for some period of time between 3 and 7 wk.