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To determine reservoir hosts for Marburg virus (MARV), we examined the fauna of a mine in northeastern Democratic Republic of the Congo. The mine was associated with a protracted outbreak of Marburg hemorrhagic fever during 1998-2000. We found MARV nucleic acid in 12 bats, comprising 3.0%-3.6% of 2 species of insectivorous bat and 1 species of fruit bat. We found antibody to the virus in the serum of 9.7% of 1 of the insectivorous species and in 20.5% of the fruit bat species, but attempts to isolate virus were unsuccessful. [ABSTRACT FROM AUTHOR]

Background Prior to this publication, the biting fly Haematobosca zuluensis (Zumpt, 1950) (Diptera, Muscidae, Stomoxyinae) was known only from a single male specimen collected in 1923 in Zululand, South Africa. Seven additional males were subsequently captured in the Kruger National Park of South Africa, one in 1984 and six in 1991, but remained unidentified until now. The genus includes species of considerable veterinary significance, but current keys for identification of species are misleading due to inadequate description of H. zuluensis. Methods External morphological features are described to enable species characterization, including intraspecific variability. Results This paper confirms the existence of H. zuluensis, expands its known range, provides a full description of males of the species, and gives an up to date set of keys for the 15 known species within the genus. Available records suggest that Haematobosca zuluensis is a low density species as yet known only from wildlife areas of South Africa. Conclusions The additional specimens of H. zuluensis have enabled an improved description of the species and an improved set of keys to identify constituent members of the genus.

This paper summarises published and unpublished data on helminths collected systematically from 424 impalas at 11 localities in eastern southern Africa, from St. Lucia in KwaZulu-Natal (KZN) to the Tuli Block in north-eastern Botswana. It includes data on collections in the Kruger National Park (KNP) in the drought of 1982, and in 1992/93 following the 1991/92 drought. Thirty-three species of nematodes, plus six taxa identified only to the generic level, three taxa of trematodes, and three species of cestodes were collected. Helminth species richness was highest in the southern KNP and lowest in the Tuli Block. The prevalence and intensity of infection of several helminths also declined from KZN and the southern KNP to the drier areas in the north and west. With the exception of St. Lucia and Nylsvley, > 80% of the helminths collected at each locality were collected in the southern KNP. St. Lucia was the most dissimilar locality; of the 20 helminths collected, five were unique. Ten of 33 species of nematodes, the paramphistomines (Trematoda) and the cestode Stilesia hepatica were collected at seven or more localities. Six of the most common nematodes, Cooperia hungi, Cooperioides hamiltoni, Impalaia tuberculata, Longistrongylus sabie, Strongyloides papillosus and Trichostrongylus deflexus are primarily parasites of impalas in the southern KNP, whereas many of the helminths collected at only one or two localities are parasites of other hosts. Nematode burdens were increased in the drought affected impalas in 1982, but helminth burdens decreased in 1992/93 following a dry cycle.

Background: Dengue is the most common and widespread mosquito-borne arboviral disease globally estimated to cause >390 million infections and >20,000 deaths annually. There are no effective preventive drugs and the newly introduced vaccines are not yet available. Control of dengue transmission still relies primarily on mosquito vector control. Although most vector control methods currently used by national dengue control programs may temporarily reduce mosquito populations, there is little evidence that they affect transmission. There is an urgent need for innovative, participatory, effective, and locally adapted approaches for sustainable vector control and monitoring in which students can be particularly relevant contributors and to demonstrate a clear link between vector reduction and dengue transmission reduction, using tools that are inexpensive and easy to use by local communities in a sustainable manner., Methods: Here we describe a cluster randomized controlled trial to be conducted in 46 school catchment areas in two townships in Yangon, Myanmar. The outcome measures are dengue cases confirmed by rapid diagnostic test in the townships, dengue incidence in schools, entomological indices, knowledge, attitudes and practice, behavior, and engagement., Conclusions: The trial involves middle school students that positions them to become actors in dengue knowledge transfer to their communities and take a leadership role in the delivery of vector control interventions and monitoring methods. Following this rationale, we believe that students can become change agents of decentralized vector surveillance and sustainable disease control in line with recent new paradigms in integrated and participatory vector surveillance and control. This provides an opportunity to operationalize transdisciplinary research towards sustainable health development. Due to the COVID-19 pandemic and political instability in Myanmar the project has been terminated by the donor, but the protocol will be helpful for potential future implementation of the project in Myanmar and/or elsewhere.Registration: This trial was registered in the ISRCTN Registry on 31 May 2022 ( https://doi.org/10.1186/ISRCTN78254298)., Competing Interests: No competing interests were disclosed., (Copyright: © 2023 Overgaard HJ et al.)

Malaria vector surveillance provides important data to inform the effective planning of vector control interventions at a local level. The aim of this study was to determine the species diversity and abundance, biting activity, and Plasmodium infectivity of Anopheles mosquitoes from a rural village in southern Mozambique. Human landing catches were performed monthly between December 2020 and August 2021. All collected Anopheles were identified to the species level and tested for the presence of malaria parasites. Eight Anopheles species were identified among the 1802 collected anophelines. Anopheles gambiae sensu lato (s.l.) were the most abundant (51.9%) and were represented by Anopheles quadriannulatus and Anopheles arabiensis . Anopheles funestus s.l. represented 4.5%. The biting activity of An. arabiensis was more pronounced early in the evening and outdoors, whereas that of An. funestus sensu stricto (s.s.) was more intense late in the night, with no significant differences in location. One An. funestus s.s. and one An. arabiensis , both collected outdoors, were infected with Plasmodium falciparum . The overall entomologic inoculation rate was estimated at 0.015 infective bites per person per night. The significant outdoor and early evening biting activity of An. arabiensis and An . funestus found in this village may negatively impact the effectiveness of current vector control interventions. Additional vector control tools that can target these mosquitoes are needed.

We collected >40,000 mosquitoes from 5 provinces in South Africa during 2011-2018 and screened for zoonotic flaviviruses. We detected West Nile virus in mosquitoes from conservation and periurban sites and potential new mosquito vectors; Banzi virus was rare. Our results suggest flavivirus transmission risks are increasing in South Africa.

Background: Malaria continues to be a leading cause of morbidity and mortality in Africa and conventional malaria control strategies, such as indoor residual spraying and insecticide-treated bed nets, have limited effectiveness for some malarial vectors. Consequently, the development of alternative or supplementary strategies is required. One potential strategy is the use of livestock-administered endectocides to control vector mosquitoes that feed outdoors on livestock. However, since this strategy requires support from local communities and livestock owners consenting for their animals to be treated, it can only be implemented if agreed to by affected communities. The aim of this study was to assess the social acceptance of the use of livestock-administered endectocides in the malaria endemic villages of Vhembe District, Limpopo Province, South Africa, where malaria incidence is high., Methods: Questionnaires were administered to 103 livestock-owning households from four villages, namely, Gumbu, Malale, Manenzhe and Bale. The assessment included questions on the acceptability of the strategy, the type and number of livestock owned, distances between houses and kraals (overnight pens) as well as previous use and awareness of endectocides. The results were analysed using descriptive statistics and multinomial logistic regression., Results: The types of livestock owned by the participants comprised, cattle, goats, sheep and donkeys, with the most dominant being goats (n = 1040) and cattle (n = 964). The majority of kraals were less than 10 m from homesteads. Most participants (72.5%) were already using chemicals to treat their livestock for parasites. All participants were amenable to the implementation of the strategy, and would give consent for their animals to be treated by endectocides., Conclusions: The use of livestock-administered endectocides appears to be a feasible and acceptable approach for control of animal-feeding malaria vector species in the malaria endemic villages of Vhembe District. This is based on a high percentage of rural residents keeping suitable livestock close to their homes and expressing willingness to use endectocides for mosquito control., (© 2022. The Author(s).)

Cambodia has one of the highest dengue infection rates in Southeast Asia. Here we report quantitative entomological results of a large-scale cluster-randomised trial assessing the impact on vector populations of a package of vector control interventions including larvivorous guppy fish in household water containers, mosquito trapping with gravid-ovitraps, solid waste management, breeding-container coverage through community education and engagement for behavioural change, particularly through the participation of school children. These activities resulted in major reductions in Container Index, House Index, Breteau Index, Pupal Index and Adult Index (all p-values 0.002 or lower) in the Intervention Arm compared with the Control Arm in a series of household surveys conducted over a follow-up period of more than one year, although the project was not able to measure the longer-term sustainability of the interventions. Despite comparative reductions in Adult Index between the study arms, the Adult Index was higher in the Intervention Arm in the final household survey than in the first household survey. This package of biophysical and community engagement interventions was highly effective in reducing entomological indices for dengue compared with the control group, but caution is required in extrapolating the reduction in household Adult Index to a reduction in the overall population of adult Aedes mosquitoes, and in interpreting the relationship between a reduction in entomological indices and a reduction in the number of dengue cases. The package of interventions should be trialled in other locations., Competing Interests: The authors have declared that no competing interests exist.

Background: Malaria control primarily depends on two vector control strategies: indoor residual spraying (IRS) and long-lasting insecticide-treated nets (LLINs). Both IRS and LLIN target indoor-biting mosquitoes. However, some of the most important malaria vectors have developed resistance against the chemical compounds used in IRS and LLINs. Insecticide-induced behavioural changes in vectors, such as increased outdoor feeding on cattle and other animals, also limit the effectiveness of these strategies. Novel vector control strategies must therefore be found to complement IRS and LLINs. A promising tool is the use of cattle-applied endectocides. Endectocides are broad-spectrum systemic drugs that are effective against a range of internal nematodes parasites and blood-feeding arthropods. The aim of this study was to investigate the effect of two endectocide drugs, injectable ivermectin and topical fipronil, on the survival and fecundity of zoophilic Anopheles arabiensis., Methods: Laboratory-reared mosquitoes were allowed to feed on cattle treated with either injectable ivermectin (0.2 mg/kg), topical fipronil (1.0 mg/kg) or saline (control) on days 0, 1, 4, 7, 13, 21 and 25 post-treatment, and mortality and egg production were recorded daily., Results: Compared to controls, the mortality of An. arabiensis increased by 3.52- and 2.43-fold with injectable ivermectin and topical fipronil, respectively. The overall fecundity of mosquitoes that fed on both ivermectin- and fipronil-treated cattle was significantly reduced by up to 90 and 60%, respectively, compared to the control group. The effects of both drugs attenuated over a period of 3 weeks. Injectable ivermectin was more effective than topical fipronil and increased mosquito mortality by a risk factor of 1.51 higher than fipronil. Similarly, both drugs significantly reduced the fecundity of An. arabiensis., Conclusions: This study demonstrates that injectable ivermectin and topical fipronil are able to suppress An. arabiensis density and could help to reduce outdoor malaria transmission. Data from the present study as well as from other similar studies suggest that current-generation endectocides have a limited duration of action and are expensive. However, new-generation, sustained-release formulations of ivermectin have a multi-week, high mortality impact on vector populations, thus holding promise of an effective reduction of outdoor malaria transmission.

Cambodia has made impressive progress in reducing malaria trends and, in 2018, reported no malaria-related deaths for the first time. However, the coronavirus disease (COVID-19) pandemic presents a potential challenge to the country's goal for malaria elimination by 2025. The path toward malaria elimination depends on sustained interventions to prevent rapid resurgence, which can quickly set back any gains achieved.Malaria Consortium supported mobile malaria workers (MMWs) to engage with target communities to build acceptance, trust, and resilience. At the start of the pandemic, Malaria Consortium conducted a COVID-19 risk assessment and quickly developed and implemented a mitigation plan to ensure MMWs were able to continue providing malaria services without putting themselves or their patients at risk. Changes in malaria intervention coverage and community uptake have been monitored to gauge the indirect effects of COVID-19. Comparisons have been made between output indicators reported in 2020 and from the same month-period of the previous year.In general, malaria service intervention coverage and utilization rates did not decline in 2020. Rather, the reported figures show there was a substantial increase in service utilization. Preliminary internal reviews and community meetings show that despite a heightened public risk perception toward COVID-19, malaria testing motivation has been well sustained throughout the pandemic. This may be attributable to proactive program planning and data monitoring and active engagement with the communities and the national authorities to circumvent the indirect effect of COVID-19 on intervention coverage in Cambodia during the pandemic., (© Feldman et al.)

Background: Anopheles species identification is essential for an effective malaria vector control programme. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been developed to identify adult Anopheles species, using the legs or the cephalothorax. The protein repertoire from arthropods can vary according to compartment, but there is no general consensus regarding the anatomic part to be used., Methods: To determine the body part of the Anopheles mosquitoes best suited for the identification of field specimens, a mass spectral library was generated with head, thorax with wings and legs of Anopheles gambiae, Anopheles arabiensis and Anopheles funestus obtained from reference centres. The MSL was evaluated using two independent panels of 52 and 40 An. gambiae field-collected in Mali and Guinea, respectively. Geographic variability was also tested using the panel from Mali and several databases containing added specimens from Mali and Senegal., Results: Using the head and a database without specimens from the same field collection, the proportion of interpretable and correct identifications was significantly higher than using the other body parts at a threshold value of 1.7 (p < 0.0001). The thorax of engorged specimens was negatively impacted by the blood meal after frozen storage. The addition of specimens from Mali into the database significantly improved the results of Mali panel (p < 0.0001), which became comparable between head and legs. With higher identification scores, the using of the head will allow to decrease the number of technical replicates of protein extract per specimen, which represents a significant improvement for routine use of MALDI-TOF MS., Conclusions: The using of the head of Anopheles may improve the performance of MALDI-TOF MS. Region-specific mass spectrum databases will have to be produced. Further research is needed to improve the standardization in order to share online spectral databases.

Background: The social-ecological systems theory, with its unique conception of resilience (social-ecological systems & resilience, SESR), provides an operational framework that currently best meets the need for integration and adaptive governance as encouraged by the Sustainable Development Goals. SESR accounts for the complex dynamics of social-ecological systems and operationalizes transdisciplinarity by focusing on community engagement, value co-creation, decentralized leadership and social innovation. Targeting Social Innovation (SI) in the context of implementation research for vector-borne diseases (VBD) control offers a low-cost strategy to contribute to lasting and contextualized community engagement in disease control and health development in low and middle income countries of the global south. In this article we describe the processes of community engagement and transdisciplinary collaboration underpinning community-based dengue management in rural primary schools and households in two districts in Cambodia., Methods: Multiple student-led and community-based interventions have been implemented focusing on empowering education, communication for behavioral change and participatory epidemiology mapping in order to engage Cambodian communities in dengue control. We describe in particular the significance of the participatory processes that have contributed to the design of SI products that emerged following iterative consultations with community stakeholders to address the dengue problem., Results: The SI products that emerged following our interaction with community members are 1) adult mosquito traps made locally from solid waste collections, 2) revised dengue curriculum with hands-on activities for transformative learning, 3) guppy distribution systems led by community members, 4) co-design of dengue prevention communication material by students and community members, 5) community mapping., Conclusions: The initiative described in this article put in motion processes of community engagement towards creating ownership of dengue control interventions tools by community stakeholders, including school children. While the project is ongoing, the project's interventions so far implemented have contributed to the emergence of culturally relevant SI products and provided initial clues regarding 1) the conditions allowing SI to emerge, 2) specific mechanisms by which it happens and 3) how external parties can facilitate SI emergence. Overall there seems to be a strong argument to be made in supporting SI as a desirable outcome of project implementation towards building adaptive capacity and resilience and to use the protocol supporting this project implementation as an operational guiding document for other VBD adaptive management in the region.

Despite the annual implementation of a robust and extensive indoor residual spraying programme against malaria vectors in Limpopo Province (South Africa), significant transmission continues and is a serious impediment to South Africa's malaria elimination objectives. In order to gain a better understanding regarding possible causes of this residual malaria, we conducted a literature review of the historical species composition and abundance of malaria vector mosquitoes in the Limpopo River Valley region of the Vhembe District, northern Limpopo Province, the region with the highest remaining annual malaria cases in South Africa. In addition, mosquito surveys were carried out in the same region between October 2017 and October 2018. A total of 2225 adult mosquitoes were collected using CO 2 -baited tent and light traps, human landing catches and cow-baited traps. Of the 1443 Anopheles collected, 516 were members of the An. gambiae complex and 511 An. funestus group. In the malaria endemic rural areas outside the Kruger National Park, one specimen each of An. gambiae s.s. and An. funestus and only three of An. arabiensis were collected. The latter species was abundant at a remote hot spring in the neighboring Kruger National Park. Eighteen other species of Anopheles were collected. Our survey results support the historical findings that An. arabiensis , the species widely held to be the prime malaria vector in South Africa, is a rare species in the malaria endemic Limpopo River Valley. The implications of the mosquito surveys for malaria transmission, elimination and vector control in northern Limpopo Province and neighboring regions are discussed.

Most data on species associations and vector potential of mosquitoes in relation to arboviral infections in South Africa date back from the 1940s to late 1990s. Contextual information crucial for disease risk management and control, such as the sampling effort, diversity, abundance, and distribution of mosquitoes in large parts of South Africa still remains limited. Adult mosquitoes were collected routinely from two horse farms in Gauteng Province; two wildlife reserves in Limpopo Province, at Orpen Gate in Kruger National Park (KNP) and Mnisi Area in Mpumalanga Province between 2014-2017, using carbon dioxide-baited light and tent traps. Mosquito diversity and richness are greater in untransformed natural and mixed rural settings. In untransformed wilderness areas, the most dominant species were Culex poicilipes, Anopheles coustani, and Aedes mcintoshi, while in mixed rural settings such as the Mnisi area, the two most abundant species were Cx. poicilipes and Mansonia uniformis. However, in peri-urban areas, Cx. theileri, Cx. univittatus, and Cx. pipiens sensu lato were the most dominant. Aedes aegypti, Ae. mcintoshi, Ae. metallicus, Ae. vittatus, Cx. pipiens s.l., Cx. theileri, and Cx. univittatus had the widest geographical distribution in northern South Africa. Also collected were Anopheles arabiensis and An. vaneedeni, both known malaria vectors in South Africa. Arbovirus surveillance and vector control programs should be augmented in mixed rural and peri-urban areas where the risk for mosquito-borne disease transmission to humans and domestic stock is greater., (© 2020 The Society for Vector Ecology.)

The Aedes aegypti mitogenome (Mt) sequences of field isolates from California and South Africa revealed a deletion between position 14,522 and 14,659 of the Mt contig of the AaegL5 reference genome. The length of the mitogenome of the California isolate was 16,659 bp and had 99.0% similarity with the AaegL5 Mt contig. The South African isolate sequence was 16,600 bp long and had 97.9% similarity with the reference. The region between 1496 and 1664 bp is similar to a nuclear pseudogene that might be a copy of a portion of the mitochondrial genome., Competing Interests: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article., (© 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Core-sheath structured fibres were developed for application as part of an alternative malaria vector control intervention aimed at reducing outdoor malaria transmission. The fibres were prepared by melt spinning of high density polyethylene (HDPE) as sheath and with a concentrate containing volatile N,N-Diethyl-m-toluamide (DEET) in poly(ethylene-co-vinyl acetate) (EVA) as core. The concentrate was prepared by a simple absorption processes to a content up to 40 wt% DEET. Scanning electron microscope imaging confirmed the formation of a bicomponent core-sheath fibre structure. Confocal Raman spectroscopy revealed the development of a concentration gradient of DEET in the sheath layer, suggesting a diffusion controlled release process. Excellent processability was demonstrated on an extrusion system melt spinning with take up speeds reaching 3000 m min -1 . Sample textiles knitted from such filaments showed high residual repellence activity even after 20 cold washes or after eight months ageing under laboratory conditions. These findings indicate that this technology offers an alternative way to prevent outdoor mosquito bites in an effective and affordable manner., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: A century of studies have described particular aspects of relatively few mosquito species in southern Africa, mostly those species involved with disease transmission, specifically malaria and arboviruses. Patterns of community composition such as mosquito abundance and species diversity are often useful measures for medical entomologists to guide broader insights and projections regarding disease dynamics and potential introduction, spread or maintenance of globally spreading pathogens. However, little research has addressed these indicators in southern Africa., Results: We collected 7882 mosquitoes from net and light traps at 11 localities comprising 66 species in 8 genera. We collected an additional 8 species using supplementary collection techniques such as larval sampling, sweep-netting and indoor pyrethrum knockdown catches. Highest diversity and species richness was found in the Okavango Delta of Botswana and in South Africa's Kruger National Park, while the lowest diversity and abundances were in the extreme southern tip of South Africa and in semi-desert Kalahari close to the South Africa border with Botswana. Species composition was more similar between proximal localities than distant ones (Linear model P-value = 0.005). Multiple arbovirus vector species were detected in all localities we surveyed (proportion of vector mosquito numbers were > 0.5 in all locations except Shingwedzi). Their proportions were highest (> 90%) in Vilankulo and Kogelberg., Conclusions: Multiple known arbovirus vector species were found in all study sites, whereas anopheline human malaria vector species in only some sites. The combination of net traps and light traps effectively sampled mosquito species attracted to carbon-dioxide or light, accounting for 89% of the 74 species collected. The 11% remaining species were collected using supplementary collection techniques mentioned above. The diversity of species weas highest in savanna type habitats, whereas low diversities were found in the drier Kalahari sands regions and the southern Cape fynbos regions.

Background: The Anopheles gambiae sensu lato (s.l.) and Anopheles funestus s.l. species complexes contain the most important malaria vectors in Africa. Within the An. funestus group of at least 11 African species, the vector status of all but the nominal species An. funestus appears poorly investigated, although evidence exists that Anopheles rivulorum and Anopheles vaneedeni may play minor roles. A new species, An. rivulorum-like, was described from Burkina Faso in 2000 and subsequently also found in Cameroon and Zambia. This is the first paper reporting the presence of this species in South Africa, thereby significantly extending its known range., Methods: Mosquitoes were collected using dry-ice baited net traps and CDC light traps in the Kruger National Park, South Africa. Sixty-four An. funestus s.l. among an overall 844 mosquitoes were captured and identified to species level using the polymerase chain reaction assay. All samples were also analysed for the presence of Plasmodium falciparum circumsporozoite protein using the enzyme-linked-immunosorbent assay., Results: Four members of the An. funestus group were identified: An. rivulorum-like (n = 49), An. rivulorum (n = 11), Anopheles parensis (n = 2) and Anopheles leesoni (n = 1). One mosquito could not be identified. No evidence of P. falciparum was detected in any of the specimens., Conclusion: This is the first report of An. rivulorum-like south of Zambia, and essentially extends the range of this species from West Africa down to South Africa. Given the continental-scale drive towards malaria elimination and the challenges faced by countries in the elimination phase to understand and resolve residual transmission, efforts should be directed towards determining the largely unknown malaria vector potential of members of the An. funestus group and other potential secondary vectors.

Key aspects of 36 mosquito-borne arboviruses indigenous to Africa are summarized, including lesser or poorly-known viruses which, like Zika, may have the potential to escape current sylvatic cycling to achieve greater geographical distribution and medical importance. Major vectors are indicated as well as reservoir hosts, where known. A series of current and future risk factors is addressed. It is apparent that Africa has been the source of most of the major mosquito-borne viruses of medical importance that currently constitute serious global public health threats, but that there are several other viruses with potential for international challenge. The conclusion reached is that increased human population growth in decades ahead coupled with increased international travel and trade is likely to sustain and increase the threat of further geographical spread of current and new arboviral disease.

Background: Culex univittatus and Culex perexiguus mosquitoes (Diptera: Culicidae) are competent arbovirus vectors, but with unclear morphological differentiation. In Europe, and in the Iberian Peninsula in particular, the presence of either or both species is controversial. However, in order to conduct adequate surveillance for arboviruses in this region, it is crucial to clarify whether Cx. univittatus is present or not, as well as to critically assess existing differentiation tools. This study aimed to clarify this situation, by morphological and molecular phylogenetic comparison of Iberian specimens deemed as Cx. univittatus, with others of South African origin, i.e. from the type-locality region., Methods: Thus, morphological characteristics useful to distinguish both species, such as midfemur pale line, hindfemur R ratio, seta g R 1 ratio, seta f shape, length of ventral arm of phalosome and number of setae on IX tergal abdominal segment, were observed. A phylogenetic analysis based on cox1 mtDNA, of which there were no sequences from Cx. univittatus yet available in the GenBank database, was performed., Results: This analysis showed that Iberian and South African specimens are morphologically similar, except for the length of the ventral arm of the phalosome, which was higher in the Iberian specimens. Although the Iberian specimens could not be accurately identified using BOLD Systems, phylogenetic analysis still grouped these closer to South African Cx. univittatus, than to Cx. perexiguus from Turkey and Pakistan, despite the observed segregation of both taxa as two individual monophyletic clusters with shared common ancestry., Conclusions: This survey demonstrates that the West Nile virus vector Cx. univittatus is present in the Iberian Peninsula.

In this study we tested an alternative method for collecting mosquito larvae called the sweep net catch method and compared its efficiency to that of the traditional dip method. The two methods were compared in various water bodies within Kruger National Park and Lapalala Wilderness area, South Africa. The sweep net catch method performed 5 times better in the collection of Anopheles larvae and equally as well as the dip method in the collection of Culex larvae (p =8.58 x 10 -5 ). Based on 15 replicates the collector's experience level did not play a significant role in the relative numbers of larvae collected using either method. This simple and effective sweep net catch method will greatly improve the mosquito larval sampling capacity in the field setting., Competing Interests: Competing interests: No competing interests were disclosed.

Background: Malaria control in Africa relies heavily on indoor vector management, primarily indoor residual spraying and insecticide treated bed nets. Little is known about outdoor biting behaviour or even the dynamics of indoor biting and infection risk of sleeping household occupants. In this paper we explore the preferred biting sites on the human body and some of the ramifications regarding infection risk and exposure management., Methods: We undertook whole-night human landing catches of Anopheles arabiensis in South Africa and Anopheles gambiae s.s. and Anopheles funestus in Uganda, for seated persons wearing short sleeve shirts, short pants, and bare legs, ankles and feet. Catches were kept separate for different body regions and capture sessions. All An. gambiae s.l. and An. funestus group individuals were identified to species level by PCR., Results: Three of the main vectors of malaria in Africa (An. arabiensis, An. gambiae s.s. and An. funestus) all have a preference for feeding close to ground level, which is manifested as a strong propensity (77.3% - 100%) for biting on lower leg, ankles and feet of people seated either indoors or outdoors, but somewhat randomly along the lower edge of the body in contact with the surface when lying down. If the lower extremities of the legs (below mid-calf level) of seated people are protected and therefore exclude access to this body region, vector mosquitoes do not move higher up the body to feed at alternate body sites, instead resulting in a high (58.5% - 68.8%) reduction in biting intensity by these three species., Conclusions: Protecting the lower limbs of people outdoors at night can achieve a major reduction in biting intensity by malaria vector mosquitoes. Persons sleeping at floor level bear a disproportionate risk of being bitten at night because this is the preferred height for feeding by the primary vector species. Therefore it is critical to protect children sleeping at floor level (bednets; repellent-impregnated blankets or sheets, etc.). Additionally, the opportunity exists for the development of inexpensive repellent-impregnated anklets and/or sandals to discourage vectors feeding on the lower legs under outdoor conditions at night.

Background: Knowledge of the ecology and behaviour of a target species is a prerequisite for the successful development of any vector control strategy. Before the implementation of any strategy it is essential to have comprehensive information on the bionomics of species in the targeted area. The aims of this study were to conduct regular entomological surveillance and to determine the relative abundance of anopheline species in the northern Kruger National Park. In addition to this, the impact of weather conditions on an Anopheles arabiensis population were evaluated and a range of mosquito collection methods were assessed., Methods: A survey of Anopheles species was made between July 2010 and December 2012. Mosquitoes were collected from five sites in the northern Kruger National Park, using carbon dioxide-baited traps, human landing and larval collections. Specimens were identified morphologically and polymerase chain reaction assays were subsequently used where appropriate., Results: A total of 3,311 specimens belonging to nine different taxa was collected. Species collected were: Anopheles arabiensis (n = 1,352), Anopheles quadriannulatus (n = 870), Anopheles coustani (n = 395), Anopheles merus (n = 349), Anopheles pretoriensis (n = 35), Anopheles maculipalpis (n = 28), Anopheles rivulorum (n = 19), Anopheles squamosus (n = 3) and Anopheles rufipes (n = 2). Members of the Anopheles gambiae species complex were the most abundant and widely distributed, occurring across all collection sites. The highest number of mosquitoes was collected using CO2 baited net traps (58.2%) followed by human landing catches (24.8%). Larval collections (17%) provided an additional method to increase sample size. Mosquito sampling productivity was influenced by prevailing weather conditions and overall population densities fluctuated with seasons., Conclusion: Several anopheline species occur in the northern Kruger National Park and their densities fluctuate between seasons. Species abundance and relative proportions within the An. gambiae complex varied between collection methods. There is a perennial presence of an isolated population of An. arabiensis at the Malahlapanga site which declined in density during the dry winter months, making this site suitable for a small pilot study site for Sterile Insect Technique as a malaria vector control strategy.

Asian bats have been identified as potential reservoir hosts of coronaviruses associated with severe acute respiratory syndrome (SARS-CoV). We detected antibody reactive with SARS-CoV antigen in 47 (6.7%) of 705 bat serum specimens comprising 26 species collected in Africa; thus, African bats may harbor agents related to putative group 4 CoV.

Background: An outbreak of Marburg hemorrhagic fever was first observed in a gold-mining village in northeastern Democratic Republic of the Congo in October 1998., Methods: We investigated the outbreak of Marburg hemorrhagic fever most intensively in May and October 1999. Sporadic cases and short chains of human-to-human transmission continued to occur until September 2000. Suspected cases were identified on the basis of a case definition; cases were confirmed by the detection of virus antigen and nucleic acid in blood, cell culture, antibody responses, and immunohistochemical analysis., Results: A total of 154 cases (48 laboratory-confirmed and 106 suspected) were identified (case fatality rate, 83 percent); 52 percent of cases were in young male miners. Only 27 percent of these men reported having had contact with other affected persons, whereas 67 percent of patients who were not miners reported such contact (P<0.001). Most of the affected miners (94 percent) worked in an underground mine. Cessation of the outbreak coincided with flooding of the mine. Epidemiologic evidence of multiple introductions of infection into the population was substantiated by the detection of at least nine genetically distinct lineages of virus in circulation during the outbreak., Conclusions: Marburg hemorrhagic fever can have a very high case fatality rate. Since multiple genetic variants of virus were identified, ongoing introduction of virus into the population helped perpetuate this outbreak. The findings imply that reservoir hosts of Marburg virus inhabit caves, mines, or similar habitats., (Copyright 2006 Massachusetts Medical Society.)

Two hundred and twenty-five small mammals belonging to 16 species were examined for ticks in Free State, Mpumalanga and Limpopo Provinces, South Africa, and 18 ixodid tick species, of which two could only be identified to genus level, were recovered. Scrub hares, Lepus saxatilis, and Cape hares, Lepus capensis, harboured the largest number of tick species. In Free State Province Namaqua rock mice, Aethomys namaquensis, and four-striped grass mice, Rhabdomys pumilio, were good hosts of the immature stages of Haemaphysalis leachi and Rhipicephalus gertrudae, while in Mpumalanga and Limpopo Provinces red veld rats, Aethomys chrysophilus, Namaqua rock mice and Natal multimammate mice, Mastomys natalensis were good hosts of H. leachi and Rhipicephalus simus. Haemaphysalis leachi was the only tick recovered from animals in all three provinces.

Fleas were collected from 61 wild carnivores belonging to 13 species in various nature reserves and on farms, two feral domestic cats in a nature reserve and a domestic dog in the city of Johannesburg. Eleven flea species, including two subspecies of one of these, belonging to six genera were recovered. Amongst these only Ctenocephalides felis felis and Ctenocephalides felis strongylus are considered specific parasites of carnivores. The remaining ten species normally infest the prey animals of the various carnivores.

Ectoparasites were collected from impalas, Aepyceros melampus, at four localities within the Kruger National Park, namely Skukuza, in the Biyamiti region, Crocodile Bridge and Pafuri. Animals were also examined at Skukuza during a severe drought and at Skukuza and Pafuri towards the end of a second drought. Parasite burdens were analysed in relation to locality, sex, age class, month and drought. The impalas were infested with 13 ixodid ticks species, including two that were identified only to genus level. Except for four animals at Pafuri, all were infested with Amblyomma hebraeum. The highest intensity of infestation with larvae of this tick occurred from April to June and during November and December at Skukuza and in the Biyamiti region. Infestation with nymphs was highest during late winter. All animals were infested with Boophilus decoloratus, and the intensity of infestation was highest during spring. The intensity of infestation with Rhipicephalus appendiculatus was highest at Crocodile Bridge and at Pafuri, and that of Rhipicephalus zambeziensis at Skukuza. With both the latter species the intensity of infestation of larvae was highest from April to August, of nymphs from July to September or October and of adults during February and March. Rhipicephalus kochi was present only at Pafuri. The impalas also harboured five louse species and two species of hippoboscid flies. The intensity of infestation with lice tended to be greater during late winter and spring than during other seasons and greater on lambs than on yearlings on which it was greater than on adult animals.

Ixodid ticks were collected from 104 wild carnivores belonging to 23 species in various nature reserves and on several farms in all nine provinces of South Africa. Seven feral cats in a nature reserve were also examined. Twenty-four tick species belonging to seven genera were recovered and identified. Amongst these ticks we consider the adults of Haemaphysalis leachi, Haemaphysalis spinulosa, Haemaphysalis zumpti, Ixodes rubicundus, Rhipicentor nuttalli, Rhipicephalus simus and Rhipicephalus turanicus to be true parasites of wild carnivores. Although numerous adult Rhipicephalus appendiculatus and Rhipicephalus zambeziensis were collected from some lions these were either sick or old animals. The immature stages of seven species regularly utilized wild carnivores as hosts on an opportunistic basis.

Objectives: To determine the use of antimalarial prophylaxis and the relative frequency of adverse events with different regimens in visitors to the Kruger National Park., Design: Retrospective postal survey of a cohort of 7,397 visitors during April 1996. Telephonic interviews were conducted with all respondents who reported neuropsychiatric adverse events necessitating medical attention, and their medical caregivers., Results: The most commonly used regimens were chloroquine and proguanil in combination (C&P) (35.6%) and mefloquine (18.4%). However, non-recommended regimens were also used by travellers to this chloroquine-resistant area, including chloroquine alone (15.7%). Adverse events were reported by 23.8% of travellers and were more common in the C&P group than the mefloquine group (28.9% v. 25.0%, P = 0.0100). Gastro-intestinal side-effects were significantly more common in the C&P group (nausea (P = 0.0170), diarrhoea (P = 0.0008), mouth ulcers (P = 0.0000)), while neuropsychiatric side-effects were more common in the mefloquine group (depression (P = 0.0000), light-headedness (P = 0.0009), anxiety (P = 0.0060)). Only 30% of travellers reported using antimalarial drugs both regularly as prescribed and for 4 weeks after leaving the malaria area. The most commonly reported reason for changing prophylaxis was advice from a physician or pharmacist (41.9%)., Conclusions: Health professionals providing medical advice to prospective travellers to malarious areas must tailor recommendations based on the balance between malaria risk in a specific geographical area and the benefits and tolerance of protective measures. Mosquito-avoidance measures should always be advocated, but counselling on antimalarial prophylaxis will be guided by the malaria/prophylaxis (serious adverse events) risk ratio. Where drug measures are indicated, the importance of their correct use should be emphasised.

Anopheles gambiae and An. arabiensis are mosquito species responsible for most malaria transmission in sub-Saharan Africa. They are also closely related sibling species that share chromosomal and molecular polymorphisms as a consequence of incomplete lineage sorting or introgressive hybridization. To help resolve these processes, this study examined the partitioning of mtDNA sequence variation within and between species across Africa, from both population genetic and phylogeographic perspectives. Based on partial gene sequences from the cytochrome b, ND1 and ND5 genes, haplotype diversity was high but sequences were very closely related. Within species, little or no population subdivision was detected, and there was no evidence for isolation by distance. Between species, there were no fixed nucleotide differences, a high proportion of shared polymorphisms, and eight haplotypes in common over distances as great as 6000 km. Only one of 16 shared polymorphisms led to an amino acid difference, and there was no compelling evidence for nonneutral variation. Parsimony networks constructed of haplotypes from both species revealed no correspondence of haplotype with either geography or taxonomy. This trend of low intraspecific genetic divergence is consistent with evidence from allozyme and microsatellite data and is interpreted in terms of both extensive gene flow and recent range expansion from relatively large, stable populations. We argue that retention of ancestral polymorphisms is a plausible but insufficient explanation for low interspecific genetic divergence, and that extensive hybridization is a contributing factor.

Thirty-three varieties of 24 species of plants and 19 species of vertebrates and invertebrates were experimentally inoculated with Ebola Zaire virus. Fruit and insectivorous bats supported replication and circulation of high titers of virus without necessarily becoming ill; deaths occurred only among bats that had not adapted to the diet fed in the laboratory.

Red veld rats (Aethomys chrysophilus) and bushveld gerbils (Tatera leucogaster) were trapped at monthly intervals, when possible, over a 2-year period, in the southern Kruger National Park, Mpumalanga Province. Forty-six specimens of each species were caught, euthenased and microscopically examined for fleas, lice, ticks and mites. Clear differences existed between the two rodent hosts in infestation intensity and also parasite species. The flea, Xenopsylla brasiliensis, commonly and exclusively utilized red veld rats, whereas Xenopsylla frayi was common and specific to bushveld gerbils. T. leucogaster were commonly infested with the lice Hoplopleura biseriata and Polyplax biseriata, while only a single A. chrysophilus hosted the louse, Hoplopleura patersoni. Red veld rats harboured small numbers of the immature stages of Haemaphysalis leachi/spinulosa and relatively large numbers of Rhipicephalus simus. The larvae of R. simus were irregularly collected from February to September and the nymphs from March to November. Bushveld gerbils hosted fewer ticks than did the rats, with a single specimen of H. leachi/spinulosa and low numbers of immature Hyalomma truncatum, the latter erratically present from June to October. Miles were abundant on both rodent hosts, A. chrysophilus hosting 13 species in six families, and T. leucogaster hosting 12 species representing seven families, with clear differences in mite assemblages between the two rodents. As the rats and gerbils were collected from the same trap lines at the same times, the differences in species composition and infestation intensity of their parasites, suggest that immunological behavioural or other segregating mechanisms are in operation to maintain discrete parasite assemblages.

Objectives: To identify possible public health consequences of Schistosoma mansoni infections in migrants entering north-eastern KwaZulu-Natal and Mpumalanga and Northern Province from southern Moçambique., Design: (i) Intestinal parasite surveys, one sample per person, within a 4-month period; (ii) temperature recordings and snail collections in an irrigation system., Setting: North-eastern KwaZulu-Natal and Kruger National Park, Mpumalanga and Northern Province., Participants: Ninety-seven migrants (1-68 years) from north-eastern KwaZulu-Natal and 47 from the Kruger Park (1 - 70 years)., Main Outcome Measures: The relatively high S. mansoni prevalence may cause problems on the rice paddy scheme in north-eastern KwaZulu-Natal. Ranges and prevalence rates of other parasites were recorded., Results: S. mansoni prevalence in Moçambicans entering north-eastern KwaZulu-Natal was 14.4%, seven times more than in local residents. S. mansoni is not endemic to this region because most bodies of water become too warm for either the parasite or its snail host, Biomphalaria pfeifferi, to develop. Recordings in paddies suggest, however, that the rice modifies the temperature pattern as it grows and provides a suitable habitat for transmission. Migrants entering the Transvaal lowveld where S. mansoni is endemic may become more severely infected. Thirteen other parasite species were recorded from migrants entering KwaZulu-Natal and 8 from those entering Mpumalanga and Northern Province., Conclusions: The relatively high S. mansoni prevalence among migrants entering north-eastern KwaZulu-Natal may have public health implications as it could encourage transmission in a non-endemic area. The range of parasites carried by migrants entering KwaZulu-Natal, Mpumalanga and Northern Province was similar to that harboured by local people but prevalence rates were generally lower.

Ixodid ticks were collected from scrub hares (Lepus saxatilis) at three localities. Nine tick species were recovered from 24 hares examined at Pafuri, Kruger National Park, Northern Transvaal. The most abundant and prevalent species were Hyalomma truncatum, Rhipicephalus kochi and a Rhipicephalus species (near R. pravus). Twelve tick species were collected from 120 scrub hares examined around Skukuza, Kruger National Park, Eastern Transvaal. The immature stages of Hyalomma truncatum were most abundant and those of Amblyomma hebraeum most prevalent on the hares. No haematozoa were found on blood smears made from these hares. Thirty-four scrub hares on mixed cattle and game farms near Hluhluwe, north-eastern KwaZulu-Natal harboured 12 tick species. The most abundant and prevalent of these were the immature stages of Rhipicephalus muehlensi. Piroplasms, tentatively identified as Babesia leporis, were present on blood smears of eight of these hares. The host status of scrub hares for 18 ixodid tick species or subspecies found in South Africa is tabulated.