Aedes species (Diptera: Culicidae) ecological and host feeding patterns in the north-eastern parts of South Africa, 2014-2018.

BACKGROUND: There is a paucity of recent data and knowledge on mosquito diversity and potential vectors of arboviruses in South Africa, with most of the available data dating back to the 1950s-1970s. Aedes and Culex species are the major vectors of some of the principal arboviruses which have emerged and re-emerged in the past few decades. METHODS: In this study we used entomological surveillance in selected areas in the north-eastern parts of South Africa from 2014 to 2018 to assess mosquito diversity, with special emphasis on the Aedes species. The impact of trap types and environmental conditions was also investigated. Identification of the blood meal sources of engorged females collected during the study period was carried out, and DNA barcodes were generated for selected species. RESULTS: Overall, 18.5% of the total Culicidae mosquitoes collected belonged to the genus Aedes, with 14 species recognised or suspected vectors of arboviruses. Species belonging to the Neomelaniconion subgenus were commonly collected in the Bushveld savanna at conservation areas, especially Aedes mcintoshi and Aedes circumluteolus. Aedes aegypti was present in all sites, albeit in low numbers. Temperature was a limiting factor for the Aedes population, and they were almost exclusively collected at temperatures between 18 °C and 27 °C. The cytochrome oxidase subunit I (COI) barcode fragment was amplified for 21 Aedes species, and for nine of these species it was the first sequence information uploaded on GenBank. CONCLUSION: This study provides a better understanding of the diversity and relative abundance of Aedes species in the north-east of South Africa. The information provided here will contribute to future arboviral research and implementation of efficient vector control and prevention strategies.

Studies of reservoir hosts for Marburg virus.

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.

Small mammals as hosts of immature ixodid ticks.

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.

Parasites of domestic and wild animals in South Africa. XLIV. Fleas (Insecta: Siphonaptera: Pulicidae) collected from 15 carnivore species.

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.

Parasites of domestic and wild animals in South Africa. XLI. Arthropod parasites of impalas, Aepyceros melampus, in the Kruger National Park.

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.

Repellent effects on Anopheles arabiensis biting humans in Kruger Park, South Africa.

Distribution of biting sites on the human body by the malaria vector Anopheles arabiensis Patton (Diptera: Culicidae) was investigated near a source of mosquitoes in the Kruger National Park, South Africa. Eight adult male volunteers (2 teams x 2 pairs of subjects) conducted human bait collections while seated on camp chairs in the open-air, wearing only short trousers (no shirt, socks or shoes). Mosquito collections during 18.30-22.30 hours on five consecutive nights in April 1998 yielded a total of 679 An. arabiensis females biting subjects with or without their ankles and feet treated with deet insect repellent (15% diethyl-3-methylbenzamide, Tabard lotion). On subjects whose feet and ankles were smeared with repellent, 160 An. arabiensis females were captured biting in 60 manhours: 88.1% on the legs, 1.4% on the arms and 1.2% on other parts of the body, but none on the repellent-treated feet or ankles. On subjects without repellent treatment, 519 An. arabiensis were caught biting in 60 man-hours: 81.1% on feet and ankles, 16.4% on legs, 1.4% on arms and 1.2% on the rest of the body. For individual subjects, the reduction of An. arabiensis bites ranged from 36.4 to 78.2% (mean protection 69.2%). Results of this study confirm previous findings that, in this part of South Africa - inhabited only by wildlife - when people sit outside during the evening An. arabiensis prefers to bite their lower limbs: 97.5% below the knees. Overall, the number of bites by the malaria vector An. arabiensis was reduced more than three-fold (from 26 to 8/person/evening), simply by treating ankles and feet with a consumer brand of deet repellent. Whether or not this provides a satisfactory degree of protection against malaria risk would depend on the malaria sporozoite rate in the malaria vector population.

Parasites of domestic and wild animals in South Africa. XXXVIII. Ixodid ticks collected from 23 wild carnivore species.

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.

Malaria protection measures used by in-flight travelers to South African game parks.

Malaria prevention in travelers depends upon dissemination of accurate information about malaria risk, prevention of mosquito bites, appropriate chemoprophylaxis use and knowledge of the symptoms of malaria. A study was undertaken of travelers to the Kruger National Park and private game parks in Mpumalanga Province, South Africa to investigate travelers knowledge, of malaria, chemoprophylaxis use, and experience of adverse events. In-flight self administered questionnaires were distributed and completed by travelers on flights returning to Johannesburg International Airport, from the malaria areas. The study was conducted during the highest malaria risk period during 1996. The Mpumalanga game parks are those most visited in South Africa and are found in the extreme northeast of the country, which adjoins Mozambique in the east and Zimbabwe in the north. This area is classified by the South African health authorities as being a high risk Malaria area.10 Chloroquine-resistant Plasmodium falciparum malaria has been described in this area.2,3 The Department of Health in South Africa recommends the use of mefloquine alone or the combination of chloroquine and proguanil, (doxycycline is prescribed for travelers in which the former antimalarials cannot be utilized), for visitors to this area during the high risk period for malaria, which extends from October to May.4 For the remainder of the year mosquito avoidance measures are recommended. Little is known about travelers' compliance with these recommendations and their knowledge of malaria. A study to explore these factors was undertaken as a joint initiative between the SAIMR travel clinic, Mpumalanga Department of Health, and the South African National Parks.

Antimalarial prophylaxis--use and adverse events in visitors to the Kruger National Park.

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.

Risk of malaria in visitors to the Kruger National Park, South Africa.

BACKGROUND: The risk of malaria to visitors to South Africa's nature reserves is unknown. Current recommendations for travelers are based on malaria incidence rates in local communities. These rates may well overestimate travelers' malaria risk and lead to unnecessary use of antimalarial prophylaxis with associated drug-related adverse events. This study investigated the incidence of malaria in visitors to South Africa's Kruger National Park (KNP). METHODS: Postal questionnaires were sent to the cohort of visitors staying in the KNP during April 1996, 6 to 8 weeks after they returned to their homes. Nonrespondents received duplicate questionnaires 8 weeks later. Responses from 3267 groups, representing 11,107 visitors (56. 8%) allowed determination of the malaria attack rate in this group of travelers and assessment of associations between malaria risk and a number of factors, including type of accommodation used during the visit; duration of stay; use of chemoprophylaxis; and use of personal protection measures. All travelers indicating that they had suffered an episode of malaria following their visit were telephonically contacted and their health providers traced to determine the basis of diagnosis and clinical outcome. RESULTS: One clinically suspected and four confirmed cases of Plasmodium falciparum malaria among the visitors suggest a low attack rate of 4. 5 cases per 10,000 visitors during April, the highest risk month for malaria in South Africa. No association was found between malaria risk and accommodation type, duration of stay, use and type of chemoprophylaxis, and use and number of personal protection measures. CONCLUSIONS: These findings confirm the importance of careful assessment of local malaria risk before individualized advice is provided to travelers.

Patterns of mitochondrial variation within and between African malaria vectors, Anopheles gambiae and An. arabiensis, suggest extensive gene flow.

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.

Experimental inoculation of plants and animals with Ebola virus.

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.

The comparative host status of red veld rats (Aethomys chrysophilus) and bushveld gerbils (Tatera leucogaster) for epifaunal arthropods in the southern Kruger National Park, South Africa.

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.

Schistosoma mansoni in migrants entering South Africa from Moçambique--a threat to public health in north-eastern KwaZulu-Natal?

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.

Fleas, lice and mites on scrub hares (Lepus saxatilis) in northern and eastern Transvaal and in KwaZulu-Natal, South Africa.

Fleas, lice and mites were collected from 24 and 120 scrub hares at Pafuri and Skukuza, Northern and Eastern Transvaal, respectively, in the Kruger National Park, and from 34 scrub hares in the Hluhluwe region, north-eastern KwaZulu-Natal. Ctenocephalides felis damarensis, the only flea recovered, reached peak burdens on the hares at each locality during late winter or spring. Juvenile hares harboured significantly fewer fleas than did adult animals. The lice Haemodipsus lyriocephalus and Haemodipsus setoni were collected from hares at each locality, with H. setoni generally being the most abundant. Listrophorus leporicolus was found on hares at Skukuza and Hluhluwe, and mites of the genus Cheyletiella were collected from hares at Skukuza.

Parasites of domestic and wild animals in South Africa. XXXIII. Ixodid ticks on scrub hares in the north-eastern regions of northern and eastern Transvaal and of KwaZulu-Natal.

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.

An outbreak of encephalomyocarditis-virus infection in free-ranging African elephants in the Kruger National Park.

A cluster of four deaths in late December 1993, marked the onset of an outbreak of disease of African elephants (Loxodonta africana) in the Kruger National Park (KNP) in South Africa, which has an estimated population of 7,500 elephants. Mortalities peaked in January 1994, with 32 deaths, and then declined steadily to reach pre-outbreak levels by September, but sporadic losses continued until November. During the outbreak altogether 64 elephants died, of which 53 (83%) were adult bulls. Archival records revealed that, in addition to the usual losses from known causes such as poaching and intraspecific fighting, sporadic deaths from unexplained causes had, in fact, occurred in widely scattered locations from at least 1987 onwards, and from that time until the perceived outbreak of disease there had been 48 such deaths involving 33 (69%) adult bulls. Carcases had frequently become decomposed or had been scavenged by the time they were found, but seven of eight elephants examined early in 1994 had lesions of cardiac failure suggestive of encephalomyocarditis (EMC)-virus infection, and the virus was isolated from the heart muscles of three fresh carcases. The results of tests for neutralizing antibody on 362 elephant sera collected for unrelated purposes from 1984 onwards and kept frozen, indicated that the virus had been present in the KNP since at least 1987. Antibody prevalences of 62 of 116 (53%) 18 of 139 (13%) and seven of 33 (21%) were found in elephants in three different regions of the KNP in 1993 and 1994. Studies had been conducted on myomorph rodents in the KNP for unrelated purposes since 1984, and trapping attempts were increased during the perceived outbreak of disease in elephants. There was a striking temporal correlation between the occurrence of a population explosion (as evidenced by markedly increased catch rates per trap-night) and a surge in prevalence of antibody to EM virus in rodents, and the occurrence of the outbreak of disease in elephants.

African horsesickness epidemiology: five species of Culicoides (Diptera: Ceratopogonidae) collected live behind the ears and at the dung of the African elephant in the Kruger National Park, South Africa.

During the culling of elephants (Loxodonta africana) at five sites in the Kruger National Park, South Africa, a total of 682 Culicoides of five species of the subgenus Avaritia were found live either behind the ears of elephants or attracted to the freshly disembowelled intestinal dung of elephants. The species are Culicoides tororoensis Khamala & Kettle, 1971; C. kanagai Khamala & Kettle, 1971; C. loxodontis Meiswinkel, 1992, and two undescribed species, i.e. Culicoides sp. #50 and Culicoides sp. #54 pale form (p.f.). Of 511 female midges found behind ears, 39.9% were nulliparous, 57.3% empty parous, 2.5% freshly bloodfed and 0.2% gravid. The age composition of this subpopulation indicates that the Culicoides were behind the ears to suck blood and, furthermore, would do so in broad daylight. The age composition of 171 Culicoides of three species attracted to dung was entirely different: 1.8% nulliparous, 14.6% empty parous, and 83.0% gravid, indicating that the great majority of midges captured at dung were about to oviposit or had just oviposited. Immediately after culling, light-traps were operated at two of the sites. Of 4,023 Culicoides of 21 species captured, 93% were of the same five species found on the ears and at the dung of elephants. Using these and other unpublished data pertaining to the rearing of these five Avaritia species from elephant dung over the past seven years, we broadly sketch the life cycle of these Culicoides, the first for any Afrotropical species of the genus. We also discuss the implications the close association between elephant and midge has for the dispersal and geographic distribution of the latter, and how it may influence the involvement of midges in the transmission of diseases such as African horsesickness. Owing to difficulties in identifying species of the subgenus Avaritia in the Afrotropical Region, the taxonomy of each of the five above-mentioned species is briefly appraised. Of the remaining 16 species (7%) captured in light traps 15 (6%) belong to that sector of the genus Culicoides whose immature stages develop in groundwater habitats and include C. imicola, which comprised only 2% of the light-trap collections. The large disparity in the adult abundance patterns of the "dung" and "groundwater" species in the middle of dry bushveld, is probably the result of differences in host and larval habitat preferences, and is briefly discussed. Finally, the few reports extant on the wild-host preferences of Afrotropical Culicoides are reviewed. Five tables and five figures accompany the text.