Pollution shapes the microbial communities in river water and sediments from the Olifants River catchment, South Africa.

Human activities such as agriculture and mining are leading causes of water pollution worldwide. Individual contaminants are known to negatively affect microbial communities. However, the effect of multifaceted pollution on these communities is less well understood. We investigated, using next-generation sequencing of the 16S rRNA genes, the effects of multisource (i.e., fertilizer industry and mining) chronic pollution on bacterial and archaeal communities in water and sediments from the Olifants River catchment, South Africa. Water samples showed less microbial species diversity than sediments and both habitats displayed different microbial communities. Within each of these habitats, pollution had no effect on alpha diversity but shaped the microbial composition and taxonomy-based predicted functions. Certain prokaryotic taxa and functional groups were indicative of different degrees of pollution. Heterotrophic taxa (e.g., Flavobacterium sp.) and sulphur-oxidizing bacteria (i.e., Thiobacillus sp.) were indicators of pollution in water and sediments, respectively. Ultimately, this information could be used to develop microbial indicators of water quality degradation.

Bacterial diversity in the waterholes of the Kruger National Park: an eDNA metabarcoding approach 1.

Bacteria are essential components of natural environments. They contribute to ecosystem functioning through roles as mutualists and pathogens for larger species, and as key components of food webs and nutrient cycles. Bacterial communities respond to environmental disturbances, and the tracking of these communities across space and time may serve as indicators of ecosystem health in areas of conservation concern. Recent advances in DNA sequencing of environmental samples allow for rapid and culture-free characterization of bacterial communities. Here we conduct the first metabarcoding survey of bacterial diversity in the waterholes of the Kruger National Park, South Africa. We show that eDNA can be amplified from waterholes and find strongly structured microbial communities, likely reflecting local abiotic conditions, animal ecology, and anthropogenic disturbance. Over timescales from days to weeks we find increased turnover in community composition, indicating bacteria may represent host-associated taxa of large vertebrates visiting the waterholes. Through taxonomic annotation we also identify pathogenic taxa, demonstrating the utility of eDNA metabarcoding for surveillance of infectious diseases. These samples serve as a baseline survey of bacterial diversity in the Kruger National Park, and in the future, spatially distinct microbial communities may be used as markers of ecosystem disturbance, or biotic homogenization across the park.

Nematodes found in Nile crocodiles in the Kruger National Park, South Africa, with redescriptions of Multicaecum agile (Wedl, 1861) (Heterocheilidae) and Camallanus kaapstaadi Southwell & Kirshner, 1937 (Camallanidae).

Sixteen Nile crocodiles were collected in the Kruger National Park, South Africa and vicinity during 2010 and 2011. A total of 11 nematode species representing six families were recovered. Heterocheilids were the dominant group, comprising five species, with Dujardinascaris madagascariensis (Chabaud & Caballero, 1966) being the most prevalent (75%), followed by Ingwenascaris sprenti Junker & Mutafchiev, 2017 (68.8%), which was also the second most numerous nematode. While less prevalent (31.3%), Typhlophoros kwenae Junker & Mutafchiev, 2017 was the most abundant species. Micropleura huchzermeyeri Junker & Mutafchiev, 2017 (Micropleuridae) was collected from five crocodiles and Crocodylocapillaria sp. (Capillariidae) occurred in a single host. Three nematodes, Camallanus kaapstaadi Southwell & Kirshner, 1937, Spirocamallanus sp. (both Camallanidae) and Ascarophis sp. (Cystidicolidae), are considered accidental infections, likely ingested with the hosts' prey. Our findings of D. dujardini (Travassos, 1920), D. madagascariensis and Multicaecum agile (Wedl, 1861) in South Africa constitute new geographical records. Crocodylocapillaria sp. represents a new host and geographical record, while T. kwenae, I. sprenti and M. huchzermeyeri have been described as new species during the course of this survey. Multicaecum agile is here redescribed based on light and scanning electron microscopy. Previously undescribed morphological characters of C. kaapstaadi, typically a parasite of Xenopus spp. (Amphibia: Pipidae), but here found in two Nile crocodiles, are also presented.

Viruses as indicators of contemporary host dispersal and phylogeography: an example of feline immunodeficiency virus (FIVPle ) in free-ranging African lion (Panthera leo).

Measuring contemporary dispersal in highly mobile terrestrial species is challenging, especially when species are characterized by low levels of population differentiation. Directly transmitted viruses can be used as a surrogate for traditional methods of tracking host movement. Feline immunodeficiency virus (FIV) is a species-specific lentivirus, which has an exceptionally high mutation rate and circulates naturally in wild felids. Using samples derived from 35 lion (Panthera leo) prides, we tested the prediction that FIV in lions (FIVPle ) can be used to track the dispersal of individuals between prides. As FIVPle subtypes are geographically structured throughout Africa, we predicted that this marker could be used to detect phylogeographic structure of lions at smaller spatial scales. Phylogenetic analyses of FIVPle pol-RT sequences showed that core pride members (females and subadults) shared evolutionary close viral lineages which differed from neighbouring core prides, whereas sequences from sexually mature males associated with the same pride were always the most divergent. In six instances, natal pride associations of divergent male lions could be inferred, on the assumption that FIVPle infections are acquired during early life stages. Congruence between the genetic pattern of FIV and pride structure suggests that vertical transmission plays an important role in lion FIV dynamics. At a fine spatial scale, significant viral geographic structuring was also detected between lions occurring north of the Olifants River within the Kruger National Park (KNP) and those occupying the southern and central regions. This pattern was further supported by phylogenetic analyses and the confinement of FIVPle subtype E to the northern region of KNP. The study provides new insights into the use of retroviral sequences to predict host dispersal and fine-scale contemporary geographic structure in a social felid species.

BRIDGING GAPS BETWEEN ZOO AND WILDLIFE MEDICINE: ESTABLISHING REFERENCE INTERVALS FOR FREE-RANGING AFRICAN LIONS (PANTHERA LEO).

The International Species Information System has set forth an extensive database of reference intervals for zoologic species, allowing veterinarians and game park officials to distinguish normal health parameters from underlying disease processes in captive wildlife. However, several recent studies comparing reference values from captive and free-ranging animals have found significant variation between populations, necessitating the development of separate reference intervals in free-ranging wildlife to aid in the interpretation of health data. Thus, this study characterizes reference intervals for six biochemical analytes, eleven hematologic or immune parameters, and three hormones using samples from 219 free-ranging African lions ( Panthera leo ) captured in Kruger National Park, South Africa. Using the original sample population, exclusion criteria based on physical examination were applied to yield a final reference population of 52 clinically normal lions. Reference intervals were then generated via 90% confidence intervals on log-transformed data using parametric bootstrapping techniques. In addition to the generation of reference intervals, linear mixed-effect models and generalized linear mixed-effect models were used to model associations of each focal parameter with the following independent variables: age, sex, and body condition score. Age and sex were statistically significant drivers for changes in hepatic enzymes, renal values, hematologic parameters, and leptin, a hormone related to body fat stores. Body condition was positively correlated with changes in monocyte counts. Given the large variation in reference values taken from captive versus free-ranging lions, it is our hope that this study will serve as a baseline for future clinical evaluations and biomedical research targeting free-ranging African lions.

Pentastome assemblages of the Nile crocodile, Crocodylus niloticus Laurenti (Reptilia: Crocodylidae), in the Kruger National Park, South Africa.

Thirty-two specimens of the Nile crocodile, Crocodylus niloticus Laurenti (Reptilia: Crocodylidae), from the Kruger National Park, South Africa, and its vicinity were examined for pentastomid parasites during 1995 to 1999 and 2010 to 2011. Pentastomid parasites occurred throughout the year and were widespread in the study area with an overall prevalence of 97% and an overall mean abundance of 23.4 (0-81). Pentastome assemblages comprised six species in three sebekid genera: Alofia nilotici Riley et Huchzermeyer, 1995, A. simpsoni Riley, 1994, Leiperia cincinnalis Sambon, 1922, Sebekia cesarisi Giglioli in Sambon, 1922, S. minor (Wedl, 1861) and S. okavangoensis Riley et Huchzermeyer, 1995. The possible influence of host age, gender and geographic location (river system) on pentastome prevalence, abundance and species richness was investigated. Generally, neither host age, gender nor locality did affect infracommunities, likely because all hosts examined were adult or subadult and displayed comparable foraging behaviour, resulting in similar exposure pathways to fish intermediate hosts. Additionally, the longevity of pentastomids would contribute to accumulative infections as hosts mature. Structuring of pentastome assemblages was observed in as far as S. minor was the dominant species based on overall prevalence and abundance, followed by the equally common species S. cesarisi and L. cincinnalis. With an overall prevalence ranging from 34% to 41% and relatively low abundances, A. nilotici, A. simpsoni and S. okavangoensis form the rarer component of pentastome communities.

Field study site selection, species abundance and monthly distribution of anopheline mosquitoes in the northern Kruger National Park, South Africa.

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.

Chlorinated, brominated, and fluorinated organic pollutants in Nile crocodile eggs from the Kruger National Park, South Africa.

Repeated annual episodes of Nile crocodile deaths in two isolated areas of the Kruger National Park prompted the investigation of possible organohalogen pollutant involvement. Crocodile eggs were collected close to one of the mortality sites (Gorge) as well as from a crocodile farm (CF) as reference. ∑DDT was significantly higher in Gorge (450ng/g wm) than in CF eggs (85ng/g wet mass). Percentage DDT of ∑DDT was significantly higher in CF (14 per cent) than in Gorge eggs (5 per cent). Mean ∑DDT was almost 70 times higher than mean ∑PCB in Gorge eggs. HCB, ß-HCH, mirex, brominated flame retardants (BFRs), and perfluorinated compounds (PFCs) occurred at lower concentrations. We believe that the BFR and PFCs data represent the first published results for any crocodile egg. Thickening of the outer eggshell layer of Gorge eggs was significantly associated with higher concentrations of ∑DDT. Concentrations of ∑DDT and other pollutants were in the same range as eggs from elsewhere, where there were no mortalities. Concentrations of ∑DDT in eggs from healthy Australian crocodiles were of the same orders of magnitude as the current study, making it highly unlikely that the concentrations of pollutants measured in the present study would have caused or substantially contributed towards the mortalities observed. Concerns about reproduction and behaviour remain. As large predators, crocodilians are at the apex of the freshwater aquatic food web. More research is needed to guide measures to manage African freshwater systems so that it will also sustainably accommodate these large, long-lived animals.

Pansteatitis of unknown etiology associated with large-scale Nile crocodile (Crocodylus niloticus) mortality in Kruger National Park, South Africa: pathologic findings.

Annual mortality events in Nile crocodiles (Crocodylus niloticus) in the Olifants River Gorge in Kruger National Park, South Africa, were experienced between 2008 and 2012, during which at least 216 crocodiles died. Live crocodiles were lethargic. Necropsy examination of 56 affected crocodiles showed dark yellow-brown firm nodules in both somatic fat and the abdominal fat body. In all of the 11 crocodiles submitted for histology, degenerative, necrotic, and inflammatory changes supported a diagnosis of steatitis in both fat types. Crocodiles are apex predators in this anthropogenically changed aquatic ecosystem that is used by humans upstream and downstream from the park for domestic, agricultural, fishing, and recreational purposes. This pathologic review of pansteatitis in crocodiles in the Olifants River system was part of a broad multidisciplinary research program. To date, no definitive causative agent has been identified. Epidemiologic evidence suggests that this event may have been a one-time event with long-standing repercussions on the health of the crocodiles. Pathologic findings are rarely documented in wild crocodilians. This study also reports on other conditions, including the presence of coccidian oocysts, capillarid and filaroid nematodes, digenetic trematodes, and pentastomes.

The microscopic five of the big five: Managing zoonotic diseases within and beyond African wildlife protected areas.

African protected areas strive to conserve the continent's great biodiversity with a targeted focus on the flagship 'Big Five' megafauna. Though often not considered, this biodiversity protection also extends to the lesser-known microbes and parasites that are maintained in these diverse ecosystems, often in a silent and endemically stable state. Climate and anthropogenic change, and associated diversity loss, however, are altering these dynamics leading to shifts in ecological interactions and pathogen spill over into new niches and hosts. As many African protected areas are bordered by game and livestock farms, as well as villages, they provide an ideal study system to assess infection dynamics at the human-livestock-wildlife interface. Here we review five zoonotic, multi-host diseases (bovine tuberculosis, brucellosis, Rift Valley fever, schistosomiasis and cryptosporidiosis)-the 'Microscopic Five'-and discuss the biotic and abiotic drivers of parasite transmission using the iconic Kruger National Park, South Africa, as a case study. We identify knowledge gaps regarding the impact of the 'Microscopic Five' on wildlife within parks and highlight the need for more empirical data, particularly for neglected (schistosomiasis) and newly emerging (cryptosporidiosis) diseases, as well as zoonotic disease risk from the rising bush meat trade and game farm industry. As protected areas strive to become further embedded in the socio-economic systems that surround them, providing benefits to local communities, One Health approaches can help maintain the ecological integrity of ecosystems, while protecting local communities and economies from the negative impacts of disease.

Decision rules for determining terrestrial movement and the consequences for filtering high-resolution global positioning system tracks: a case study using the African lion (Panthera leo).

The combined use of global positioning system (GPS) technology and motion sensors within the discipline of movement ecology has increased over recent years. This is particularly the case for instrumented wildlife, with many studies now opting to record parameters at high (infra-second) sampling frequencies. However, the detail with which GPS loggers can elucidate fine-scale movement depends on the precision and accuracy of fixes, with accuracy being affected by signal reception. We hypothesized that animal behaviour was the main factor affecting fix inaccuracy, with inherent GPS positional noise (jitter) being most apparent during GPS fixes for non-moving locations, thereby producing disproportionate error during rest periods. A movement-verified filtering (MVF) protocol was constructed to compare GPS-derived speed data with dynamic body acceleration, to provide a computationally quick method for identifying genuine travelling movement. This method was tested on 11 free-ranging lions (Panthera leo) fitted with collar-mounted GPS units and tri-axial motion sensors recording at 1 and 40 Hz, respectively. The findings support the hypothesis and show that distance moved estimates were, on average, overestimated by greater than 80% prior to GPS screening. We present the conceptual and mathematical protocols for screening fix inaccuracy within high-resolution GPS datasets and demonstrate the importance that MVF has for avoiding inaccurate and biased estimates of movement.

Equal contributions of feline immunodeficiency virus and coinfections to morbidity in African lions.

Feline immunodeficiency virus (FIV) is a pathogenic lentivirus related to human and simian immunodeficiency viruses that has been associated with AIDS-like pathologies in domestic and wild cats, as well as in hyenas. Despite known pathologies, progressive immunosuppression and ill health effects driven by these lentiviruses in association with other secondary infections remain understudied in free-ranging species. Here, the role of coinfections by gastrointestinal parasites and tick-borne hemoparasites for FIV disease progression was explored in 195 free-ranging African lions (Panthera leo) living in Kruger National Park (KNP), South Africa. Using statistical methodology, we evaluated the effects of FIV on a range of health indicators to explore how direct and indirect effects of FIV and associated coinfections align to determine lion health outcomes. Findings show direct negative effects of FIV on host immunity and nutritional status, and exacerbation of aggressive behaviors, conditions which may increase exposure/susceptibility to other secondary infections. When taken together, the contribution of coinfecting parasites to morbidity in lions is of similar magnitude as direct effects of FIV infection alone, suggesting that the particular coinfection assemblage may play a role in mediating disease progression within natural lion populations.

First report of cystic echinococcosis in rhinos: A fertile infection of Echinococcus equinus in a Southern white rhinoceros (Ceratotherium simum simum) of Kruger National Park, South Africa.

Despite being a parasitic disease known since ancient times, some epidemiological aspects of cystic echinococcosis (CE) remain unclear. Many studies describe its prevalence and genotyping in populations of domestic animals and livestock, but data regarding wildlife are often scarce and incomplete. The available literature suggests that CE has never been reported in African rhinos. Considering the fragile conservation status of these species due to continued poaching, this study tries to clarify some neglected epidemiological aspects. In February 2020, an adult female of the Southern white rhinoceros, Ceratotherium simum simum (Burchell, 1817), was killed by poachers. The subsequent necropsy performed by the state veterinary team revealed the presence of seven cysts within the pulmonary tissue (four cysts in the right medio-caudal lobe and three cysts in the left medio-caudal lobe) with a diameter of between 1.5 and 2.3 cm. Given the state of decomposition of the carcass, only two of these were suitable for microscopic examination. Specimens were examined under 10x and 40x microscopic magnification for the confirmation of fertility of the cysts, based on the presence of numerous protoscoleces in different stages of maturation. A histopathological examination was also performed to describe the relationship between parasite and host tissue reaction. Cyst samples were subjected to PCR. The primers successfully amplified the expected fragments of the cox-1 and the nad-1 gene from the isolated genomic DNA, revealing high sequence identity with published sequences of Echinococcus equinus Williams & Sweatman, 1963 isolate G4 and E. equinus isolate SLG5-G4.

Levels of DDTs and other organochlorine pesticides in healthy wild Nile crocodiles (Crocodylus niloticus) from a flagship conservation area.

Mass Nile crocodile (Crocodylus niloticus) deaths in Africa's premier conservation area, the Kruger National Park (KNP), prompted numerous studies to determine possible causes of the sudden die-offs. The majority highlighted the involvement of the anthropocene to crocodile mass mortalities. One of the potential causative agents was identified as legacy compounds such as organochlorine pesticides (OCPs) present in the various aquatic ecosystems of the KNP. Thus, this study aimed to quantify OCP levels in wild crocodiles from the aquatic systems associated with the KNP. As part of a larger study, muscle tissue samples were collected from tails of 12 wild crocodiles. Results indicated that eight of the 19 OCPs analysed for were quantifiable. These included DDTs (2130-167968 ng/g lw), chlordanes (BD - 7583 ng/g lw) and cyclodeines (BD - 872 ng/g lw). Measured concentrations indicated spatial- and sex-related differences in accumulation patterns. DDT and its metabolites, paricularly p,p'-DDE accumulated to the highest concentrations of the OCP groups. Levels of the different groups of OCPs were the highest recorded in recent history from any crocodilian tissue. The different measured OCP residues were between 2.5 and 120 times higher than levels reported in literature, depending on the compound. These results are of ecotoxicological significance and have several management implications. In view of individuals being sampled from a conservation area it emphasizes the fact that pollutants know no boundaries and that premier conservation areas such as the world renowned KNP may be some of the most contaminated areas of the world.

How often should dead-reckoned animal movement paths be corrected for drift?

BACKGROUND: Understanding what animals do in time and space is important for a range of ecological questions, however accurate estimates of how animals use space is challenging. Within the use of animal-attached tags, radio telemetry (including the Global Positioning System, 'GPS') is typically used to verify an animal's location periodically. Straight lines are typically drawn between these 'Verified Positions' ('VPs') so the interpolation of space-use is limited by the temporal and spatial resolution of the system's measurement. As such, parameters such as route-taken and distance travelled can be poorly represented when using VP systems alone. Dead-reckoning has been suggested as a technique to improve the accuracy and resolution of reconstructed movement paths, whilst maximising battery life of VP systems. This typically involves deriving travel vectors from motion sensor systems and periodically correcting path dimensions for drift with simultaneously deployed VP systems. How often paths should be corrected for drift, however, has remained unclear. METHODS AND RESULTS: Here, we review the utility of dead-reckoning across four contrasting model species using different forms of locomotion (the African lion Panthera leo, the red-tailed tropicbird Phaethon rubricauda, the Magellanic penguin Spheniscus magellanicus, and the imperial cormorant Leucocarbo atriceps). Simulations were performed to examine the extent of dead-reckoning error, relative to VPs, as a function of Verified Position correction (VP correction) rate and the effect of this on estimates of distance moved. Dead-reckoning error was greatest for animals travelling within air and water. We demonstrate how sources of measurement error can arise within VP-corrected dead-reckoned tracks and propose advancements to this procedure to maximise dead-reckoning accuracy. CONCLUSIONS: We review the utility of VP-corrected dead-reckoning according to movement type and consider a range of ecological questions that would benefit from dead-reckoning, primarily concerning animal-barrier interactions and foraging strategies.

Comparison of the lipid properties of captive, healthy wild, and pansteatitis-affected wild Nile crocodiles (Crocodylus niloticus).

The results presented describe and compare the fatty acid composition and melting properties of captive, healthy wild, and pansteatitis-affected wild crocodiles (Crocodylus niloticus). Differences in fatty acid composition between intramuscular and adipose fat is noted in captive crocodiles, and the latter differs from wild crocodiles as a result of different diets. Adipose fat of healthy wild crocodiles differs minimally from diseased ones, respectively with 37.3+/-2.6% vs. 43.2+/-2.3% monounsaturated fatty acids, and 43.2+/-2.9% in dead crocodiles, while polyunsaturated fatty acids decrease from 27.3+/-1.9% to as low as 21.9+/-3.6% respectively. Of the unsaturated fatty acids 18:2n-6 decreased from 6.5+/-2.6% in unaffected crocodiles to 3.5+/-0.6% in highly affected and 3.2+/-0.4% in dead crocodiles, and 22:5n-3 from 2.8+/-0.6% to 1.8+/-0.3% and 2.2+/-0.3% respectively. The melting properties as determined by differential scanning calorimetry show that extracted adipose fat is a small degree softer in pansteatitis-affected tissue, specifically in the temperature range 7-36 degrees C, and does not contribute to the hard texture noted for adipose fat tissue of pansteatitis-affected animals. A high moisture content of 51.0+/-19.7% of the fat tissue of pansteatitis-affected animals vs.17.1+/-8.0% of healthy ones, suggests that physiological changes due to interstitial inflammation may contribute to the hard texture.

Human practices promote presence and abundance of disease-transmitting mosquito species.

Humans alter the environment at unprecedented rates through habitat destruction, nutrient pollution and the application of agrochemicals. This has recently been proposed to act as a potentially significant driver of pathogen-carrying mosquito species (disease vectors) that pose a health risk to humans and livestock. Here, we use a unique set of locations along a large geographical gradient to show that landscapes disturbed by a variety of anthropogenic stressors are consistently associated with vector-dominated mosquito communities for a wide range of human and livestock infections. This strongly suggests that human alterations to the environment promote the presence and abundance of disease vectors across large spatial extents. As such, it warrants further studies aimed at unravelling mechanisms underlying vector prevalence in mosquito communities, and opens up new opportunities for preventative action and predictive modelling of vector borne disease risks in relation to degradation of natural ecosystems.

Identification of toxigenic Microcystis strains after incidents of wild animal mortalities in the Kruger National Park, South Africa.

The eutrophic process potentially caused by a high urine and faecal load resulting from an unusually high hippopotamus (Hippopotamus amphibious) density in the Nhlanganzwane Dam, Kruger National Park, South Africa, triggered a chain of events characterised by an increase in the growth of primary producers (Microcystis aeruginosa). This increase in M. aeruginosa biomass was followed by bio-intoxication incidents in wild animals. In this study, we determine if a M. aeruginosa bloom with a total microcystin level of 23,718microgl(-1) have been responsible for mortalities of megaherbivores in the Nhlanganzwane Dam. We further use microcystin molecular markers derived from the mcy gene cluster to identify potentially toxigenic environmental Microcystis strains in the dam during the occurrence of animal intoxications. The estimated total microcystin-LR daily intake by an adult male white rhinoceros (Ceratotherium simum) from cyanobacterial-contaminated water of the dam during the toxic event was an order of magnitude higher (754.29microgkg(-1)bw) in comparison with the lowest observed adverse effecting level (LOAEL) value measured for pigs in a previous study by other authors. In this study the presence of toxic cyanobacterial strains was confirmed with the use of molecular markers that detected the presence of the mcy gene cluster responsible for the production of toxin by M. aeruginosa.

A comparative assessment of adult mosquito trapping methods to estimate spatial patterns of abundance and community composition in southern Africa.

BACKGROUND: Assessing adult mosquito populations is an important component of disease surveillance programs and ecosystem health assessments. Inference from adult trapping datasets involves comparing populations across space and time, but comparisons based on different trapping methods may be biased if traps have different efficiencies or sample different subsets of the mosquito community. METHODS: We compared four widely-used trapping methods for adult mosquito data collection in Kruger National Park (KNP), South Africa: Centers for Disease Control miniature light trap (CDC), Biogents Sentinel trap (BG), Biogents gravid Aedes trap (GAT) and a net trap. We quantified how trap choice and sampling effort influence inferences on the regional distribution of mosquito abundance, richness and community composition. RESULTS: The CDC and net traps together collected 96% (47% and 49% individually) of the 955 female mosquitoes sampled and 100% (85% and 78% individually) of the 40 species or species complexes identified. The CDC and net trap also identified similar regional patterns of community composition. However, inference on the regional patterns of abundance differed between these traps because mosquito abundance in the net trap was influenced by variation in weather conditions. The BG and GAT traps collected significantly fewer mosquitoes, limiting regional comparisons of abundance and community composition. CONCLUSIONS: This study represents the first systematic assessment of trapping methods in natural savanna ecosystems in southern Africa. We recommend the CDC trap or the net trap for future monitoring and surveillance programs.

Eutrophication governs predator-prey interactions and temperature effects in Aedes aegypti populations.

BACKGROUND: Mosquito population dynamics are driven by large-scale (e.g. climatological) and small-scale (e.g. ecological) factors. While these factors are known to independently influence mosquito populations, it remains uncertain how drivers that simultaneously operate under natural conditions interact to influence mosquito populations. We, therefore, developed a well-controlled outdoor experiment to assess the interactive effects of two ecological drivers, predation and nutrient availability, on mosquito life history traits under multiple temperature regimes. METHODS: We conducted a temperature-controlled mesocosm experiment in Kruger National Park, South Africa, with the yellow fever mosquito, Aedes aegypti. We investigated how larval survival, emergence and development rates were impacted by the presence of a locally-common invertebrate predator (backswimmers Anisops varia Fieber (Notonectidae: Hemiptera), nutrient availability (oligotrophic vs eutrophic, reflecting field conditions), water temperature, and interactions between each driver. RESULTS: We observed that the effects of predation and temperature both depended on eutrophication. Predation caused lower adult emergence in oligotrophic conditions but higher emergence under eutrophic conditions. Higher temperatures caused faster larval development rates in eutrophic but not oligotrophic conditions. CONCLUSIONS: Our study shows that ecological bottom-up and top-down drivers strongly and interactively govern mosquito life history traits for Ae. aegypti populations. Specifically, we show that eutrophication can inversely affect predator-prey interactions and mediate the effect of temperature on mosquito survival and development rates. Hence, our results suggest that nutrient pollution can overrule biological constraints on natural mosquito populations and highlights the importance of studying multiple factors.