Diel activity and preferred landing sites in Culicoides biting midges attacking Fjord horses

In the summer of 2014, in the central part of The Netherlands, Culicoides spp. (Diptera: Ceratopogonidae) attack rates, biting rates, and preferred landing sites were determined for a pair of Fjord horses maintained permanently at pasture in an area devoid of cattle. Eleven body regions of the horses were screened for midges, each region sampled randomly for 5 min using a handheld mouth aspirator (pooter). Observations were confined to the hour immediately before and after sunset. Culicoides spp. were obtained from every body region, of which the four most abundant species – Culicoides chiopterus (Meigen), Culicoides punctatus (Meigen), the species complex Culicoides obsoletus (Meigen), and Culicoides dewulfi Goetghebuer – all were proven or potential vectors for arboviral diseases in livestock. Culicoides spp. activity was distinctly bimodal across the day, surging at sunset and 1 h after sunrise. Midges were inactive between 11:00 and 16:00 hours, these hours marking the time of day when horses can be pastured most safely but, thereafter, to avoid escalating attacks, would have to be stabled protectively. Around sunset, the mean attack rate of the four most abundant species ranged from 3.0 to 11.7 midges per min; of these, C. dewulfi and C. chiopterus were reared out of the dung of experimental horses. The Netherlands is home to the world's densest horse population (11 per km²), of which half are estimated to stay outdoors permanently with no access to protective housing. In the absence of a preventive vaccination policy, it is difficult to envisage how horses in northern Europe will be protected from infection during an outbreak of a Culicoides‐transmitted disease like African horse sickness.     

Culicoides species as potential vectors of African horse sickness virus in the southern regions of South Africa

African horse sickness (AHS), a disease of equids caused by the AHS virus, is of major concern in South Africa. With mortality reaching up to 95% in susceptible horses and the apparent reoccurrence of cases in regions deemed non‐endemic, most particularly the Eastern Cape, epidemiological research into factors contributing to the increase in the range of this economically important virus became imperative. The vectors, Culicoides (Diptera: Ceratopogonidae), are considered unable to proliferate during the unfavourable climatic conditions experienced in winter in the province, although the annual occurrence of AHS suggests that the virus has become established and that vector activity continues throughout the year. Surveillance of Culicoides within the province is sparse and little was known of the diversity of vector species or the abundance of known vectors, Culicoides imicola and Culicoides bolitinos. Surveillance was performed using light trapping methods at selected sites with varying equid species over two winter and two outbreak seasons, aiming to determine diversity, abundance and vector epidemiology of Culicoides within the province. The research provided an updated checklist of Culicoides species within the Eastern Cape, contributing to an increase in the knowledge of AHS vector epidemiology, as well as prevention and control in southern Africa.     

The distribution and species characteristics of the Culicoides biting midge fauna of South Africa

Abstract.  1. Monthly light trap collections over 2 years at 39 sites across South Africa captured over 3 000 000 Culicoides of 86 species, some of them known vectors of African horse sickness and bluetongue viruses.
2. To aid disease control and risk evaluation, cluster analysis and association measurements were carried out on species present at each site. Six species occurred at almost all sites. From the remaining 80 species, eight clusters were identified at > 48% similarity.
3.  Culicoides citroneus and C. krameri were characteristic of cluster 8, C. loxodontis of cluster 4, and two numbered, but un-named, species, C. #119 and C. #89 , of cluster 5.
4. Multiple anova and multiple discriminant analysis on the climatic and geographic parameters of sites in each cluster gave two significant discriminant functions that explained 91.5% of the variance between clusters. Function 1 contrasted sites with high cold cloud duration and high vegetation index with sites with high wind speeds. Function 2 contrasted upland clusters that also had high cold cloud duration with hotter, lower altitude sites.
5. Species associations were explained in terms of biotic provinces, based on ecogeographic factors that have previously been useful to describe communities of birds.
6. None of the clusters was ideal for siting an African horse sickness-vector free quarantine station as C. imicola and C. bolitinos , the main vectors, were found in all clusters but in very low abundance in cluster 5. The analysis provides a firm basis for studying the changing patterns of species distribution under varying climatic influences.     

Preliminary note on the distribution and ecology of Culicoides imicola in Portugal

Abstract. Data on Culicoides imicola were obtained during studies carried out during the recent outbreak of African horse sickness in Portugal. The previous most northerly published record of C.imicola in Portugal was 38^o40'N (Pégöes). In the present work the geographical distribution of this species is extended to the parallel of 41^o17'N. We have also confirmed the continuous presence of adult C.imicola in Southern Portugal (Alentejo and Algarve) throughout the year. In the laboratory we obtained this species from a sample of cattle faeces and from another of soil contaminated with animal excreta. In relation to host association 57.37% of C.imicola were trapped in the vicinity of pigsties. Finally, we collected 11 ,463 Culicoides of which 12.47% were C. imicola.     

Control techniques for Culicoides biting midges and their application in the U.K. and northwestern Palaearctic

The recent emergence of bluetongue virus (Reoviridae: Orbivirus) (BTV) in northern Europe, for the first time in recorded history, has led to an urgent need for methods to control the disease caused by this virus and the midges that spread it. This paper reviews various methods of vector control that have been employed elsewhere and assesses their likely efficacy for controlling vectors of BTV in northern Europe. Methods of controlling Culicoides spp. (Diptera: Ceratopogonidae) have included: (a) application of insecticides and pathogens to habitats where larvae develop; (b) environmental interventions to remove larval breeding sites; (c) controlling adult midges by treating either resting sites, such as animal housing, or host animals with insecticides; (d) housing livestock in screened buildings, and (e) using repellents or host kairomones to lure and kill adult midges. The major vectors of BTV in northern Europe are species from the Culicoides obsoletus (Meigen) and Culicoides pulicaris (L.) groups, for which there are scant data on breeding habits, resting behaviour and host-oriented responses. Consequently, there is little information on which to base a rational strategy for controlling midges or for predicting the likely impact of interventions. However, data extrapolated from the results of vector control operations conducted elsewhere, combined with some assessment of how acceptable or not different methods may be within northern Europe, indicate that the treatment of livestock and animal housing with pyrethroids, the use of midge-proofed stabling for viraemic or high-value animals and the promotion of good farm practice to at least partially eliminate local breeding sites are the best options currently available. Research to assess and improve the efficacy of these methods is required and, in the longer term, efforts should be made to develop better bait systems for monitoring and, possibly, controlling midges. All these studies will need better methods of analysing the ecology and behaviour of midges in the field than are currently in use. The paucity of control options and basic knowledge serve to warn us that we must be better prepared for the possible emergence of other midge-borne diseases, particularly African horse sickness.     

Phylogenetic status and matrilineal structure of the biting midge, Culicoides imicola, in Portugal, Rhodes and Israel

The biting midge Culicoides imicola Kieffer (Diptera: Ceratopogonidae) is the most important Old World vector of African horse sickness (AHS) and bluetongue (BT). Recent increases of BT incidence in the Mediterranean basin are attributed to its increased abundance and distribution. The phylogenetic status and genetic structure of C. imicola in this region are unknown, despite the importance of these aspects for BT epidemiology in the North American BT vector. In this study, analyses of partial mitochondrial cytochrome oxidase subunit I gene (COI) sequences were used to infer phylogenetic relationships among 50 C. imicola from Portugal, Rhodes, Israel, and South Africa and four other species of the Imicola Complex from southern Africa, and to estimate levels of matrilineal subdivision in C. imicola between Portugal and Israel. Eleven haplotypes were detected in C. imicola, and these formed one well-supported clade in maximum likelihood and Bayesian trees implying that the C. imicola samples comprise one phylogenetic species. Molecular variance was distributed mainly between Portugal and Israel, with no haplotypes shared between these countries, suggesting that female-mediated gene flow at this scale has been either limited or non-existent. Our results provide phylogenetic evidence that C. imicola in the study areas are potentially competent AHS and BT vectors. The geographical structure of the C. imicola COI haplotypes was concordant with that of BT virus serotypes in recent BT outbreaks in the Mediterranean basin, suggesting that population subdivision in its vector can impose spatial constraints on BT virus transmission.     

Host use patterns of Culicoides spp. biting midges at a big game preserve in Florida,U.S.A., and implications for the transmission of orbiviruses

Culicoides spp. biting midges (Diptera: Ceratopogonidae) are vectors of pathogens that have a significant economic impact on the livestock industry. White‐tailed deer (Odocoileus virginianus), a farmed species in the U.S.A., are susceptible to two Culicoides spp. borne orbiviruses: bluetongue virus and epizootic haemorrhagic disease virus. Elucidating host–vector interactions is an integral step in studying disease transmission. This study investigated the host range of Culicoides spp. present on a big game preserve in Florida on which a variety of Cervidae and Bovidae freely roam. Culicoides were captured with Centers for Disease Control and Prevention (CDC) miniature light traps run twice weekly on the preserve for 18 consecutive months (July 2015–December 2016). Host preference was quantified through forage ratios, based upon PCR‐based bloodmeal analysis of Culicoides spp. and overall animal relative abundance on the preserve. Culicoides stellifer preferentially fed on Cervus spp. and fallow deer (Dama dama) and displayed a relative avoidance of Bovidae and white‐tailed deer. Culicoides debilipalpis preferred white‐tailed deer and avoided all Bovidae. Culicoides pallidicornis and Culicoides biguttatus showed preferences for white‐tailed deer and Père David's deer (Elaphurus davidianus), respectively. These results add to current knowledge of preferred hosts of Florida Culicoides spp. and have implications for the spread of orbiviruses. Copyright © 2018 John Wiley & Sons, Ltd.     

Benthic macroinvertebrate assemblage composition and distribution pattern in the upper Mthatha River,Eastern Cape,South Africa

Benthic macroinvertebrates were sampled and environmental variables measured at Langeni Forest and Kambi Valley, two sites of varying degrees of human impact in the upper Mthatha River, South Africa, in 2010–2011. Four species, Simulium adersi Pomeroy, Baetis harrisoni Barnard, Pseudocloeon sp. and Tricorythus sp., occurred at all site sampling units. Specimen counts increased from the rainy months of January to April/May, and higher numbers of individuals occurred at Langeni. Species occurrences per site were directly or indirectly affected by season, substrate and habitat heterogeneity. Canonical correspondence analysis results showed that conductivity, pH, flow, substrate and marginal vegetation were important in determining species distribution patterns. Both rare/habitat-restricted and common/widespread benthic macroinvertebrates identified in this study can be used as indicators for assessing the ecological integrity of the Mthatha River. Habitat management along the river should aim at preserving indigenous vegetation, especially during the summer season when habitat requirements are optimal for growth stages of most benthic macroinvertebrate fauna. The practicality of using species-level surveys on a routine basis can be challenging, given the lack of taxonomic expertise and time constraints. It might be advisable to select a few indicator species and to monitor them as surrogates for the whole community.     

Angulate and leopard tortoises in the Thicket Biome, Eastern Cape, South Africa: populations and biomass estimates

The population parameters (morphometrics, population size and biomass) were determined for leopard and angulate tortoises in the Thicket Biome, Eastern Cape, South Africa. Leopard tortoises had a mean total length of 306.47mm (SD = 94.65mm, range 59–496mm, n = 89). Males were smaller (mean total length 277.31mm, SD = 71.85mm, range 73–377mm, n = 36) than females (mean total length = 332.33mm, SD = 97.64mm, range 59–496mm, n = 53). Angulate tortoises had a mean total length of 155.17mm (SD = 33.89mm, range 59–194mm, n = 18). Males were larger (mean total length = 186mm, SD = 8.40mm, range 174–194mm, n = 5) than females (mean total length = 143.31mm, SD = 32.48mm, range 59–178mm, n = 13). The estimated population density for leopard tortoises was 0.85 tortoises ha⁻¹ with an estimated biomass of 6.02kg ⁻¹. Angulate tortoise population density was estimated at 0.12 tortoises ha⁻¹ with a mean biomass of 0.06kg ha⁻¹. The possible consequences of this high density and biomass are discussed.     

Seasonal abundance, reproduction and hunting of common quail Coturnix coturnix in the Eastern Cape Province, South Africa

This study investigated the potential overlap between breeding and hunting seasons for a population of common quail Coturnix coturnix in the Alexandria District. Seasonal abundance (assessed as the number of birds flushed per unit effort), sex ratios and reproductive condition (based on gonad sizes) of the quail were measured over a period of 16 months, including two hunting seasons. Hunting impacts were assessed in terms of the number of birds killed per unit of hunting effort. Quail exhibited a spring peak in abundance and reproduction, which coincided with the legal hunting season, and bird numbers varied markedly between years. Sex ratio varied, with males dominating early in spring. Hunters killed nearly 60% of the birds flushed, and hunter efficiency was positively correlated with the abundance of quail. These results confirm the dilemma for managers, as quail are currently hunted during the main breeding season and offtakes are high. The development of management strategies is hampered by a lack of understanding of the population biology and ecology, and impacts of hunting. A number of possible hunting strategies are discussed. Since current harvests may not be sustainable, further research is necessary.     

Modelling expected trout ranges under current and future water temperature regimes in the Eastern Cape,South Africa

Different values have resulted in conflicts between anglers and conservation lobbies in the management of trout in South Africa. Key to the conflict is the demarcation of boundaries to areas in which brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss currently occur, or are likely to establish following stocking for angling. To provide a longer-term perspective on these areas, we developed models to link salmonid biological thermal thresholds to elevation. These, when applied spatially using a digital elevation model with a probability of occurrence model, provided the basis for estimating potentially available thermal habitat for these two cold water species. Here, we acknowledge that other variables (stocking history; river connectivity) also play a role in understanding trout distributions. Using a simple scenario of an increase in mean daily water temperatures of 2 °C, we demonstrated that both brown and rainbow trout are likely to exhibit considerable range reductions in the future. Because it is possible that these range restrictions will result in an increasing desire to introduce trout into areas above their current distribution limits for the maintenance of angling opportunities, conservation managers should prioritise these areas, with management interventions seeking to understand what will help to limit introductions.     

Aspects of population dynamics and feeding by piscivorous birds in the intermittently open Riet River estuary,Eastern Cape,South Africa

Aspects of the population dynamics and feeding activity of piscivorous birds in the small (c. 5 ha) intermittently open Riet River estuary, on the south-eastern coastline of South Africa, were investigated monthly from August 2005 to July 2006. A total of 188 birds of 13 species were recorded, of which six were wading piscivores, four aerial divers and three were pursuit swimmers. The Reed Cormorant (Phalacrocorax africanus) was the numerically dominant species, with a mean of 8.25 (SD ± 7.90) individuals per count. Mean numbers of the Pied Kingfisher (Ceryle rudis) and Giant Kingfisher (Megaceryle maximus) were 3.42 (SD ± 1.20) and 1.17 (SD ± 0.60) individuals per count, respectively. The remaining 10 species revealed mean values <0.5 individuals per count. Breaching events were associated with a change in feeding groups from waders to pursuit feeders, and a decrease in total bird numbers, most likely due to loss of potential littoral zone foraging habitat for waders resulting from reduced water levels. The highest bird numbers were recorded in winter reflecting the migration of large numbers of Reed Cormorant into the system. Monthly food consumption by all piscivorous birds showed large temporal variability, ranging from 26.35 to 140.58 kg per month.     

Home range size, spatial distribution and habitat use of elephants in two enclosed game reserves in the Eastern Cape Province, South Africa

We studied space use and habitat selection by elephants in two enclosed reserves in the Eastern Cape Province (South Africa) that conserve components of the regionally important and poorly conserved Thicket Biome. Home ranges of bulls and herds occupied most of the reserves, and core areas were significantly smaller and centred on permanent water. There was no significant difference in home range size of bulls and herds and both were significantly smaller in winter than summer. Components of the Thicket Biome occurred in the home ranges and core areas of all elephants where it was either used according to its abundance or avoided. However, this should not be interpreted as indicating that elephants will not affect plants of the Thicket Biome since change will be cumulative and monitoring should be ongoing to ensure the conservation of this regionally important Biome.     

Continued decline in tree euphorbias (Euphorbia tetragona and E. triangularis) on the Great Fish River Reserve,Eastern Cape,South Africa

A dramatic decline averaging 43% over a 4‐year period has occurred in tree Euphorbia (Euphorbia tetragona and Euphorbia triangularis) populations on the Great Fish River Reserve, South Africa. These changes are evident from data gathered by general vegetation monitoring methods as well as from a focused study of four tree Euphorbia populations. The decline from 2003 to 2007 was more marked for E. triangularis than for E. tetragona and was accompanied by a general absence of seedlings and a reduced presence of younger age classes of both species, decreasing the proportion of younger trees in the populations. The role of megaherbivores, specifically the black rhinoceros, in these changes is well established. However, the impact of baboon activity, leading to damage to tree crowns and upper branches, is also substantial, especially on E. triangularis populations. Damaged crowns were recorded significantly more often for E. triangularis than for E. tetragona, and the damage frequency increased with decreasing tree height. Thus, our work provides the first evidence that these two closely related Euphorbia species may be affected differently by herbivory.     

A survey of fruit-feeding insects and their parasitoids occurring on wild olives,Olea europaea ssp. cuspidata,in the Eastern Cape of South Africa

Fruits of wild olives, Olea europaea ssp. cuspidata (Wall. ex G. Don) Cif., were collected in the Eastern Cape, South Africa, during 2003–2005 to quantify levels of fruit-infesting pests and their parasitoids. Two species of Tephritidae, Bactrocera oleae (Rossi) and B. biguttula (Bezzi), were the most abundant insects recovered and were reared from most samples. Fruit infestation rates by the Bactrocera spp. were generally below 8% and over half of the infestations were under 1%. When parasitism occurred in samples with flies, levels ranged from 7 to 83%. Several species of opiine braconid wasps, Psyttalia concolor (Szépligeti), Psyttalia lounsburyi (Silvestri), and Utetes africanus (Szépligeti) and one braconine wasp, Bracon celer Szépligeti, were reared from fruits containing B. oleae and/or B. biguttula. Chalcidoid parasitoids and seed wasps included seven species of Eurytomidae (Eurytoma oleae, Eurytoma sp., and Sycophila sp.), Ormyridae (Ormyrus sp.), Torymidae (Megastigmus sp.), and Eupelmidae (Eupelmus afer and E. spermophilus). One species of moth, Palpita unionalis (Hübner) (Crambidae), was recovered in very low numbers and without parasitoids. The survey results indicate that fruit flies might not become economic pests of the nascent commercial olive industry in the Eastern Cape, and the small numbers present may be controlled to a considerable level by natural enemies.     

Natural variation in macroinvertebrate assemblages and the development of a biological banding system for interpreting bioassessment data—a preliminary evaluation using data from upland sites in the south-western Cape,South Africa

The variability of macroinvertebrate assemblages was investigated at 27 upland reference sites in the south-western Cape, South Africa. Multivariate analyses showed that sites did not group on the basis of geomorphological zonation, i.e. mountain stream and foothill-cobble bed. When separate analyses were undertaken for mountain stream (n = 21) and foothill-cobble bed sites (n = 6), assemblages formed three and two groups, respectively. Similarity amongst groups ranged from 47% to 52%, while within-group similarity was between 54% and 67%. Environmental variables shown to contribute to this variability included distance from source, cation ratio ([Na⁺]+[K⁺]/([Na⁺]+[K⁺]+[Ca²⁺]+[Mg²⁺]), pH, longitude and stream width. Whilst overall variability in the metrics of the biotic index, SASS (South African Scoring System), is high at reference sites, the interpretation of monitoring-site data using biological bands derived from a range of reference sites, ensured that variability was taken into account and that detection of disturbance at a monitoring site was not impeded. A biological banding system has been developed for upland sites in the south-western Cape, together with a list of reference or expected SASS-taxa. This list includes details pertaining to seasonality and biotope preferences. The ability to define reference conditions that take intrinsic variability amongst reference sites into account is important for the accurate interpretation of bioassessment data. Handling editor: D. Dudgeon     

Sea-level changes, river capture and the evolution of populations of the Eastern Cape and fiery redfins (Pseudobarbus afer and Pseudobarbus phlegethon, Cyprinidae) across multiple river systems in South Africa

Aim The phylogeography of the two closely related species Pseudobarbus afer and Pseudobarbus phlegethon was investigated to assess the association of evolutionary processes, inferred from mitochondrial DNA (mtDNA) sequence variation, with hypothetical palaeoriver systems and other climatic and landscape changes. Location One western and several southern river systems in South Africa. Methods We sampled known populations and confirmed known distribution gaps. This was followed by an assessment of mtDNA control region sequence variation for 31 localities across 17 river systems across the range of the species complex. A map of possible offshore drainage patterns during the last major regression event was constructed based on bathymetry and geological studies. Results The genetic distinction of four major lineages of P. afer strongly correspond with proposed palaeoriver systems. However, a western ‘Forest’ lineage, is widespread across two such proposed systems and is closely related to P. phlegethon on the west coast of South Africa. Both the ‘Krom’ and ‘St Francis’ lineages were identified in the single palaeoriver system proposed for St Francis Bay. A fourth ‘Mandela’ lineage is restricted to the one or two palaeoriver systems proposed for Nelson Mandela Bay. Four minor lineages were identified within the Forest lineage and two within the Mandela lineage. Main conclusions The close relationship between P. phlegethon and the Forest lineage of P. afer can only be explained by a series of river captures. We suggest the Gourits River system as a historical link that could account for this relationship. On the south coast, lower sea levels than at present allowed confluence between currently isolated river systems, offering opportunities for dispersal among these populations. At present, isolation between different river systems rather than dispersal appears to have a dominant influence on mtDNA diversity.     

Using a trait‐based approach for assessing the vulnerability and resilience of hillslope seep wetland vegetation cover to disturbances in the Tsitsa River catchment,Eastern Cape,South Africa

Hill slope seep wetlands are ecologically and economically important ecosystems as they supply a variety of ecosystem services to society. In South Africa, livestock grazing is recognized as one of the most important disturbance factors changing the structure and function of hill slope seep wetlands. This study sought to investigate the potential effect of livestock grazing on the resilience and vulnerability of hillslope seep wetland vegetation cover using a trait‐based approach (TBA). Changes in vegetation cover were used as a surrogate for indicating grazing intensity. The degree of human disturbances was assessed using the Anthropogenic Activity Index. A TBA was developed using seven plant traits, resolved into 27 trait attributes. Based on the developed approach, plant species were grouped into vulnerable and resilient groups in relation to grazing pressure. It was then predicted that species belonging to the vulnerable group would be less dominant at the highly disturbed sites, as well as in the winter season when grazing pressure is at its peak. The approach developed enabled accurate predictions of the responses of hillslope plant species to grazing pressure seasonally, but spatially, only for the summer season. The predicted responses during the winter season across sites did not match the observed results, which could be attributed to the difficulty in species identification and accurate estimation of vegetation cover during winter. Overall, the approach developed here provides a general framework for applying the TBA and can thus be tested and applied elsewhere.     

福寿螺的寄主植物及其对福寿螺体重的影响

为了明确福寿螺对寄主植物的选择性,进一步掌握其可能的危害及入侵暴发机制,对稻田水域及周边的24科43种植物进行了室外取食试验,分别在取食后3、5d对受试样品及成螺称重,比较其无选择条件下的取食偏好性及其对成螺增长率的影响。结果表明:福寿螺对受试的24科43种植物(样品)的根、茎、叶、果实以及块根(茎)均能取食,对水生(湿生)和陆生植物没有偏好性;在取食过程中,优先取食细嫩的植株和部分,硬老部分需经水浸泡软化后才能被取食;辣椒、苦瓜等果实的辣苦味对福寿螺无驱避作用;取食率与体重增长率呈显著正相关,取食越多生长越快;但体重与植物的总酚含量呈负相关。福寿螺的广食性、取食量、植物总酚含量、植物丰盛度等种群和生态系统特征是影响其快速扩散的重要原因,同时可以借助福寿螺对植物的不同利用类型防范其扩散和危害。     

A lonely dot on the map: Exploring the climate signal in tree-ring density and stable isotopes of clanwilliam cedar,South Africa

Clanwilliam cedar (Widdringtonia cedarbergensis; WICE), a long-lived conifer with distinct tree rings in Cape Province, South Africa, has potential to provide a unique high-resolution climate proxy for southern Africa. However, the climate signal in WICE tree-ring width (TRW) is weak and the dendroclimatic potential of other WICE tree-ring parameters therefore needs to be explored. Here, we investigate the climatic signal in various tree-ring parameters, including TRW, Minimum Density (MND), Maximum Latewood Density (MXD), Maximum Latewood Blue Intensity (MXBI), and stable carbon and oxygen isotopes (δ¹⁸O and δ¹³C) measured in WICE samples collected in 1978. MND was negatively influenced by early spring (October-November) precipitation whereas TRW was positively influenced by spring November-December precipitation. MXD was negatively influenced by autumn (April-May) temperature whereas MXBI was not influenced by temperature. Both MXD and MXBI were negatively influenced by January-March and January-May precipitation respectively. We did not find a significant climate signal in either of the stable isotope time series, which were measured on a limited number of samples. WICE can live to be at least 356 years old and the current TRW chronology extends back to 1564 CE. The development of full-length chronologies of alternative tree-ring parameters, particularly MND, would allow for an annually resolved, multi-century spring precipitation reconstruction for this region in southern Africa, where vulnerability to future climate change is high.