Comparative performance of microscopy, nested PCR, and real-time PCR for screening avian haemosporidian parasites in Afrotropical starlings (family Sturnidae).

Prevalence studies of avian haemosporidian parasites frequently use microscopy and the nested polymerase chain reaction (PCR) protocols for detecting infections. Newer PCR protocols to detect parasites are being developed, with the distinct advantage of reducing screening cost and time, as well as increasing efficiency and sensitivity. The detection ability of microscopy and nested PCR was compared against a real-time PCR (qPCR) protocol using genomic DNA extracted from 240 bird blood samples collected from three starling species (Cape Starling, the Greater Blue-eared Starling, and the Wattled Starling; family Sturnidae) in the Kruger national park, South Africa. All three protocols successfully detected avian haemosporidian parasites with the qPCR having a considerable edge against the other two methods. Fifteen unique cytochrome b lineages were identified of which seven were new lineages. Microscopy and nested PCR recorded similar prevalence (32.92% and 35.42% respectively). The qPCR protocol used here, although more sensitive (52.92% prevalence), is not able to differentiate between parasite genera but provides the opportunity to screen a large number of samples in large-scale studies within a specific region. This study recommends the development and adoption of new molecular protocols with increased sensitivity and accuracy in prevalence studies. Nevertheless, microscopy remains essential for the morphological description of parasites and for distinguishing between abortive and successful chronic infections. The PCR-based method displays the detection of the parasitic genome but does not reveal whether parasites have or will develop into a successful infection.

Use of Epidemiology Surge Support to Enhance Robustness and Expand Capacity of SARS-CoV-2 Pandemic Response, South Africa.

As COVID-19 cases increased during the first weeks of the pandemic in South Africa, the National Institute of Communicable Diseases requested assistance with epidemiologic and surveillance expertise from the US Centers for Disease Control and Prevention South Africa. By leveraging its existing relationship with the National Institute of Communicable Diseases for >2 months, the US Centers for Disease Control and Prevention South Africa supported data capture and file organization, data quality reviews, data analytics, laboratory strengthening, and the development and review of COVID-19 guidance This case study provides an account of the resources and the technical, logistical, and organizational capacity leveraged to support a rapid response to the COVID-19 pandemic in South Africa.

Avian haemosporidia in native and invasive sparrows at an Afrotropical region.

Bio-invasions are a major threat to biodiversity and ecosystems globally and may contribute to the proliferation of emerging infectious diseases. We examined the prevalence and phylogenetic diversity of avian haemosporidian parasites infecting the non-native house sparrows (Passer domesticus) and the native southern grey-headed sparrows (Passer diffusus). Blood samples from 104 sparrows (74 house sparrows and 30 southern grey-headed sparrows) mist-netted inside and around the Kruger National Park were used. Genomic DNA was extracted from each blood sample and subjected to nested PCR analyses, Sanger sequencing and phylogenetic analyses. Overall, 35.57% (37/104) of the birds sampled were infected with at least one haemosporidian parasites. Southern grey-headed sparrows had a higher parasite prevalence (60%) than house sparrows (24.3%). A total of 16 parasite lineages were identified, of which eight were novel lineages. Whereas Haemoproteus spp. showed the highest lineage diversity, Leucocytozoon spp. were the most prevalent parasites, albeit with significant differences between sparrow species. A single Plasmodium sp. infection was recorded in a southern grey-headed sparrow. In support of the enemy release hypothesis, we found that prevalence on non-native house sparrows was lower than prevalence recorded in their region of origin and also that they were infected only by indigenous parasites lineages.

Plant shade enhances thermoregulation of internal environments in Trinervitermes trinervoides mounds.

Microhabitats may be crucial in buffering organisms from temperature extremes, particularly given increases in maximum temperature associated with global climate change. For example, thermoregulation in termite mounds is influenced by prevailing ambient conditions, and plant canopies may reduce external temperatures, in turn lowering internal temperatures. This buffering may be crucial during heat waves. Whether this happens, and to what extent, remains equivocal, however. We tracked internal temperatures in eight inhabited and six uninhabited Trinervitermes trinervoides mounds, half of each group of which were shaded by vegetation. T. trinervoides seek to maintain internal mound temperatures at c. 20 °C in winter and c. 30 °C in summer. Temperatures were logged for 72 h in winter, and again in summer. Internal temperatures of uninhabited mounds mirrored those of external temperatures, with temperatures varying by c. 15 °C, although shading was associated with some buffering of internal temperatures. Internal temperatures within inhabited mounds were far less variable, varying by c. 6 °C over the course of our study. In summer, exposed inhabited mounds maintained temperatures c. 29.5 °C, whilst shaded inhabited mounds were c. 27.5 °C. In winter, mean internal temperatures of exposed and shaded inhabited mounds were very similar, at 21.8 and 22.0 °C, respectively. Internal mound temperature varied significantly with external (ambient) temperature, mound activity, temperature, shading, and to a small extent, mound volume. The buffering effect of shade was evident in summer (c. 2 °C) but not in winter, suggesting that the benefit of such temperature modulation may be most important when ambient temperatures reach heat extremes, e.g. during heat waves.

Water for African elephants (Loxodonta africana): faecal microbial loads affect use of artificial waterholes.

In semi-arid protected areas, artificial waterholes ensure that water is locally available to animals for extended periods. However, artificial waterholes may limit animal movement, which contributes towards habitat deterioration. Challenges of artificial water provisioning worsen in the presence of ecosystem engineers like African elephants Loxodonta africana, capable of transforming environments. Camera traps were used to monitor elephant visitation at 21 artificial waterholes in the Kruger National Park, South Africa. We also assessed if water quality parameters influenced elephant preference for certain waterholes. There were no significant correlations between elephant abundance and water physicochemical properties. However, there was a strong negative correlation between elephant abundance and levels of Escherichia coli in water. Our findings suggest that elephants avoid drinking water with high levels of faecal microbial loads. Whereas most studies addressing animal management in protected areas consider waterholes as homogeneous units, we posit that water quality could also determine local landscape use and movement patterns of key species like elephants, a finding with relevant implications in reserve management practices.

Diversity and Host Specificity of Avian Haemosporidians in an Afrotropical Conservation Region.

Afrotropical regions have high bird diversity, yet few studies have attempted to unravel the prevalence of avian haemosporidia in conservation areas. The diversity and host specificity of parasites in biodiversity hotspots is crucial to understanding parasite distribution and potential disease emergence. We test the hypothesis that biodiverse regions are associated with highly diverse parasites. By targeting the cytochrome b (Cytb) gene, we molecularly screened 1035 blood samples from 55 bird species for avian haemosporidia infections to determine its prevalence and diversity on sites inside and adjacent to the Kruger National Park. Overall infection prevalence was 28.41%. Haemoproteus, Leucocytozoon, and Plasmodium presented prevalences of 17.39%, 9.24%, and 4.64%, respectively. One hundred distinct parasite lineages were detected, of which 56 were new lineages. Haemoproteus also presented the highest diversity compared to Leucocytozoon and Plasmodium with varying levels of specificity. Haemoproteus lineages were found to be specialists while Plasmodium and Leucocytozoon lineages were generalists. We also found a positive relationship between avian host diversity and parasite diversity, supporting an amplification effect. These findings provide insight data for host-parasite and co-evolutionary relationship models.

Pediatric and adolescent HIV viral load coverage and suppression rates in the context of the COVID-19 pandemic in 12 PEPFAR-supported sub-Saharan African countries in 2019 and 2020.

The early period of the COVID-19 pandemic limited access to HIV services for children and adolescents living with HIV (C/ALHIV). To determine progress in providing care and treatment services, we describe viral load coverage (VLC) and suppression (VLS) (<1000 copies/ mL) rates during the COVID-19 pandemic in 12 United States President's Emergency Plan for AIDS Relief (PEPFAR)-supported countries. Data for children (0-9 years) and adolescents (10-19 years) on VLC and VLS were analyzed for 12 sub-Saharan African (SSA) countries between 2019 (pre-COVID-19) and 2020 (during COVID-19). We report the number of viral load (VL) tests, and percent change in VLC and VLS for patients on ART. For 12 countries, 181,192 children had a VL test during the pre-COVID-19 period compared with 177,683 December 2020 during COVID-19. VLC decreased from 68.8% to 68.3% overall. However, 9 countries experienced an increase ranging from a 0.7%-point increase for Tanzania and Zimbabwe to a 15.3%-point increase for Nigeria. VLS increased for all countries from 71.2% to 77.7%. For adolescents the number with a VL test increased from 377,342 to 402,792. VLC decreased from 77.4% to 77.1%. However, 7 countries experienced an increase ranging from 1.8% for Mozambique to 13.8% for Cameroon. VLS increased for all countries from 76.8% to 83.8%. This analysis shows variation in HIV VLC across 12 SSA countries. VLS consistently improved across all countries demonstrating resilience of countries during 2020. Countries should continue to improve clinical outcomes from C/ALHIV despite service disruptions that may occur during pandemic response.

Host associations, biogeography, and phylogenetics of avian malaria in southern African waterfowl.

The relevance of spatial variation in the environment and host communities for parasite community composition is poorly documented, creating a need for additional case studies from which general principles can be developed. Avian malaria in southern African waterfowl has not previously been studied. As a first step towards documenting and understanding its biogeography, we used PCR and molecular sequencing techniques to analyse 454 blood samples from Afrotropical ducks from 5 different locations (spread around the subregion) for avian malaria. Fifty-five blood samples were positive for one or more genera of haematozoa. The regional infection rate across all sites and sampling periods was 12·1%. Nine individuals carried dual infections containing multiple haematozoa. Fifteen different cytochrome b haplotypes among 52 positives (3 samples failed to sequence) and 61 total sequences were found. Eleven haplotypes closely matched Plasmodium, whereas 4 were more similar to Haemoproteus. Five distinct haematozoan clades were identified. Haemoproteus parasites appeared to be more host-specific than Plasmodium, which occurred at every sampling location and in every host species examined. There were no significant differences in overall parasite prevalence attributable to either site or species, although Plasmodium and Haemoproteus occurrences differed by site-species combination and the borderline significance of our test for between-site variation (P < 0·06) implied that with a larger sample size, differences in parasite prevalence among locations might be detectable.

Linking avian communities and avian influenza ecology in southern Africa using epidemiological functional groups.

The ecology of pathogens, and particularly their emergence in multi-host systems, is complex. New approaches are needed to reduce superficial complexities to a level that still allows scientists to analyse underlying and more fundamental processes. One promising approach for simplification is to use an epidemiological-function classification to describe ecological diversity in a way that relates directly to pathogen dynamics. In this article, we develop and apply the epidemiological functional group (EFG) concept to explore the relationships between wild bird communities and avian influenza virus (AIV) in three ecosystems in southern Africa. Using a two year dataset that combined bird counts and bimonthly sampling for AIV, we allocated each bird species to a set of EFGs that captured two overarching epidemiological functions: the capacity of species to maintain AIV in the system, and their potential to introduce the virus. Comparing AIV prevalence between EFGs suggested that the hypothesis that anseriforms (ducks) and charadriiforms (waders) drive AIV epidemiology cannot entirely explain the high prevalence observed in some EFGs. If anseriforms do play an important role in AIV dynamics in each of the three ecosystems, the role of other species in the local maintenance of AIV cannot be ruled out. The EFG concept thus helped us to identify gaps in knowledge and to highlight understudied bird groups that might play a role in AIV epidemiology. In general, the use of EFGs has potential for generating a range of valuable insights in epidemiology, just as functional group approaches have done in ecology.

Birdcall lures improve passerine mist-net captures at a sub-tropical African savanna.

Field research techniques are constantly evolving to meet the needs of the scientific community. There is a growing need for field biology studies to shift towards increasing efficiency and quality of results while simultaneously decreasing cost in both the researcher's time and resources. I tested the efficacy of using multiple recorded birdcall lures (n = 172 species) to improve mist-net captures at a subtropical African savanna setting. Capture success was compared between passive and birdcall enhanced mist-nets during winter and summer seasons. Results suggest that the use of birdcalls does significantly increase the total number of birds caught in both seasons and also increases the diversity of passerine species. Conventional passive mist-nets without an audio lure were initially productive but their capture rate subsequently decreased as sampling days progressed. Birdcall lure enhanced mist-nets had a constant capture output during the summer season. The most responsive birds to audio lures were gregarious species (e.g. Pycnonotus barbatus, Dryoscopus cubla, Prionops plumatus, Phoeniculus purpureus, Turdoides jardineii and Lamprotornis chalybaeus) and the aggressive Dicrurus adsimilis and Acridotheres tristis. I conclude that birdcall lures can be used in summer and winter seasons to improve mist-net captures especially for studies focusing on gregarious and aggressive passerine species in a sub-tropical African savanna setting.

Geographic variation in factors that influence timing of moult and breeding in waterfowl.

Waterfowl flight-feather moult is expected to occur when energy is not needed for breeding and when a suitable safe habitat is available. Flight-feather regrowth is an energetically costly stage in the annual cycle of waterfowls. In this study, we tested the hypothesis that moult will coincide with the time of year when food and aquatic habitats are most abundant. We investigated how the timing of rainfall relates to the timing of breeding and flight-feather moult in five common southern African waterfowl at two sites in South Africa with opposite rainfall regimes (one summer, one winter). We then incorporated published data to compare and contrast the relative timing of breeding and moult in southern hemisphere (southern African and Australian) waterfowl with northern hemisphere (European and North American) species. Our results showed that southern African waterfowl breed in the wet season and moult during the dry season. Tadorna cana was an exception, breeding in the dry season and moulting during the wet season in the summer-rainfall area. There was also a long lag period between peak breeding and peak moult in southern hemisphere waterfowl species, the longest lag being that of birds in the summer-rainfall area. By comparison, northern hemisphere waterfowl species breed and moult during the warm season, with a shorter lag period between peak breeding and peak moult compared to southern hemisphere species. We concluded that waterfowl in southern Africa (with the exception of T. cana), southeast Australia, Europe and North America time their breeding period to coincide with peaks in the availability of both food and breeding sites. Northern hemisphere species moult where chances of predation are low, when temperatures are warm, and before food and aquatic habitats approach their winter minima. By contrast, southern hemisphere waterfowl delay the onset of moult until the dry season, opting to moult when both food and aquatic moulting habitats are scarce.

Investigating avian influenza infection hotspots in old-world shorebirds.

Heterogeneity in the transmission rates of pathogens across hosts or environments may produce disease hotspots, which are defined as specific sites, times or species associations in which the infection rate is consistently elevated. Hotspots for avian influenza virus (AIV) in wild birds are largely unstudied and poorly understood. A striking feature is the existence of a unique but consistent AIV hotspot in shorebirds (Charadriiformes) associated with a single species at a specific location and time (ruddy turnstone Arenaria interpres at Delaware Bay, USA, in May). This unique case, though a valuable reference, limits our capacity to explore and understand the general properties of AIV hotspots in shorebirds. Unfortunately, relatively few shorebirds have been sampled outside Delaware Bay and they belong to only a few shorebird families; there also has been a lack of consistent oropharyngeal sampling as a complement to cloacal sampling. In this study we looked for AIV hotspots associated with other shorebird species and/or with some of the larger congregation sites of shorebirds in the old world. We assembled and analysed a regionally extensive dataset of AIV prevalence from 69 shorebird species sampled in 25 countries across Africa and Western Eurasia. Despite this diverse and extensive coverage we did not detect any new shorebird AIV hotspots. Neither large shorebird congregation sites nor the ruddy turnstone were consistently associated with AIV hotspots. We did, however, find a low but widespread circulation of AIV in shorebirds that contrast with the absence of AIV previously reported in shorebirds in Europe. A very high AIV antibody prevalence coupled to a low infection rate was found in both first-year and adult birds of two migratory sandpiper species, suggesting the potential existence of an AIV hotspot along their migratory flyway that is yet to be discovered.

The ecology of influenza A viruses in wild birds in southern Africa.

Avian influenza viruses (AIVs) are pathogens of global concern, but there has been little previous research on avian influenza in southern Africa and almost nothing is known about the dynamics of AIVs in the region. We counted, captured and sampled birds regularly at five sites, two in South Africa (Barberspan and Strandfontein) and one in each of Botswana (Lake Ngami), Mozambique (Lake Chuali) and Zimbabwe (Lakes Manyame and Chivero) between March 2007 and May 2009. The South African and Zimbabwean sites were visited every 2 months and the sites in Botswana and Mozambique every 4 months. During each visit we undertook 5-7 days of standardised bird counts followed by 5-10 days of capturing and sampling water-associated birds. We sampled 4,977 birds of 165 different species and completed 2,503 half-hour point counts. We found 125 positive rRT-PCR cases of avian influenza across all sites. Two viruses (H1N8 and H3N8) were isolated and additional H5, H6 and H7 strains were identified. We did not positively identify any highly pathogenic H5N1. Overall viral prevalence (2.51%) was similar to the lower range of European values, considerable spatial and temporal variation occurred in viral prevalence, and there was no detectable influence of the annual influx of Palearctic migrants. Although waterbirds appear to be the primary viral carriers, passerines may link wild birds and poultry. While influenza cycles are probably driven by the bird movements that result from rainfall patterns, the epidemiology of avian influenza in wild birds in the subregion is complex and there appears to be the possibility for viral transmission throughout the year.