Applying environmental DNA (eDNA) metabarcoding to samples from waterholes and their surroundings offers a promising approach for monitoring terrestrial vertebrates in semi-arid and arid ecosystems, such as the southern African savannas. However, minimal guidance exists on key sampling design parameters for terrestrial ecosystems, which can significantly influence species detection. This study investigated the effects of sampled substrate, sampling season, and metabarcoding primer pair on species richness and taxonomic group detection in terrestrial vertebrates, with a focus on mammals, using eDNA samples from waterholes in Botsalano Game Reserve, South Africa. A total of 725 eDNA samples were collected from 94 sampling events across wet and dry seasons, detecting 95 species (45 birds, 42 mammals, 4 amphibians, 3 reptiles, and 1 fish). Sediment samples provided more reliable detection of abundant taxa, whereas water samples had higher detection frequencies of rare taxa. A mixed sampling approach yielded the highest species richness. Sampling during the wet season yielded higher species richness overall, while more mammal species were detected from dry season sampling. Overlap in species detection between the two metabarcoding primers tested was low (47%). We formulate recommendations for future eDNA metabarcoding study designs in similar systems, including remote sampling logistics and discuss potential sources of false positives in eDNA metabarcoding, including (1) secondary eDNA input, (2) incomplete genetic reference databases, and (3) the low genetic resolution of metabarcoding markers.

The role of Muscidae flies as vectors of bacterial pathogens with veterinary and public health significance necessitates accurate species identification and distribution mapping. This review analysed records from the Barcode of Life Data Systems (BOLD) and the South African National Biodiversity Institute (SANBI) national Diptera checklist, the BOLD Arthropoda progress report and literature, to assess South African Muscidae diversity, distribution and pathogen transmission. Through comparative analysis, we documented a total of 276 Muscidae fly species within the South African region. Three genera (n = 3) of the 38 genera, namely Coenosia, Lispe and Musca, demonstrated notable barcoding percentage gaps of 99%, 98% and 98%, respectively, indicating substantial genetic identification challenges. For the remaining 35 genera, a complete absence of barcode information was recorded, representing a 100% barcoding data gap. The striking disparity in Barcode Index Number (BIN) records across the three predominant fly genera (Coenosia, Lispe and Musca) reflects a concerning pattern in South African dipteran molecular taxonomy. The species Musca domestica demonstrated the most extensive pathogen diversity, carrying 22 distinct bacterial species, which include the ESKAPE group of pathogens. The results reveal critical gaps in taxonomic data alongside substantial pathogen transmission risks, necessitating integrated molecular-taxonomic approaches and targeted disease control strategies.

The global decline in biodiversity, driven by habitat loss, overexploitation, climate change, biological invasions, and illegal trade, poses significant challenges for conservation management. Although many South African ecosystems and species are under threat, effective conservation efforts are hindered by incomplete foundational biodiversity data and assessments, caused by taxonomic gaps and unverified distributions. DNA barcoding has emerged as an invaluable tool for species identification and classification of biodiversity. While substantial barcoding progress has been made, for many taxa, others remain underrepresented in sequence databases. This study evaluates the status and progress of DNA barcoding in South Africa through a gap analysis, comparing verified species checklists with barcoded sequences from the Barcode of Life Database (BOLD) and GenBank to assess taxonomic and geographic representation. A literature review (2003-2023) highlights applications across terrestrial, freshwater, and marine habitats. Of the 931,476 South African species barcode records, 52% were publicly available. Although the insects dominated with the highest number of records and BINs, reptiles had the highest taxonomic representation. Plants and fungi were underrepresented (16.1% and 2.8%, respectively). Regionally, Mpumalanga and Limpopo provinces showed the highest BIN counts, while North-West and Free State provinces had the lowest. The majority of barcode records were for mtDNA genes such as cytochrome c oxidase subunit I (COI) and were contributed by both local and international institutions. Discrepancies between GenBank records and those mined by BOLD indicated that many GenBank sequences for South Africa have poor quality metadata, including geographic sampling locality information. While significant progress has been made across taxa, further efforts are needed to expand species and geographic coverage, enhance sequence quality, improve species metadata, and resolve inconsistencies in BIN assignments, particularly for underrepresented groups such as plants and fungi. These advances would strengthen biodiversity assessments and support conservation efforts in South Africa.

Haemosporidian parasites from the genera Haemoproteus, Plasmodium, and Leucocytozoon are significant avian pathogens. This study aimed to identify and characterize these parasites in cranes (family Gruidae), using combined morphological and molecular methods. The results confirmed the presence of Haemoproteus balearicae, redescribed here from Balearica regulorum and associated with cytb lineage hBAREGI210. This lineage, previously assigned to Haemoproteus antigonis, is reassigned to H. balearicae, suggesting possible cryptic speciation within the H. antigonis complex. The findings broaden the known host range and geographic distribution of H. balearicae, detected in captive-born cranes in France and captive cranes housed in conservation facilities in South Africa. Phylogenetic analyses revealed three distinct Haemoproteus clades in Gruidae, corresponding to at least three species, including H. balearicae and lineages representing H. antigonis. These crane-specific parasites may require taxonomic revision as a separate subgenus or genus, pending further studies on their life cycles and vectors. Additionally, several novel cytb lineages of Plasmodium and Leucocytozoon were detected, many unassigned to morphospecies. Notably, the pCATUS05 lineage, a member of the Plasmodium lutzi group previously reported only in the Americas, was detected for the first time in South African cranes, along with Leucocytozoon aff. californicus (lCIAE02), a widespread lineage lacking morphological description. Together, these findings reveal underestimated genetic diversity of haemosporidian parasites in cranes and highlight the importance of combining morphological and molecular data to clarify parasite taxonomy and host associations. This study advances our understanding of avian parasite ecology and systematics, with implications for crane conservation and disease management.

Ticks transmit pathogens of veterinary and public health importance. Understanding their diversity is critical as infestations lead to significant economic losses globally. To date, over 90 species across three families have been identified in South Africa. However, the taxonomy of most species has not been resolved due to morphological identification challenges. DNA barcoding through the Barcode of Life Data Systems (BOLD) is therefore a valuable tool for species verifications for biodiversity assessments. This study conducted an analysis of South African tick COI barcodes on BOLD by verifying species on checklists, literature, and other sequence databases. The compiled list represented 97 species, including indigenous (59), endemics (27), introduced (2), invasives (1), and eight that could not be classified. Analyses indicated that 31 species (32%) from 11 genera have verified COI barcodes. These are distributed across all nine provinces with the Eastern Cape having the highest species diversity, followed by Limpopo, with KwaZulu-Natal having the least diversity. Rhipicephalus, Hyalomma, and Argas species had multiple barcode index numbers, suggesting cryptic diversity or unresolved taxonomy. We identified 21 species of veterinary or zoonotic importance from the Argasidae and Ixodidae families that should be prioritised for barcoding. Coordinating studies and defining barcoding targets is necessary to ensure that tick checklists are updated to support decision-making for the control of vector-borne diseases and alien invasives.

Combating wildlife crimes in South Africa requires accurate identification of traded species and their products. Diagnostic morphological characteristics needed to identify species are often lost when specimens are processed and customs officials lack the expertise to identify species. As a potential solution, DNA barcoding can be used to identify morphologically indistinguishable specimens in forensic cases. However, barcoding is hindered by the reliance on comprehensive, validated DNA barcode reference databases, which are currently limited. To overcome this limitation, we constructed a barcode library of cytochrome c oxidase subunit 1 and cytochrome b sequences for threatened and protected mammals exploited in southern Africa. Additionally, we included closely related or morphologically similar species and assessed the database's ability to identify species accurately. Published southern African sequences were incorporated to estimate intraspecific and interspecific variation. Neighbor-joining trees successfully discriminated 94%-95% of the taxa. However, some widespread species exhibited high intraspecific distances (>2%), suggesting geographic sub-structuring or cryptic speciation. Lack of reliable published data prevented the unambiguous discrimination of certain species. This study highlights the efficacy of DNA barcoding in species identification, particularly for forensic applications. It also highlights the need for a taxonomic re-evaluation of certain widespread species and challenging genera.

The Southern Ground-hornbill (SGH) (Bucorvus leadbeateri) is considered an umbrella species for biodiversity conservation in savannah biomes since they require large territories and significant protection measures that help to conserve a wide range of biodiversity with similar savanna and grassland requirements. Declines of the species are attributed to low reproductive rates coupled with multiple anthropogenic threats, including secondary poisoning, and persecution. Little is known about connectivity and population structure of SGH populations in Africa, south of the equator. Knowledge of population differentiation is needed to ensure that targeted conservation management plans can be implemented to slow population declines and ensure survival of the species. To inform a long-term conservation strategy, we investigated the broad-scale population structure of Southern Ground-hornbill across their sub-equatorial range. Our study based on 16 microsatellite loci identified moderate variation (average of 5.889 alleles per locus and a mean observed heterozygosity of 0.546) similar to other long-lived avian species. In contrast, mitochondrial DNA sequences analysis identified low diversity (Hd = 0.3313, π = 0.0015). A Bayesian assignment approach, principal component analysis, analysis of molecular variance and phylogenetic analysis identified weak to moderate population structuring across long distances and mitochondrial data showed a shallow phylogeny. Restriction to long-distance dispersal was detected that could not be attributed to isolation by distance, suggesting that other factors, such as their dispersal biology, are shaping the observed genetic differentiation. Although our study does not support the designation of populations as independent conservation units, we advocate that population management should continue to follow the Precautionary Principle (mixing founders from the same range state, rather than allowing mixing of founders from the extremes of the range) until there is scientific certainty. Following further research, if no independent conservation units are detected, then the global captive population can contribute to reintroductions across the range. In the wild, populations at the edge of the species range may need additional management strategies and gene flow should be promoted between neighbouring populations.

The African BioGenome Project (AfricaBP) Open Institute for Genomics and Bioinformatics aims to overcome barriers to capacity building through its distributed African regional workshops and prioritizes the exchange of grassroots knowledge and innovation in biodiversity genomics and bioinformatics. In 2023, we implemented 28 workshops on biodiversity genomics and bioinformatics, covering 11 African countries across the 5 African geographical regions. These regional workshops trained 408 African scientists in hands-on molecular biology, genomics and bioinformatics techniques as well as the ethical, legal and social issues associated with acquiring genetic resources. Here, we discuss the implementation of transformative strategies, such as expanding the regional workshop model of AfricaBP to involve multiple countries, institutions and partners, including the proposed creation of an African digital database with sequence information relating to both biodiversity and agriculture. This will ultimately help create a critical mass of skilled genomics and bioinformatics scientists across Africa. The African BioGenome Project (AfricaBP) Open Institute for Genomics and Bioinformatics established a series of regional workshops in 2023 to exchange knowledge and overcome barriers, which could serve as a model for other scientific communities.

Ticks are arachnid blood-feeding parasites, which infest livestock, wildlife, and humans, transmitting medically and veterinary significant pathogens. Their biodiversity and distribution in wild animals remains complex. This study analysed archived tick samples (n = 48) from the South African Biodiversity Institute (SANBI) Wildlife Biobank utilizing morphology and genetic analyses of the 16S rRNA and COI (DNA barcoding) mitochondrial genes to identify ticks collected among 13 vertebratesavian, reptilian, and mammalian host species. The specimens came from nine localities including nature reserves and captive facilities (zoological garden) in South Africa, Namibia, and Botswana. These ticks were also assessed for associated pathogens with the reverse line blot (RLB) hybridization assay. Seven tick genera, Amblyomma, Hyalomma, Haemaphysalis, Ixodes, Rhipicephalus, Rhipicentor, and Otobius were identified, with Amblyomma being the most prevalent (22.9 %) in our sample set. Obtained sequences were 95-100 % similar to published records of tick species collected from wild and domestic animals, as well as those collected from vegetation, from different southern African areas. However, tick specimens (n = 3) identified morphologically as Hyalomma truncatum, Rhipicephalus e. evertsi, and R. simus, were, on a molecularly level, more closely related to their sister taxa (H. glabrum, R. e. mimeticus, and R. gertrudae, respectively) suggesting a need for taxonomic verification. With the RLB hybridization assay, six samples reacted with the Ehrlichia/Anaplasma genus-specific probe, while two reacted with the Theileria/Babesia genus-specific probe. Sequencing of the RLB amplicons targeting the 18S rRNA gene (n = 2) indicated 100 % similarity to Hepatozoon fitzsimonsi, while one was closely related to He. ingwe with 99.39 % similarity. The results show that wildlife harbour different tick species, and pathogen detection identified novel genotypes, indicating wildlife as potential pathogens reservoirs. This study enhances our understanding of tick biodiversity, distribution and highlights wildlife's role in harbouring diverse tick species and novel pathogens.

Environmental DNA (eDNA) is used for biodiversity assessments in a variety of ecosystems across the globe, whereby different eDNA concentration, preservation and extraction methods can outperform others depending on the sampling conditions and environment. Tropical and subtropical ecosystems in Africa are among the less studied systems concerning eDNA-based monitoring. Waterholes in arid parts of southern Africa represent important agglomeration points for terrestrial mammals, and the eDNA shed into such waterbodies provides a powerful source of information for monitoring mammalian biodiversity in the surrounding area. However, the applied methods for eDNA sampling, preservation and filtering in different freshwater systems vary greatly, and rigorous protocol testing in African freshwater systems is still lacking. This study represents the first attempt to examine variations in eDNA concentration, preservation and extraction methods under remote field conditions using waterborne eDNA in a savanna system. Collected samples were heavily affected by microalgal and bacterial growth, impeding eDNA capture and PCR success. We demonstrate clear effects of the methodological choices, which also depend on the state of eDNA. A preliminary metabarcoding run showed little taxonomic overlap in mammal species detection between two metabarcoding primers tested. We recommend water filtering (using filters with pore sizes >1 mu m) over centrifugation for eDNA concentration, Longmire's solution for ambient temperature sample preservation and Qiagen's DNeasy PowerSoil Pro Kit for DNA extraction of these inhibitor-prone samples. Furthermore, at least two independent metabarcoding markers should be utilized in order to maximize species detections in metabarcoding studies.

The blue crane (Anthropoides paradiseus), wattled crane (Bugeranus carunculatus), and grey-crowned crane (Balearica regulorum) are species of concern as their populations are declining and they face several threats including habitat loss, disturbance and illegal trade. In South Africa, these species are bred in captivity for trade purposes which is permitted and regulated globally under the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Legal sustainable trade through captive breeding of endangered wildlife species such as cranes has been promoted to counteract the illegal trade of individuals from the wild. Captive breeding independent of wild populations may reduce the harvest pressures on wild bird populations which in turn benefit the recovery of exploited species. This approach is considered to be controversial by some individuals. Although captive breeding of endangered species, for both population sustainability and commercial purposes, is promoted to aid in conserving species, concerns have been raised with regards to breeding facilities being used for laundering of animals. To monitor the legal trade of cranes in South Africa a short tandem repeat (STR) assay following recommendations of the International Society for Forensic Genetics (ISFG) was developed and validated. An STR assay comprising of four multiplexes that include 16 STR markers and two gender determination markers was proven to be highly informative with average polymorphic information content (PIC) values of 0.806, 0.646 and 0.725 for A. paradiseus, B. regulorum and B. carunculatus respectively. In addition, the assay showed sufficient discriminatory power for parentage assignment of closely related individuals in all three species (A. paradiseus: PI = 1.7×10-24, PIsibs = 4.7×10-08, and B. carunculatus: PI = 1.4×10-19, PIsibs = 2.9×10-07 and B. regulorum: PI = 1.7×10-12, PIsibs = 5.0×10-05). Analysis of 251 samples suggested that the validated multiplex assay ensures reliability, reproducibility, and repeatability for applications in forensic case work where illegal trade of offspring is suspected through verifying parentage of captive birds in breeding facilities.

In 2022, around 54 % of African students were denied student visas to study in the United States (US), compared to 36 % of Asian students and 9 % of European students, despite African immigrants in the US often being more highly educated than the US native-born population. This issue cannot be attributed solely to the dichotomy between the Global North and South in visa regimes, but it is also evident among African nations across regional economic blocs. The African BioGenome Project (AfricaBP) Open Institute for Genomics and Bioinformatics, which aims to overcome barriers to capacity building through its distributed African regional workshops, prioritizes grassroots knowledge exchange and innovation in biodiversity genomics and bioinformatics. In 2023, we orchestrated the implementation of 27 capacity building workshops on biodiversity genomics and bioinformatics, covering 10 African countries across 5 African geographical regions. The AfricaBP Open Institute regional workshops raised awareness of biodiversity genomics and bioinformatics among 3788 registered participants, and trained 408 African scientists in hands-on molecular biology, genomics, and bioinformatics techniques. Here, we discuss the implementation of transformative strategies by deploying the AfricaBP Open Institute multi-country, multi-institution, and multi-partner hybrid regional workshop model, including the proposed creation of an African digital database containing sequence information relating to biodiversity and agriculture.

The Diederik Cuckoo Chrysococcyx caprius is an African species widely distributed south of the Sahara, which migrates seasonally between breeding and nonbreeding sites. It is currently unknown whether the species consists of a single panmictic population or if it is genetically structured. To investigate this, we analysed sequence variation in three mitochondrial and two nuclear gene regions in combination with morphological measurements in specimens from four localities. Phylogenetic relationships were estimated using maximum-likelihood methods and included samples of Klaas’s Cuckoo Chrysococcyx klaas, Red-chested Cuckoo Cuculus solitarius, and African Cuckoo Cuculus gularis. Haplotype networks and analysis of molecular variance were used to characterise the spatial distribution of genetic diversity. A principal component analysis was performed to investigate morphological variation among localities. Molecular analysis identified two divergent mitochondrial lineages, which were found to occur in sympatry in one South African locality (Limpopo Province). The magnitude of divergence between versus within these lineages was low (0.4–1%) yet significant (FST: 0.84–0.88). Lack of apparent phylogeographic structure provides support for the absence of physical barriers to gene flow in this species. The divergent mitochondrial lineages did not differ in morphological measurements. The emergence and persistence of shallow mitochondrial divergence among sympatric lineages in the Diederik Cuckoo could be linked to maternal divergence in host selection of these brood parasites — a hypothesis requiring additional data to be tested.

The Woodland Kingfisher Halcyon senegalensis is widely distributed throughout sub-Saharan Africa and occupies a wide variety of woodland and savannah habitat. Thus far, three subspecies have been described based on morphological variation. In the present study, using western, eastern and southern African populations, we examined the relationship between morphological and genetic divergence among two named subspecies, H. s. cyanoleuca and H. s. senegalensis, using three mitochondrial markers (CO1, Cytb, 16S) and two nuclear markers (FIB5 and RAG1). Southern birds showed clear evidence for morphological divergence, with a longer wing and tail length, when compared with eastern and western birds. Phylogenetic analyses using Bayesian methods identified two well-characterised genetic clusters, representing the two subspecies. We determined that H. s. senegalensis and H. s. cyanoleuca are closely related subspecies that split recently, approximately 0.66–1.31 MYA in the Pleistocene. Furthermore, genetic substructure was evident within H. s. senegalensis, with three distinct genetic clusters in each region. The separation between the Ghana+Gabon and Uganda lineages of H. s. senegalensis occurred approximately 0.12–0.57 MYA. Nuclear–mitochondrial discordance was detected, however, wherein the pattern of divergence was not detected in the RAG1 and FIB5 sequences. Our results suggest that climate change, biogeographic barriers and local adaptation has played a role in the diversification of Woodland Kingfishers in Africa.

Currently little is known about intra-African bird migrants. Migratory connectivity between populations may be affected by past climatic fluctuations as well as contemporary threats that affect habitat connectivity resulting in genetic differentiation. Here, integrated molecular and morphological data was used to examine genetic diversity, elucidate patterns of differentiation and assess evolutionary history of the African Pygmy Kingfisher (Ispidina picta). Three subspecies have been described namely: I. p. picta, I. p. ferrugina and I. p. natalensis. Here, molecular analysis was performed for two subspecies; I. p. natalensis (South Africa) and I. p. picta (Uganda, Ghana and Nigeria) using mitochondrial and nuclear genes. Our results provided evidence of two genetic lineages corresponding to subspecies designation: I. p. natalensis, and I. p. picta that diverged 1.74 Mya ago in the Pleistocene. Lack of differentiation was observed within the two subspecies indicating that connectivity between the populations has been maintained. Morphometric variation identified differences between the subspecies and further identified sexual dimorphism within I. p. natalensis. This study provides for the first time a genetic and morphometric appraisal of African Pygmy Kingfishers. We recommend that this or similar approaches be applied to other widespread African bird species that are often overlooked in a global conservation context.

Genetic diversity is a fundamental measure of a populations ability to adapt to future environmental change. Subpopulations may carry unique genetic lineages that contribute to fitness and genetic diversity of species across their distribution range. Therefore, considerations, or lack thereof, of genetic diversity in wildlife management practices may result in either population persistence or extinction over time. Some management tools may pose a greater risk to a species' survival than others when populations are impacted. In South Africa, there has been great interest to translocate animals, sometimes with little consideration to the potential impacts on the species and/or populations survival. Thus, there is a need to collate scientific information to better inform decision-making and review these management practices and their effects on populations. Here, we focus on three antelope species, the blue duiker (Philantomba monticola), oribi (Ourebia ourebi), and tsessebe (Damaliscus lunatus). We review the genetic status of each species across South Africa, with regards to taxonomy, genetic diversity and population structure, threats that may compromise the genetic diversity within species and across populations, conservation management actions and how they may compromise or benefit the genetic status and lastly make recommendations on possible alternative management actions and future research to inform conservation policy and sustainable management practice.

A southwestern Indian Ocean (SWIO) percoid fish Serranus knysnaensis Gilchrist, 1904, was long synonymised with the comber, Serranus cabrilla (Linnaeus, 1758), from the eastern Atlantic Ocean, Mediterranean and Black Sea. However, when the species was brought out of synonymy by Heemstra Heemstra (2004), reasons for this decision were not given. This study aims to revalidate the present taxonomic status of S. knysnaensis using morphological and molecular assessments. The two species are distinguished by the number of circumpeduncular scales (2634 in S. knysnaensis versus 3438 in S. cabrilla) and total gill rakers (1822 versus 2224). Serranus knysnaensis is also distinct from S. novemcinctus Kner, 1864, the other SWIO species of Serranus, based on total gill raker counts (1822 versus 3135). Genetic analysis of mitochondrial DNA barcode (COI) sequences for 17 Serranus species revealed three closely-related monophyletic clusters corresponding to S. cabrilla, S. novemcinctus and S. knysnaensis that were supported (P 0.001) by species delimitation methods. Even though the genetic distances among the three species were the lowest in the genus (1.60-1.99%), these species may be ecomorphs or lineages that have only recently diverged from each other. These three species also have allopatric distributions and our morphological and molecular data thus confirm that S. knysnaensis is a valid species.

A rapid allelic discrimination real-time polymerase chain reaction (qPCR) assay has been developed for the confirmation of lion (Panthera leo) or tiger (P. tigris) DNA. The method includes a low cost allele specific PCR assay designed for each SNP variant where the genotype is determined by incorporation and subsequent fluorescence of FAM or HEX following PCR. The described method was able to accurately and rapidly distinguish between the two species, and all three mitochondrial regions (16S, 12S and control region) performed equally well. The proposed real-time PCR assay can be applied for the accurate confirmation of either lion or tiger DNA that could be used by law enforcement agencies around the world as a tool to monitor illegal trade of tiger bones.

The National Zoological Gardens (NZG) is a facility of the South African National Biodiversity Institute (SANBI) and the largest zoo in southern Africa. Among the 9000 captive animals kept by the NZG, is the endangered African penguin (Spheniscus demersus). There have been several post-mortem reports on deaths of penguins in the NZG due to haemosporidian infections, however, the haemosporidian lineages involved and possible insect vector are unknown. Haemosporidians are apicomplexan parasites that infect vertebrates through blood-sucking dipteran insects. Therefore, the current study aimed to identify mosquitoes that are potential vectors found within the African penguin enclosure as well as to detect the haemosporidian parasites from these insects using nested-PCR and real-time PCR (qPCR) analyses. Mosquito samples were collected using an overnight UV-light trap setup for 3 months. From the 65 pooled samples representing 325 mosquitoes, morphological and molecular analysis showed that Culex pipiens (52.31%) was the dominant species followed by Cx.theileri (30.77%) and Cx. quinquefasciatus (16.92%). Nested-PCR detected parasite DNA of Leucocytozoon sp. and Plasmodium sp. The Cx. pipiens had the highest minimum infection rate (MIR) of 5.88% by nested-PCR and 9.41% by qPCR whilst Cx. quinquefasciatus had MIR of 3.64% in both assays and no haemosporidian parasites were detected from Cx.theileri. One Cx. pipiens sample had a co-infection of both Plasmodium sp. and Leucocytozoon sp. detected by nested-PCR. These findings suggest that effective control measures for blood-sucking dipteran insects is required at the NZG and more studies should be conducted to determine the actual prevalence of these haemosporidian parasites among other bird species within NZG.

Patterns of genetic structure and connectivity of the monotypic cigar wrasse Cheilio inermis within western Indian Ocean (WIO) are poorly understood. Whether the species exists as a single panmictic population across the WIO is unclear. Sequence data were generated from two mitochondrial genes (cytochrome b and ATPase 6) and one nuclear intron (S7 intron I). High levels of haplotype and allelic diversity (h = 0.88–0.98; A = 0.95–0.98), along with low nucleotide diversities were observed across all markers. The pairwise ΦSTvalues indicated differentiation of Tanga from the four WIO localities (Inhaca, Nosy Bé, Gazi, and Shimoni), as well as differentiation between the northernmost WIO localities. AMOVAs indicated high differentiation among defined locality groups, whereas nuclear gene analysis found little differentiation among groups. The observed genetic differentiation in C. inermis could be caused by oceanic barriers, and by limited larval dispersal with the pelagic larvae possibly settling near their parental origin and promoting differentiation.