Genetic identification of three CITES-listed sharks using a paper-based Lab-on-a-Chip (LOC).

Threatened shark species are caught in large numbers by artisanal and commercial fisheries and traded globally. Monitoring both which shark species are caught and sold in fisheries, and the export of CITES-restricted products, are essential in reducing illegal fishing. Current methods for species identification rely on visual examination by experts or DNA barcoding techniques requiring specialist laboratory facilities and trained personnel. The need for specialist equipment and/or input from experts means many markets are currently not monitored. We have developed a paper-based Lab-on-a-Chip (LOC) to facilitate identification of three threatened and CITES-listed sharks, bigeye thresher (Alopias superciliosus), pelagic thresher (A. pelagicus) and shortfin mako shark (Isurus oxyrinchus) at market source. DNA was successfully extracted from shark meat and fin samples and combined with DNA amplification and visualisation using Loop Mediated Isothermal Amplification (LAMP) on the LOC. This resulted in the successful identification of the target species of sharks in under an hour, with a working positive and negative control. The LOC provided a simple "yes" or "no" result via a colour change from pink to yellow when one of the target species was present. The LOC serves as proof-of-concept (PoC) for field-based species identification as it does not require specialist facilities. It can be used by non-scientifically trained personnel, especially in areas where there are suspected high frequencies of mislabelling or for the identification of dried shark fins in seizures.

Taxonomic and geographic bias in 50 years of research on the behaviour and ecology of galagids.

Identifying knowledge gaps and taxonomic and geographic bias in the literature is invaluable for guiding research towards a more representative understanding of animal groups. Galagids are nocturnal African primates and, for many species, detailed information on their behaviour and ecology is unavailable. To identify gaps and bias in the literature we reviewed published peer-reviewed research articles on galagid behaviour and ecology over a 50-year period from January 1971 to December 2020. Using the Web of Science and Google Scholar databases, we identified 758 articles, assessed 339 full texts for eligibility and included 211 in the review. Species of Otolemur have been extensively researched in comparison to other genera (78.2% of studies; Euoticus: 13.3% of studies; Galago: 66.4% of studies; Galagoides: 20.9% of studies; Paragalago: 22.3% of studies; Sciurocheirus: 15.2% of studies). The most common category of research was physiology (55.0% of studies), followed by behavioural ecology (47.4% of studies), and fewer studies were on genetics and taxonomy (16.1% of studies) and habitat and distribution (14.2% of studies). Text mining revealed that the word 'behaviour' was the most common word used in abstracts and keywords, and few words were related to ecology. Negative binomial regression revealed that mean body mass and geographic range size were significant positive predictors of the total number of scientific outputs on each species. Research on wild populations was carried out in only 24 (60%) of the 40 countries galagids are thought to inhabit. Studies were undertaken in locations with lower mean annual temperatures and higher human population densities over warmer and less populated areas. We encourage a more equal sampling effort both taxonomically and geographically that in particular addresses the paucity of research on smaller species and those with restricted ranges. Research on in situ populations, especially in warmer and remote areas, is urgently needed, particularly in West, Central and some Southern African countries.

Hunting territories and land use overlap in sedentarised Baka Pygmy communities in southeastern Cameroon.

A significant number of Baka Pygmies in Cameroon have been sedentarised in roadside villages, in contrast to their nomadic hunter-gatherer existence of the past. Although this change in lifestyle has had important consequences on health, most Baka villages still supplement their diets from forest products, especially wild meat. We used a combination of participatory methods and monitoring of individual hunters to map hunting territories in 10 Baka villages in southeastern Cameroon. From these, we determined whether wild meat extraction levels per village were related to the size of hunting territories, measured habitat use by hunters and finally defined the overlap between hunting territories and extractive industries in the region. Mapped village hunting areas averaged 205.2 ± 108.7 km2 (range 76.8-352.0 km2); all villages used a total of 2052 km2. From 295 tracks of 51 hunters, we showed that hunters travelled an average of 16.5 ± 13.5 km (range 0.9-89.8 km) from each village. Home ranges, derived from kernel utilization distributions, were correlated with village offtake levels, but hunter offtake and distance travelled were not significantly related, suggesting that enough prey was available even close to the villages. Hunters in all village areas exhibited a clear bias towards certain habitats, as indicated by positive Ivlev's index of selectivity values. We also showed that all village hunting territories and hunter home ranges fall within mining and logging concessions. Our results are important for local understanding of forest land uses and to reconcile these with the other land uses in the region to better inform decisions concerning land use policy and planning.

On-site genetic analysis for species identification using lab-on-a-chip.

This paper presents a microfluidic device capable of performing genetic analysis on dung samples to identify White Rhinoceros (Ceratotherium simum). The development of a microfluidic device, which can be used in the field, offers a portable and cost-effective solution for DNA analysis and species identification to aid conservation efforts. Optimization of the DNA extraction processes produced equivalent yields compared to conventional kit-based methods within just 5 minutes. The use of a color-changing loop-mediated isothermal amplification reaction for simultaneous detection of the cytochrome B sequence of C. simum enabled positive results to be obtained within as little as 30 minutes. Field testing was performed at Knowsley Safari to demonstrate real-world applicability of the microfluidic device for testing of biological samples.

Are fission-fusion dynamics consistent among populations? A large-scale study with Cape buffalo.

Fission-fusion dynamics allow animals to manage costs and benefits of group living by adjusting group size. The degree of intraspecific variation in fission-fusion dynamics across the geographical range is poorly known. During 2008-2016, 38 adult female Cape buffalo were equipped with GPS collars in three populations located in different protected areas (Gonarezhou National Park and Hwange National Park, Zimbabwe; Kruger National Park, South Africa) to investigate the patterns and environmental drivers of fission-fusion dynamics among populations. We estimated home range overlap and fission and fusion events between Cape buffalo dyads. We investigated the temporal dynamics of both events at daily and seasonal scales and examined the influence of habitat and distance to water on event location. Fission-fusion dynamics were generally consistent across populations: Fission and fusion periods lasted on average between less than one day and three days. However, we found seasonal differences in the underlying patterns of fission and fusion, which point out the likely influence of resource availability and distribution in time on group dynamics: During the wet season, Cape buffalo split and associated more frequently and were in the same or in a different subgroup for shorter periods. Cape buffalo subgroups were more likely to merge than to split in open areas located near water, but overall vegetation and distance to water were very poor predictors of where fission and fusion events occurred. This study is one of the first to quantify fission-fusion dynamics in a single species across several populations with a common methodology, thus robustly questioning the behavioral flexibility of fission-fusion dynamics among environments.

Rapid isolation and characterization of microsatellites in the critically endangered mountain bongo (Tragelaphus eurycerus isaaci).

High-throughput sequencing tools promise to revolutionize many aspects of genetic research, e.g. by allowing the identification of functional adaptive genetic variation. However, the expense and expertise required to apply these tools to basic conservation questions is a challenge for applications outside academia, resulting in a so-called 'conservation genomics gap' (Shafer et al. 2015). The conservation genetics paradigm is that, basic information about inbreeding and gene flow are often critical to inform conservation management of small populations (Ouborg et al. 2010). This information is often needed quickly and ideally should be accessible to workers without special expertise in genomics (DeSalle and Amato 2004). While the inferential power of highthroughput sequencing to interrogate the genome is profound, the cost for population analysis is higher (though decreasing) than for traditional neutral markers. Thus, the use of neutral markers is still relevant in conservation applications. However, this assumes that neutral markers have been discovered and characterized for a given species of conservation concern, which is often untrue for nonmodel organisms. Here, we use a fast, cost-efficient, high-throughput sequencing method (Illumina MiSeq) to rapidly identify and characterize microsatellites in the mountain bongo (Tragelaphus eurycerus isaaci), which has a clear and timely conservation imperative but lacks any described neutral markers.

Sex-biased inbreeding effects on reproductive success and home range size of the critically endangered black rhinoceros.

A central premise of conservation biology is that small populations suffer reduced viability through loss of genetic diversity and inbreeding. However, there is little evidence that variation in inbreeding impacts individual reproductive success within remnant populations of threatened taxa, largely due to problems associated with obtaining comprehensive pedigree information to estimate inbreeding. In the critically endangered black rhinoceros, a species that experienced severe demographic reductions, we used model selection to identify factors associated with variation in reproductive success (number of offspring). Factors examined as predictors of reproductive success were age, home range size, number of nearby mates, reserve location, and multilocus heterozygosity (a proxy for inbreeding). Multilocus heterozygosity predicted male reproductive success (p< 0.001, explained deviance >58%) and correlated with male home range size (p < 0.01, r(2) > 44%). Such effects were not apparent in females, where reproductive success was determined by age (p < 0.01, explained deviance 34%) as females raise calves alone and choose between, rather than compete for, mates. This first report of a 3-way association between an individual male's heterozygosity, reproductive output, and territory size in a large vertebrate is consistent with an asymmetry in the level of intrasexual competition and highlights the relevance of sex-biased inbreeding for the management of many conservation-priority species. Our results contrast with the idea that wild populations of threatened taxa may possess some inherent difference from most nonthreatened populations that necessitates the use of detailed pedigrees to study inbreeding effects. Despite substantial variance in male reproductive success, the increased fitness of more heterozygous males limits the loss of heterozygosity. Understanding how individual differences in genetic diversity mediate the outcome of intrasexual competition will be essential for effective management, particularly in enclosed populations, where individuals have restricted choice about home range location and where the reproductive impact of translocated animals will depend upon the background distribution in individual heterozygosity.