Genetic analysis of Octopus cyanea reveals high gene flow in the South-West Indian Ocean.

Octopus cyanea (Gray, 1849), abundant in the South-West Indian Ocean (SWIO), constitutes a vital resource for both subsistence and commercial fisheries. However, despite this socioeconomic importance, and recent indications of overfishing, little is known about the population structure of O. cyanea in the region. To inform sustainable management strategies, this study assessed the spatio-temporal population structure and genetic variability of O. cyanea at 20 sites in the SWIO (Kenya, Tanzania, Mozambique, Madagascar, Mauritius, Rodrigues, and the Seychelle Islands) by complementary analysis of mitochondrial DNA (mtDNA) noncoding region (NCR) sequences and microsatellite markers. MtDNA analysis revealed a shallow phylogeny across the region, with demographic tests suggesting historic population fluctuations that could be linked to glacial cycles. Contrary to expectations, NCR variation was comparable to other mtDNA regions, indicating that the NCR is not a hypervariable region. Both nuclear and mtDNA marker types revealed a lack of genetic structure compatible with high gene flow throughout the region. As adults are sedentary, this gene flow likely reflects connectivity by paralarval dispersal. All samples reported heterozygote deficits, which, given the overall absence of structure, likely reflect ephemeral larval recruitment variability. Levels of mtDNA and nuclear variability were similar at all locations and congruent with those previously reported for harvested Octopodidae, implying resilience to genetic erosion by drift, providing current stock sizes are maintained. However, as O. cyanea stocks in the SWIO represent a single, highly connected population, fisheries may benefit from additional management measures, such as rotational closures aligned with paralarval ecology and spanning geopolitical boundaries.

Seasonal development of a tidal mixing front drives shifts in community structure and diversity of bacterioplankton.

Bacterioplankton underpin biogeochemical cycles and an improved understanding of the patterns and drivers of variability in their distribution is needed to determine their wider functioning and importance. Sharp environmental gradients and dispersal barriers associated with ocean fronts are emerging as key determinants of bacterioplankton biodiversity patterns. We examined how the development of the Celtic Sea Front (CF), a tidal mixing front on the Northwest European Shelf affects bacterioplankton communities. We performed 16S-rRNA metabarcoding on 60 seawater samples collected from three depths (surface, 20 m and seafloor), across two research cruises (May and September 2018), encompassing the intra-annual range of the CF intensity. Communities above the thermocline of stratified frontal waters were clearly differentiated and less diverse than those below the thermocline and communities in the well-mixed waters of the Irish Sea. This effect was much more pronounced in September, when the CF was at its peak intensity. The stratified zone likely represents a stressful environment for bacterioplankton due to a combination of high temperatures and low nutrients, which fewer taxa can tolerate. Much of the observed variation was driven by Synechococcus spp. (cyanobacteria), which were more abundant within the stratified zone and are known to thrive in warm oligotrophic waters. Synechococcus spp. are key contributors to global primary productivity and carbon cycling and, as such, variability driven by the CF is likely to influence regional biogeochemical processes. However, further studies are required to explicitly link shifts in community structure to function and quantify their wider importance to pelagic ecosystems.

Barriers to help-seeking among music festival attendees in New South Wales, Australia.

INTRODUCTION: Prompt help-seeking behaviour by music festival attendees can reduce risks associated with drug use; however, little is known about perceived barriers to help-seeking when experiencing or witnessing illness at music festivals. We explored potential barriers and their association with festivalgoer characteristics. METHODS: We conducted an on-site cross-sectional survey of attendees at New South Wales music festivals in 2019/2020. Perceived barriers to help-seeking in the hypothetical event of the respondent or a friend becoming unwell at the festival were assessed, and regression analyses were conducted to identify characteristics associated with these barriers. RESULTS: Across six festivals, 1229 people were surveyed and four-fifths (83.2%) reported ≥1 barrier: 32.7% fear of getting in trouble with the police, 20.6% not knowing where to find help, 17.2% not knowing how unwell someone might be and 15.3% concern about friends or relatives finding out. In multivariable analyses, people of diverse sexuality and people using drugs that day had greater odds of reporting fear of trouble with the police. People reporting drug use that day had lower odds of reporting not knowing where to find help. Men, gender-diverse people and people using drugs that day had greater odds of reporting concern about friends or relatives finding out. DISCUSSION AND CONCLUSIONS: Our data substantiate concerns regarding policing strategies and their impact on festivals. Initiatives to support conversations about drugs with friends and families may be best targeted to younger people and those from gender-diverse backgrounds.

Temporally consistent predominance and distribution of secondary malaria vectors in the Anopheles community of the upper Zambezi floodplain.

Regional optimisation of malaria vector control approaches requires detailed understanding both of the species composition of Anopheles mosquito communities, and how they vary over spatial and temporal scales. Knowledge of vector community dynamics is particularly important in settings where ecohydrological conditions fluctuate seasonally and inter-annually, such as the Barotse floodplain of the upper Zambezi river. DNA barcoding of anopheline larvae sampled in the 2019 wet season revealed the predominance of secondary vector species, with An. coustani comprising > 80% of sampled larvae and distributed ubiquitously across all ecological zones. Extensive larval sampling, plus a smaller survey of adult mosquitoes, identified geographic clusters of primary vectors, but represented only 2% of anopheline larvae. Comparisons with larval surveys in 2017/2018 and a contemporaneous independent 5-year dataset from adult trapping corroborated this paucity of primary vectors across years, and the consistent numerical dominance of An. coustani and other secondary vectors in both dry and wet seasons, despite substantial inter-annual variation in hydrological conditions. This marked temporal consistency of spatial distribution and anopheline community composition presents an opportunity to target predominant secondary vectors outdoors. Larval source management should be considered, alongside prevalent indoor-based approaches, amongst a diversification of vector control approaches to more effectively combat residual malaria transmission.

Wastewater analysis for psychoactive substances at music festivals across New South Wales, Australia in 2019-2020.

INTRODUCTION: Implementation of wastewater surveillance at music festivals has been limited to date. We aimed to use wastewater analysis and a self-report survey to determine the range of psychoactive substances being used during a music festival season in New South Wales, Australia. METHODS: We sampled six single-day music festivals requiring a music festival license in New South Wales from March 2019 to March 2020; between 15% and 100% of portaloos (temporary, un-fixed toilet facilities) were sampled at each festival. Samples were screened for 98 psychoactive substances and/or their metabolites with results qualitatively expressed as detection frequencies for each substance at each festival and across all festivals. We compared these data with the results of surveys of self-reported drug use at four of the six festivals. RESULTS: Festival attendance ranged from 6200 to 14,975 people. Amphetamine, cocaine, ketamine, methylone, MDMA, MDA, alprazolam, diazepam, etizolam, oxazepam and temazepam were found in almost all samples from all festivals. Ethylone, mephedrone and methcathinone were also found in over 50% of festivals. A norfentanyl (a fentanyl metabolite) and n-ethylpentylone were found at 2/6 and 1/6 festivals. No festival survey participant reported intentionally taking cathinones. DISCUSSION: The detection frequency for cathinones was higher than expected relative to recent other data sources and this may represent adulteration or substitution. Similarly, the appearance of etizolam may be related to the use of counterfeit alprazolam. The detection of highly toxic substances such as N-ethylpentylone and norfentanyl may warrant public health alerts. CONCLUSION: If provided close to real time, wastewater analysis at festivals could be complemented with information sources such as drug checking, on-site surveys, medical presentations and intelligence from peer networks to feed into early warning systems, public health alerts and peer-based harm reduction education during the festival season.

Correlates of higher-risk drug-related behaviours at music festivals in New South Wales, Australia.

INTRODUCTION: There are few contemporary data on illicit drug use at music festivals. We describe drug use patterns and prevalence of specific higher-risk drug-related behaviours, and their associations with festivalgoer characteristics. METHODS: We approached attendees at six major music festivals in New South Wales, Australia, from November 2019 to March 2020. Participants self-completed an anonymous survey on prior and intended drug use and associated higher-risk behaviours; double dropping; higher-volume ethanol alongside drug use; higher quantity 3,4-methylenedioxymethamphetamine (MDMA); mixing stimulants; and preloading. Logistic regression and UpSet analyses were performed to identify festivalgoer characteristics and the intersection of high-risk behaviours, respectively. RESULTS: Of 1229 participants, 372 (30.3%) used or planned to use drugs at the festival. In multivariable analyses, men and those purchasing drugs both inside and outside the venue had greater odds of engaging in higher-risk behaviours. Of those using MDMA, 47.9% reported double dropping. People using drugs for the first time had 3.3 (95% confidence interval 1.2-8.7) greater odds of higher-volume ethanol alongside drug use. People reporting that police/police dog presence influenced their decision to take drugs had 2.2 (95% confidence interval 1.4-3.6) greater odds of preloading. In UpSet analysis, preloading was the most common intersection (17% of those using drugs). DISCUSSIONS AND CONCLUSIONS: Engagement in the five higher-risk drug behaviours was common, particularly amongst males and those using drugs for the first time, while police/police dog presence appeared to influence higher-risk behaviours amongst festival attendees. This information can be used to inform harm reduction advice, public health and law enforcement strategies.

Geographically extensive larval surveys reveal an unexpected scarcity of primary vector mosquitoes in a region of persistent malaria transmission in western Zambia.

BACKGROUND: The Barotse floodplains of the upper Zambezi River and its tributaries are a highly dynamic environment, with seasonal flooding and transhumance presenting a shifting mosaic of potential larval habitat and human and livestock blood meals for malaria vector mosquitoes. However, limited entomological surveillance has been undertaken to characterize the vector community in these floodplains and their environs. Such information is necessary as, despite substantial deployment of insecticide-treated nets (ITNs) and indoor residual spraying (IRS) against Anopheles vectors, malaria transmission persists across Barotseland in Zambia's Western Province. METHODS: Geographically extensive larval surveys were undertaken in two health districts along 102 km of transects, at fine spatial resolution, during a dry season and following the peak of the successive wet season. Larvae were sampled within typical Anopheles flight range of human settlements and identified through genetic sequencing of cytochrome c oxidase I and internal transcribed spacer two regions of mitochondrial and nuclear DNA. This facilitated detailed comparison of taxon-specific abundance patterns between ecological zones differentiated by hydrological controls. RESULTS: An unexpected paucity of primary vectors was revealed, with An. gambiae s.l. and An. funestus representing < 2% of 995 sequenced anophelines. Potential secondary vectors predominated in the vector community, primarily An. coustani group species and An. squamosus. While the distribution of An. gambiae s.l. in the study area was highly clustered, secondary vector species were ubiquitous across the landscape in both dry and wet seasons, with some taxon-specific relationships between abundance and ecological zones by season. CONCLUSIONS: The diversity of candidate vector species and their high relative abundance observed across diverse hydro-ecosystems indicate a highly adaptable transmission system, resilient to environmental variation and, potentially, interventions that target only part of the vector community. Larval survey results imply that residual transmission of malaria in Barotseland is being mediated predominantly by secondary vector species, whose known tendencies for crepuscular and outdoor biting renders them largely insensitive to prevalent vector control methods.

Deep phylogeographic structure may indicate cryptic species within the Sparid genus Spondyliosoma.

Two geographically nonoverlapping species are currently described within the sparid genus Spondyliosoma: Spondyliosoma cantharus (Black Seabream) occurring across Mediterranean and eastern Atlantic waters from NW Europe to Angola and S. emarginatum (Steentjie) considered endemic to southern Africa. To address prominent knowledge gaps this study investigated range-wide phylogeographic structure across both species. Mitochondrial DNA sequences revealed deep phylogeographic structuring with four regionally partitioned reciprocally monophyletic clades, a Mediterranean clade and three more closely related Atlantic clades [NE Atlantic, Angola and South Africa (corresponding to S. emarginatum)]. Divergence and distribution of the lineages reflects survival in, and expansion from, disjunct glacial refuge areas. Cytonuclear differentiation of S. emarginatum supports its validity as a distinct species endemic to South African waters. However, the results also indicate that S. cantharus may be a cryptic species complex wherein the various regional lineages represent established/incipient species. A robust multilocus genetic assessment combining morphological data and detailing interactions among lineages is needed to determine the full diversity within Spondyliosoma and the most adequate biological and taxonomic status.

Genetic analysis provides insights into species distribution and population structure in East Atlantic horse mackerel (Trachurus trachurus and T. capensis).

Two sister species of horse mackerel (Trachurus trachurus and T. capensis) are described that are intensively harvested in East Atlantic waters. To address long-standing uncertainties as to their respective geographical ranges, overlap and intraspecific population structure this study combined genetic (mitochondrial DNA and microsatellite) analysis and targeted sampling of the hitherto understudied West African coast. mtDNA revealed two reciprocally monophyletic clades corresponding to each species with interspecies nuclear differentiation supported by FST values. The T. trachurus clade was found across the north-east Atlantic down to Ghana but was absent from Angolan and South African samples. The T. capensis clade was found only in South Africa, Angola and a single Ghanaian individual. This pattern suggests that both species may overlap in the waters around Ghana. The potential for cryptic hybridization and/or indiscriminate harvesting of both species in the region is discussed. For T. capensis mtDNA supports high gene flow across the Benguela upwelling system, which fits with the species' ecology. The data add to evidence of a lack of significant genetic structure throughout the range of T. trachurus though the assumption of demographic panmixia is cautioned against. For both species, resolution of stock recruitment heterogeneity relevant to fishery management, as well as potential hybridization, will require more powerful genomic analyses.

The complete mitochondrial genome of record-breaking migrant Arctic tern (Sterna paradisaea).

The analysis of mitochondrial DNA (mtDNA) base composition, codon usage, and genome arrangement patterns can provide insight into metabolic pathways and evolutionary history. Here, we report on the complete mitochondrial genome (mitogenome) of Arctic tern (Sterna paradisaea) a species notable for undertaking the longest migrations of any species as well as breeding in sub-polar habitats and capable of enduring extreme altitude. The complete mitogenome was 16,708 bp long and was typical of other avian mitogenomes in size and content. The phylogenetic position of the Arctic tern within Charadriiformes based on the coding region on the mtDNA corresponded closely to that based on nuclear loci. The sequence will provide a useful resource for investigations of metabolic adaptations of this remarkable species.

Cryptic species and parallel genetic structuring in Lethrinid fish: Implications for conservation and management in the southwest Indian Ocean.

Analysis of genetic variation can provide insights into ecological and evolutionary diversification which, for commercially harvested species, can also be relevant to the implementation of spatial management strategies and sustainability. In comparison with other marine biodiversity hot spots, there has been less genetic research on the fauna of the southwest Indian Ocean (SWIO). This is epitomized by the lack of information for lethrinid fish, which support socioeconomically important fisheries in the region. This study combines comparative phylogeographic and population genetic analyses with ecological niche modeling to investigate historical and contemporary population dynamics of two species of emperor fish (Lethrinus mahsena and Lethrinus harak) across the SWIO. Both species shared similarly shallow phylogeographic patterns and modeled historical (LGM) habitat occupancies. For both species, allele frequency and kinship analyses of microsatellite variation revealed highly significant structure with no clear geographical pattern and nonrandom genetic relatedness among individuals within samples. The genetic patterns for both species indicate recurrent processes within the region that prevent genetic mixing, at least on timescales of interest to fishery managers, and the potential roles of recruitment variability and population isolation are discussed in light of biological and environmental information. This consistency in both historical and recurrent population processes indicates that the use of model species may be valuable in management initiatives with finite resources to predict population structure, at least in cases wherein biogeographic and ecological differences between taxa are minimized. Paradoxically, mtDNA sequencing and microsatellite analysis of samples from the Seychelles revealed a potential cryptic species occurring in sympatry with, and seemingly morphologically identical to, L. mahsena. BLAST results point to the likely misidentification of species and incongruence between voucher specimens, DNA barcodes, and taxonomy within the group, which highlights the utility and necessity of genetic approaches to characterize baseline biodiversity in the region before such model-based methods are employed.