Development of a non-invasive technique to determine reproductive hormones in cetaceans

Reproductive physiology plays a vital role in population growth and vitality. Baseline data on reproductive physiology and a comprehensive knowledge of breeding biology are essential to conservation management. Great whales have been hunted from the 16th century to the present day. Although many populations are increasing there are populations with low or declining reproductive rates. In 2001 it was recommended to the International Whaling Commission that new techniques be developed to assess the internal physiology of great whales. This study, based on this recommendation, aims to develop analytical methods to assess reproductive hormones in cetacean blow samples and determine the feasibility of its use with free-swimming great whales. A method for the assessment of steroid hormone concentrations using liquid chromatography-mass spectrometry (LC-MS) was developed and validated. These methods were then used to determine testosterone and progesterone concentrations in saliva and blow of bottlenose dolphins. The stability of testosterone and progesterone was found to be a major issue. Without inhibitors, hormone concentrations increased by up to 65% over three hours at 21oC. Storing samples at low temperatures (-20oC or -80oC) slowed but did not cease the rate of change. The addition of inhibitors, manganese chloride and amoxycillin potassium/clavulanate, improved the stability of testosterone and progesterone. It is proposed that when using dolphin saliva and blow samples to measure reproductive hormones the samples are extracted as soon as possible after collection to prevent degradation. This study highlighted the need to address steroid hormone stability prior to any longterm biological program, to ensure that changes seen in hormone concentration are due to biological activity rather than storage. A technique to collect blow samples from free-swimming great whales was developed. This technique, in conjunction with the specially developed LC-MS methods allowed for the determination of testosterone and progesterone concentrations in humpback whale blow. The techniques developed in this study to determine reproductive hormones in cetacean saliva and blow have applications for both captive and wild population studies. In captive institutions, saliva and/or blow can be used to monitor reproductive cycling in both females and males. As it is noninvasive it can be used on a daily basis with minimal stress to the animals. The use of blow sampling has the capacity to improve our understanding of reproductive cycling in great whales as it can be used to sample animals in both the breeding and feeding areas. This technique may allow us to now examine whether reproductive dysfunction is playing a role in the slow recovery of critically endangered species such as the North Atlantic right whale

Phylogeography of New Zealand’s coastal benthos

During the past 30 years, 42 molecular studies have been undertaken in New Zealand to examine the phylogeography of coastal benthic invertebrates and plants. Here, we identify generalities and/or patterns that have emerged from this research and consider the processes implicated in generating genetic structure within populations. Studies have used various molecular markers and examined taxonomic groups with a range of life histories and dispersal strategies. Genetic disjunctions have been identified at multiple locations, with the most frequently observed division occurring between northern and southern populations at the top of the South Island. Although upwelling has been implicated as a cause of this disjunction, oceanographic evidence is lacking and alternative hypotheses exist. A significant negative correlation between larval duration and genetic differentiation (r2 = 0.39, P < 0.001, n = 29) across all studies suggests that larval duration might be used as a proxy for dispersal potential. However, among taxa with short larval durations (<10 days) there was greater variability in genetic differentiation than among taxa with longer pelagic periods. This variability implies that when larval duration is short, other factors may determine dispersal and connectivity among populations. Although there has been little congruence between the phylogeographic data and recognised biogeographic regions, recent research has resolved population subdivision at finer spatial scales corresponding more closely with existing biogeographic classifications. The use of fast-evolving and ecologically significant molecular markers in hypothesis-driven research could further improve our ability to detect population subdivision and identify the processes structuring marine ecosystems

Disentangling the mechanisms of mate choice in a captive koala population

Successful captive breeding programs are crucial to the long-term survival of many threatened species. However, pair incompatibility (breeding failure) limits sustainability of many captive populations. Understanding whether the drivers of this incompatibility are behavioral, genetic, or a combination of both, is crucial to improving breeding programs. We used 28 years of pairing data from the San Diego Zoo koala colony, plus genetic analyses using both major histocompatibility complex (MHC)-linked and non-MHC-linked microsatellite markers, to show that both genetic and non-genetic factors can influence mating success. Male age was reconfirmed to be a contributing factor to the likelihood of a koala pair copulating. This trend could also be related to a pair’s age difference, which was highly correlated with male age in our dataset. Familiarity was reconfirmed to increase the probability of a successful copulation. Our data provided evidence that females select mates based on MHC and genome-wide similarity. Male heterozygosity at MHC class II loci was associated with both pre- and post-copulatory female choice. Genome-wide similarity, and similarity at the MHC class II DAB locus, were also associated with female choice at the post-copulatory level. Finally, certain MHC-linked alleles were associated with either increased or decreased mating success. We predict that utilizing a variety of behavioral and MHC-dependent mate choice mechanisms improves female fitness through increased reproductive success. This study highlights the complexity of mate choice mechanisms in a species, and the importance of ascertaining mate choice mechanisms to improve the success of captive breeding programs

Charity registration and reporting:a cross-Jurisdictional and theoretical analysis of regulatory impact

Increasingly governments worldwide regulate charities, seeking to restrict the number of organizations claiming taxation exemptions, reduce abuse of state support and fraud. Under public interest theory governments may increase philanthropy through public trust and confidence in charities. Under public choice theory regulators will maximize political returns, ‘manage’ charity-government relationships, and avoid regulatory capture. Phillips and Smith (2014) suggest that charities’ regulatory regimes should coalesce, despite jurisdictional diversity. We analyse charity regulatory regimes against underlining theories of regulation, and assess regulatory costs and benefits. Thus regulators can reduce regulatory inefficiency, and balance accountability and transparency demands with charities’ abilities to deliver

Immunization strategies producing a humoral IgG immune response against devil facial tumor disease in the majority of Tasmanian devils destined for wild release

Devil facial tumor disease (DFTD) is renowned for its successful evasion of the host immune system. Down regulation of the major histocompatabilty complex class I molecule (MHC-I) on the DFTD cells is a primary mechanism of immune escape. Immunization trials on captive Tasmanian devils have previously demonstrated that an immune response against DFTD can be induced, and that immune-mediated tumor regression can occur. However, these trials were limited by their small sample sizes. Here, we describe the results of two DFTD immunization trials on cohorts of devils prior to their wild release as part of the Tasmanian Government's Wild Devil Recovery project. 95% of the devils developed anti-DFTD antibody responses. Given the relatively large sample sizes of the trials (N = 19 and N = 33), these responses are likely to reflect those of the general devil population. DFTD cells manipulated to express MHC-I were used as the antigenic basis of the immunizations in both trials. Although the adjuvant composition and number of immunizations differed between trials, similar anti-DFTD antibody levels were obtained. The first trial comprised DFTD cells and the adjuvant combination of ISCOMATRIX™, polyIC, and CpG with up to four immunizations given at monthly intervals. This compared to the second trial whereby two immunizations comprising DFTD cells and the adjuvant combination ISCOMATRIX™, polyICLC (Hiltonol®) and imiquimod were given a month apart, providing a shorter and, therefore, more practical protocol. Both trials incorporated a booster immunization given up to 5 months after the primary course. A key finding was that devils in the second trial responded more quickly and maintained their antibody levels for longer compared to devils in the first trial. The different adjuvant combination incorporating the RNAase resistant polyICLC and imiquimod used in the second trial is likely to be responsible. The seroconversion in the majority of devils in these anti-DFTD immunization trials was remarkable, especially as DFTD is hallmarked by its immune evasion mechanisms. Microsatellite analyzes of MHC revealed that some MHC-I microsatellites correlated to stronger immune responses. These trials signify the first step in the long-term objective of releasing devils with immunity to DFTD into the wild.Ruth Pye, Amanda Patchett, Elspeth McLennan, Russell Thomson, Scott Carver ... A. Bruce Lyons ... et al

Extant and extinct bilby genomes combined with Indigenous knowledge improve conservation of a unique Australian marsupial

Ninu (greater bilby, Macrotis lagotis) are desert-dwelling, culturally and ecologically important marsupials. In collaboration with Indigenous rangers and conservation managers, we generated the Ninu chromosome-level genome assembly (3.66 Gbp) and genome sequences for the extinct Yallara (lesser bilby, Macrotis leucura). We developed and tested a scat single-nucleotide polymorphism panel to inform current and future conservation actions, undertake ecological assessments and improve our understanding of Ninu genetic diversity in managed and wild populations. We also assessed the beneficial impact of translocations in the metapopulation (N = 363 Ninu). Resequenced genomes (temperate Ninu, 6; semi-arid Ninu, 6; and Yallara, 4) revealed two major population crashes during global cooling events for both species and differences in Ninu genes involved in anatomical and metabolic pathways. Despite their 45-year captive history, Ninu have fewer long runs of homozygosity than other larger mammals, which may be attributable to their boom-bust life history. Here we investigated the unique Ninu biology using 12 tissue transcriptomes revealing expression of all 115 conserved eutherian chorioallantoic placentation genes in the uterus, an XYY sex chromosome system and olfactory receptor gene expansions. Together, we demonstrate the holistic value of genomics in improving key conservation actions, understanding unique biological traits and developing tools for Indigenous rangers to monitor remote wild populations

The Earth BioGenome Project 2020: Starting the clock

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Barriers to antiretroviral therapy adherence in rural Mozambique

<p>Abstract</p> <p>Background</p> <p>HIV is treated as a chronic disease, but high lost-to-follow-up rates and poor adherence to medication result in higher mortality, morbidity, and viral mutation. Within 18 clinical sites in rural Zambézia Province, Mozambique, patient adherence to antiretroviral therapy has been sub-optimal.</p> <p>Methods</p> <p>To better understand barriers to adherence, we conducted 18 community and clinic focus groups in six rural districts. We interviewed 76 women and 88 men, of whom 124 were community participants (CP; 60 women, 64 men) and 40 were health care workers (HCW; 16 women, 24 men) who provide care for those living with HIV.</p> <p>Results</p> <p>While there was some consensus, both CP and HCW provided complementary insights. CP focus groups noted a lack of confidentiality and poor treatment by hospital staff (42% CP vs. 0% HCW), doubt as to the benefits of antiretroviral therapy (75% CP vs. 0% HCW), and sharing medications with family members (66% CP vs. 0%HCW). Men expressed a greater concern about poor treatment by HCW than women (83% men vs. 0% women). Health care workers blamed patient preference for traditional medicine (42% CP vs. 100% HCW) and the side effects of medication for poor adherence (8% CP vs. 83% CHW).</p> <p>Conclusions</p> <p>Perspectives of CP and HCW likely reflect differing sociocultural and educational backgrounds. Health care workers must understand community perspectives on causes of suboptimal adherence as a first step toward effective intervention.</p