scholarly journals Using Noninvasive Genetic Sampling to Survey Rare Butterfly Populations

Insects ◽  
2019 ◽  
Vol 10 (10) ◽  
pp. 311
Author(s):  
Caroline Storer ◽  
Jaret Daniels ◽  
Lei Xiao ◽  
Kristin Rossetti

Advances in nondestructive genetic sampling techniques continue to offer new opportunities for studying organisms, particularly those of conservation concern where more traditional invasive sampling methods are often not available. As part of a proof-of-concept, we investigated the effectiveness of using the chorion from residual butterfly egg debris as a source of viable genetic material for analysis. Laboratory material from a captive breeding population of the federally endangered Miami blue butterfly (Cyclargus thomasi bethunebakeri) was used to test efficacy and refine the methodology. The resulting best practices were subsequently evaluated using field-collected material from extant north Florida populations of the at-risk frosted elfin butterfly (Callophyrs irus). Our results demonstrated that it is possible to extract DNA of sufficiently high quantity and quality for successful gene sequencing. We additionally describe a simple, low-cost, and reliable method of collecting and storing egg debris samples that can be consistently adopted for field or laboratory work as well as deployed with projects that have a larger geographic scope and/or involve citizen scientists. Potential limitations related to field sample collection are discussed as well as needs for future evaluation.

Diversity ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 570
Author(s):  
Miriam A. Zemanova ◽  
Daniel Ramp

Dispersal is a key process for population persistence, particularly in fragmented landscapes. Connectivity between habitat fragments can be easily estimated by quantifying gene flow among subpopulations. However, the focus in ecological research has been on endangered species, typically excluding species that are not of current conservation concern. Consequently, our current understanding of the behaviour and persistence of many species is incomplete. A case in point is the eastern grey kangaroo (Macropus giganteus), an Australian herbivore that is subjected to considerable harvesting and population control efforts. In this study, we used non-invasive genetic sampling of eastern grey kangaroos within and outside of the Mourachan Conservation Property to assess functional connectivity. In total, we genotyped 232 samples collected from 17 locations at 20 microsatellite loci. The clustering algorithm indicated the presence of two clusters, with some overlap between the groups within and outside of the reserve. This genetic assessment should be repeated in 10–15 years to observe changes in population structure and gene flow over time, monitoring the potential impact of the planned exclusion fencing around the reserve.


2021 ◽  
Author(s):  
◽  
Gemma Bowker-Wright

<p>Pateke/brown teal (Anas chlorotis) have experienced a severe population crash leaving only two remnant wild populations (at Great Barrier Island and Mimiwhangata, Northland). Recovery attempts over the last 35 years have focused on an intensive captive breeding programme which breeds pateke, sourced almost exclusively from Great Barrier Island, for release to establish re-introduced populations in areas occupied in the past. While this important conservation measure may have increased pateke numbers, it was unclear how much of their genetic diversity was being retained. The goal of this study was to determine current levels of genetic variation in the remnant, captive and re-introduced pateke populations using two types of molecular marker, mitochondrial DNA (mtDNA) and microsatellite DNA. Feathers were collected from pateke at Great Barrier Island, Mimiwhangata, the captive breeding population and four re-introduced populations (at Moehau, Karori Wildlife Sanctuary, Tiritiri Matangi Island and Mana Island). DNA was extracted from the base of the feathers, the mitochondrial DNA control region was sequenced, and DNA microsatellite markers were used to genotype individuals. The Great Barrier Island population was found to have only two haplotypes, one in very high abundance which may indicate that historically this population was very small. The captive breeding population and all four re-introduced populations were found to contain only the abundant Great Barrier Island haplotype as the vast majority of captive founders were sourced from this location. In contrast, the Mimiwhangata population contained genetic diversity and 11 haplotypes were found, including the Great Barrier Island haplotype which may have been introduced by captive-bred releases which occurred until the early 1990s. From the microsatellite results, a loss of genetic diversity (measured as average alleles per locus, heterozygosity and allelic richness) was found from Great Barrier Island to captivity and from captivity to re-introduction. Overall genetic diversity within the re-introduced populations (particularly the smaller re-introduced populations at Karori Wildlife Sanctuary, Tiritiri Matangi Island and Mana Island) was much reduced compared with the remnant populations, most probably as a result of small release numbers and small population size. Such loss of genetic diversity could render the re-introduced populations more susceptible to inbreeding depression in the future. Suggested future genetic management options are included which aim for a broader representation of genetic diversity in the pateke captive breeding and release programme.</p>


2020 ◽  
Author(s):  
Jana von Freyberg ◽  
Julia L. A. Knapp ◽  
Andrea Rücker ◽  
Bjørn Studer ◽  
James W. Kirchner

Abstract. Automated field sampling of streamwater or precipitation for subsequent analysis of stable water isotopes (2H and 18O) is often conducted with off-the-shelf automated samplers. However, water samples stored in the field for days and weeks in open bottles inside autosamplers undergo isotopic fractionation and vapor mixing, thus altering their isotopic signature. We therefore designed an evaporation protection method which modifies autosampler bottles using a syringe housing and silicone tube, and tested whether this method reduces evaporative fractionation and vapor mixing in water samples stored for up to 24 days in ISCO autosamplers (Teledyne ISCO., Lincoln, US). Laboratory and field tests under different temperature and humidity conditions showed that water samples in bottles with evaporation protection were far less altered by evaporative fractionation and vapor mixing than samples in conventional open bottles. Our design is a cost-efficient approach to upgrade the 1-litre sample bottles of ISCO 6712 Full-size Portable Samplers, allowing secure water sample collection in warm and dry environments. Our design can be readily adapted (e.g., by using a different syringe size) to fit the bottles used by many other field autosamplers.


2011 ◽  
Vol 14 (3) ◽  
Author(s):  
Thomas McDade

The Health and Retirement Study (HRS) is an important national resource for policy makers and investigators across a wide range of disciplines, and it is critical that the study collects the best information possible on the health status of its participants within the constraints of the survey design, and without compromising the integrity of the sample. Potential directions for the collection and analysis of biomarker data in future waves of HRS are discussed, with a primary focus on blood-based biomarkers. Advantages and disadvantages of various methods for collecting blood in the home are considered, with particular attention given to the strengths and weaknesses of dried blood spot (DBS) sampling. DBS sampling has been widely applied in recent biosocial surveys due to the low cost and burden associated with sample collection, but these benefits need to be weighed against challenges associated with quantification in the laboratory. Attention is also given to additional biomarkers that may be of relevance to HRS, and that would expand the survey’s current focus on obesity and metabolic syndrome. Measures of inflammation, pathogen exposure, reproductive function, stress, and epigenetic modifications are suggested as potentially productive future directions for the study. In addition, the analysis concludes with the following recommendations for HRS: Continue to collect DBS samples, but consider alternatives; implement enhanced procedures for quality control; calibrate DBS results against plasma values, and invest in methods development.


Acrocephalus ◽  
2012 ◽  
Vol 33 (152-153) ◽  
pp. 25-104 ◽  
Author(s):  
Dejan Bordjan

Between the end of February 2007 and mid-February in 2008, 73 systematic surveys of waterbirds and raptors were carried out during 10-day periods at Cerknica polje (Southern Slovenia). The main objectives of our research were: to ascertain the abundance and temporal dynamics of the species occurring here, to specify their status, to make a comparison with the past period of systematic data gathering in the 1991-1992 period, to present estimates of the breeding, migrating and wintering populations, as well as to make, on these bases, a nature-conservancy evaluation of the area. Other ornithological data, acquired till the end of 2010, were gathered as well. Between the beginning of May and the beginning of September 2007, the water in the area virtually ran dry (it was retained only in the channels of the streams), while the surface itself was partially or fully icebound from mid-November 2007 till the beginning of February 2008. By the end of 2010, a total of 129 waterbird and raptor species were recorded at Cerknica polje, while in the 2007-2008 period 83 were registered. Dynamics of the species occurrence with more than nine observations made in the area is presented in a greater detail in the species overview. The breeding status was held by 27 bird species, while further seven were forage guests that bred in the vicinity of the study area. Most of the species (118) had the status of passage visitor, whereas those with the status of summer visitor (34) and winter visitor (40) were fewer. Among the recorded species, 34 were accidental and 21 rare visitors. There were 16 winter and seven summer residents, while year-round residents were six. In all 10-day periods of the research period, four species were recorded: Mallard Anas plathyrhynchos, Grey Heron Ardea cinerea, Buzzard Buteo buteo and Kestrel Falco tinnunculus. In more than 90% of 10-day periods, the Great Egret Ardea alba was recorded as well. In the 2007-2008 period, two species were eudominant (Mallard 27.2%, Buzzard 10.1%), whereas another two were dominant (Garganey Anas querquedula 7.4%, Coot Fulica atra 6.4%). Most individuals were registered at the end of March and in early April (up to 1,978 ind.), whereas the greatest numbers of species (48) were recorded in mid-April. Between May and August, the numbers of individuals and species were low owing to the dried up lake. The smallest area of occurrence was occupied by the Cormorant Phalacrocorax carbo and a group of ducks of the genera Aythya, Bucephala and Mergellus. Gulls, egrets, herons, waders, harriers Circus sp. and the Red-footed Kestrel Falco vespertinus occurred in the greater part of the research area. 10 breeders fulfilled the criteria of the species of the greatest conservation importance, two of which (Ferruginous Duck Aythya nyroca and Curlew Numenius arquata) are species of global conservation concern, whereas seven are of conservation concern on a European scale. Among non-breeders, 14 waterbird species and raptors occurred in significant numbers (> 0.1% biogeographical population), five of which occurred at least occasionally with more than 1% of their biographical population. Two species (Red-necked Grebe Podiceps grisegena and Snipe Gallinago gallinago) breed in Slovenia only at Cerknica polje, while further six species have here at least 40% of their Slovenian breeding population


2021 ◽  
Author(s):  
Nicole E Adams ◽  
Madeleine A Becker ◽  
Suzanne Edmands

Abstract Background With developing understanding that host-associated microbiota play significant roles in individual health and fitness, taking an interdisciplinary approach combining microbiome research with conservation science is increasingly favored. Here we establish the scat microbiome of the imperiled Channel Island fox (Urocyon littoralis) and look at the effects of geography and captivity on the variation in bacterial communities. Results Using high throughput 16S rRNA gene amplicon sequencing, we discovered distinct bacterial communities in each island fox subspecies. Weight, timing of the sample collection, and sex contributed to the geographic patterns. We uncovered significant taxonomic differences and an overall decrease in bacterial diversity in captive versus wild foxes. Conclusions Understanding the drivers of microbial variation in this system provides a valuable lens through which to evaluate the health and conservation of these genetically depauperate foxes. The island-specific bacterial community baselines established in this study can make monitoring island fox health easier and understanding the implications of inter-island translocation clearer. The decrease in bacterial diversity within captive foxes could lead to losses in the functional services normally provided by commensal microbes and suggests that zoos and captive breeding programs would benefit from maintaining microbial diversity.


2017 ◽  
Author(s):  
Natalie Emma Calatayud ◽  
Amanda Kathryn Mullen ◽  
Cecilia Jane Langhorne

ABSTRACTLoss of reproductive viability, physiologically and/or behaviorally, can have profound effects on the fitness of a captive population and conservation efforts. The southern rocky mountain (SRM) population of the boreal toad has declined over the past 35 years, making captive breeding necessary to protect and augment the species in the wild. In recent years, a notable reduction in the incidence of amplexus and viable offspring from the captive breeding population has been observed. Hormone treatment protocols to stimulate gamete release in males and females are established in this species and in vitro fertilization has been performed successfully. However, successful hormone stimulation of reproductive behaviors and natural fertilization has not been well documented. During the breeding season of 2012, 24 males and 24 female toads were selected from a population of over 600 captive animals. Both sexes were treated with Human chorionic gonadotropin (hCG) and Gonadotropin Releasing Hormone (GnRH) or phosphate buffered saline (PBS). Females were primed twice with 3.7IU/g hCG and then injected with an ovulatory dose (OvD) of 13.5 IU/ g BW (Body weight) hCG and 0.4 μg/ g BW GnRH. Males were injected a single time with 10 IU/g BW hCG and 0.4 μg/ g BW GnRH, 12 h after females received their OvD. In 2013, knowing the approximate time when females oviposited after hormone treatments, we tested the best time to induce amplexus and spermiation. Males were divided into 4 groups and injected at 4 different times: (a) 12 h before females OvD; (b) at the same time as OvD; (c) 12 h after OvD; (d) control injected with PBS. Results from 2012 indicated that oviposition was solely dependent on females receiving hormone treatments not males. However, in 2013 we found that the duration of amplexus significantly influenced oviposition (P>0.05), and males injected 12 h prior to females spent more time in amplexus than males injected at the same time or 12 h after the females received hormones. Promoting reproductive behaviors and synchronizing gamete deposition continues to be imprecise and may require more than exogenous hormones. The complexity of promoting breeding behaviors may require a closer assessment of the captive environment.


2020 ◽  
Vol 24 (12) ◽  
pp. 5821-5834
Author(s):  
Jana von Freyberg ◽  
Julia L. A. Knapp ◽  
Andrea Rücker ◽  
Bjørn Studer ◽  
James W. Kirchner

Abstract. Automated field sampling of streamwater or precipitation for subsequent analysis of stable water isotopes (2H and 18O) is often conducted with off-the-shelf automated samplers. However, when water samples are stored in the field for days and weeks in open bottles inside autosamplers, their isotopic signatures can be altered by evaporative fractionation and vapor mixing. We therefore designed an evaporation protection method which modifies autosampler bottles using a syringe housing and silicone tube, and we tested whether this method reduces evaporative fractionation and vapor mixing in water samples stored for up to 24 d in 6712 full-size portable samplers (Teledyne ISCO, Lincoln, USA). Laboratory and field tests under different temperature and humidity conditions showed that water samples in bottles with evaporation protection were far less altered by evaporative fractionation and vapor mixing than samples in conventional open bottles. Our design is a cost-efficient approach to upgrade the 1 L sample bottles of the ISCO autosamplers, allowing secure water sample collection in warm and dry environments. Our design can be readily adapted (e.g., by using a different syringe size) to fit the bottles used by many other field autosamplers.


2021 ◽  
Author(s):  
◽  
Gemma Bowker-Wright

<p>Pateke/brown teal (Anas chlorotis) have experienced a severe population crash leaving only two remnant wild populations (at Great Barrier Island and Mimiwhangata, Northland). Recovery attempts over the last 35 years have focused on an intensive captive breeding programme which breeds pateke, sourced almost exclusively from Great Barrier Island, for release to establish re-introduced populations in areas occupied in the past. While this important conservation measure may have increased pateke numbers, it was unclear how much of their genetic diversity was being retained. The goal of this study was to determine current levels of genetic variation in the remnant, captive and re-introduced pateke populations using two types of molecular marker, mitochondrial DNA (mtDNA) and microsatellite DNA. Feathers were collected from pateke at Great Barrier Island, Mimiwhangata, the captive breeding population and four re-introduced populations (at Moehau, Karori Wildlife Sanctuary, Tiritiri Matangi Island and Mana Island). DNA was extracted from the base of the feathers, the mitochondrial DNA control region was sequenced, and DNA microsatellite markers were used to genotype individuals. The Great Barrier Island population was found to have only two haplotypes, one in very high abundance which may indicate that historically this population was very small. The captive breeding population and all four re-introduced populations were found to contain only the abundant Great Barrier Island haplotype as the vast majority of captive founders were sourced from this location. In contrast, the Mimiwhangata population contained genetic diversity and 11 haplotypes were found, including the Great Barrier Island haplotype which may have been introduced by captive-bred releases which occurred until the early 1990s. From the microsatellite results, a loss of genetic diversity (measured as average alleles per locus, heterozygosity and allelic richness) was found from Great Barrier Island to captivity and from captivity to re-introduction. Overall genetic diversity within the re-introduced populations (particularly the smaller re-introduced populations at Karori Wildlife Sanctuary, Tiritiri Matangi Island and Mana Island) was much reduced compared with the remnant populations, most probably as a result of small release numbers and small population size. Such loss of genetic diversity could render the re-introduced populations more susceptible to inbreeding depression in the future. Suggested future genetic management options are included which aim for a broader representation of genetic diversity in the pateke captive breeding and release programme.</p>


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Shailabh Kumar ◽  
Felix J. H. Hol ◽  
Sujit Pujhari ◽  
Clayton Ellington ◽  
Haripriya Vaidehi Narayanan ◽  
...  

AbstractMosquito bites transmit a number of pathogens via salivary droplets deposited during blood-feeding, resulting in potentially fatal diseases. Little is known about the genomic content of these nanodroplets, including the transmission dynamics of live pathogens. Here we introduce Vectorchip, a low-cost, scalable microfluidic platform enabling high-throughput molecular interrogation of individual mosquito bites. We introduce an ultra-thin PDMS membrane which acts as a biting interface to arrays of micro-wells. Freely-behaving mosquitoes deposit saliva droplets by biting into these micro-wells. By modulating membrane thickness, we observe species-dependent differences in mosquito biting capacity, utilizable for selective sample collection. We demonstrate RT-PCR and focus-forming assays on-chip to detect mosquito DNA, Zika virus RNA, as well as quantify infectious Mayaro virus particles transmitted from single mosquito bites. The Vectorchip presents a promising approach for single-bite-resolution laboratory and field characterization of vector-pathogen communities, and could serve as a powerful early warning sentinel for mosquito-borne diseases.


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