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Oral | Posters


Oral Presentations

 

 

Day: 2, Session: 2, Talk: 4

Examination of Morphological Variation in Faxonius jeffersoni Populations Indicates the Presence of a Species Complex

Zachary L. Couch

Faxonius jeffersoni Rhoades is a poorly known member of the crayfish family Cambaridae currently found in six small stream systems in Jefferson, Oldham, and Bullitt counties in Kentucky, USA. Fitzpatrick (1967) was the first to report morphological variation in gonopod sculpturing, rostrum width, and areola width between populations of F. jeffersoni found in the Beargrass and Pond Creek drainages in Jefferson and Bullitt counties. During qualitative and quantitative surveys conducted as a part of this study from 2007–2010 throughout Kentucky, examination of several morphological characteristics indicate that F. jeffersoni may represent multiple species. Examined specimens of F. jeffersoni from the Beargrass (n=48) and Goose Creek (n=15) drainages possess a gonopod with terminal elements (equal in length) that are 26.51% (0.026) and 26.62% (0.019) of the total length of the gonopod, respectively, lack a shoulder below the central projection, and typically exhibit a smooth mandibular margin. Specimens of F. jeffersoni examined from the Pond Creek (n=21) and Abrams Run (n=6) drainages are typically characterized by having a gonopod with terminal elements (the central projection being longer than the mesial process) that are 31.93% (0.019) and 31.54% (0.026) of the total length of the gonopod, respectively, possess a shoulder on the gonopod below the central projection, and typically exhibit a dentate mandibular margin. A median carina is often observed in larger specimens from the Pond Creek and Abrams Run drainages but was always found to be lacking from examined specimens from the Beargrass and Goose Creek drainages. This study reports the findings of preliminary morphometric analyses collected to clarify the taxonomic status of F. jeffersoni. Additional data collection and analysis is ongoing. However, results of morphometric analyses conducted to date suggests that the population of F. jeffersoni found in the Pond Creek and Abrams Run drainages represents a new species. As the current range of the species resides in a largely urban watershed, these initial findings are encouraged for use by resource managers to begin to reassess the conservation status of the F. jeffersoni species complex and consider its need for legal protection at the state and federal levels.

 

 

Day: 2, Session: 3, Talk: 1

Assessing Crayfish Habitat Requirements: A Proposed Method for Determining Habitat Breadth in Lotic Ecosystems

CHRISTOPHER A. TAYLOR and Christopher J. Rice

Lotic ecosystems in the central and eastern United States can be highly variable in habitat structure, ranging from shallow riffles to deep pools. These same ecosystems harbor a highly endemic and threatened crayfish fauna. Understanding habitat requirements for rare species is fundamental for effective conservation, yet habitat descriptions for many of these species rely solely on data from easily accessed sampling locations. We tested a new sampling method for collecting the narrowly endemic Coldwater Crayfish (Faxonius eupunctus) in the southern Missouri Ozarks by utilizing SCUBA in deep, largely non-wadeable pools. Our methods allowed for unbiased and repeatable sampling within and across sites and for the collection of rigorous data on crayfish density and habitat associations. By combining our proposed pool sampling method with wadable riffle sampling, we expanded the known habitat requirements for the Coldwater Crayfish. Data from pool sampling increased known suitable ranges for temperature and substrate size for the species. Our work also found agreement between both habitat types for physcial habitat variables exhibiting significant relationships with Coldwater Crayfish density. Using mulitple sampling methods has long been known to increase the accuracy of community inventory sampling given differences in microhabitat use by community members. Our work demonstrates that employing multiple sampling methods can increase the accuracy of habitat requirements for rare crayfish species.

 

 

Day: 2, Session: 3, Talk: 3

Modeling Effects of Crayfish Invasion and Drought on Crayfish Population Dynamics

Leah Bayer, Robert Fournier and DANIEL D. MAGOULICK

Crayfish play a crucial ecological role and are often considered a keystone species within freshwater ecosystems. However, North American crayfish species face several environmental and ecological threats including limited natural ranges, invasive species, and intensified drought. Demographic models can allow examination of population dynamics of a targeted species under a wide variety of disturbance scenarios. Here, we model the population dynamics of crayfish species with varied theoretical life histories and assess their responses to biological invasions and drought. We used RAMAS-Metapop to construct stage-based demographic metapopulation models parameterized using vital rates from established literature sources. Our models explored the population viability of four theoretical species under eleven disturbance scenarios and calculated estimates of terminal extinction risk, median time to quasi-extinction, and metapopulation occupancy. Our models indicate that populations respond differentially to disturbance based on life history. However, both r- and K-selected species appear to be highly susceptible to decline when faced with the additive effects of reduced carrying capacity due to invasion and reduced vital rates due to drought. By constructing models that explore a broad array of life histories and disturbance regimes, we hope to provide managers with tools to develop generalized, widely-applicable conservation strategies.

 

 

Day: 2, Session: 3, Talk: 7

Introduced and Then Almost Forgotten: Invasive Crayfish (Procambarus clarkii and Cherax quadricarinatus) in Costa Rica, Central America - Preliminary Results

JUAN CARLOS AZOFEIFA SOLANO, Ingo S. Wehrtmann, Fresia Villalobos-Rojas, Raquel Romero-Chaves and Adrián García-Rodríguez

Costa Rica harbors a remarkable diversity of freshwater native decapods, including 15 crab species (Pseudothelphusidae) and more than 21 species of caridean shrimps (Atyidae and Palaemonidae). Crayfish (Astacoidea) are not part of the native freshwater decapod fauna of the country. However, during the last century, two crayfish species were introduced: Procambarus clarkii in the Reventazón basin (Caribbean slope) in the 1970's, and Cherax quadricarinatus in the Tempisque basin (Pacific slope) in the 1980's. Both species have been recognized as invasive species in many other countries, and there is broad evidence suggesting its negative impact on native species, ecosystem functioning, and even human activities. Despite the potential threat of these crayfish species for the Costa Rican freshwater environments, very limited information is available about their occurrence in the country. Therefore, it is imperative to document the current distribution of both species and to assess their possible effects on the freshwater ecosystems in Costa Rica. We initiated a study aimed to explore the presence of these invasive crayfish species in Costa Rican streams and lakes and to provide information about their feeding ecology. So far, we have collected P. clarkii in two locations, outside the Reventazón basin were it was initially introduced, and belonging to the Pacific slope. More than 160 specimens have been analyzed, ranging in size from 6.5 to 78.1 mm total length. The predominant food item found at the stomachs was unidentified plant detritus, followed by filamentous plants, fragments of chironomid and Ephemeroptera larvae, and plastic. Our collections revealed two new locations for C. quadricarinatus in Costa Rica: one of them a stream in the Caribbean slope, and the other in the Arenal Lake, the largest reservoir in Costa Rica. The size of the collected specimens (n=7) ranged from 45.1 to 92.2 mm total length. Currently, both crayfish species inhabit both Caribbean and Pacific slopes in Costa Rica. It remains to be studied whether these species dispersed naturally or if they were locally introduced by humans. Since both species have been introduced in Costa Rica and the Central American region, the results of the study aim to raise awareness about the possible impacts of these crayfish species on the native flora and fauna of the local freshwater ecosystems, and to develop recommendations for the implementation of management programs.

 

 

Day: 2, Session: 4, Talk: 3

The Life History of Cambarus veteranus Faxon 1914 (Decapoda: Cambaridae) in the Clear Fork of the Guyandotte River, WV, USA

NICOLE SADECKY and Zachary J. Loughman

Cambarus veteranus Faxon, 1914 (Guyandotte River Crayfish), is an endangered, narrow endemic, residing in just two streams in the southern coalfields of West Virginia. A life history study was initiated for C. veteranus in Clear Fork of the Guyandotte River, Wyoming County, West Virginia to gather basic life history information needed for future conservation efforts. Monthly collections began June 2017 and continued through May 2018. Two 400-meter stream reaches were designated as life history study sites with two different 100-meter sub-reaches sampled each month. Specimens, regardless of species, were collected, sexed, and molt stage determined. Preliminary results suggest an importance of water temperature on crayfish capture with December yielding considerably higher capture rates of C. veteranus in comparison to other species collected. Form I males are ever present and reach their highest density in early winter. Pre-glaired females reached their highest densities in October and December with glaired females present nearly every month. A single ovigerous female was collected, bearing just two stage 4 juveniles, during the November sampling event. Three ovigerous females bearing stage 4 juveniles were subsequently collected during the March sampling event, thus suggesting overwintering with young. Molting events were observed between September and October with pre-molting individuals present in September and freshly molted individuals present in October. Additionally, molting events occurred in March with pre-molting individuals present as well as freshly molted individuals and in May with the majority of the population observed in the soft or fresh molt state. Providing life history information for C. veteranus will assist in conservation efforts and possible repatriation of C. veteranus in the future. Additionally, life history information for C. veteranus can be compared to closely related species that are often used as a surrogate for C. veteranus in captive rearing studies.

 

 

Day: 2, Session: 4, Talk: 4

An Update on the Distribution and Conservation Status of the Crayfishes of Alabama

STUART W. McGREGOR, Guenter A. Schuster, Christopher A. Taylor, Rebecca A. Bearden and E. Anne Wynn

Each state is required to have a State Wildlife Action Plan (SWAP) to be eligible for federal funds through the Wildlife Conservation Restoration Program (WCRP) or the State Wildlife Grants Program (SWG). For Alabama to move forward in conserving its aquatic species, a first step is understanding biodiversity patterns: What species do we have and where are they found? Second, it is important to know which species need conservation action. These fundamental levels of understanding did not exist for crayfishes in Alabama before our project. Starting in 2005 Drs. Guenter Schuster and Chris Taylor performed an exhaustive literature search and visited numerous museum collections and compiled a database with over 4,600 records documenting 85 crayfish species from Alabama. Subsequently, with funding supplied on two occasions by the SWG program and independently by the U.S. Fish and Wildlife Service, Geological Survey of Alabama staff, Drs. Schuster and Taylor, and associates sampled crayfish in areas of the state that showed a dearth of records based on maps generated from the Schuster and Taylor database and surveyed for species petitioned for listing by the U.S. Fish and Wildlife Service. The first SWG project (2008-10) added over 760 collection records, documented 64 of 85 species recognized from the state at that time, tightened collection coverage gaps, and recommended a preliminary conservation priority status for each species. In 2012, a group of subject matter experts convened in Auburn, Alabama, for the Third Nongame Wildlife Symposium and provided information on each species known from Alabama at the time. The result was an updated SWAP with conservation priority status designated for each species. Crayfish were included for the first time and 12 species were found to be of Highest Conservation Priority, 30 of High Conservation Priority, 15 of Moderate Conservation Priority, 14 of Low Conservation Priority, and 12 of Lowest Conservation Priority. Another important result of the first SWG project was the need to further close coverage gaps, further address undersampled habitats, and refine species-specific distributional information. The second SWG grant was secured to address those needs (2014-17). Final results of these studies yield about 9,300 records documenting 97 species of crayfishes (94 natives), with 15 state endemics, a few species whose taxonomic status remains unclear, a few undescribed taxa awaiting formal descriptions, and 5 hypothetical species. Another result was the opportunity to make preliminary conservation priority recommendations for species added to the state list or systematically reassigned during the latter phase of the project. We recommended 1 species status be changed to Highest Conservation Priority, 6 others added to Highest Conservation Priority, 4 to High Conservation Priority, 2 to Low Conservation Priority, and 1 to Lowest Conservation Priority. The state list will very likely surpass 100 species upon further research. During our studies 94 of the 97 species known from the state were encountered, with only 3 rare troglobites unobserved (but likely extant).

 

 

Day: 3, Session: 1, Talk: 3

STURE ABRAHAMSSON MEMORIAL LECTURE: Crayfish Color Patterns: Their Overlooked Significance

Guenter A. Schuster

Crayfish colors and color patterns have not been well studied. Horton H. Hobbs, Jr. and others, in descriptions of new species, mostly relied on verbal color pattern descriptions. This began to change with the publication of Raymond Bouchard's late 1980s color poster entitled "America's Crayfish." Since then, crayfish books and color posters representing crayfishes from several states and countries have been published. Now, color photographs are usually included in new species descriptions. State and federal agencies, as well as NGOs, are commonly using color photographs of crayfishes for conservation purposes. This talk addresses the North American cambarid crayfish fauna research on vision, and how color patterns might be useful to crayfishes. It will also address how these color patterns could provide important insight into the biology, behavior, taxonomy and systematics of crayfishes.

 

 

Day: 3, Session: 2, Talk: 1

Development of Mass Production Hatchery Technology for Cherax quadricarinatus

CLIVE JONES and Colin Valverde

Aquaculture production of redclaw (Cherax quadricarinatus) in Australia, has never reached the high level projections made in the 1990’s. This can be attributed to a range of factors, but the most significant is the supply of seedstock. One of the most positive aquaculture characteristics of redclaw is its bearing of offspring in adult form, obviating the requirement for larval rearing, as is necessary for most successfully commercialised aquaculture species. Among other positive characteristics, this was seen as the most significant and methods for pond-based, mass production of juvenile redclaw were devised and implemented by industry. This relied on natural reproduction, and natural productivity in the pond to support the survival and growth of the juveniles. Mean productivity was reported as 63.2 juveniles produced per berried female, and a harvest density of 24.6 juveniles per m2. Under suitable, summer photoperiod and temperature conditions, three consecutive crops of juveniles could be produced per year. Such managed pond production of seed stock for redclaw production was applied by industry with some success, but it soon became evident that productivity was too low and the practice occupied valuable pond space that would be more profitably applied to growout. An alternative production method for the supply of lobster juvenile redclaw was conceived by an innovative redclaw farmer, Colin Valverde, utilising artificial egg incubators, that had been successfully applied to freshwater crayfish species in Europe. The putative advantages of artificial egg incubation include savings of space, water and energy, minimising egg loss, control over the period of embryogenesis, known parentage for genetic selection and prevention of transmission of disease from parent to offspring. The primary advantages were initially for its application to genetic selection and for generating specific pathogen free stock. However, an even greater benefit became apparent, that this approach could support mass production of seedstock. The system that formed the basis of the that developed in Australia for redclaw was based primarily on the Hemputin™ incubator from Finland used for Pacifastacus leniusculus and Astacus astacus. Its design was modified to suit the specific requirements of redclaw and greater production volumes. The procedures and equipment have evolved, providing a foundation for more consistent and greater production of seed. The juvenile crayfish produced by the hatchery have generally moulted twice since hatching and are referred to as stage 3 juveniles or craylings. A hatchery supply of craylings provides the opportunity for farmers to stock an exact quantity of uniform size crayfish into growout ponds, which in turn enables calculation of likely survival, growth and biomass at the end of the production cycle. The Valverde hatchery system can generate tens of thousands of craylings per batch and is scalable. However there are production issues to be resolved including year round supply of eggs to the hatchery and management of egg health. This paper will detail the development of the Valverde hatchery system, and the challenges to see it develop into fully commercial production that can support expansion of the redclaw farming industry.

 

 

Day: 3, Session: 3, Talk: 6

Body Size in Freshwater Crayfish: An Intercontinental Comparison

Alaistair M. M. Richardson

The independent evolution of the astacoidean and parastacoidean crayfish in the Northern and Southern Hemispheres provides an opportunity to compare their characteristics. The comparison can be sharpened by comparing the crayfish faunas of Australia and North America, having roughly similar areas and lying at comparable latitudes. Size data were collected from the literature for 230 North American and 125 Australian species, and were compared directly and in terms of the species' burrowing habits. The Australian fauna includes several much larger species, and while the modal body size is very similar in the two faunas, the North American modal size class, and also the entire fauna, is dominated by tertiary burrowers, the Australian by primary ones. Further, Australian primary burrowers are smaller on average. The factors influencing these differences are discussed.

 

 

Day: 3, Session: 4, Talk: 3

Epigenetic Regulation in the Marbled Crayfish

VITOR COUTINHO CARNEIRO, Anny Gatzmann, Cassandra Falckenhayn and Frank Lyko

The all-female marbled crayfish (Procambarus virginalis) attracted the attention of the scientific community due to its 'self-cloning' capacity. The obligatory parthenogenetic reproduction provided interesting research opportunities and also established a potent ecological threat. Despite its identical DNA, this model has an extraordinary variety in appearance and behaviour between isogenic batchmates reared in the same environment. This suggests that epigenetic mechanisms play a key role in marbled crayfish phenotypic variation. Our group has recently annotated the draft genome of the marbled crayfish, which revealed a conserved, functional and versatile DNA methylation system for epigenetic regulation. We have also used whole-genome bisulfite sequencing, RNA-seq and ATAC-seq for a comprehensive analysis of multiple individuals and tissues. Our results provide a novel concept for how methylation-dependent regulation of gene expression may facilitate the phenotypic adaptation and invasive spread of this animal.

 

 

Day: 5, Session: 2, Talk: 1

Crayfish Chimney Function: Airflow, Oxygen, and Pheromones

JAMES A. STOECKEL, Hisham Abdelrahman, Mary Szoka, David Blersch and Jeremiah Davis

Most burrowing crayfish build chimneys. However, chimneys are often present only seasonally, located only over a single burrow opening, or completely absent, suggesting that they are periodically constructed for specific function and purpose(s). In this study we use a combination of field and wind-tunnel experiments to investigate function and purpose of Cambarus c.f. polychromatus chimneys. Field trials with smoke bombs and natural burrows showed that air flow is a major outcome of chimney construction. In a typical burrow complex, smoke was drawn in through three non-chimney openings, traveled ~0.5 m underground to the groundwater level, and then back up to the surface and exited through the chimney. Smoke was not drawn through the burrow when the chimney was removed. Chimneys appear to drive airflow through burrows by creating temperature differentials with non-chimney openings, and/or pressure differentials related to wind blowing across chimney and non-chimney entrances (Bernouli’s Principle). Wind tunnel trials with model burrows showed that air velocity through burrows was highest when chimneys were upwind of chimneyless openings. Air velocity through burrows also increased with increasing chimney height and increasing wind speed. Ongoing field studies suggest that need for increased oxygen is not likely the primary purpose driving crayfish to build chimneys to draw air through burrows. Individuals that were freshly molted or brooding, representing two life-history stages that require the most oxygen, were only found in plugged and chimneyless burrows, respectively. Alternatively, based on results of this and previous studies, we hypothesize that an important outcome of increased airflow through and out the burrows is dispersal of pheromones to facilitate intraspecific communication between adults during the reproductive season, and subsequent recruitment of young to conspecific burrow colonies.

 

 

Day: 5, Session: 2, Talk: 6

Simultaneous eDNA Monitoring of the Host-pathogen Complex Pacifastacus leniusculus and Aphanomyces astaci Under Varying Environmental Conditions

JOHANNES C. RUSCH, David A. Strand, Charlotte Laurendz, Stein I. Johnsen, Lennart Edsman and Trude Vrålstad

In 2016 the North-American signal crayfish Pacifastacus leniusculus was included in the list of Invasive Alien Species of concern to the European Union. In Norway the species is black-listed and rated as a high-risk species, and eradication has been carried out whenever feasible. This is because it is a chronic carrier of the oomycete Aphanomyces astaci, which is lethal to all European freshwater crayfish species and listed among the 100 worst invasive species. Environmental DNA (eDNA) methodology is now being used on a wide variety of target species and on many different platforms including targeted PCR and broad spectred sequencing methods. Detection and monitoring of invasive, endangered and elusive species is commonly performed using species specific quantitative real-time PCR (qPCR) or droplet digital PCR (ddPCR), and has proved useful for targeting species of specific concern. Recently eDNA monitoring of A. astaci has been implemented in the national crayfish plague surveillance in Norway. For the carrier of the crayfish plague, the signal crayfish, a qPCR assay for eDNA detection has also been published recently. In this study, we present simultaneous eDNA monitoring of the host-pathogen complex P. leniusculus - A. astaci under varying environmental conditions both in aquarium and field experiments. We used the published qPCR assays for both species and redesigned them so they can be run as a duplex ddPCR assay. We compared water samples by means of qPCR and ddPCR from two lakes in Sweden and Norway with different signal crayfish population densities and A. astaci prevalence in the population. We also studied eDNA emission from the host-pathogen couple in aquarium-experiments with A. astaci positive signal crayfish held at different densities, temperatures and feeding regimes. Samples were obtained by filtering water (1 & 5 L) on-site through glass fibre filters. These were subsequently analysed using the species-specific qPCR and ddPCR assays for the respective targets. The concentrations of detectable eDNA copies of the two targets are influenced differently and by several factors, including population density of the crayfish, pathogen prevalence, temperature, and turbidity including microbiological activity in the water. They are therefore subject to significant fluctuation. Thus, there seems to be no straightforward correlation between eDNA copy-number and crayfish density and the probability of detecting one target rather than the other varies according to different conditions. Therefore, for a reliable monitoring of crayfish plague (A. astaci) and signal crayfish alone or together, the simultaneous monitoring concept for both targets is recommended.

 

 

Day: 5, Session: 3, Talk: 1

Geographical Variation in Size and Growth in the Giant Freshwater Crayfish, Astacopsis gouldi. Deductions from a Large Opportunistic Database

ALASTAIR M. M. RICHARDSON and Todd Walsh

A tagging program on the Tasmanian giant freshwater crayfish, Astacopsis gouldi, has been carried out since 1998, using PIT tags since 2007. Almost 600 animals, ranging from 20-215 mm carapace length, have been tagged and around 2500 captures recorded from 123 localities on 62 rivers across the species' range. Comparisons of length-weight relationships and growth are made between the discrete western and eastern ranges of the species, and between populations living in nutrient-poor and nutrient-rich waters. Animals from the eastern range are slightly lighter for a given weight than western animals. Growth is marginally slower in animals from nutrient-poor waters. Individual growth histories are available for 26 animals, some of which have been recaptured up to 6 times, over as long as 10 years. Growth rates vary considerably within river populations, suggesting that there may be fast- and slow-growing animals within the same population. Gompertz growth models showed lower growth rates and asymptotes for the nutrient-poor population, but their usefulness was affected by the limited size at known age data. Models suggested that animals of 220 mm CPL may be 60-70 years old.

 

 

Day: 5, Session: 3, Talk: 3

Observations of Chelae Injury in Two Crayfish Species in Three Sinuous Rills

CHESTER R. FIGIEL JR.

Crayfish often lose or autotomize limbs (e.g., chelae) during agonistic conflicts. This defense mechanism increases the probability of surviving the encounter, however can result in long-term functional and energetic costs. For example, chela autotomy influences crayfish competitive ability, foraging time, capacity to obtain mates, and modifies crayfish distribution and behavior. Biotic factors (e.g., population density or predators), as well as abiotic factors (e.g., refugia or habitat complexity) can influence the frequency of injury. In this study, I investigated the prevalence of chela injury (loss or partially regenerated chelae) in two crayfish species (Cambarus striatus and Procambarus spiculifer) that were collected in three sinuous rills (Mountain Creek (MC), Liberty Bell Creek (LB), and Cascade Branch (CB)) in west central Georgia, USA over a four year period. My objectives were to determine if injury was similar among sites, among crayfish species, or varied with time of year. Additionally, within a species my objectives were to examine whether injury differed in crayfish size classes or sex. Cambarus striatus were collected in each of the three streams with the percent injured as follows: MC 15.5%, n = 579; LB 16.3%, n = 1555; and CB 21.3%, n = 1159. There were significantly more injuries from crayfish collected at CB than at the other two streams (p < 0.05). This was most likely driven by the greater percent of injuries in the juvenile size classes (6.0 mm to 15.9 mm carapace length (CL)): CB (24.5%) compared to the similar size classes at LB (16.6%) and MC (15.0%). The frequency of injury between males (18.2%), females (20.0%), or juveniles (19.0%) did not differ significantly nor were there significant differences between Form I (15.6%) and Form II (18.9) males (p > 0.05). Of the 590 C. striatus crayfish having an injury, approximately 13% of these crayfish had both chelae injured. Procambarus spiculifer were collected in each of the three streams (however only 3 individuals were collected at LB so data were not used for that site). The percent injured for P. spiculifer differed significantly between MC 12.1%, n = 727; and CB 23.7%, n = 169 (p < 0.05). There were a greater percent of injuries in the larger adult size classes (> 31.0 mm CL = 26.4%) compared to smaller size classes (< 31.0 mm CL = 9.9%). The frequency of injury among males (13.8%) and females (15.9%) did not differ significantly, however differed from the percent of injured juvenile crayfish (7.3%). Of the 126 P. spiculifer crayfish having an injury, approximately 12% of these crayfish had both chelae injured. The percent of injured crayfish captured varied widely from month to month for both species and no discernable pattern emerged through time. Given the level of injury in these populations, it is vital to understand how injury influences population dynamics of these two species with differing ecological needs and lifestyles.

 

 

Day: 5, Session: 3, Talk: 4

The Development and Application of a Commercially Viable and User Friendly Environmental DNA (eDNA) Methodology for the Conservation of the Endangered White-Clawed Crayfish

CHRISTOPHER TROTH and Michael J. Sweet

Over the last thirty years, in the UK and Europe the endangered indigenous white-clawed crayfish (Austropotamobius pallipes) has suffered extensive population decline due to the introduction of competitive and disease carrying non-native species. Detection of these now isolated populations is becoming more difficult using established and often destructive or invasive traditional ecological survey techniques. The recent emergence of molecular species detection, defined by the term ‘environmental DNA – eDNA’ has proven to be a valid additional cost-effective method to traditional surveys for many aquatic species worldwide. Here we develop an eDNA based method for the non-invasive detection of A. pallipes; inferring fast species presence or absence data within freshwater habitats. Working alongside stakeholders, commercial organisations, ecologists and end-user groups in the UK we have validated this method for use as a commercially available tool, through careful design and assessment of all variables and limitations that are known to effect eDNA. This has enabled the validation of the technique in ‘real world’ conservation and commercial based settings. Primary trials into the potential quantification of white-clawed crayfish biomass using eDNA have had success with high and medium densities of crayfish in controlled environments. However, at lower natural densities quantification attempts were more varied. We also present the analysis of two different, commonly used sample collection methods to identify the most suitable and commercially applicable technique for effective detection of A. pallipes which may depend on the goal of each individual study. In this paper we report our findings on A. pallipes distribution and abundance in both lotic and lentic habitats using eDNA, incorporating two different approaches to sample collection. We also explore the variation of eDNA detection rates through the seasons, to determine if eDNA could be used for the detection of A. pallipes all year round, or during the more active summer season of crayfish activity. Coupled with further research into the rate of eDNA decay over time, and sample collection methodology choice we can we can now apply these methods in the field successfully taking all major limitations into account to get the most out of A. pallipes detection using non-invasive eDNA techniques. Using feedback from end-users we can now provide a basis for the commercial development and use of eDNA for crayfish, providing a more accessible detection method which will allow for more citizen science within this field to contribute to a greater conservation effort of the white-clawed crayfish.

 

 

Day: 5, Session: 4, Talk: 3

Crustacyanin Genes of Cambarus Crayfish

PAUL R. CABE, Morgan Trimas, Jeronimo Reyes-Olmedo and Christian Kim

Decapod crustaceans, including crayfish, exhibit a tremendous range of coloration. These various colors are all produced by a combination of diet-derived carotenoid pigments with proteins coded by the crustacyanin genes, a crustacean-specific evolutionary innovation. In general, peptides from two different crustacyanin genes, crustacyanin A and crustacyanin C, combine in multi-unit proteins with carotenoids, shifting the absorption spectra of carotenoids to produce a range of colors. Despite their evolutionary and ecological importance, little is known about these genes in crayfish. We attempted to determine the sequence of these genes in Cambarus longulus and Cambarus bartonii (family Cambaridae) using an RNA-seq approach. RNA was extracted from tissues on the inner surface of the carapace (endocuticle, epithelium, hypodermis) and converted to cDNA. The cDNA pool was sequenced on the Illumina platform, yielding more than 7 million paired-end reads per species, which were assembled into transcripts. These transcripts were searched using crustacyanin sequences from other decapods. The sequence data suggest there are three different crustacyanin A genes in these species which differ primarily in non-coding regions. Using primers designed from these sequences, we were able to amplify these genes directly from genomic DNA, which confirmed the transcript sequences and revealed the presence of short introns. Likewise, the data also suggest both species have several distinct crustacyanin C genes. Direct knowledge of the sequence of these genes opens the possibility for comparative study of these evolutionary important genes in crayfish of the family Cambaridae.


 

Poster Presentations

 

 

POSTER 55

Development of a Captive Rearing Protocol for Threatened & Endangered Appalachian Crayfish

CHRISTOPHER VOPAL, Emmy Delekta and Zachary J. Loughman

In 2016, two Appalachian endemic species were federally listed by the United States Fish & Wildlife Service: Cambarus callainus (Big Sandy Crayfish) and Cambarus veteranus (Guyandotte River Crayfish), which are listed as threatened and endangered respectively. Both species were listed due to limited and declining ranges caused by various anthropogenic activities, especially those causing stream sedimentation. Captive propagation can be used as a tool for crayfish conservation by helping to restore the native range of a species or improving their fecundity within their current range. Over a ten week period, 120 young-of-the-year (YOY) Cambarus chasmodactylus (New River Crayfish), a surrogate species for C. callainus and C. veteranus, were raised in individual cells to compare (1) growth and (2) survival on two different diets. They were fed every other day, with half (60) raised on trout diet pellets (TD) and the other half raised on blood worms (BW). Results showed more YOY growth with BW (17.3% growth) than with TD (13.6% growth). Increased survival was also observed with BW (84.5% survival) when compared to the TD (70.5% survival). Our results may be influenced by the ease and ability for the crayfish to forage on the blood worms and may also have a higher nutritional value in comparison to the TD. Our findings suggests a BW diet may be more effective in the captive rearing of Camabrus crayfish. Using information gathered from this study, a modified protocol will be used for a new study beginning July 2018 for C. callainus and Cambarus smilax (Greenbrier Crayfish). This modified protocol will compare three diets (bloodworm, detritus, and bloodworm/detritus) in the growth and survival of YOY C. callainus and C. smilax in a six month period.

 

 

POSTER 57

Water Quality Analysis and Habitat Threats Concerning Cambarus cracens on Sand Mountain in Northeast Alabama

Rebecca A. Bearden, E. Anne Wynn, Patrick E. O’Neil, STUART W. McGREGOR, Guenter A. Schuster and Christopher A. Taylor

Understanding habitat threats for species of concern is paramount for establishing effective conservation strategies. Although the Slenderclaw Crayfish, Cambarus cracens, was found in the 1970s at five sites in Scarham, Short, and Town creeks on Sand Mountain in northeast Alabama, surveys in 2011 found the species at only a single site in Scarham Creek. Our goals were to determine the current range of this species and identify any water quality issues or habitat threats that may be causing its decline. We conducted status surveys for C. cracens at 71 sites in northeastern Alabama and northwestern Georgia from 2015 to 2017 and collected the species in low abundance at five sites in Scarham and Town creeks. We also collected the invasive Virile Crayfish, Faxonius virilis, in Short Creek, revealing a possible recent threat to the status of C. cracens. Our water quality surveys in Scarham, Short, and Town creeks in 2015 and 2016 revealed elevated levels of ammonia, nitrate and phosphorus, concentrations of lead and zinc that exceeded aquatic life criteria, the presence of pesticides, and concentrations of bacteria that exceeded established limits. Our land use analysis confirmed intense poultry production and high levels of human disturbance in Scarham, Short, and Town creeks. In order to conserve remaining populations of C. cracens, we recommend continued efforts at establishing watershed projects to reduce pollutant loads, promoting best management practices for agriculture, and monitoring future water-quality trends to help assure the integrity of water quality in these tributaries and assist in improving habitat quality throughout the Scarham, Short, and Town Creek watersheds.

 

 

POSTER 58

An Assessment of Cambarus spicatus, Broad River Spiny Crayfish

RILEY W. AULICK and Zachary J. Loughman

The Broad River Spiny crayfish, Cambarus spicatus, is endemic to the Broad River and some of its tributaries. Few life history studies of C. spicatus have caused the IUCN to list it as data deficient. The goal of this study was to determine the impacts of land development on the distribution of C. spicatus. In the summer of the 2017, the West Liberty University Crayfish Conservation Research Lab surveyed the Catawba watershed in North Carolina and the Broad and Saluda watersheds in South Carolina in search of C. spicatus. A standard protocol of ten seine hauls per riffle was implemented in one hundred and twenty-three streams. Dip nets were used in addition to seines to survey the banks of the streams. ArcMap, an application of ArcGIS, was utilized by adding layers such as land cover and a buffer around each collection area which provided land type percentages for each survey site. Six individuals from four sites in North Carolina and one individual from South Carolina were collected out of a total of one-hundred and twenty-three sites. According to the models, C. spicatus was least likely to be found in areas developed for agriculture and urban development. This study provides strong evidence that land development is negatively impacting C. spicatus distribution. Additional studies are needed throughout the species range to make a final determination that land development has a negative impact on C. spicatus.

 

 

POSTER 61

Are Soil Properties Good Predictors in Distribution Modelling for Three European Crayfish?

CONSTAN?A MIHAELA ION, Andrei Dornik and Lucian Pârvulescu

Niche-based species distribution models (SDMs) using different algorithms (like generalized linear model, MaxEnt, random forest) are largely used in various applications for many species of plants and animals. By far, the most used variables for modelling are bioclimatic variables, but also slope, land use, vegetation cover and soil are sometimes considered. Several soil properties are now available at regional and global level, with suitable spatial resolution for SDMs. We therefore aim to increase our understanding on how these soil properties affect spatial distribution of crayfish species. We used a database consisting of 470 presence and absence locations in Romania for three European species of crayfish (Austropotamobius torrentium, Astacus leptodactylus and A. astacus), to extract information from several soil properties layers (grid type at 250 m resolution at seven standard depths) obtained from ISRIC - World Soil Information. Analyzing soil texture, 3000 randomly selected points from the study area were grouped mainly into five soil textural classes (silty clay, clay loam, silty clay loam, loam and silt loam), while crayfish presence points fell only in loam and clay loam. Using SDM we found that soil properties are good predictors for the current distributions of the three investigated crayfish species. For A. leptodactylus, the predicted distribution covers low plains up to the hills, while for A. astacus it ranges from higher plains, over foothills and tablelands and into lower altitude units of the Carpathian Mountains. A. torrentium predicted distribution is clearly restricted to the foothill region. The analysis based on crayfish abundance pointed out that there is a positive response to clay content and soil bulk density, and a negative response to sand content as well as to coarse fragments for all investigated crayfish species. We speculate that the burrows integrity against the erosion along the shorelines might be the explanation of these results, challenging new perspectives in further ecological approaches.

 

 

POSTER 63

Exploring the Limit and Beyond of Hypoxia: Behavioural-driven Conservation of an Ancestral Legacy of Freshwater Crayfish

LUCIAN PÂRVULESCU, Adrian Neculae, Eva Kaslik, Claudia Zaharia, Zanethia Barnett, Marcelo M. Dalosto, James M. Furse, Tadashi Kawai, Sandro Santos and Ovidiu I. Sîrbu

Freshwater crayfish burrowing is not simply sheltering, but an active and conscious behavior in which the animal invests considerable time and energy. As aerobic organisms, crayfish are often recorded as being related to high levels of dissolved oxygen. Approaches considering the in-burrow requirements of oxygen are scarce. We monitored the respiratory behavior and survival under acute hypoxia under controlled conditions in the laboratory of ten ecologically and phylogenetically dissimilar species of crayfish from different geographical locations (5 species of Cambaridae, 3 of Astacidae and 2 of Parastacidae). We found that primary burrowing species (Parastacus brasiliensis and Cambarus striatus) cannot tolerate severe hypoxia, whereas secondary and tertiary burrowing species (Faxonius limosus, F. etnieri, Procambarus vioscai, Cambaroides japonicus, Austropotamobius torrentium, Astacus leptodactylus, A. astacus and Cherax quadricarinatus) were not only able to withstand prolonged anoxia, but also able to remain active for up to 40 hours after reaching zero-oxygen conditions. Using nonlinear regression tools applied to the available experimental data, we estimated the critical values of the dissolved oxygen levels which characterize the transition from aerobic to anaerobic respiration for each species found tolerating the anoxia. Based on the diffusion-convection transport and the experimentally determined oxygen consumption function, we developed a mathematical model describing the time-dependent changes of the dissolved oxygen concentration which takes into account both aerobic and anaerobic respiratory processes for A. leptodactylus and O. limosus in a virtual burrow filled with water. We further validated our models by comparing numerical simulations with laboratory measurements for different geometries of burrows. Excluding a region at the entrance, the mathematical predictions for a normal day-night cycle of a crayfish inside a (virtual) burrow show that the water-dissolved oxygen inside the burrow reaches anoxia levels within hours. We speculate that the ability of crayfish to cope with oxygen shortages might be a phylogenetic legacy from their ancestors, lobsters, known to encounter low levels of oxygen in deep waters. Most probably, the primary burrowing species lost this ability since the oxygen diffusion is much faster in fossorial burrows, and thus leading to weaker conservation of the specific mechanisms during evolution. These results challenge the current behavioral and physiological knowledge of crayfish, and might drive new perspectives on the ecology, conservation and even evolutionary processes.

 

 

POSTER 64

Influence of Climate Warming on the Ecological Impacts of Invasive Crayfishes

VICTORIA CHICATUN and Anthony Ricciardi

Aquatic systems in temperate regions are particularly sensitive to temperature change, which can cause seasonal stress for cold-water adapted species and hospitable conditions for warm-water invaders. Altered thermal regimes may mediate the ecological impacts of non-native species by affecting their abundance and per capita effects, causing shifts in predator-prey dynamics and competitive dominance over native species. High-impact invaders tend to exhibit higher functional responses (maximum feeding rates) than functionally-similar native taxa. It has also been shown that individuals' maximum feeding rates are inversely proportional to the deviation from their environmental optima and could potentially be used a performance metric for invasive species across a thermal gradient. My research investigates the effects of water temperature and population latitude on prey consumption and competitive dominance by invasive (Faxonius rusticus) and native (F. virilis) crayfishes in the Great Lakes basin. For this, I have planned a series of lab experiments comparing functional responses and outcomes of competitive interactions across temperatures (based on projected warming scenarios for the lower Great Lakes) to test the prediction that native species will exhibit lower maximum feeding rates than invaders and that increasing temperatures will result in competitive dominance of southern species over northern species.

 

 

POSTER 65

Analysis of Species-environmental Relationships with Variance Partitioning and Distance-based Moran Eigenvector Maps: Application for Crayfish Distribution and Community Models

WILLIAM R. BUDNICK, Sophia I. Passy and Michael D. Kaller

Advances in numerical ecology have developed robust modeling techniques that can include spatial information in analyses of species-environmental relationships. We demonstrate how variance partitioning and distance-based Moran eigenvector maps (dbMEM) can determine which spatial scales that environmental factors structure crayfish communities and distributions. We sampled 56 streams from 5 major Louisiana river drainages from 2013-2014. Variance partitioning with redundancy analyses of environmental factors and geographic spatial distances produced a poor model fit and great environmental-spatial covariance, which confounded interpretation. However, including orthogonal spatial variables obtained from dbMEM not only improved model fits, but elucidated which environmental variables constrained community composition across spatial scales, namely among drainages (broad scale), within drainages (intermediate scale) and within stream (small scale). Presence of sand, specific conductance, and stream depth were important community drivers across scales, but presence of clay and grassy banks were more locally important. Temperature, a climatic factor, was important at broad scales. Our methods provided valuable insight into the relevant scales of environmental influence on crayfish and it is our hope that we see wider adoption of these methods for future work.

 

 

POSTER 70

Historical and Current Distribution of Western Pennsylvania’s Appalachian Primary Burrowing Crayfishes: A Century of Change or Stasis?

KATIE SCOTT, Zachary W. Dillard, Nicole M. Sadecky, Zachary J. Loughman and David A. Lieb

Astacological efforts in Pennsylvania have increased over the past decade. However, the distribution and conservation standing of Western Pennsylvania (WPA) burrowing crayfish represents the greatest void in knowledge regarding the state’s crayfish fauna. To rectify this situation, burrowing crayfish surveys were initiated across WPA in 2014-2016 using Ortmann’s (1906) historical records as a guide. 61 historic sites were resampled, and 19.6% maintained burrowing crayfish populations. 57 new sites were sampled, of which 71.9% supported burrowing crayfish populations. Overall, burrowing crayfish were detected at 44.9% of the 118 sites sampled. Ortmann documented Cambarus dubius, Cambarus monongalensis, and Cambarus thomai in WPA. All three species were found during this survey, with each taxa allied to a physiographic region. Urbanization has negatively impacted burrowing crayfish over the past century, and greenspaces proved to be important islands of habitation in the presence of urbanization. Comprehensively, our survey results indicate that WPA burrowing crayfish taxa are currently stable.


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