Considerations for fatty acids in standardized reference diet for parthenogenetic marbled crayfish Procambarus virginalis model organism.

Fatty acid accumulation was studied in the parthenogenetic all-female marbled crayfish Procambarus virginalis using six arbitrarily designed experimental feeds and related to individuals with glair glands (sexual maturity) after 100 days of ad libitum feeding at 21 °C, including gravid females from the wild as a reference. Fatty acids 16:0 and 18:1n-9 comprised 40% of the total amount of fatty acids and tended to up-concentrate in bodies. Shorter chain 14:0 depleted from feed to body. Across diets, there was a concomitant decrease in precursor fatty acid and increase in product fatty acid, such as reinforcements in monounsaturated fatty acid (18:1n-9), eicosanoid precursors 20:4n-6 (arachidonic acid, ARA) and 20:5n-3 (eicosapentaenoic acid, EPA) in-vivo, but not 22:6n-3 (docosahexaenoic acid, DHA) except when deficient in CHI or CHI + SPI diets. Saturation kinetics modeling (R2 0.7-0.9, p < 0.05) showed that when the ARA share is ~ 1%, the EPA share is ~ 8%, and the DHA share is ~ 2% in the food lipids, the accumulation of fatty acids in body lipids levels off. The lowest DHA in the CHI (0% glair glands) or CHI + SPI (0-3.9% glair glands) diets, and the lowest ARA in SER (0% glair glands) or SER + SPI (0-3% glair glands) diets, were synchronous with negligible sexual maturity despite a wide range of observed specific growth rates (2.77-3.60% per day), body size (0.44-0.84 g), ≤ 5% crude lipid and 40-46% crude protein feed. The FISH and SHRIMP diets (56% protein, 11-14% lipid) with the highest ARA, EPA, and DHA together seem to be the most conducive diets for sexual maturity (up to 20% of individuals with glair glands). We propose a fatty acid profile mimicking the FISH or SHRIMP diets as a starting point for designing the lipid content required in the marbled crayfish standardized reference diet.

Considerations for protein and amino acids in standardized reference diet for parthenogenetic marbled crayfish Procambarus virginalis model organism.

The concept of a standardized reference diet (SRD) is used in laboratory model organisms to ensure nutritional control between studies and laboratories. Although models using the genetically identical, all female parthenogenetic marbled crayfish (Procambarus virginalis) are growing in popularity, research into nutrition in this species still has many knowledge gaps. To fast track the development of a SRD in terms of protein and amino acids (SRDprotein) for this species, we first analyzed the composition of its body amino acids to determine the ideal protein concept (IPC) of indispensable amino acids in wild-caught P. virginalis (which had an unusually high preponderance of leucine and arginine). Then, we strategically evaluated three common clusters of types of fish feed: (1) ornamental fish feed (SER) fortified with a naturally occurring alga (Spirulina). This type of feed was protein-high in arginine and leucine (SER + SPI) that fulfils the species' IPC for iso-protein (~ 40%), iso-phosphorus (~ 0.8%) and near iso-energetic (~ 475 kcal 100 g-1); (2) freeze-dried live feed consisting of chironomid larvae (CHI) fortified with Spirulina (CHI + SPI) that fulfils the IPC for iso-protein (~ 46%), iso-phosphorus (~ 0.7%) and near iso-energetic (~ 405 kcal 100 g-1); and (3) a commercially standardized 'starter diet' for carnivorous fish larvae (FISH) and post-larval shrimps (SHRIMP) with iso-protein (~ 56%) and iso-phosphorus (~ 1.6%). A total of six diets, embracing a diverse range of proteinaceous feeds, were used in a 100-day ad libitum feeding and growth trial. The FISH group outperformed all the other groups (p < 0.05) and our exploratory multivariate analysis revealed an ideal demand of > 44% protein (tailored to deliver high arginine 3% and leucine 4%, followed by the usual lysine > 3.5% and methionine 1.2%) but also the lowest carbohydrate level (21%). For SRDprotein, our findings show that the FISH diet is ideal and suggest the possibilities of using a CHI + SPI diet for further optimization (more economic use of protein and phosphorus).

Assessing the role of non-native species and artificial water bodies on the trophic and functional niche of Mediterranean freshwater fish communities.

Habitat alterations and the introduction of non-native species have many ecological impacts, including the loss of biodiversity and a deterioration of ecosystem functioning. The effects of these combined stressors on the community trophic web and functional niche are, however, not completely clear. Here, we investigated how artificial ecosystems (i.e. reservoirs) and non-native species may influence the trophic and functional niche space of freshwater fish communities. To do so, we used carbon and nitrogen stable isotope and abundance data to compute a set of isotopic, trait, and functional metrics for 13 fish communities sampled from 12 distinct ecosystems in Türkiye. We show that in reservoirs, fish were more similar in their trophic niche compared to lakes, where the trophic niche was more variable, due to higher habitat complexity. However, there were no differences in the trait and functional metrics between the two ecosystem types, suggesting a higher prey diversity than assumed in reservoirs. We also found that the number of non-native species did not affect the trophic niche space, nor the trait or functional space occupied by the fish community. This indicates that non-native species tended to overlap their trophic niche with native species, while occupying empty functional niches in the recipient community functional space. Similarly, the proportion of non-native species did not affect any trophic, trait, or functional metric, suggesting that changes in community composition were not reflected in changes in the community niche space. Moreover, we found that trait richness, but not functional richness, was positively related to the isotopic niche width and diversity, indicating that a wider occupied trait niche space corresponded with a wider occupied trophic niche and lesser interspecific similarity. Our findings underscore the complexity of ecological relationships within freshwater ecosystems and highlight the need for comprehensive management strategies to mitigate the impacts of human activities and biological invasions.

Biological invasions are a population-level rather than a species-level phenomenon.

Biological invasions pose a rapidly expanding threat to the persistence, functioning and service provisioning of ecosystems globally, and to socio-economic interests. The stages of successful invasions are driven by the same mechanism that underlies adaptive changes across species in general-via natural selection on intraspecific variation in traits that influence survival and reproductive performance (i.e., fitness). Surprisingly, however, the rapid progress in the field of invasion science has resulted in a predominance of species-level approaches (such as deny lists), often irrespective of natural selection theory, local adaptation and other population-level processes that govern successful invasions. To address these issues, we analyse non-native species dynamics at the population level by employing a database of European freshwater macroinvertebrate time series, to investigate spreading speed, abundance dynamics and impact assessments among populations. Our findings reveal substantial variability in spreading speed and abundance trends within and between macroinvertebrate species across biogeographic regions, indicating that levels of invasiveness and impact differ markedly. Discrepancies and inconsistencies among species-level risk screenings and real population-level data were also identified, highlighting the inherent challenges in accurately assessing population-level effects through species-level assessments. In recognition of the importance of population-level assessments, we urge a shift in invasive species management frameworks, which should account for the dynamics of different populations and their environmental context. Adopting an adaptive, region-specific and population-focused approach is imperative, considering the diverse ecological contexts and varying degrees of susceptibility. Such an approach could improve and refine risk assessments while promoting mechanistic understandings of risks and impacts, thereby enabling the development of more effective conservation and management strategies.

Taming the terminological tempest in invasion science.

Standardised terminology in science is important for clarity of interpretation and communication. In invasion science - a dynamic and rapidly evolving discipline - the proliferation of technical terminology has lacked a standardised framework for its development. The result is a convoluted and inconsistent usage of terminology, with various discrepancies in descriptions of damage and interventions. A standardised framework is therefore needed for a clear, universally applicable, and consistent terminology to promote more effective communication across researchers, stakeholders, and policymakers. Inconsistencies in terminology stem from the exponential increase in scientific publications on the patterns and processes of biological invasions authored by experts from various disciplines and countries since the 1990s, as well as publications by legislators and policymakers focusing on practical applications, regulations, and management of resources. Aligning and standardising terminology across stakeholders remains a challenge in invasion science. Here, we review and evaluate the multiple terms used in invasion science (e.g. 'non-native', 'alien', 'invasive' or 'invader', 'exotic', 'non-indigenous', 'naturalised', 'pest') to propose a more simplified and standardised terminology. The streamlined framework we propose and translate into 28 other languages is based on the terms (i) 'non-native', denoting species transported beyond their natural biogeographic range, (ii) 'established non-native', i.e. those non-native species that have established self-sustaining populations in their new location(s) in the wild, and (iii) 'invasive non-native' - populations of established non-native species that have recently spread or are spreading rapidly in their invaded range actively or passively with or without human mediation. We also highlight the importance of conceptualising 'spread' for classifying invasiveness and 'impact' for management. Finally, we propose a protocol for classifying populations based on (i) dispersal mechanism, (ii) species origin, (iii) population status, and (iv) impact. Collectively and without introducing new terminology, the framework that we present aims to facilitate effective communication and collaboration in invasion science and management of non-native species.

Assessing the potential phytosanitary threat of the house cricket Acheta domesticus.

Phytosanitary threats can pose substantial risks to global agriculture and ecological systems, affecting biodiversity, human well-being, and food security. Meanwhile, global warming is projected to exacerbate these threats in the future. One in Europe already widely distributed potential phytosanitary threat that may benefit from global warming is the house cricket Acheta domesticus. This study explored the potential of A. domesticus as a relevant non-native phytosanitary threat under changing climatic conditions by conducting a series of functional response experiments across a temperature gradient (20, 25, and 30 °C). Acheta domesticus exhibited comparable patterns of seed consumption and functional responses. Seed type (millet seeds, wheat grains) and temperature increase influenced the damage inflicted on seeds, with softer and smaller seeds being more susceptible to damage, further amplified by warmer temperatures. The study's outcomes underline the phytosanitary threat that A. domesticus may pose. Considering the species' established presence and adaptable nature in urban environments exacerbates the potential for A. domesticus to transition to rural and agricultural areas. Its increasing production as a food item, paired with the here-identified potential to damage seeds, emphasizes the need for proactive and science-based strategies to address emerging phytosanitary threats driven by non-native species under changing climatic conditions. As global temperatures continue to rise, the assessment and management of potential pest species like A. domesticus will be crucial for safeguarding agriculture productivity and ecological balance.

Diversity and distribution of Aphanomyces astaci in a European hotspot of ornamental crayfish introductions.

Ornamental trade has become an important introduction pathway of non-native aquatic species worldwide. Correspondingly, there has been an alarming increase in the number of established crayfish of aquarium origin in Europe over the previous decade. The oomycete Aphanomyces astaci, the pathogen causing crayfish plague responsible for serious declines of European crayfish populations, is dispersed with introduced North American crayfish. The role of ornamental taxa in introducing and spreading different genotypes of this pathogen in open waters remains unclear. We investigated the distribution, prevalence, and diversity of A. astaci in Budapest, Hungary, which became a hotspot of aquarium crayfish introductions. Their establishment in this area was facilitated by locally abundant thermal waters. We screened for A. astaci in six host taxa from 18 sites sampled between 2018 and 2021: five cambarids (Cambarellus patzcuarensis, Faxonius limosus, Procambarus alleni, P. clarkii, P. virginalis) and one native astacid (Pontastacus leptodactylus). The pathogen was confirmed at five sampled sites in four host taxa: P. virginalis, P. clarkii, F. limosus, and for the first time in European open waters also in P. alleni. Genotyping was successful only in individuals from two different brooks where multiple host species coexisted but revealed unexpected patterns. Mitochondrial B-haplogroup of A. astaci, previously usually reported from Pacifastacus leniusculus or infected European species, was detected in P. virginalis at both sites, and in both F. limosus and P. virginalis sampled from a thermally stable tributary of Barát brook in 2018. In contrast, A-haplogroup of A. astaci was detected in coexisting F. limosus, P. virginalis and P. clarkii sampled in the same watercourse just a few hundred meters downstream in 2020. Additional genotyping methods indicated that a previously unknown A. astaci strain was associated with the latter haplogroup. One P. virginalis individual from 2020 was apparently co-infected by strains representing both mitochondrial haplogroups. The results indicated multiple sources of A. astaci in Budapest, likely directly associated with the introduction of ornamental species, interspecific transmission of this pathogen among ornamental hosts, and potential for a quick spatial or temporal turnover of dominant A. astaci strains at a certain locality. This highlights that in regions with high richness of potential A. astaci hosts, host taxon/pathogen genotype combinations become unpredictable, which might prevent reliable genotyping of pathogen sources in local crayfish mass mortalities.

The wild cost of invasive feral animals worldwide.

Invasive non-native species are a growing burden to economies worldwide. While domesticated animals (i.e. livestock, beasts of burden or pets) have enabled our ways of life and provide sustenance for countless individuals, they may cause substantial impacts when they escape or are released (i.e. become feral) and then become invasive with impacts. We used the InvaCost database to evaluate monetary impacts from species in the Domestic Animal Diversity Information System database. We found a total cost of $141.95 billion from only 18 invasive feral species. Invasive feral livestock incurred the highest costs at $90.03 billion, with pets contributing $50.93 billion and beasts of burden having much lower costs at $0.98 billion. Agriculture was the most affected sector at $80.79 billion, followed by the Environment ($43.44 billion), and Authorities-Stakeholders sectors ($5.52 billion). Damage costs comprised the majority ($124.94 billion), with management and mixed damage-management costs making up the rest ($9.62 and $7.38 billion, respectively). These economic impacts were observed globally, where Oceania, North America and Europe were the most impacted regions. Islands recorded a higher economic burden than continental areas, with livestock species dominating costs more on islands than mainlands compared to other feral species. The costs of invasive feral animals were on average twice higher than those of wild species. The management of invasive feral populations requires higher investment, updated regulations, and comprehensive risk assessments. These are especially complex when considering the potential conflicts arising from interventions with species that have close ties to humans. Effective communication to raise public awareness of the impacts of feral populations and appropriate legislation to prevent or control such invasive feral populations will substantially contribute to minimizing their socioeconomic and environmental impacts.

Toxicological effects of diclofenac on signal crayfish (Pacifastacus leniusculus) as related to weakly acidic and basic water pH.

The widespread use and continuous discharge of pharmaceuticals to environmental waters can lead to potential toxicity to aquatic biota. Pharmaceuticals and their metabolites are often complex organic and environmentally persistent compounds that are bioactive at low doses. This study aimed to investigate the effects of diclofenac (DCF) on the antioxidant defence system and neurotoxicity biomarkers in signal crayfish (Pacifastacus leniusculus) under weakly acidic and basic conditions. Crayfish were exposed to 200 µg/L of DCF at pH 6 and 8 for 96 h and subsequently underwent the depuration phase for 96 h. Gills, hepatopancreas, and muscle were sampled after the exposure and depuration phases to assess the toxicological biomarker responses of DCF in crayfish by evaluating lipid peroxidation (LPO) levels, activities of antioxidant enzymes and acetylcholinesterase. After the exposure phase, the hemolymph DCF concentration was detected one order higher at pH 6 than at pH 8. The DCF was subsequently fully eliminated from the hemolymph during the depuration phase. Our results showed that DCF caused alteration in the activities of six of the seven tested biomarkers in at least one crayfish tissue. Although exposure to DCF caused imbalances in the detoxification system on multiple tissue levels, it was regenerated to a balanced state after the depuration phase. Integrated biomarker response (IBRv2) showed that the highest toxicological response to DCF exposure was elicited in the gills, whereas the hepatopancreas was the highest-responding tissue after the depuration phase. Exposure to DCF at pH 6 caused higher toxicological effects than at pH 8; however, crayfish antioxidant mechanisms recovered more quickly at pH 6 than at pH 8 after the depuration phase. Our results showed that water pH influenced the toxicological effects of DCF, an ionisable compound in crayfish.

The neglect of nonnative orthopterans as potential invaders: A call for awareness.

Despite the potential ecological and economic impacts of invasive species, there is a dearth of data on the presence, impacts, and management implications of potentially invasive Orthoptera species. This lack of research and inconsistent data, including risk screenings and impact assessments, is especially evident in Europe. Consequently, assessing the status, distribution, and potential threats of nonnative Orthoptera in Europe remains challenging, impeding the development of effective management strategies. To address this gap, we call for increased efforts to collect and curate data on non-native and possibly invasive Orthoptera in Europe. Such efforts will improve our understanding of this order's invasion dynamics, facilitate the identification of priority areas for conservation, and support the development of effective management policies and preventive measures.

Protein from whole-body crayfish homogenate may be a high supplier of leucine or branched-chain amino acids - A call for validation on genus Procambarus sp.

Essential proteinogenic branched-chain amino acids (BCAA), particularly leucine (Leu) have been investigated for their role in enhancing human myofibrillar protein synthesis and biomedical research on tumor models. However, only a few protein sources in our current food system have high enough BCAA or Leu coefficients (% of total amino acids) to be considered as supplements for food, sport, or biomedical research. Mostly dairy-sourced proteins such as casein and whey or rarely plant source such as maize gluten are typically regarded as the gold standards. This study hypothesized that protein isolates derived from the whole-body homogenate (including the chitinous exoskeleton) of procambarid crayfish might exhibit unusually high BCAA and Leu content. The study provides open-access data on the amino acid compositions of two procambarid crayfish (Procambarus virginalis and P. clarkii), as well as a comparison with casein. The mentioned crayfish species could offer 6.36-7.39 g Leu 100 g-1 dry matter (at 43-48% protein only). Crayfish whole-body protein isolates exhibit a Leu coefficient (18.41±2.51% of total amino acids) and a BCAA coefficient (28.76±2.39% of total amino acids), which is comparable to or higher than of casein (Leu coefficient 8.65±0.08%; BCAA coefficient 20.03±0.73%). However, it is important to interpret these results with caution, due to the challenges associated with leucine and isoleucine separation, as well as potential interactions within the sample matrices. Hence, international validation of these findings is recommended. NOVELTY STATEMENT: Protein isolates from whole-body homogenate (including chitinous exoskeleton) of P. virginalis and/or P. clarkii are hypothesized to be dense in BCAA and Leu. For potential use in biomedical research or as additives in supplements for BCAA and Leu.

Feeding in predator naïve crayfish is influenced by cues from a fish predator.

In this study, we experimentally evaluated how the feeding behaviour of marbled crayfish Procambarus virginalis is influenced by cues from conspecifics and the round goby Neogobius melanostomus, a fish predator, in tanks that permitted chemical communication but not visual recognition. We used four experimental groups with different combinations in two sub-tanks. The first sub-tank always contained a crayfish and prey (40 individuals of the water louse Asellus aquaticus). The other sub-tanks were set up as follows: (i) empty, serving as a control (C); (ii) with a conspecific crayfish (Cr); (iii) with a round goby (G) to simulate predator-only odour; and (iv) a round goby and three small conspecific crayfish (G + Cr) to simulate the presence of a predator and/or the alarm odour. Two sub-treatments were defined for the fourth group, categorised as 'injured' or 'not injured' depending on whether prey crayfish were visibly injured or not, respectively. We observed a significant decline in the consumption of water lice in the G and G + Cr treatments compared to the C and Cr treatments (up to 47% on average). There were no significant differences in consumption between the G and G + Cr treatments, or C and Cr treatments. No significant differences in food consumption parameters were detected between sub-treatments with 'injured' and 'not injured' conspecific crayfish. Knowledge of modifications in the feeding behaviour of marbled crayfish in the presence of round goby (and fish predators in general) is essential for ecologists attempting to understand the changes and impacts occurring in invaded freshwater ecosystems.

Unveiling the hidden economic toll of biological invasions in the European Union.

Background: Biological invasions threaten the functioning of ecosystems, biodiversity, and human well-being by degrading ecosystem services and eliciting massive economic costs. The European Union has historically been a hub for cultural development and global trade, and thus, has extensive opportunities for the introduction and spread of alien species. While reported costs of biological invasions to some member states have been recently assessed, ongoing knowledge gaps in taxonomic and spatio-temporal data suggest that these costs were considerably underestimated. Results: We used the latest available cost data in InvaCost (v4.1)-the most comprehensive database on the costs of biological invasions-to assess the magnitude of this underestimation within the European Union via projections of current and future invasion costs. We used macroeconomic scaling and temporal modelling approaches to project available cost information over gaps in taxa, space, and time, thereby producing a more complete estimate for the European Union economy. We identified that only 259 out of 13,331 (~ 1%) known invasive alien species have reported costs in the European Union. Using a conservative subset of highly reliable, observed, country-level cost entries from 49 species (totalling US$4.7 billion; 2017 value), combined with the establishment data of alien species within European Union member states, we projected unreported cost data for all member states. Conclusions: Our corrected estimate of observed costs was potentially 501% higher (US$28.0 billion) than currently recorded. Using future projections of current estimates, we also identified a substantial increase in costs and costly species (US$148.2 billion) by 2040. We urge that cost reporting be improved to clarify the economic impacts of greatest concern, concomitant with coordinated international action to prevent and mitigate the impacts of invasive alien species in the European Union and globally. Supplementary Information: The online version contains supplementary material available at 10.1186/s12302-023-00750-3.

Strong temporal variation of consumer δ13C value in an oligotrophic reservoir is related to water level fluctuation.

Using stable carbon and nitrogen isotope analysis (δ13C and δ15N) to assess trophic interactions in freshwater ecosystems is a well established method, providing insight into ecosystem functioning. However, the spatial and temporal variability of isotope values, driven by environmental fluctuation is poorly understood and can complicate interpretations. We investigated how the temporal variation of stable isotopes in consumers (fish, crayfish and macrozoobenthos) of a canyon-shaped oligotrophic reservoir is associated with environmental factors such as water temperature, transparency, flooded area, and water quality measures. Consumers and their putative food sources were sampled and analyzed for carbon and nitrogen stable isotopes annually, and environmental parameters were measured monthly from 2014 to 2016. Results revealed significant differences in δ13C and δ15N values in each consumer among studied years. Over the years, fish and crayfish expressed differences in δ13C between 3 and 5‰, whereas in zoobenthos differences were 12‰. Variability in δ15N was similar across all consumers (2-4‰). Moreover, results suggest that the flooded area of the reservoir was a major driver of δ13C stable isotope values variation in consumers, while variation in δ15N was not linked to any of the studied environmental factors. Bayesian mixing models further showed significant changes in the origin of detritivorous zoobenthos carbon sources (reversal shift from terrestrial detritus to algae origin) between years with low water level to years with the standard water level. Other species showed only slight differences in food source utilization among years. Our study highlights the importance of environmental factors as sources of variation in consumer's stable isotope values which should be considered especially when studied ecosystem strongly fluctuate in some environmental factor.

Alien species and climate change drive shifts in a riverine fish community and trait compositions over 35 years.

Alien fish substantially impact aquatic communities. However, their effects on trait composition remain poorly understood, especially at large spatiotemporal scales. Here, we used long-term biomonitoring data (1984-2018) from 31 fish communities of the Rhine river in Germany to investigate compositional and functional changes over time. Average total community richness increased by 49 %: it was stable until 2004, then declined until 2010, before increasing until 2018. Average abundance decreased by 9 %. Starting from 198 individuals/m2 in 1984 abundance largely declined to 23 individuals/m2 in 2010 (-88 %), and then consequently increased by 678 % up to 180 individuals/m2 until 2018. Increases in abundance and richness starting around 2010 were mainly driven by the establishment of alien species: while alien species represented 5 % of all species and 0.1 % of total individuals in 1993, it increased to 30 % (7 species) and 32 % of individuals in 2018. Concomitant to the increase in alien species, average native species richness and abundance declined by 26 % and 50 % respectively. We identified increases in temperature, precipitation, abundance and richness of alien fish driving compositional changes after 2010. To get more insights on the impacts of alien species on fish communities, we used 12 biological and 13 ecological traits to compute four trait metrics each. Ecological trait dispersion increased before 2010, probably due to diminishing ecologically similar native species. No changes in trait metrics were measured after 2010, albeit relative shares of expressed trait modalities significantly changing. The observed shift in trait modalities suggested the introduction of new species carrying similar and novel trait modalities. Our results revealed significant changes in taxonomic and trait compositions following alien fish introductions and climatic change. To conclude, our analyses show taxonomic and functional changes in the Rhine river over 35 years, likely indicative of future changes in ecosystem services.

Long-term trends in crayfish invasions across European rivers.

Europe has experienced a substantial increase in non-indigenous crayfish species (NICS) since the mid-20th century due to their extensive use in fisheries, aquaculture and, more recently, pet trade. Despite relatively long invasion histories of some NICS and negative impacts on biodiversity and ecosystem functioning, large spatio-temporal analyses of their occurrences are lacking. Here, we used a large freshwater macroinvertebrate database to evaluate what information on NICS can be obtained from widely applied biomonitoring approaches and how usable such data is for descriptions of trends in identified NICS species. We found 160 time-series containing NICS between 1983 and 2019, to infer temporal patterns and environmental drivers of species and region-specific trends. Using a combination of meta-regression and generalized linear models, we found no significant temporal trend for the abundance of any species (Procambarus clarkii, Pacifastacus leniusculus or Faxonius limosus) at the European scale, but identified species-specific predictors of abundances. While analysis of the spatial range expansion of NICS was positive (i.e. increasing spread) in England and negative (significant retreat) in northern Spain, no trend was detected in Hungary and the Dutch-German-Luxembourg region. The average invasion velocity varied among countries, ranging from 30 km/year in England to 90 km/year in Hungary. The average invasion velocity gradually decreased over time in the long term, with declines being fastest in the Dutch-German-Luxembourg region, and much slower in England. Considering that NICS pose a substantial threat to aquatic biodiversity across Europe, our study highlights the utility and importance of collecting high resolution (i.e. annual) biomonitoring data using a sampling protocol that is able to estimate crayfish abundance, enabling a more profound understanding of NICS impacts on biodiversity.

Hemocyte coagulation and phagocytic behavior in early stages of injury in crayfish (Arthropoda: Decapoda) affect their morphology.

Crustacean hemocytes are important mediators of immune functions such as coagulation and phagocytosis. We employed an in situ approach to investigate the ultrastructural behavior of hemocytes during coagulation and phagocytosis in the early stages after injury caused by leg amputation, using transmission electron microscopy technique in marbled crayfish Procambarus virginalis. Hemocytes underwent drastic morphological changes during coagulation. The morphology of the cytoplasmic granules changed from electron-dense to electron-lucent forms in an expanding manner. The transformed granules containing amorphous electron-lucent material were observed to merge and discharge their contents into extracellular space for coagulation. We also observed that the contents of the nucleus participate in the process of coagulation. In addition, leg amputation induced extensive muscle degeneration and necrotic tissues were avidly taken up by the phagocytic hemocytes containing distinct phagosomes. Interestingly, we observed for the first time how the digested contents of phagocytized necrotic tissues are incorporated into granules and other cellular components that change the cell morphology by increasing the granularity of the hemocytes. Nevertheless, the degranulation of hemocytes during coagulation can also reduce their granularity. Given that morphological traits are important criteria for hemocyte classification, these morphological changes that occur during coagulation and phagocytosis must be taken into account.

The pet trade as a source of non-native decapods: the case of crayfish and shrimps in a thermal waterbody in Hungary.

Ornamental aquaculture and the related pet industry are known to be important sources of non-native species worldwide. In the temperate zone, thermal waterbodies are attractive places for irresponsible owners to release unwanted freshwater pets including decapod crustaceans. Several non-native ornamental species have been reported in the thermal locality of Miskolctapolca (a suburb of Miskolc, Hungary). So we surveyed this site in March 2019-November 2021 to update local occurrence records and detect potentially newly released species. A well-established population of Neocaridina denticulata and the occurrence of Caridina cf. babaulti had previously been noted. However, for the first time at this site, we found the shrimps Atyopsis moluccensis, Caridina gracilirostris and C. multidentata, as well as the crayfish Procambarus virginalis, P. clarkii, Cherax quadricarinatus, C. boesemani and C. snowden, and several formally undescribed Cherax species originating from New Guinea. Furthermore, in most species, gravid females carrying eggs were also noticed. Three shrimps, A. moluccensis, C. gracilirostris and C. multidentata, were recorded for the first time in European wild. Further monitoring of this locality and better education of the general public regarding the risks associated with the release of non-native species are strongly recommended.

Integrated biomarker response in signal crayfish Pacifastacus leniusculus exposed to diphenhydramine.

Diphenhydramine (DPH) is a pharmaceutical with multiple modes of action, primarily designed as an antihistamine therapeutic drug. Among antihistamines, DPH is a significant contaminant in the environment, frequently detected in surface waters, sediments, and tissues of aquatic biota. In the present study, signal crayfish Pacifastacus leniusculus was used as a model organism because of their prominent ecological roles in freshwater ecosystems. The biochemical effects were investigated in crayfish exposed to the environmental (low: 2 µg L-1), ten times elevated (medium: 20 µg L-1), and the sublethal (high: 200 µg L-1) nominal concentrations of DPH in water for 96 h. Lipid peroxidation, antioxidant enzyme activities, and acetylcholinesterase activity were assessed as toxicological biomarkers in crayfish hepatopancreas, gills, and muscles. Low and medium DPH exposure caused imbalances only in glutathione-like enzyme activities. Integrated biomarker response showed the absolute DPH toxicity effects on all tested tissues under high exposure. This study identified that high, short-term DPH exposure induced oxidative stress in crayfish on multiple tissue levels, with the most considerable extent in muscles.

Global economic costs of herpetofauna invasions.

Biological invasions by amphibian and reptile species (i.e. herpetofauna) are numerous and widespread, having caused severe impacts on ecosystems, the economy and human health. However, there remains no synthesised assessment of the economic costs of these invasions. Therefore, using the most comprehensive database on the economic costs of invasive alien species worldwide (InvaCost), we analyse the costs caused by invasive alien herpetofauna according to taxonomic, geographic, sectoral and temporal dimensions, as well as the types of these costs. The cost of invasive herpetofauna totaled at 17.0 billion US$ between 1986 and 2020, divided split into 6.3 billion US$ for amphibians, 10.4 billion US$ for reptiles and 334 million US$ for mixed classes. However, these costs were associated predominantly with only two species (brown tree snake Boiga irregularis and American bullfrog Lithobates catesbeianus), with 10.3 and 6.0 billion US$ in costs, respectively. Costs for the remaining 19 reported species were relatively minor (< 0.6 billion US$), and they were entirely unavailable for over 94% of known invasive herpetofauna worldwide. Also, costs were positively correlated with research effort, suggesting research biases towards well-known taxa. So far, costs have been dominated by predictions and extrapolations (79%), and thus empirical observations for impact were relatively scarce. The activity sector most affected by amphibians was authorities-stakeholders through management (> 99%), while for reptiles, impacts were reported mostly through damages to mixed sectors (65%). Geographically, Oceania and Pacific Islands recorded 63% of total costs, followed by Europe (35%) and North America (2%). Cost reports have generally increased over time but peaked between 2011 and 2015 for amphibians and 2006 to 2010 for reptiles. A greater effort in studying the costs of invasive herpetofauna is necessary for a more complete understanding of invasion impacts of these species. We emphasise the need for greater control and prevention policies concerning the spread of current and future invasive herpetofauna.