Pacifastacus Leniusculus Research Articles

Evaluating the Effectiveness of Sodium Hypochlorite for Genomic DNA Decontamination

ABSTRACTEnvironmental DNA (eDNA) is an increasingly popular, sensitive, and cost‐efficient method for studying biodiversity and detecting species. This noninvasive approach involves collecting environmental samples that contain genetic material shed by organisms into their surroundings. Due to the method's sensitivity, robust decontamination strategies are crucial, with sodium hypochlorite, commonly known as bleach, frequently employed. Despite its widespread use, there is no consensus on the most effective bleach concentration, leading to inconsistencies in how the chemical is used in research. This study aimed to determine the minimum concentration of bleach needed for effective decontamination. Genomic DNA of signal crayfish was treated with various concentrations of bleach, ranging from 0.01% to 5% (w/w). Results were observed using Qubit High Sensitivity reagents, quantitative PCR, agarose gel electrophoresis, and the Agilent TapeStation. Our results indicate that a minimum concentration of 0.5% (w/w) bleach is sufficient to prevent the detection of genomic DNA by the techniques tested. These results provide important insights into the use of bleach for decontamination in eDNA research. Establishing a standard bleach concentration for decontamination protocols will help to reduce inconsistencies and enhance the reliability of eDNA studies.

English

Abstract Pacifastacus leniusculus (Dana 1852), the signal crayfish, is a freshwater astacid native to the Pacific Northwest and introduced in western Europe and Japan. Though P. leniusculus populations have been well-studied abroad, where they degrade local habitat and biodiversity, their ecology and natural history are less understood within their native range, where they fill key roles and risk out-competition by exotic crayfish. The present study describes density, morphology, and behavior of resident crayfish in a fourthorder Oregon stream, Tryon Creek, via surveys of nine sites from June–September 2022. Across sites, there was an estimated average P. leniusculus population density of 0.08 m-2, and no exotic crayfish were detected. Amongst crayfish collected (n = 162), total body length (TL) ranged from 13–90 mm, μ = 53 mm. Branchiobdellidans colonized one in four crayfish throughout Tryon Creek, damage to appendages occurred at a rate of 0.45, and the likelihood of both increased with TL. In a case study of Tryon’s Hwy- 43 Culvert, crayfish showed potential signs of stress; across numerous sites, young juveniles resided gregariously. Despite the present study’s novel strides toward native-range P. leniusculus population characterization, further research is encouraged to establish a well-rounded regional baseline.

PHARMACOKINETICS OF CEFTAZIDIME IN SIGNAL CRAYFISH (PACIFASTACUS LENIUSCULUS) FOLLOWING A SINGLE DOSE.

Crustaceans are housed in zoos and aquariums and have also gained importance in the private sector and food industry. Shell lesions are common and often attributed to bacterial infections. However, few controlled studies have been performed evaluating antibiotics in crustaceans. This study assessed the pharmacokinetics of a single dose of ceftazidime (22 mg/kg) given by IM or IV injection in wild-caught signal crayfish (Pacifastacus leniusculus). Pharmacokinetic parameters were calculated by noncompartmental analysis of sparse data. Maximum ceftazidime hemolymph concentration following IM administration was 124.6 ± 14.7 µg/mL and Tmax was 5 min, with 80% bioavailability. Following IV administration, the extrapolated maximum concentration of ceftazidime, (C [0]), was 581.4 µg/mL. Ceftazidime was last detected at 72 hr and 120 hr post IM and IV administration, respectively. Terminal half-life was 8.03 hr and 10.3 hr after IM and IV administration, respectively. Results suggest that both routes have the capacity to reach a maximum hemolymph concentration quickly in signal crayfish. Moreover, ceftazidime was maintained above a concentration of 4 µg/mL, a published minimal inhibitory concentration for Vibrio spp., for 24 hr for both IM and IV routes. Therefore, ceftazidime may be useful for infections with susceptible Vibrio spp. in signal crayfish.

Invasive dynamics of the signal crayfish Pacifastacus leniusculus in a protected area

Invasive species have been drivers of biodiversity loss and functional changes in aquatic ecosystems, including in protected areas. Therefore, monitoring population invasion dynamics and biological traits is fundamental to better understand their ecological and economic impacts and for management actions development. We followed signal crayfish (Pacifastacus leniusculus) invasion in Rabaçal River upper reach at Montesinho Natural Park, Portugal. We collected information on the spread and biological traits (abundance, size, weight, physical condition, sex ratio, and aggressiveness) to assess differences between invasion core and front areas and among years. Signal crayfish population remained restricted since first reports in 2013 in the invasion core until 2017. After 2019, signal crayfish population has been spreading downstream, decreasing abundance at invasion core but increasing at invasion front. Significant higher number of crayfish with claw loss indicate potential higher signs of aggressiveness in the invasion front. Results also demonstrate a significant dominance of females although sex ratio is closer to 1:1 at the invasion front. Overall, results indicate signal crayfish is spreading and increasing their abundance at Rabaçal River highlighting the need for immediate management actions to hold dispersion and mitigate possible impacts.

Insect responses to seasonal time constraints under global change are facilitated by warming and counteracted by invasive alien predators.

In seasonal environments, organisms with complex life cycles not only contend with seasonal time constraints (TC) but also increasingly face global change stressors that may interfere with responses to TC. Here, we tested how warming and predator stress imposed during the egg and larval stages shaped life history and behavioural responses to TC in the temperate damselfly Ischnura elegans. Eggs from early and late clutches in the season were subjected to ambient and 4°C warming temperature and the presence or absence of predator cues from perch and signal crayfish. After hatching, larvae were retained at the same thermal regime, and the predator treatment was continued or not up to emergence. The late eggs decreased their development time, especially under warming and when not exposed to predator cues. However, the late eggs increased their development time when exposed to predator cues, especially to crayfish cues. The TC decreased survival of late larvae that were as eggs exposed to crayfish cues, indicating a carry-over effect. The TC and warming additively reduced late larvae development time to emergence. Independent of the TC, predator cue effects on development time were stronger during the egg than during the larval stage. The late individuals expressed lower mass at emergence, which mirrored the size difference between field-collected mothers. Warming caused a higher mass at emergence. The late individuals increased their boldness and showed a higher number of moves, whereas warming caused a decreased boldness. There was no predator cue effect on larval behaviour. The results indicate that late individuals compensate for late season egg laying, which is facilitated under warming but counteracted under predation risk, especially when imposed by the crayfish.

Osmoregulatory Capacity and Non-Specific Food Preferences as Strengths Contributing to the Invasive Success of the Signal Crayfish Pacifastacus leniusculus: Management Implications

Osmoregulatory Capacity and Non-Specific Food Preferences as Strengths Contributing to the Invasive Success of the Signal Crayfish Pacifastacus leniusculus: Management Implications

Escape rooms: behavioural response of two invasive crayfish species under water decline scenarios

Climate change and invasive alien species pose a significant threat to biodiversity and the survival of native species. This study considered the impact of drought conditions on the escape behaviour through terrestrial dispersal ability of two invasive freshwater crayfish species, the red swamp crayfish (Procambarus clarkii) and signal crayfish (Pacifastacus leniusculus). Using an experimental design simulating drought conditions and Mediterranean summer temperatures, we tested the hypothesis that P. clarkii exhibits a greater tendency to terrestrial migration and a higher land-walking speed than the P. leniusculus. The results indicated that both species demonstrated strong escape behaviour, with P. clarkii showing escape success higher than P. leniusculus, and increasing with decreasing mean night temperature and crayfish weight. Although drought conditions did not trigger escape, invasive species showed increased resistance and the ability to move to more favourable environments, suggesting that drought is not an effective geographic barrier against the spread of these species. These results underline the importance of considering invasive species' ability to escape and climb as part of management and control strategies.

An i-type lysozyme from a crustacean, Pacifastacus leniusculus, functions as a clot-destabilising enzyme

Lysozymes are hydrolytic enzymes, and they are ubiquitous among all living organisms. They are mostly associated with antibacterial properties through their muramidase activity, while other properties such as iso-peptidase activity are also common. Invertebrate-type (i-type) lysozymes include the enzyme Destabilase, which is present in the salivary secretions of the medicinal leach Hirundo medicinalis. Destabilase has the ability to hydrolyse the ε-(γ-glutamyl)-lysine iso-peptide bonds formed by transglutaminase in fibrin of vertebrate blood, thereby destabilising blood clots.We have identified an i-type lysozyme from the hemocytes of the freshwater crayfish Pacifastacus leniusculus, which was found to be upregulated at the protein level in response to an injection of the β-1,3-glucan laminarin. Based on its sequence we predicted that this lysozyme would lack muramidase activity, and therefore we decided to determine its putative immune function. The P. leniusculus i-type lysozyme (Pl-ilys), is a protein with 159 amino acid residues, including a 29 residue signal peptide, with a predicted molecular weight of 16 kDa and a predicted pI of 5.6. It is expressed primarily in the hemocytes and to a lesser extent in the hematopoietic tissue. A recombinant mature Pl-ilys using an E. coli expression system was produced, and we could ascertain that this enzyme was deficient of muramidase activity. Moreover, no iso-peptidase activity could be detected against the substrate l-γ-glutamine-p-nitroanilide. Analysis of the conserved domains in Pl-ilys showed a putative destabilase domain, and thus we tested the clot dissolving activity of this enzyme. We could show that the purified P. leniusculus clotting protein which had been coagulated and clotted with transglutaminase was dissolved by the addition of Pl-ilys. Taken together our results indicate that Pl-ilys has a clot dissolving or destabilising activity in crustacean blood.

Prevalence of the crayfish plague pathogen in red swamp crayfish populations in western France: How serious is the risk for the native white-clawed crayfish?

The crayfish plague pathogen Aphanomyces astaci has been implicated in a number of mass mortalities and irreversible population declines of native crayfish across Europe. At present, the reservoirs of the pathogen in Europe are mainly populations of invasive North American crayfish species. In southwestern Europe, including France, a particularly widespread invader is the red swamp crayfish Procambarus clarkii. Recent distribution data confirm that P. clarkii is present in at least 75 French departments, i.e. more than 78% of those in metropolitan France. We analysed the prevalence and pathogen load of A. astaci in 42 populations of this species in western France (Nouvelle Aquitaine region), where the species is most densely distributed, particularly in a wide range of environments around the Gironde estuary. The pathogen was detected by two different quantitative PCR assays in more than three quarters of the populations studied (34 out of 42); 163 out of 480 analysed crayfish individuals tested positive for the presence of A. astaci. In most cases, individual infection levels were very low, detectable with quantitative PCR but not sufficient for pathogen genotyping. In seven P. clarkii individuals from four populations, however, we were able to assess A. astaci variation by microsatellite markers and sequencing of mitochondrial markers. All these host specimens carried A. astaci genotype group D, haplotype d1, which has caused the majority of crayfish plague outbreaks in neighbouring Spain. In contrast, the French outbreaks genotyped to date (including eight newly analysed in this study) were mostly caused by strains of genotype group B, specific to the signal crayfish Pacifastacus leniusculus. Haplotype d1 found in P. clarkii was involved in one of the newly characterised outbreaks. Our study confirms that P. clarkii is a potentially important reservoir of the crayfish plague pathogen in France, but not the main source of the pathogen in mass mortalities of A. pallipes, probably due to different ecological requirements of the different invasive host crayfish. However, as P. clarkii continues to spread, the threat posed by this species to native crayfish is likely to increase.

Microcystins in the benthic food-web of the Sacramento-San Joaquin Delta, California

Harmful cyanobacteria blooms are a growing threat in estuarine waters as upstream blooms are exported into coastal environments. Cyanobacteria can produce potent toxins, one of which—hepatotoxic microcystins (MCs)—can persist and accumulate within the food web. Filter-feeding invertebrates may biomagnify toxins up to 100× ambient concentrations. As such, bivalves can be used as an environmentally relevant and highly sensitive sentinel for MC monitoring. To date there has been little research on cyanotoxin bioaccumulation in estuaries. The Sacramento-San Joaquin Delta (Delta) aquatic food web has undergone a profound change in response to widespread colonization of aquatic invasive species such as Asian clams (Corbicula fluminea) in the freshwater portion of the Delta. These clams are prolific—blanketing areas of the Delta at densities up to 1000 clams/m2 and are directly implicated in the pelagic organism decline of threatened and endangered fishes. We hypothesized that Asian clams accumulate MCs which may act as an additional stressor to the food web and MCs would seasonally be in exceedance of public health advisory levels. MCs accumulation in Delta Asian clams and signal crayfish (Pacifastacus leniusculus) were studied over a two-year period. ELISA and LC-MS analytical methods were used to measure free and protein-bound MCs in clam and crayfish tissues. We describe an improved MC extraction method for use when analyzing these taxa by LC-MS. MCs were found to accumulate in Asian clams across all months and at all study sites, with seasonal maxima occurring during the summer. Although MC concentrations rarely exceeded public health advisory levels, the persistence of MCs year-round still poses a chronic risk to consumers. Crayfish at times also accumulated high concentrations of MCs. Our results highlight the utility of shellfish as sentinel organisms for monitoring in estuarine areas.

Environmental DNA can inform the trade‐off between proactive and reactive strategies for crayfish conservation

AbstractThe introduction of the signal crayfish Pacifastacus leniusculus to British rivers has led to ecological degradation and the decline of the native white‐clawed crayfish Austropotamobius pallipes. To manage and mitigate the impact of the signal crayfish, conservation agencies and government bodies employ multiple conservation strategies. These take the form of proactive native crayfish breeding and stocking programs and reactive invasive crayfish control programs. Here, we used eDNA to assess the populations of native and invasive crayfish species across 50 sites in 10 river catchments in Norfolk, United Kingdom (UK). The sites were chosen to enable assessment of the potential of eDNA to inform proactive and reactive crayfish conservation strategies. Three of the catchments sampled were selected to assess the success of recent A. pallipes reintroduction, whereas the remaining seven were selected to better understand the distribution of each species at the landscape scale. Combining results of eDNA‐based methods with net searches within an occupancy model enabled us to confidently determine the presence of P. leniusculus at eight sites, and A. pallipes at three sites, which was more than visual searches alone (five and two study sites, respectively). Neither eDNA nor net searches detected A. pallipes at sites where A. pallipes had been reintroduced. We recommend that practitioners using eDNA‐based surveys for management and conservation of crayfish should consider: (1) designing eDNA surveys with an emphasis on large spatial scales to comprehensively describe the distributions of native and invasive crayfish in a region of interest; (2) work with local conservation organizations and/or government bodies to inform the selection of study sites to generate results that are meaningful to real‐world conservation actions; and (3) use results from eDNA‐based crayfish surveys to target limited conservation resources to appropriate proactive and/or reactive conservation actions.

Human Consumption of Non-Native Species in a Circular Economy: Determination of Persistent Organic Pollutants in the Invasive Signal Crayfish from a Baltic Coastal River and Its Assessment for Consumption

A circular economy aims at decoupling value creation from waste generation and resource use. The signal crayfish Pacifastacus leniusculus is kept worldwide in aquaculture and after escaping into the wild, may further be used for human consumption rather than eradicated and used for purposes such as fertilizing fields. The level of contamination by two groups of persistent organic pollutants (POPs), polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), in non-native signal crayfish from a Baltic coastal river, was measured in two locations (under anthropogenic pressure and semi-natural pressure) to understand whether its consumption might be a threat to human health. Concentrations of indicators and total compounds of POPs in the edible parts of crayfish (muscular tissue of crayfish) with potential human health risks were determined. The levels of concentrations of POPs meet the requirements of Regulations (EU) No. 836/2011 and No. 1259/2011 for the consumption of crustaceans. Our results also indicate no significant public health risk caused by consumption of the signal crayfish (hazard quotients (HQ) < 1). The results show that the bioaccumulation of POPs depends on the species’ traits and environment.

Accounting for the power of nature: Using flume and field studies to compare the capacities of bio‐energy and fluvial energy to move surficial gravels

AbstractRiver channels, riparian and floodplain forms and dynamics are all influenced strongly by biological processes. However, the influence of macroinvertebrates on entrainment and transport of river sediments remains poorly understood. We use an energy‐based approach to explore the capacity of benthic animals to move surficial, gravel‐bed particles in field and laboratory settings and use the results to assess the relative significance of biological and physical benthic processes. Our results showed that in 11 British gravel‐bed rivers, the maximum energy content (i.e., calorific content) of macroinvertebrate communities generally matched the flow energy associated with median discharges and, at multiple sites, exceeded that of the 10‐year return interval flood. A series of laboratory experiments used to estimate the minimum energy expended by signal crayfish (Pacifastacus leniusculus) when performing geomorphic work established that crayfish move gravel particles at energy levels below that expected of the flow, complicating direct comparisons of the capacity for macroinvertebrates and fluvial flows to influence bed mobility. Our findings suggest that the influence of macroinvertebrate communities in either promoting or suppressing, the mobilisation of the bed may be large compared to equivalent values of fluvial energy. Based on these findings, we conclude that in the gravel‐bed rivers studied, the macroinvertebrate community's potential to perform geomorphic work matches or exceeds the stream power during most of the year. Although our study examined biological and fluvial energy systems separately, it is important to recognise that in nature, these systems are highly interactive. It follows that utilising the energy framework presented in this paper could lead to rapid advances in both fluvial biogeomorphology and river management and restoration.

Plastic Debris in the Stomach of the Invasive Signal Crayfish Pacifastacus leniusculus from a Baltic Coastal River

The signal crayfish Pacifastacus leniusculus is one of the most widespread non-indigenous crayfish in Europe and is of interest to aquaculture in many countries. Once they escape into the wild, they disperse and become a potential source of food exploited for consumption by local inhabitants. The ingestion of plastics by the invasive signal crayfish in the Wieprza River, a tributary to the Baltic Sea, was determined by the FTIR identification of plastic compounds found in stomachs. The occurrence of plastic debris in the stomachs of P. leniusculus is reported for the first time. Plastic particles with the size range between 70 and 450 µm were observed only among crayfish inhabiting the lower, urbanized part of the river (7.3% of specimens). The presence of PTFE (polytetrafluoroethylene), cellophane, PP (polypropylene), PE (polyethylene), PMMA (polymethyl methacrylate), and nylon was confirmed with the highest frequency of PTFE in the stomachs of crayfish. Fibres were the predominant type of microplastics in crayfish stomachs. The results indicated the size-selective uptake and ingestion of plastics depending on the traits of the species and environment.

Microgeographic differentiation in thermal and antipredator responses and their carry-over effects across life stages in a damselfly.

Global warming and invasive species, separately or combined, can impose a large impact on the condition of native species. However, we know relatively little about how these two factors, individually and in combination, shape phenotypes in ectotherms across life stages and how this can differ between populations. We investigated the non-consumptive predator effects (NCEs) imposed by native (perch) and invasive (signal crayfish) predators experienced only during the egg stage or during both the egg and larval stages in combination with warming on adult life history traits of the damselfly Ischnura elegans. To explore microgeographic differentiation, we compared two nearby populations differing in thermal conditions and predator history. In the absence of predator cues, warming positively affected damselfly survival, possibly because the warmer temperature was closer to the optimal temperature. In the presence of predator cues, warming decreased survival, indicating a synergistic effect of these two variables on survival. In one population, predator cues from perch led to increased survival, especially under the current temperature, likely because of predator stress acclimation phenomena. While warming decreased, predator cues increased larval development time with a proportionally stronger effect of signal crayfish cues experienced during the egg stage, indicating a negative carry-over effect from egg to larva. Warming and predator cues increased mass at emergence, with the predator effect driven mainly by exposure to signal crayfish cues during the egg stage, indicating a positive carry-over effect from egg to adult. Notably, warming and predator effects were not consistent across the two studied populations, suggesting a phenotypic signal of adaptation at a microgeographic scale to thermal conditions and predator history. We also observed pronounced shifts during ontogeny from synergistic (egg and early larval stage) toward additive (late larval stage up to emergence) effects between warming and predator stress. The results point out that population- and life-stage-specific responses in life-history traits to NCEs are needed to predict fitness consequences of exposure to native and invasive predators and warming in prey at a microgeographic scale.

A reevaluation and redescription of the branchiobdellidan Sathodrilus tetrodonta (Pierantoni, 1906) new comb. (Annelida: Clitellata).

The questionable status of Branchiobdella tetrodonta Pierantoni, 1906 is resolved and the species is transferred to the correct genus. The original description was made from specimens removed from signal crayfish collected in California, USA, unfortunately, Pierantoni (1906c) did not designate any type specimens nor where the preparations were deposited; they are now presumed lost. Holt (1967) believed B. tetrodonta possessed unique penial hooks and a chitinous sheath which was due to his mistranslation of the original Italian description. As a result, Sathodrilus attenuatus Holt, 1981, was described even though specimens came from the same area and host and possessed very similar jaw characteristics to B. tetrodonta. In addition, the jaw characteristics of both endemic species are unique in the Pacific Ocean drainage of the USA. A reassessment of the literature and re-examination of Holts type specimens has resulted in the species name becoming a new combination, Sathodrilus tetrodonta (Pierantoni, 1906), with Sathodrilus attenuatus Holt, 1981, as its junior synonym.

Host shift to non-native species or ecological dead end? Endangered branchiobdellidans (Annelida: Clitellata) found on introduced Signal Crayfish in Japan

Symbionts, including parasites, are usually small and thus have been overlooked for their existence and scientific research. In the context of biological invasion, host shifts between native and non-native species occur. Since symbionts often affect host behaviour and survival, it is important to reveal the cryptic invasions and host shifts of such small symbionts. Here, we report the first evidence of the potential host shift of a native endangered branchiobdellidans, ectosymbiotic crayfish worms, to a non-native Signal Crayfish, Pacifastacus leniusculus (Dana 1852), in Hokkaido, Japan. Pacifastacus leniusculus is native to North America and was recently introduced to the Atsubetsu River system in Sapporo. In addition to the North American ectosymbiont, Sathodrilus tetrodonta (Pierantoni, 1906), we found an endangered Japanese species, Cirrodrilus cirratus complex on P. leniusculus. However, C. cirratus complex as well as S. tetrodonta were not found on the native Japanese Crayfish, Cambaroides japonicus (De Haan, 1841), in the same river system, indicating a host shift of the ectosymbiont to invasive crayfish. It remains unclear whether C. cirratus complex will persist for a long time with the non-native host or if this is only a transient leading to local extinction.

Long-term decline of whitefish (Coregonus lavaretus) population in the boreal lake Pyhäjärvi, southwest Finland, relative to simultaneous abiotic and biotic changes

In recent decades many lake fish populations have faced complex pressures particularly due to climate warming and eutrophication. In Lake Pyhäjärvi, southwest Finland, profound changes have occurred in the commercial winter fisheries targeting the coregonids vendace, Coregonus albula, and whitefish, Coregonus lavaretus. The responses of vendace have been examined earlier, and here the detailed analyses are extended to the whitefish. In the 1970s and 1980s, the coregonid fishery was sustainable, but in 1991–1998, intense exploitation led to recruitment overfishing of vendace. Despite temporarily improved growth and recruitment of whitefish during vendace decline, whitefish population biomass declined throughout the study period, while perch (Perca fluviatilis), roach (Rutilus rutilus) and ruffe (Gymnocephalus cernuus) increased and vendace and smelt (Osmerus eperlanus) maintained their abundance. As many other northern lakes, Pyhäjärvi seems to be changing from coregonid to perch dominance due to climate warming and eutrophication. Whitefish decline was associated with decreases in fecundity, winter survival of embryos, and larval density. The spawning stock was kept small by intensive fishery. The decline of the relative fecundity was likely due to food competition with vendace and especially the perch and roach populations that increased with eutrophication and more frequent hot summers. The winter survival of whitefish embryos decreased when the introduced signal crayfish (Pacifastacus leniusculus) increased. Despite decreasing fishing effort in the 2000s the larval numbers continued to decline. In the prevailing circumstances in Pyhäjärvi, whitefish will with high probability remain as a minor and still declining species of the fish assemblage.

A new alternative technique for sterilising invasive crayfish: removing female pleopods did not alter courtship pheromone release in signal crayfish

A new alternative technique for sterilising invasive crayfish: removing female pleopods did not alter courtship pheromone release in signal crayfish

Host-related traits influence the microbial diversity of the invasive signal crayfish Pacifastacus leniusculus

The microbiome influences a variety of host-environment interactions, and there is mounting evidence of its significant role in biological invasions. During invasion, shifts in microbial diversity and function can occur due to both changing characteristics of the novel environment and physiological condition of the host. The signal crayfish (Pacifastacus leniusculus) is one of the most successful crayfish invaders in Europe. During range expansion, its populations often exhibit differences in many traits along the invasion range, including sex-composition, size-structure and aggressiveness, but to date it was not studied whether crayfish traits can also drive changes in the host microbiome. Thus, we used 16S rRNA gene amplicon sequencing to examine the effects of host-related traits, namely total length (TL), body condition index (FCF), hepatosomatic index (HSI) and sex on the microbial diversity of the signal crayfish. We examined both external (exoskeletal) and internal (intestinal, hepatopancreatic, hemolymph) microbiomes of 110 signal crayfish individuals from four sites along its invasion range in the Korana River, Croatia. While sex did not exhibit a significant effect on the microbial diversity in any of the examined tissues, exoskeletal, intestinal and hemolymph microbial diversity significantly decreased with increasing crayfish size. Additionally, significant effects of signal crayfish condition (FCF, HSI) on microbial diversity were recorded in the hepatopancreas, a main energy storage organ in crayfish that supports reproduction and growth and also regulates immune response. Our findings provide a baseline for evaluating the contribution of microbiome to an invader's overall health, fitness and subsequent invasion success.