A Comparison of Trace Metal Accumulation in Indigenous and Alien Freshwater Macro-Decapods

Concentrations of Cd, Cu, Fe, Mg, Mn, Ni, Pb and Zn were determined in the exoskeleton and hepatopancreas of three freshwater macro-decapods widely distributed in Tuscany, Italy. The species were the indigenous Austropotamobius pallipes (the white-clawed crayfish) and Potamon fluviatile (the river crab) and the alien Procambarus clarkii (the red swamp crayfish, in Italy aquacultured since 1977 and invasive since 1989). Water and sediments from the three environments in which they occur were analysed for trace metals to evaluate the degree of contamination of the habitats. Procambarus clarkii showed a significantly higher accumulation of most of the metals analysed, including the non-essential cadmium, nickel and lead. Interspecific differences were not related to exposure to higher concentrations of metals, but indigenous and alien macro-decapods possibly differ in their physiological requirements for essential trace metals and/or in their rates of essential and non-essential metal uptake, accumulation, detoxification and excretion.     

Early assessment of the impact of alien species: differential consequences of an invasive crayfish on adult and larval amphibians

Aim Early assessment of the impact of invasive alien species is crucial to set up timely management, but often the impact is evident when it is too late for action. We evaluated relationships between the alien crayfish, Procambarus clarkii, the distribution of native amphibians, and the abundance of their larvae. We assessed whether considering measures of reproductive success provide a more prompt measure of impact than considering just species distribution. Location One hundred and twenty‐five wetlands in Northern Italy, in an area recently invaded by P. clarkii. Methods We surveyed wetlands to assess the presence of breeding activity of amphibians and the distribution of P. clarkii. We measured the abundance of amphibian larvae before metamorphosis through pipe sampling. We built models analysing the relationships between amphibian and crayfish distribution, while taking into account spatial autocorrelation and environmental features. Analyses were performed at both the species level (generalized linear models and spatial eigenvector mapping) and community level (constrained redundancy analysis). Results In terms of breeding site distribution, only two amphibians (Lissotriton vulgaris and Hyla intermedia) were negatively associated with P. clarkii, while the relationships between other the species and P. clarkii were positive or not significant. However, larval abundance for all amphibian species was negatively associated with the alien crayfish. Analyses performed at community and single species levels yielded consistent results. Main conclusions Procambarus clarkii impacts amphibians through different processes. Newts probably avoid invaded wetlands for breeding. Other species attempt breeding in wetlands with crayfish, but suffer very low success. Considering distribution data alone would not provide a correct picture of the impact of this alien species; measures of reproductive success may allow a more accurate assessment of the impact.     

The North American crayfish <Emphasis Type="Italic">Procambarus clarkii</Emphasis> is the carrier of the oomycete <Emphasis Type="Italic">Aphanomyces astaci</Emphasis> in Italy

Aphanomyces astaci (Saprolegniales, Oomycetes) is classified among the 100 world’s worst invasive species. This species is endemic to North America and has been introduced into Europe by imports of their hosts, the North American crayfish species. As a consequence, extensive mass mortalities involved several populations of the European crayfish. Here, we checked its occurrence in four Italian populations of Procambarus clarkii, the most widespread alien crayfish in Italy. Digital image analyses and image processing techniques were used to select micro-melanized areas in the subabdominal cuticle of 2–10 crayfish per population. All the selected areas tested positive for A. astaci ITS nrDNA specific primers; moreover, the obtained sequences clearly corresponded to A. astaci, thus revealing that P. clarkii is an active carrier of this oomycete in Italy. Decisions are to be urgently made to control the spread of both P. clarkii and A. astaci for the conservation of the indigenous crayfish biodiversity.     

Physiological biomarkers of hypoxic stress in red swamp crayfish Procambarus clarkii from field and laboratory experiments

The crayfish industry in Louisiana is the largest in the United States, with crayfish frequently harvested from waters that experience episodic or chronic hypoxia (dissolved oxygen [DO]≤ 2 mg/l). We examined physiological biomarkers (hemolymph lactate, glucose, and protein concentrations) of hypoxic stress in the red swamp crayfish Procambarus clarkii from chronically hypoxic natural habitats and laboratory hypoxia experiments. P. clarkii from normoxic and hypoxic areas in the Atchafalaya River Basin were sampled monthly from April to July 2010. Laboratory experiments subjected P. clarkii to severe hypoxia (1 mg/l DO), moderate hypoxia (2 mg/l DO), or normoxic conditions (control: DO>7.5 mg/l) for 12, 24, and 48 h. P. clarkii from normoxic and hypoxic natural habitats did not display significantly different hemolymph lactate or glucose concentrations; however, mean hemolymph protein concentration was significantly lower in crayfish from hypoxic areas. P. clarkii exposed to severe hypoxia in laboratory experiments had significantly higher hemolymph lactate and glucose concentrations for all three exposure times, whereas large differences in protein concentrations were not observed. These results suggest that elevated hemolymph lactate and glucose concentrations are responses to acute hypoxia in P. clarkii, while differences in protein concentrations are the result of chronic hypoxic exposure.     

Pike and red swamp crayfish: a new case on predator-prey relationship between aliens in central Spain

Pike in the Ruidera Lakes (central Spain) chiefly ate the recently introduced crayfish Procambarus clarkii . It was the dominant prey in occurrence, number and biomass for every size class and season. Likewise, number (up to 27) and total weight of ingested crayfish were directly related with pike size. The principal prey fish were all introduced species (except the endangered Blennius fluviatilis ), most native species having disappeared since pike were introduced in 1953. (c) 1996 The Fisheries Society of the British Isles     

Macro-invertebrate predators of freshwater pulmonate snails in Africa,with particular reference to Appasus grassei (Heteroptera) and Procambarus clarkii (Decapoda)

A range of African and alien macro-invertebrates has been reported preying on freshwater pulmonate snails, including those that serve as intermediate hosts for bloodflukes of the genus Schistosoma. Predation by five molluscivorous taxa is reviewed here: indigenous leeches (Glossiphoniidae), marsh fly larvae (Sciomyzidae), waterbugs (Belostomatidae), crabs (Potamonautidae) and invasive crayfish (Astacidae). Common features are a lack of prey specificity but clear prey-size specificity. Attention is drawn to the ability of invasive snail species (Physidae and Lymnaeidae) to avoid predation by several of these taxa. Evidence suggests that only the alien invasive crayfish Procambarus clarkii has potential as a snail biocontrol agent, but that its use should not be encouraged.     

Serotonin-like immunoreactivity in the stomatogastric nervous systems of crayfishes from four genera

We used whole-mount immunocytochemistry to characterize the distribution of serotonin in the stomatogastric nervous systems of seven species of crayfish representing three genera from the family Cambaridae (Orconectes, Cambarus, and Procambarus) and one from the family Astacidae (Pacifastacus). In all species, we observed serotonin-like immunoreactivity in four gastropyloric receptor (GPR) neurons located in the lateral ventricular nerves, with one pair of neurons in each nerve. As in other crustaceans, the GPR axons project to the stomatogastric ganglion and to the bilateral commissural ganglia. In three crayfishes, we observed the GPR axons crossing the commissural ganglia, and extending toward the thoracic nervous system. This feature was most clearly and consistently seen in Pacifastacus leniusculus. The number of stained somata in the commissural ganglia varied among crayfish species from two (in Procambarus clarkii) to five (in Pacifastacus leniusculus). The largest soma (the L cell) displayed both serotonin- and tyrosine hydroxylase-like immunoreactivity in all species, suggesting that serotonin and dopamine are cotransmitters in this cell. The inferior esophageal nerve and a branch of this nerve (the inner labral nerve) contained several axons with serotonin-like immunoreactivity. These axons were clearly present in only one species (Procambarus clarkii). Serotonin acts as a neuromodulator of rhythms produced by circuits in the crab and lobster stomatogastric ganglion, and is likely to play a similar role in crayfish. Differences are apparent in the distribution of serotonin among crayfish species and between crayfish and other crustaceans, and could result in differences in the physiological action of this modulator.     

One century away from home: how the red swamp crayfish took over the world

Reviews in Fish Biology and Fisheries - The red swamp crayfish (Procambarus clarkii) (hereafter RSC), native to the southern United States and north-eastern Mexico, is currently the most widely...     

家蚕蛹表达的重组Vp28疫苗对克氏原螯虾的抗病毒保护效应

对虾白斑综合症病毒(WSSV)的致病性强、危害性大、地域分布和宿主范围广泛,目前还不能有效地控制疫情。将含有WSSV囊膜蛋白Vp28基因的重组杆状病毒HyNPV-Vp28感染家蚕(Bombyx mori)蛹,对发病蚕血淋巴进行SDS-PAGE和Western blotting分析,结果表明Vp28在家蚕体内得到了表达。将重组病毒囊膜蛋白rVp28疫苗配制成药饵,持续口服免疫75天,对克氏原螯虾进行预防WSSV,实验虾分为2%重组Vp28疫苗、2%普通蚕蛹组织匀浆(阳性对照)和普通饵料(阴性对照)3个处理组。免疫35天后进行口服攻毒,20天内rVp28疫苗组的累积存活率为63.33%,与阳性和阴性对照比差异显著(P<0.05),PRP分别达54.16%和59.26%;注射攻毒后20 天内rVp28疫苗组的累积存活率与阳性和阴性对照组比差异不显著(P>0.05),PRP分别为46.12% 和49.99%。第55天对存活虾再口服攻毒,20天内rVp28疫苗组与阳性和阴性对照组比累积存活率差异显著(P<0.05),PRP分别为55.80%和63.16%;二次注射攻毒后,rVp28疫苗组的PRP均为31.25%。对vVp28疫苗组存活虾的胃、肠和肝胰腺组织进行病毒的原位杂交检测均呈阴性反应,而对照组死亡虾组织都呈阳性反应。本研究表明,口服免疫家蚕蛹表达的病毒囊膜蛋白Vp28能诱导螯虾产生抗病毒保护作用,对应用疫苗预防对虾的病毒性疾病具有重要意义。     

家蚕蛹表达的重组Vp28疫苗对克氏原螯虾的抗病毒保护效应

The vaccine made of recombinant envelope protein (rVp28) of white spot syndrome virus (WSSV) expressed in silkworm (Bombyx mori) pupae using a baculovirus vector was used to investigate the efficacy of oral administration on WSSV disease resistance of Procambarus clarkii. Vaccine was mixed with diet at a ratio of 2% (w/w), and Procambarus clarkii were orally administered throughout 75 days. Vaccination with rVP28 showed the significantly higher cumulative survival compared with positive and negative control (P < 0.05) following an oral challenge on the 35th day post-vaccination (dpv), with PRP values 54.16% and 59.26%, respectively. rVP28 induced higher resistance via IM (intramuscular) injection challenge with WSSV stock, with PRP value of 46.12% and 49.99%, respectively. The survivors were subsequently re-challenged on the 55th dpv. rVP28 induced the significantly higher resistance to oral re-challenge (P < 0.05), with both PRP values 55.80% and 63.16%, respectively. rVP28 induced higher resistance to IM injection re-challenge, with both PRP values 31.25%. A DIG labeled WSSV DNA probe was used to detect WSSV by in situ hybridization. The positive cells were observed in epithelial cells of stomach, hepatopancreas and gut of the infected control crayfish, while negative reaction were observed in the tissues of survivors-vaccinated. These results indicated that vaccination of crayfish with recombinant protein had significant effect on oral infection, and had higher resistance against intramuscular injection challenge. This suggested the protection against WSSV could be induced in crayfish by recombinant protein rVp28 expressed in silkworm pupae.     

Effect of variation in photoperiod and light intensity on oxygen consumption, lactate concentration and behavior in crayfish Procambarus clarkii and Procambarus digueti

The effects of light intensity and duration on metabolic and behavioral parameters of two species of crayfish, Procambarus clarkii and Procambarus digueti, were studied. Sixty animals of each species were submitted to high irradiance conditions of two different photoperiod lengths, one normal light/dark (LD) 12:12 and one extreme LD 20:4 for 2 weeks. Hemolymph, lactate and oxygen consumption were determined throughout the experimental period. Simultaneously in 18 additional animals of each species, motor activity was individually recorded under the same control and experimental conditions. Both species showed a decrease in oxygen uptake and an increase in hemolymph lactate concentration. The statistical significance of this finding was higher for LD 20:4. This extreme condition evoked a significant decrease of motor activity in P. clarkii and a high mortality rate in P. digueti. P. digueti did not survive after the experiment, whereas P. clarkii survived and adapted to the laboratory conditions. Changes in metabolic and behavioral parameters could indicate different adaptation abilities in these species.     

White spot syndrome virus (WSSV) transmission from rotifer inoculum to crayfish

To test the possibility that shrimp pond rotifer resting eggs and hatched rotifers could transmit white spot syndrome virus (WSSV) to crayfish (Procambarus clarkii), we injected crayfish with rotifer and resting egg inocula that were WSSV-positive only by dot-blot analysis of PCR products. No crayfish became WSSV-positive after challenge with the resting egg inoculum. However, 1/15 crayfish became WSSV-positive after challenge with the rotifer inoculum. The results demonstrated that rotifers constitute a potential risk for WSSV transmission to crayfish and other cultivated crustaceans. However, the actual quantitative risk of transmission in an aquaculture setting depends on many variables that remain untested.     

Causes and consequences of crayfish extinction: Stream connectivity,habitat changes,alien species and ecosystem services

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Bioenergetics and phenology of reproduction of the introduced red swamp crayfish, Procambarus clarkii, in Do·ana National Park, Spain, and implications for species management

1. The red swamp crayfish, Procambarus clarkii (Girard), is a North American species which was introduced into the Lower Guadalquivir, Spain, in 1974. Procambarus clarkii has become a key species since it has greatly altered the functioning and structure of the ecosystems it has invaded.
2. Since P. clarkii spawn spontaneously throughout the year, it is to be expected that their ecological impact on the ecosystem may vary depending of the timing of habitat flooding (e.g. temporary freshwater marsh, and temporary or permanent pond). The objective of the present study was to understand the reproductive phenology and bioenergetics of P. clarkii in Do·ana National Park, Spain, freshwater ecosystems, and consequently, propose some management plans for their populations.
3. Five stages of ovarian development were detected. Final ovarian maturation occurred a few days before spawning and spawning was synchronized among mature females of all populations. Egg numbers hatched in pleopods ranged from 450 to 550. Timing of juvenile release from pleopods varied greatly (10–35 days).
4. Gravid females support most of their metabolic needs by catabolizing carbohydrates (overall mean of O:N = 30.39), and to a lesser extent, lipids. The aerobic and excretory metabolisms of gravid females were lower than those of immature animals of similar weight.
5. Analysis suggested that several factors influence the spawning of crayfish at the monitored sites. The major factors are water regime and temperature. Red swamp crayfish appear to be a spontaneous spawning species which may adjust their reproductive cycle to the timing of flooding, enhancing the utilization of available resources.
6. Populations of P. clarkii in Do·ana National Park could be controlled by the management of the water regime and by devising a fishing scheme which would maintain an appropriate population structure.     

Environment and host-related factors modulate gut and carapace bacterial diversity of the invasive red swamp crayfish (Procambarus clarkii)

Hydrobiologia - The invasive red swamp crayfish Procambarus clarkii is present in most of the Portuguese rivers with well-known economic and environmental impacts, being also an important reservoir...     

Crayfish co-introduced symbiotic ostracod found on native crab in Japan: The first record of epibiont ostracod found a new host

Ankylocythere sinuosa (Rioja, 1942), a symbiotic ostracod native to North America, was found from the Japanese mitten crab Eriocheir japonica (De Haan, 1835), a species native to Japan, collected from a pond in Shizuoka City, Shizuoka Prefecture, central Japan. Introduced North American crayfish Procambarus clarkii (Girard, 1852), which is a host of A. sinuosa in their native range, inhabits ponds sympatrically with Japanese mitten crabs, and it is thought that the ostracods transferred from the exotic crayfish to the native crabs. In recent years, along with the artificial transportation of crayfish around the world, their symbiotic ostracods also have been found on the body surfaces of exotic crayfish in Europe and Japan. However, no studies have confirmed the infestation of exotic ostracods on native crustaceans in the field. A wide range of developmental stages of A. sinuosa from juveniles to adults were found in Japanese mitten crabs, and mating individuals were also found. This strongly suggests that they can reproduce on the body surface of Japanese mitten crabs. In the future, it will be necessary to strengthen measures against alien species to prevent these exotic symbionts from infestating native ecosystems, and we also need to investigate the exact impact of this symbiont on Japanese mitten crabs.     

Managing invasive crayfish: use of X-ray sterilisation of males

1. In the last decade, many attempts have been made to contain the spread of the North American crayfish Procambarus clarkii , an invader of many European water bodies, but none has been successful.
2. This study investigates the effects that ionising irradiation has on the reproduction of P. clarkii males by analysing the behaviour of the treated individuals, their reproductive output, the vitality and survival of their offspring and the damages induced to their gonads.
3. We found that a dose of 20 Gy X-rays did not compromise either the survival or the mating ability of males, but reduced the size of their testes and significantly altered spermatogenesis. The number of aborted eggs was larger in clutches sired by treated males, with a consequent reduction (by 43%) in the number of offspring.
4. These results foster hopes in our efforts to control invasive crayfish, showing the potential effectiveness of ionising irradiation to reduce male fertility in the perspective of adopting the Sterile Male Release Technique to control P. clarkii populations.     

Eleven polymorphic microsatellites isolated from red swamp crayfish, Procambarus clarkii

Eleven microsatellites were isolated from a genomic DNA library enriched for CA- and GA-repeats, and were characterized in 48 individuals of red swamp crayfish Procambarus clarkii. All 11 microsatellites were polymorphic with an average of allele number of 6.6 per locus. Genotypic distributions of these 11 markers were found to be in conformance with the expectations of Hardy-Weinberg equilibrium. All 11 markers were unlinked. These markers are being used to study the invasion routine, genetic diversity and population structure of the species P. clarkii.     

Effect of hyperthermia on the replication of white spot syndrome virus (WSSV) in Procambarus clarkii

The effect of hyperthermia on the development of white spot syndrome virus (WSSV) in the crayfish Procambarus clarkii was studied by competitive PCR. Crayfish were exposed to different temperatures (24 +/- 1 and 32 +/- 1 degrees C) after WSSV injection. No mortality was observed when crayfish were held at 32 +/- 1 degrees C, but mortality reached 100% when crayfish were transferred to 24 +/- 1 degrees C. Competitive PCR showed that viral levels at 32 +/- 1 degrees C remained at 10(5) copies mg(-1) tissue, while at 24 +/- 1 degrees C levels were significantly higher, rising from 10(4) to 10(10) copies mg(-1) tissue. These results suggest that hyperthermia reduces viral replication, but does not eliminate viral particles from WSSV-infected crayfish.     

Identification of troponin-I of crayfish myofibrils

Crayfish (Procambarus clarkii) myofibrils contain two basic proteins of molecular weights of 25,000 and 23,000. Both of the two proteins inhibit actomyosin ATPase as the vertebrate troponin-I does. These results differ from the previous one that troponin-I of crayfish (Astacus leptodactylus) showed a single band on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE).