Evaluation of the exploitation potential of red swamp crayfish (Procambarus clarkii) in Portugal

Dissertação para obtenção do Grau de Mestre em Engenharia do Ambiente, perfil de Ordenamento do Território e Impactes Ambientais

Eradication d'une population d'écrevisses de Louisiane, Procambarus clarkii, à l'étang de Vidy et situation des populations d'écrevisses à Lausanne, Suisse.

En juillet 2007, des écrevisses de Louisiane, Procambarus clarkii, ont été observées, pour la première fois en Suisse romande, dans l'étang artificiel de Vidy à Lausanne. Au printemps 2008, l'étang a été asséché pendant 2 mois pour tenter d'éradiquer les écrevisses. En juillet 2008, et au printemps 2009, des écrevisses de Louisiane furent à nouveau observées dans l'étang: des individus avaient survécu au traitement et/ou des écrevisses avaient à nouveau été introduites. En 2010, une nouvelle campagne d'éradication a été mise en place: l'étang fut asséché, son lit chaulé et les interstices entre les enrochements bétonnés. Lors du suivi 2011, plus aucune écrevisse ne fut capturée. L'opération semble donc finalement avoir été un succès. Les observations effectuées sur 20 plans d'eau dans un rayon de 4.5 km autour de l'étang de Vidy n'ont pas mis en évidence d'autres populations d'écrevisses de Louisiane. Plusieurs populations d'écrevisses américaines, orconectes limosus, et signal ̧ Pacifastacus lenuisculus, ont été localisées, ainsi qu'une population d'écrevisses à pattes rouges, Astacus astacus.

Isolation of DNA sequences with homology to a degenerate FaRP oligonucleotide from the crayfish Procambarus clarkii

The neuropeptide Th1RFamide with the sequence Phe-Met-Arg-Phe-amide was originally isolated in the clam Macrocallista nimbosa (price and Greenberg, 1977). Since its discovery, a large family ofFl\1RFamide-related peptides termed FaRPs have been found to be present in all major animal phyla with functions ranging from modulation of neuronal activity to alteration of muscular contractions. However, little is known about the genetics encoding these peptides, especially in invertebrates. As FaRP-encoding genes have yet to be investigated in the invertebrate Malacostracean subphylum, the isolation and characterization ofFaRP-encoding DNA and mRNA was pursued in this project. The immediate aims of this thesis were: (1) to amplify mRNA sequences of Procambarus clarkii using a degenerate oligonucleotide primer deduced from the common amino acid sequence ofisolated Procambarus FaRPS, (2) to determine if these amplification products encode FaRP gene sequences, and (3) to create a selective cDNA library of sequences recognized by the degenerate oligonucleotide primer. The polymerase chain reaction - rapid amplification of cDNA ends (PCR-RACE) is a procedure in which a single gene-specific primer is used in conjunction with a generalized 3' or 5' primer to amplify copies ofthe region between a single point in the transcript and the 3' or 5' end of cDNA of interest (Frohman et aI., 1988). PCRRACE reactions were optimized with respect to primers used, buffer composition, cycle number, nature ofgenetic substrate to be amplified, annealing, extension and denaturation temperatures and times, and use of reamplification procedures. Amplification products were cloned into plasmid vectors and recombinant products were isolated, as were the recombinant plaques formed in the selective cDNA library. Labeled amplification products were hybridized to recombinant bacteriophage to determine ligated amplification product presence. When sequenced, the five isolated PCR-RACE amplification products were determined not to possess FaRP-encoding sequences. The 200bp, 450bp, and 1500bp sequences showed homology to the Caenorhabditis elegans cosmid K09A11, which encodes for cytochrome P450; transfer-RNA; transposase; and tRNA-Tyr, while the 500bp and 750bp sequences showed homology with the complete genome of the Vaccinia virus. Under the employed amplification conditions the degenerate oligonucleotide primer was observed to bind to and to amplify sequences with either 9 or 10bp of 17bp identity. The selective cDNA library was obselVed to be of extremely low titre. When library titre was increased, white. plaques were isolated. Amplification analysis of eight isolated Agt11 sequences from these plaques indicated an absence of an insertion sequence. The degenerate 17 base oligonucleotide primer synthesized from the common amino acid sequence ofisolated Procambarus FaRPs was thus determined to be non-specific in its binding under the conditions required for its use, and to be insufficient for the isolation and identification ofFaRP-encoding sequences. A more specific primer oflonger sequence, lower degeneracy, and higher melting temperature (TJ is recommended for further investigation into the FaRP-encoding genes of Procambarlls clarkii.

Tamaño poblacional, morfometría y crecimiento de Procambarus clarkii (Girard) (Crustácea: cambaridae) del área central de Nuevo León, México

Tesis (Maestría en Ciencias con Especialidad en Ecología Acuática y Pesca) U.A.N.L.

Evaluación del crecimiento y sobrevivencia del acocil rojo procambarus clarkii (GIRARD, 1852) con un alimento natura y una dieta formulada bajo condiciones de laboratorio.

Tesis (Maestría en Ciencias con Especialidad en Recursos Alimenticios y Producción Agrícola) UANL, 2012.

Fisiología reproductiva del acocil rojo Procambarus clarkii (Crustacea: decapoda) : establecimiento del ciclo de maduración gonadal y evaluación de su potencial reproductivo

Tesis (Doctorado en Ciencias Biológicas con Especialidad en Acuacultura) UANL

Morfometría, distribución actual y potencial en el norte de México del acocil rojo procambarus clarkii (Girard, 1852) (crustacea: cambaridae)

Tesis (Doctor en Ciencias con acentuación en Manejo de Vida Silvestre y Desarrollo Sustentable) UANL, 2014.

Assessment of Donana National Park contamination in Procambarus clarkii: Integration of conventional biomarkers and proteomic approaches

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biochemical and proteomic effects in Procambarus clarkii after chlorpyrifos or carbaryl exposure under sublethal conditions

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Response of crayfish, Procambarus clarkii, haemocytes infected by white spot syndrome virus

White spot syndrome virus ( WSSV) is a serious pathogen of aquatic crustaceans. Little is known about its transmission in vivo and the immune reaction of its hosts. In this study, the circulating haemocytes of crayfish, Procambarus clarkii, infected by WSSV, and primary haemocyte cultures inoculated with WSSV, were collected and observed by transmission electron microscopy and light microscopy following in situ hybridization. In ultrathin sections of infected haemocytes, the enveloped virions were seen to be phagocytosed in the cytoplasm and no viral particles were observed in the nuclei. In situ hybridization with WSSV-specific probes also demonstrated that there were no specific positive signals present in the haemocytes. Conversely, strong specific positive signals showed that WSSV replicated in the nuclei of gill cells. As a control, the lymphoid organ of shrimp, Penaeus monodon, infected by WSSV was examined by in situ hybridization which showed that WSSV did not replicate within the tubules of the lymphoid organ. In contrast to previous studies, it is concluded that neither shrimp nor crayfish haemocytes support WSSV replication.White spot syndrome virus (WSSV) is a serious pathogen of aquatic crustaceans. Little is known about its transmission in vivo and the immune reaction of its hosts. In this study, the circulating haemocytes of crayfish, Procambarus clarkii, infected by WSSV, and primary haemocyte cultures inoculated with WSSV, were collected and observed by transmission electron microscopy and light microscopy following in situ hybridization. In ultra-thin sections of infected haemocytes, the enveloped virions were seen to be phagocytosed in the cytoplasm and no viral particles were observed in the nuclei. In situ hybridization with WSSV-specific probes also demonstrated that there were no specific positive signals present in the haemocytes. Conversely, strong specific positive signals showed that WSSV replicated in the nuclei of gill cells. As a control, the lymphoid organ of shrimp, Penaeus monodon, infected by WSSV was examined by in situ hybridization which showed that WSSV did not replicate within the tubules of the lymphoid organ. In contrast to previous studies, it is concluded that neither shrimp nor crayfish haemocytes support WSSV replication.

The red swamp crayfish Procambarus clarkii in Europe: Impacts on aquatic ecosystems and human well-being

Procambarus clarkii is currently recorded from 16 European territories. On top of being a vector of crayfish plague, which is responsible for large-scale disappearance of native crayfish species, it causes severe impacts on diverse aquatic ecosystems, due to its rapid life cycle, dispersal capacities, burrowing activities and high population densities. The species has even been recently discovered in caves. This invasive crayfish is a polytrophic keystone species that can exert multiple pressures on ecosystems. Most studies deal with the decline of macrophytes and predation on several species (amphibians, molluscs, and macroinvertebrates), highlighting how this biodiversity loss leads to unbalanced food chains. At a management level, the species is considered as (a) a devastating digger of the water drainage systems in southern and central Europe, (b) an agricultural pest in Mediterranean territories, consuming, for example, young rice plants, and (c) a threat to the restoration of water bodies in north-western Europe. Indeed, among the high-risk species, P. clarkii consistently attained the highest risk rating. Its negative impacts on ecosystem services were evaluated. These may include the loss of provisioning services such as reductions in valued edible native species of regulatory and supporting services, inducing wide changes in ecological communities and increased costs to agriculture and water management. Finally, cultural services may be lost. The species fulfils the criteria of the Article 4(3) of Regulation (EU) No 1143/2014 of the European Parliament (species widely spread in Europe and impossible to eradicate in a cost-effective manner) and has been included in the “Union List”. Particularly, awareness of the ornamental trade through the internet must be reinforced within the European Community and import and trade regulations should be imposed to reduce the availability of this high-risk species.

Palatability of the Asian clam Corbicula fluminea (Müller 1774) in an invaded system

Dissertação de mestrado em Ecology

Pla de gestió i conservació dels Estanyets de Pals i les Closes Fondes (Pals, Baix Empordà)

Es destaca la presència de tres espècies bioinvasores, tals com la gambúsia(Gambusiaaffinis), la canya (Arundodonax) i el cranc vermell americà (Procambarusclarkii). S’ha detectat que la qualitat de l’aigua ha disminuït en alguns paràmetres, especialment en l’Estanyet del Safareig. En els tres estanyets els sòlids dissolts totalsa TSD) estan al voltant del límit màxim recomanat per la EPA (Agència de Protecció Ambiental d’Estats Units). S’ha observat que la Cladophora, indicadora de concentracions elevades de nitrogen a l’aigua, és un cloròfit molt abundant. S’han identificat dos hàbitats d’interès comunitari no prioritari, segons la Directiva Hàbitats: les closes i les freixenedes termòfiles de Fraxinusangustifolia. En funció dels resultats obtinguts s’han elaborat les propostes de gestió i conservació per aquest espai.

Microphallus fonti sp. n. (Digenea: Microphaliidae) from the red swamp crawfish in southern United States

A new species of digenean, Microphallus fonti, is described from the red swamp crawfish in Lousiania, U.S.A. It has a small pharynx and a rudimentary gut like M. opacus and a possibly related species from crayfishes, but it differs from them by its relatively large male copulatory papilla and a conspicuous metraterm.