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Giuseppe Nascetti - One of the best experts on this subject based on the ideXlab platform.
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species specific real time pcr primers probe systems to identify fish parasites of the genera Anisakis pseudoterranova and hysterothylacium nematoda ascaridoidea
Fisheries Research, 2017Co-Authors: Arne Levsen, Michela Paoletti, Simonetta Mattiucci, Alessandra Colantoni, Giuseppe NascettiAbstract:Abstract Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.) , P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum , a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency ( E ) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.
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Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis, Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)
Fisheries Research, 2017Co-Authors: Michela Paoletti, Arne Levsen, Simonetta Mattiucci, Alessandra Colantoni, Giuseppe NascettiAbstract:Abstract Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.) , P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum , a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency ( E ) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.
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Additional records of metazoan parasites from Caribbean marine mammals, including genetically identified anisakid nematodes
Parasitology Research, 2009Co-Authors: Marlene M. Colón-llavina, Antonio A. Mignucci-giannoni, Michela Paoletti, Simonetta Mattiucci, Giuseppe Nascetti, Ernest H WilliamsAbstract:Studies of marine mammal parasites in the Caribbean are scarce. An assessment for marine mammal endo- and ectoparasites from Puerto Rico and the Virgin Islands, but extending to other areas of the Caribbean, was conducted between 1989 and 1994. The present study complements the latter and enhances identification of anisakid nematodes using molecular markers. Parasites were collected from 59 carcasses of stranded cetaceans and manatees from 1994 to 2006, including Globicephala macrorhynchus , Kogia breviceps , Kogia sima , Lagenodelphis hosei , Mesoplodon densirostris , Peponocephala electra , Stenella longirostris , Steno bredanensis , Trichechus manatus . Tursiops truncatus , and Ziphius cavirostris . Sixteen species of endoparasitic helminthes were morphologically identified, including two species of acanthocephalans ( Bolbosoma capitatum , Bolbosoma vasculosum ), nine species of nematodes ( Anisakis sp., Anisakis brevispiculata , Anisakis paggiae , Anisakis simplex , Anisakis typica , Anisakis ziphidarium , Crassicauda anthonyi , Heterocheilus tunicatus , Pseudoterranova ceticola ), two species of cestodes ( Monorygma grimaldi , Phyllobothrium delphini ), and three species of trematodes ( Chiorchis groschafti , Pulmonicola cochleotrema , Monoligerum blairi ). The nematodes belonging to the genus Anisakis recovered in some stranded animals were genetically identified to species level based on their sequence analysis of mitochondrial DNA (629 bp of mtDNA cox 2 ). A total of five new host records and six new geographic records are presented.
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advances and trends in the molecular systematics of anisakid nematodes with implications for their evolutionary ecology and host parasite co evolutionary processes
Advances in Parasitology, 2008Co-Authors: Simonetta Mattiucci, Giuseppe NascettiAbstract:Abstract The application of molecular systematics to the anisakid nematodes of the genera Anisakis, Pseudoterranova and Contracaecum, parasites of aquatic organisms, over the last two decades, has advanced the understanding of their systematics, taxonomy, ecology and phylogeny substantially. Here the results of this effort on this group of species from the early genetic works to the current status of their revised taxonomy, ecology and evolutionary aspects are reviewed for each of three parasitic groups. It has been shown that many anisakid morphospecies of Anisakis, Contracaecum and Pseudoterranova include a certain number of sibling species. Molecular genetic markers provided a rapid, precise means to screen and identify several species that serve as definitive and intermediate and or/paratenic hosts of the so far genetically characterized species. Patterns of differential distribution of anisakid nematodes in various definitive and intermediate hosts are presented. Differences in the life history of related species can be due both to differential host—parasite co-adaptation and co-evolution, and/or to interspecific competition, that can reduce the range of potential hosts in sympatric conditions. Phylogenetic hypotheses attempted for anisakid nematodes and the possible evolutionary scenarios that have been proposed inferred from molecular data, also with respect to the phylogeny of their hosts are presented for the parasite—host associations Anisakis-cetaceans and Contracaecum-pinnipeds, showing that codivergence and host-switching events could have accompanied the evolution of these groups of parasites. Finally, examples in which anisakid nematodes recognized genetically at the species level in definitive and intermediate/paratenic hosts from various geographical areas of the Boreal and Austral regions and their infection levels have been used as biological indicators of fish stocks and food-web integrity in areas at high versus low levels of habitat disturbance (pollution, overfishing, by-catch) are presented.
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The metazoan parasites of swordfish from Atlantic tropical-equatorial waters
'Wiley', 2008Co-Authors: A. Garcia, Giuseppe Nascetti, N. Santosm, S. Damiano, Simonetta MattiucciAbstract:The metazoan parasite fauna composition of the swordfish Xiphias gladius is reported from 145 (72 females and 73 males) specimens caught in Atlantic tropical-equatorial waters during June to July 2006 and August 2007. Seventeen metazoan taxa (two monogeneans, two digeneans, one acanthocephalan, four cestodes, seven nematodes and one copepod) were identified by morphological and genetic methodologies. Among those identified, Anisakis brevispiculata, Anisakis paggiae and the acanthocephalan Bolbosoma vasculosum are new host records. Dominant species were: the anisakid nematode larvae Anisakis physeteris, Anisakis brevispiculata, Hysterothylacium incurvum and Hysterothylacium corrugatum (s.l.), the adult cestode Fistulicola plicatus, the plerocercoid Hepatoxylon squali and the monogeneans Tristoma integrum and Tristoma coccineum, which showed high prevalences and intensities. The relationship between metazoan parasites and host size and sex was investigated for the first time. No statistically significant correlation was found between host sex and parasite burden. Infection levels with T. integrum and T. coccineum were observed to be significantly negatively correlated to host size. Infection levels by endoparasites observed in X. gladius from Atlantic tropical-equatorial area may be related to host feeding behaviour and the availability of the parasites' intermediate and final hosts in the area studied. Anisakids of the genus Anisakis, were identified genetically to species level, and were considered to be promising biological tags for the characterization of X. gladius stocks
Michela Paoletti - One of the best experts on this subject based on the ideXlab platform.
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species specific real time pcr primers probe systems to identify fish parasites of the genera Anisakis pseudoterranova and hysterothylacium nematoda ascaridoidea
Fisheries Research, 2017Co-Authors: Arne Levsen, Michela Paoletti, Simonetta Mattiucci, Alessandra Colantoni, Giuseppe NascettiAbstract:Abstract Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.) , P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum , a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency ( E ) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.
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Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis, Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)
Fisheries Research, 2017Co-Authors: Michela Paoletti, Arne Levsen, Simonetta Mattiucci, Alessandra Colantoni, Giuseppe NascettiAbstract:Abstract Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.) , P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum , a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency ( E ) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.
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Anisakis nascettii n. sp. (Nematoda: Anisakidae) from beaked whales of the southern hemisphere: morphological description, genetic relationships between congeners and ecological data
Systematic Parasitology, 2009Co-Authors: Simonetta Mattiucci, Michela Paoletti, Stephen C WebbAbstract:A new anisakid nematode, Anisakis nascettii n. sp., is described from beaked whales Mesoplodon spp. off the coast of New Zealand and South Africa. Morphological and molecular (allozymes and mtDNA cox2 sequence) data were used for diagnostic and identification purposes. Among the 19 allozymes studied, 10 were found to be unique and characteristic for A. nascettii n. sp. Analysis of allozymes demonstrated reproductive isolation from A. ziphidarum Paggi, Nascetti, Webb, Mattiucci, Cianchi & Bullini, 1998 and mtDNA cox2 sequences depict this Anisakis species as a distinct and unique entity. Key morphological diagnostic traits for A. nascettii with respect to the genetically closely related species A. ziphidarum include: spicule length, the spicule/body length ratio, the arrangement of the caudal papillae and the shape of the plectanes of the adult males. Genetic data confirmed that Anisakis sp. A of Pontes et al. ( 2005 ), which was partly described by Iglesias et al. ( 2008 ), and Anisakis sp. of Valentini et al. ( 2006 ) are conspecific with A. nascettii . Both molecular and morphological data indicate that the new species belongs to the ‘ ziphidarum- group’; however, it is genetically very distinct from A. ziphidarum ( D _ Nei = 0.69, K2P = 0.09), as well as from all of the previously genetically characterised Anisakis spp. All tree topologies inferred by different methods (MP, NJ and Bayesian) support the finding that A. nascettii n. sp. and A. ziphidarum are sister-species. It is also confirmed that A. nascettii n. sp. is, at the adult stage, a parasite of beaked whales of the genus Mesoplodon , whereas, as a larva, it has been identified from the squid Moroteuthis ingens Smith. Furthermore, Mesoplodon bowdoini Andrews represents a new host record for A. ziphidarum . The parallelism between the clade formed by these two anisakine taxa, i.e. A. ziphidarum and A. nascettii , and that formed by their definitive hosts further supports the hypothesis of host–parasite co-evolutionary relationships, as previously suggested for Anisakis spp. and their cetacean hosts.
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Additional records of metazoan parasites from Caribbean marine mammals, including genetically identified anisakid nematodes
Parasitology Research, 2009Co-Authors: Marlene M. Colón-llavina, Antonio A. Mignucci-giannoni, Michela Paoletti, Simonetta Mattiucci, Giuseppe Nascetti, Ernest H WilliamsAbstract:Studies of marine mammal parasites in the Caribbean are scarce. An assessment for marine mammal endo- and ectoparasites from Puerto Rico and the Virgin Islands, but extending to other areas of the Caribbean, was conducted between 1989 and 1994. The present study complements the latter and enhances identification of anisakid nematodes using molecular markers. Parasites were collected from 59 carcasses of stranded cetaceans and manatees from 1994 to 2006, including Globicephala macrorhynchus , Kogia breviceps , Kogia sima , Lagenodelphis hosei , Mesoplodon densirostris , Peponocephala electra , Stenella longirostris , Steno bredanensis , Trichechus manatus . Tursiops truncatus , and Ziphius cavirostris . Sixteen species of endoparasitic helminthes were morphologically identified, including two species of acanthocephalans ( Bolbosoma capitatum , Bolbosoma vasculosum ), nine species of nematodes ( Anisakis sp., Anisakis brevispiculata , Anisakis paggiae , Anisakis simplex , Anisakis typica , Anisakis ziphidarium , Crassicauda anthonyi , Heterocheilus tunicatus , Pseudoterranova ceticola ), two species of cestodes ( Monorygma grimaldi , Phyllobothrium delphini ), and three species of trematodes ( Chiorchis groschafti , Pulmonicola cochleotrema , Monoligerum blairi ). The nematodes belonging to the genus Anisakis recovered in some stranded animals were genetically identified to species level based on their sequence analysis of mitochondrial DNA (629 bp of mtDNA cox 2 ). A total of five new host records and six new geographic records are presented.
Simonetta Mattiucci - One of the best experts on this subject based on the ideXlab platform.
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Anisakis simplex s s larvae nematoda anisakidae hidden in the mantle of european flying squid todarodes sagittatus cephalopoda ommastrephidae in ne atlantic ocean food safety implications
International Journal of Food Microbiology, 2021Co-Authors: Paolo Cipriani, Simonetta Mattiucci, Marialetizia Palomba, Lucilla Giulietti, Miguel Bao, Arne LevsenAbstract:Abstract Few reports exist upon the occurrence and localization of zoonotic anisakid nematodes in T. sagittatus, especially in the mantle of the squid. The occurrence and site of infection of larval anisakids in 98 T. sagittatus caught West off St. Kilda, NE Atlantic Ocean, were investigated. Squids were examined for anisakids using the UV-Press method. In total, 689 nematodes were detected in the viscera and mantle. According to morphology, all the larvae (L3) were assigned to genus Anisakis. Diagnostic allozymes and mtDNA cox2 sequence analysis permitted to genetically identify all larvae as Anisakis simplex (s.s.) (N = 100). Overall prevalence (P = 81%) and mean intensity (mI = 8.6) of infection with A. simplex are provided. Most of the larvae present in the mantle cavity were embedded in the stomach wall or attached in the outer layer of the stomach and caecum (49%). Over a third of squids (37%) hosted A. simplex (s.s.) larvae in the mantle. A novel schematized representation of larvae distribution in the mantle is provided, showing where they were mostly located. According to the results obtained, the risk of anisakiasis associated with consumption of raw or undercooked T. sagittatus should be considered.
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molecular identification of zoonotic parasites of the genus Anisakis nematoda anisakidae from fish of the southeastern pacific ocean off peru coast
Pathogenetics, 2020Co-Authors: Renato Aco Alburqueque, Marialetizia Palomba, Mario Santoro, Simonetta MattiucciAbstract:The study aims to perform, for the first time, the molecular identification of anisakid larvae in commercial fish from the Southeastern Pacific Ocean off the Peru coast, and to provide data on their infection level by fishing ground, fish host, and site of infection. Fish specimens (N = 348) from the northern and the central coast of Peru were examined for parasites. The fish fillets were examined by the UV-press method. Anisakis spp. larvae (N = 305) were identified by mtDNA cox2 sequences analysis and by the ARMS-PCR of the locus nas10 nDNA. Two hundred and eighty-eight Anisakis Type I larvae corresponded to Anisakis pegreffii, whereas 17 Anisakis Type II larvae clustered in a phylogenetic lineage distinct from Anisakis physeteris deposited in GenBank, and corresponding to a phylogenetic lineage indicated as Anisakis sp. 2, previously detected in fish from both Pacific and Atlantic waters. Anisakis pegreffii was found to infect both the flesh and viscera, while Anisakis sp. 2 occurred only in the viscera. The average parasitic burden with A. pegreffii in the examined fish species from the two fishing grounds was significantly higher than that observed with Anisakis sp. 2. The results obtained contribute to improve the knowledge on the distribution and occurrence of Anisakis species in Southeastern Pacific waters and their implications in seafood safety for the local human populations.
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species specific real time pcr primers probe systems to identify fish parasites of the genera Anisakis pseudoterranova and hysterothylacium nematoda ascaridoidea
Fisheries Research, 2017Co-Authors: Arne Levsen, Michela Paoletti, Simonetta Mattiucci, Alessandra Colantoni, Giuseppe NascettiAbstract:Abstract Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.) , P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum , a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency ( E ) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.
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Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis, Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)
Fisheries Research, 2017Co-Authors: Michela Paoletti, Arne Levsen, Simonetta Mattiucci, Alessandra Colantoni, Giuseppe NascettiAbstract:Abstract Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.) , P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum , a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency ( E ) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.
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Anisakis nascettii n. sp. (Nematoda: Anisakidae) from beaked whales of the southern hemisphere: morphological description, genetic relationships between congeners and ecological data
Systematic Parasitology, 2009Co-Authors: Simonetta Mattiucci, Michela Paoletti, Stephen C WebbAbstract:A new anisakid nematode, Anisakis nascettii n. sp., is described from beaked whales Mesoplodon spp. off the coast of New Zealand and South Africa. Morphological and molecular (allozymes and mtDNA cox2 sequence) data were used for diagnostic and identification purposes. Among the 19 allozymes studied, 10 were found to be unique and characteristic for A. nascettii n. sp. Analysis of allozymes demonstrated reproductive isolation from A. ziphidarum Paggi, Nascetti, Webb, Mattiucci, Cianchi & Bullini, 1998 and mtDNA cox2 sequences depict this Anisakis species as a distinct and unique entity. Key morphological diagnostic traits for A. nascettii with respect to the genetically closely related species A. ziphidarum include: spicule length, the spicule/body length ratio, the arrangement of the caudal papillae and the shape of the plectanes of the adult males. Genetic data confirmed that Anisakis sp. A of Pontes et al. ( 2005 ), which was partly described by Iglesias et al. ( 2008 ), and Anisakis sp. of Valentini et al. ( 2006 ) are conspecific with A. nascettii . Both molecular and morphological data indicate that the new species belongs to the ‘ ziphidarum- group’; however, it is genetically very distinct from A. ziphidarum ( D _ Nei = 0.69, K2P = 0.09), as well as from all of the previously genetically characterised Anisakis spp. All tree topologies inferred by different methods (MP, NJ and Bayesian) support the finding that A. nascettii n. sp. and A. ziphidarum are sister-species. It is also confirmed that A. nascettii n. sp. is, at the adult stage, a parasite of beaked whales of the genus Mesoplodon , whereas, as a larva, it has been identified from the squid Moroteuthis ingens Smith. Furthermore, Mesoplodon bowdoini Andrews represents a new host record for A. ziphidarum . The parallelism between the clade formed by these two anisakine taxa, i.e. A. ziphidarum and A. nascettii , and that formed by their definitive hosts further supports the hypothesis of host–parasite co-evolutionary relationships, as previously suggested for Anisakis spp. and their cetacean hosts.
Alessandra Colantoni - One of the best experts on this subject based on the ideXlab platform.
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species specific real time pcr primers probe systems to identify fish parasites of the genera Anisakis pseudoterranova and hysterothylacium nematoda ascaridoidea
Fisheries Research, 2017Co-Authors: Arne Levsen, Michela Paoletti, Simonetta Mattiucci, Alessandra Colantoni, Giuseppe NascettiAbstract:Abstract Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.) , P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum , a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency ( E ) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.
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Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis, Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)
Fisheries Research, 2017Co-Authors: Michela Paoletti, Arne Levsen, Simonetta Mattiucci, Alessandra Colantoni, Giuseppe NascettiAbstract:Abstract Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.) , P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum , a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency ( E ) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers.
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anisakidae nematodes isolated from the flathead grey mullet fish mugil cephalus of buenaventura colombia
International journal for parasitology. Parasites and wildlife, 2017Co-Authors: Jenniffer Alejandra Castellanos, Andrés Ricardo Tangua, Liliana SalazarAbstract:Anisakiasis is a parasitic infection caused by larval stages of nematodes of the genus Anisakis, Pseudoterranova and Contracaecum, of the Anisakidae family. The lifecycle of these nematodes develops in aquatic organisms and their final hosts are marine mammals. However, humans can act as accidental hosts and become infected with infective stage larvae (L3) by consuming raw or undercooked fish or shellfish carrying the parasite. Of this group of parasites, the genus Anisakis is the most studied: its presence in humans is associated with non-specific gastrointestinal symptoms or allergic responses that can trigger anaphylactic shock. The lack of studies in anisakiasis and Anisakis in Colombia has resulted in this infection being little-known by medical practitioners and therefore potentially underreported. The objective of this study was to identify anisakid nematodes in the flathead grey mullet fish (Mugil cephalus), caught by artisanal fishing methods and commercialized in Buenaventura. Morphological identification was carried out by classical taxonomy complemented by microscopy study using the histochemical technique Hematoxylin-Eosin. Nematodes of the genus Anisakis were found in the host M. cephalus. The Prevalence of Anisakis larvae in flathead grey mullet fish was 33%. The findings confirm the presence of Anisakis sp. in fish for human consumption in the Colombian Pacific region, a justification for further investigation into a possible emerging disease in this country.
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Anisakidae nematodes isolated from the flathead grey mullet fish (Mugil cephalus) of Buenaventura, Colombia
Elsevier, 2017Co-Authors: Jenniffer Alejandra Castellanos, Andrés Ricardo Tangua, Liliana SalazarAbstract:Anisakiasis is a parasitic infection caused by larval stages of nematodes of the genus Anisakis, Pseudoterranova and Contracaecum, of the Anisakidae family. The lifecycle of these nematodes develops in aquatic organisms and their final hosts are marine mammals. However, humans can act as accidental hosts and become infected with infective stage larvae (L3) by consuming raw or undercooked fish or shellfish carrying the parasite. Of this group of parasites, the genus Anisakis is the most studied: its presence in humans is associated with non-specific gastrointestinal symptoms or allergic responses that can trigger anaphylactic shock. The lack of studies in anisakiasis and Anisakis in Colombia has resulted in this infection being little-known by medical practitioners and therefore potentially underreported. The objective of this study was to identify anisakid nematodes in the flathead grey mullet fish (Mugil cephalus), caught by artisanal fishing methods and commercialized in Buenaventura. Morphological identification was carried out by classical taxonomy complemented by microscopy study using the histochemical technique Hematoxylin-Eosin. Nematodes of the genus Anisakis were found in the host M. cephalus. The Prevalence of Anisakis larvae in flathead grey mullet fish was 33%. The findings confirm the presence of Anisakis sp. in fish for human consumption in the Colombian Pacific region, a justification for further investigation into a possible emerging disease in this country. Keywords: Anisakis, Allergy, Anisakiasis, Mullet fish, Emerging disease