Genetic markers in ribosomal DNA for the identification of members of the genus Anisakis (Nematoda: Ascaridoidea) defined by polymerase-chain-reaction-based restriction fragment length polymorphism

D’Amelio, Stefano; Mathiopoulos, Kostas D.; Santos, Cláudia Portes; Pugachev, O.; Webb, Stephen C.; Picanço, Marilucia R. A.; Paggi, L.

doi:10.1016/s0020-7519(99)00178-2

Cited by 292 publications

(186 citation statements)

References 12 publications

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“…Internal transcribed spacer ribosomal DNA (ITSrDNA) was amplified as described (Zhu et al 1998), and the resultant PCR products were purified by MinElute Gel Purification (Qiagen) and sequenced on a CEQ 8800 using the GenomeLab DTCS Quick Start kit (Beckman Coulter). Restriction fragment length polymorphism (RFLP) analysis was performed on purified ITS PCR products using HinfI and HhaI (D'Amelio et al 2000, Abollo et al 2003, Pontes et al 2005, Umehara et al 2007). PCR products were cloned using the TOPO TA cloning vector (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

“…larvae from both cavity and vent as A. simplex s.s., based on HinfI and HhaI RFLP patterns (Fig. 14) (D'Amelio et al 2000) and sequence data. RFLP patterns were identical for all parasites from both vent and cavity.…”

Section: Molecular Analysismentioning

confidence: 99%

“…Type I or Type II (sensu Berland 1961) , Mattiucci et al 2005, Valentini et al 2006. Moreover, genetic markers from multilocus allozyme electrophoresis have shown that A. simplex (Davey 1971) falling in the first clade is a complex of sibling species comprising A. simplex sensu stricto (s.s.), A. pegreffii and A. simplex C , Mattiucci et al 1997, D'Amelio et al 2000, They not only differ in their genetic structure, but also in their lifehistory and geographic distribution, with A. simplex s.s. being reported to be widespread between 35°N and the Arctic polar circle of both the western and eastern waters of both the Atlantic and the Pacific oceans, with a southern limit of the eastern North Atlantic in the waters of the Gibraltar area (Mattiucci et al 1997, D'Amelio et al 2000, Abollo et al 2001. The current taxonomy and ecology of the recognised species have been recently reviewed by Mattiucci & Nascetti (2008).…”

Section: Introductionmentioning

confidence: 99%

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Red vent syndrome in wild Atlantic salmon Salmo salar in Scotland is associated with Anisakis simplex sensu stricto (Nematoda: Anisakidae)

Noguera

Collins²,

Bruno³

et al. 2009

Dis. Aquat. Org.

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Simultaneous reports were received between June and July 2007 of wild Atlantic salmon Salmo salar with red, swollen, bloody vents returning to geographically diverse rivers in Scotland. By the end of September the condition, colloquially known as 'red vent syndrome' (RVS), was reported from > 50 rivers across Scotland. Fish were generally in good overall condition but the vent area showed mild to severe lesions. External characteristics of the syndrome included a swollen, raised, haemorrhagic vent and surrounding tissues, with erosion of the skin, scale loss and moderate to severe bleeding in more advanced cases. Predominantly, the fish affected were 1-sea-winter grilse; however, RVS was also recorded in 2-sea-winter salmon and sea trout S. trutta. High numbers of the nematode Anisakis Type I larvae were found infesting the discrete region of the vent, a localisation that is reported as novel for the parasite. The hypothesis that this is a different species than that commonly found in the body cavity and viscera was investigated through molecular studies. These studies failed to show evidence that the parasites infesting the vent were different from those in the body cavity, i.e. all were identified as A. simplex sensu stricto. No other disease agent was found associated with the lesions or was isolated systemically, and no mortality or prevention of spawning was recorded during the 2007 season. Possible causes, including warming environments in the North Atlantic, are hypothesised as playing a role in the development of RVS in Atlantic salmon.

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Section: Methodsmentioning

confidence: 99%

Section: Molecular Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Red vent syndrome in wild Atlantic salmon Salmo salar in Scotland is associated with Anisakis simplex sensu stricto (Nematoda: Anisakidae)

Noguera

Collins²,

Bruno³

et al. 2009

Dis. Aquat. Org.

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“…The ITS-1 region, lying between the genes coding for the 18S and the 5.8S rRNA, allows classification of the parasites studied because of the differences in interspecific sequences and the low level of intraspecific variation. 18 This region does not encode any product and has less impact on organism viability, which permits it to evolve at a faster rate than the ribosomal coding regions. 19 Recently, the evaluation of the 7SL rRNA gene for the identification of Leishmania spp.…”

Section: Discussionmentioning

confidence: 99%

The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species

Tomás-Pérez

Fisa

Riera

2013

The American Society of Tropical Medicine and Hygiene

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Abstract. Leishmaniasis is a disease caused by different species belonging to the genus Leishmania. It presents different epidemiological and clinical features and requires the development of rapid, sensitive techniques to improve specific diagnosis. In this study, we compared the traditional technique of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with PCR-fluorescent fragment length analysis (PCR-FFL). Fluorescently tagged primers, designed in the rRNA fragment ITS-1 and 7SL region, were used to amplify fragments, which were later digested and whose sizes were accurately determined using an automated DNA sequencer. We validated the technique using 19 Leishmania strains from five cutaneous Leishmania species before testing 36 clinical samples: 23 skin biopsies and 13 skin scrapings/lesion exudates on filter paper. In real diagnostic, PCR-FFL has proved to be quick, accurate, and more sensitive (83.3% testing the ITS-1 fragment and 94.4% testing the 7SL) than PCR-RFLP analysis (75% and 80.6%). Filter papers improved the specific diagnosis in both techniques using non-invasive samples.

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“…Nematodes were cut into three parts: the head and caudal part were used for morphological identification and cleared in lactophenol and embedded in glycerine-gelatine. The midpart of the nematodes was used for PCR-RLFP analysis and sequencing of the nuclear rDNA containing both ITS (Internal Transcribed Spacers) and gene 5.8S (D'Amelio et al 2000, Kijewska et al 2002, Pontes et al 2005.…”

Section: Parasite Counts and Identificationmentioning

confidence: 99%

Occurrence of anisakid nematodes in Atlantic cod (Gadus morhua) and Greenland cod (Gadus ogac), West Greenland

Mouritsen

Hedeholm

Schack

et al. 2010

Acta Parasitologica

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Anisakid nematodes commonly infect gadids, and are of economic and aesthetic importance to the commercial fishing industry in Greenland as some species are pathogenic to humans. However, very little is known about the occurrence of these parasites and their impact on the hosts in Greenland waters. During a survey in 2005, stomach sample of 227 Atlantic cod (Gadus morhua) and 64 Greenland cod (Gadus ogac) was collected in Godthaab and Sisimiut fiord systems in West Greenland waters. All cod were dissected for stomach contents and anisakid nematodes were removed from the visceral cavity. Third stage larvae (L3) of three anisakid species were found, including Contracaecum osculatum (Rudolphi, 1802), Anisakis simplex (Rudolphi, 1809) and Hysterothylacium aduncum (Rudolphi, 1802). Molecular identification by PCR-RFLP indicated the presence of A. simplex s.s. and the sibling species C. osculatum B and C. The prevalence of infection by C. osculatum was higher in Greenland cod (84.3%) than in Atlantic cod (73.9%) whereas the prevalence of A. simplex showed an opposite pattern (Greenland cod 8.3%; Atlantic cod 24.2%). Only one G. morhua (1.0%) was infected by H. aduncum. No gender specific difference in both nematode species regarding prevalence of infection and mean infection intensity was evident, and there was no relationship between fish condition and the intensity of nematode infections. Standardised for size, capelin-eating cod were in better condition and more heavily infected than fish subsisting on alternative prey at the point of collection. Hence, nematode infections in the two gadids seem governed in part by feeding behaviour, and capelin appears a significant source of larval anisakids.

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Genetic markers in ribosomal DNA for the identification of members of the genus Anisakis (Nematoda: Ascaridoidea) defined by polymerase-chain-reaction-based restriction fragment length polymorphism

Cited by 292 publications

References 12 publications

Red vent syndrome in wild Atlantic salmon Salmo salar in Scotland is associated with Anisakis simplex sensu stricto (Nematoda: Anisakidae)

Red vent syndrome in wild Atlantic salmon Salmo salar in Scotland is associated with Anisakis simplex sensu stricto (Nematoda: Anisakidae)

The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species

Occurrence of anisakid nematodes in Atlantic cod (Gadus morhua) and Greenland cod (Gadus ogac), West Greenland

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