Fundamentals of Methods for Studying Fish Blood Parasites

Losev, Alexander; Овчаренко, Н. А.

doi:10.1615/hydrobj.v40.i2.110

Cited by 4 publications

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“…For the other two Czech isolates, the available laboratory cultures were used for this purpose. As for the material from Ukraine, trypanosomes were isolated from the fresh blood of infected frogs by the microhaematocrite method [ 30 ] and used for DNA extraction as described elsewhere [ 31 ].…”

Section: Methodsmentioning

confidence: 99%

Diversity and evolution of anuran trypanosomes: insights from the study of European species

et al. 2018

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BackgroundAmphibian trypanosomes were the first ever described trypanosomatids. Nevertheless, their taxonomy remains entangled because of pleomorphism and high prevalence of mixed infections. Despite the fact that the first species in this group were described in Europe, virtually none of the trypanosomes from European anurans was analyzed using modern molecular methods.MethodsIn this study, we explored the diversity and phylogeny of trypanosomes in true frogs from Europe using light microscopy and molecular methods.ResultsA comparison of observed morphotypes with previous descriptions allowed us to reliably identify three Trypanosoma spp., whereas the remaining two strains were considered to represent novel taxa. In all cases, more than one morphotype per blood sample was observed, indicating mixed infections. One hundred and thirty obtained 18S rRNA gene sequences were unambiguously subdivided into five groups, correspondent to the previously recognized or novel taxa of anuran trypanosomes.ConclusionsIn this work we studied European frog trypanosomes. Even with a relatively moderate number of isolates, we were able to find not only three well-known species, but also two apparently new ones. We revealed that previous assignments of multiple isolates from distant geographical localities to one species based on superficial resemblance were unjustified. Our work also demonstrated a high prevalence of mixed trypanosome infections in frogs and proposed a plausible scenario of evolution of the genus Trypanosoma.

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Section: Methodsmentioning

confidence: 99%

Diversity and evolution of anuran trypanosomes: insights from the study of European species

et al. 2018

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“…Fish specimens were between 0.5 and 5 years old. The presence of trypanosomes in the blood and the level of parasitaemia were established on smears and recalculated per millilitre of blood as described before (Losev and Ovcharenko 2004). The 'seasonal' parasitaemia and infection prevalence were assayed in the following conditions: water temperature rising from +5 °C to +17 °C (spring); water temperature between +17 °C and +28 °C (summer); water temperature falling from +17 °C to +7 °C (autumn).…”

Section: Methodsmentioning

confidence: 99%

“…Parasites were isolated from fresh blood using a protocol described elsewhere (Losev and Ovcharenko 2004). Concentrated cells were stored in SDS-EDTA solution .…”

Section: Methodsmentioning

confidence: 99%

High prevalence of trypanosome co-infections in freshwater fishes

Grybchuk-Ieremenko¹,

Losev²,

Kostygov³

et al. 2014

FOLIA PARASIT

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Abstract:One thousand three hundred seventy three fish specimens of eight different species from the vicinity of Kyiv, Ukraine, were examined for the presence of trypanosomes and 921 individuals were found to be infected. The prevalence of infection ranged from 24% in freshwater bream, Abramis brama (Linnaeus), to 100 % in spined loach, Cobitis 'taenia' Linnaeus. The level of parasitaemia also varied significantly between generally mild infections in pikeperch, Sander lucioperca (Linnaeus), and heavy ones in C. 'taenia'. In most cases the infections with trypanosomes were asymptomatic. Cases of co-infection with species of Trypanoplasma Laveran et Mesnil, 1901 were documented for five out of eight examined host species. Molecular analysis of the 18S rDNA sequences revealed that four hosts, namely northern pike, Esox lucius Linnaeus, freshwater bream, spined loach and European perch, Perca fluviatilis Linnaeus, were simultaneously infected with two different trypanosome species. Our findings advocate the view that to avoid the risk posed by mixed infections, subsequent molecular taxonomic studies should be performed on clonal lines derived from laboratory cultures of fish trypanosomes.

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Host specificity, pathogenicity, and mixed infections of trypanoplasms from freshwater fishes

et al. 2014

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This work summarizes the results of the 8-year study focused on Trypanoplasma sp. parasitizing freshwater fishes in the vicinity of Kyiv, Ukraine. Out of 570 fish specimens of 2 different species analyzed, 440 individuals were found to be infected. The prevalence of infection ranged from 24 % in Abramis brama Linnaeus (freshwater bream) to 100 % in Cobitis taenia Linnaeus (spined loach). The level of parasitemia also varied between moderate in freshwater bream and very high in spined loach. Interestingly, no clinical manifestations of trypanoplasmosis were observed even in extremely heavily infected C. taenia. We hypothesize that different species may differ in evolutionary timing allowing for reciprocal adaptation of the members of the "host-parasite" system. Molecular analysis of the 18S rRNA sequences revealed that several specimens were simultaneously infected with at least two different trypanoplasm species. To the best of our knowledge, this is the first report of the mixed infection with fish trypanoplasms.

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Fundamentals of Methods for Studying Fish Blood Parasites

Cited by 4 publications

References 0 publications

Diversity and evolution of anuran trypanosomes: insights from the study of European species

Diversity and evolution of anuran trypanosomes: insights from the study of European species

High prevalence of trypanosome co-infections in freshwater fishes

Host specificity, pathogenicity, and mixed infections of trypanoplasms from freshwater fishes

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