Cercarial trematodes in freshwater snails from Bangkok, Thailand: prevalence, morphological and molecular studies and human parasite perspective

We investigated the prevalence, morphological characters and molecular classifications of trematode cercariae in freshwater snails randomly collected from 59 sampling localities in Bangkok from May 2018 to March 2019. We used a crushing technique to observe the cercarial stage inside each snail body and amplified the internal transcribed spacer 2 regions of cercarial DNA using polymerase chain reaction methodology. The associated phylogenetic tree was reconstructed using Bayesian inference analyses. A total of 517 of 15 621 examined snails were infected with trematode cercariae, and the infected snails were classified into 11 species of seven families with a 3.31% overall prevalence of the infection. The Bithynia siamensis siamensis snail displayed the highest prevalence of infection (16.16%), whereas the Physella acuta snail exhibited the lowest prevalence (0.08%) of infection. Eight morphological types of cercariae were observed. The highest prevalence of infection was observed in mutabile cercaria (1.86%). Based on molecular investigations, the phylogram revealed eight cercaria types assigned to at least nine digenean trematode families, of which five belong to groups of human intestinal flukes. Although, with the exception of schistosome cercaria, trematode cercariae are not known to directly damage humans, understanding the general biology of trematode cercariae (including diversity, distribution, infection rates and host range) is important and necessary for the prevention and control of parasitic transmission that impacts aquatic cultivations, livestock farming and human health.

An efficient photograph-based quantitative method for assessing castrating trematode parasites in bivalve molluscs

Parasitic castration of bivalves by trematodes is common, and may significantly reduce the reproductive capacity of ecologically important species. Understanding the intensity of infection is desirable, as it can indicate the time that has passed since infection, and influence the host's physiological and reproductive response. In addition, it is useful to know the developmental stage of the trematode, to understand trematode population trends and reproductive success. However, most existing methods (e.g. visually estimating the degree of infection) to assess intensity are approximate only and not reproducible. Here, we present a method to accurately quantify the percentage of bivalve gonad filled with digenean trematode tissue, based on small squashes of gonad tissue rapidly photographed under light microscopy. A maximum of 15 photographs is required to determine the percentage of the whole gonad occupied by trematodes with a minimum of 90% confidence, with smaller mussels requiring fewer. In addition, the stage of trematode infection can be assessed because full sporocysts, spent sporocysts and free cercariae are clearly distinguishable. Although variation exists in the distribution of trematodes in gonad tissue, and thus in the estimate of percentage of the gonad filled with trematodes, this method represents a marked improvement on current coarse assessments of infection which typically focus on binary presence/absence measures. This technique can be used to facilitate a more sophisticated understanding of host–parasite interactions in bivalves, and can inform the conservation and reproductive biology of environmentally crucial species.

First Report of Clinostomum complanatum (Trematoda: Digenea) in European Perch (Perca fluviatilis) from an Italian Subalpine Lake: A Risk for Public Health?

Clinostomum complanatum, a digenean trematode of the Clinostomidae family, is a fish-borne zoonotic parasite responsible for Halzoun syndrome in humans and is transmitted through the consumption of raw or undercooked freshwater fish. Of the total of 112 specimens of European perch (Perca fluviatilis) sampled from a subalpine lake (Lake Endine) in North Italy in 2019, 21 (18.75%) tested positive for encysted metacercariae in the fillet. This study reports the first isolation of C. complanatum in P. fluviatilis and highlights the possible zoonotic risk for consumers, since P. fluviatilis is a food fish used in the traditional local cuisine.

Digenean trematode cysts within the heads of threatened Galaxiella species (Teleostei : Galaxiidae) from south-eastern Australia

The dwarf galaxias (Galaxiella pusilla) and little galaxias (Galaxiella toourtkoourt) are both threatened freshwater fish from south-eastern Australia. Occasionally populations have been found with enlarged heads associated with the accumulation of ‘white balls’, but the cause of these deformities has not previously been investigated. In this study, histopathology and molecular techniques were employed to identify cysts extracted from the heads of Galaxiella species across six populations. Histopathology and DNA sequences from both mitochondrial (cytochrome c oxidase subunit I) and nuclear (ribosomal internal transcribed spacer 1) regions identified the cysts as metacercariae of Apatemon gracilis (Rudolphi, 1819), a cosmopolitan digenean trematode species. Heavy infestations of trematode metacercariae within Galaxiella populations are of concern due to the potential to cause increased mortality associated with altered behaviour of the fish host that increases the likelihood of predation. Direct mortality from infestations is also possible, but not quantified in this study.

Helminths from the red-eared slider Trachemys scripta elegans (Chelonia: Emydidae) in marshes from the eastern Iberian Peninsula: first report of Telorchis attenuata (Digenea: Telorchiidae)

The present work describes the presence of a digenean in the red-eared turtle Trachemys scripta elegans (Wied-Neuwied, 1839) in marshes of the Valencian Community. The faeces and intestinal tract of 105 animals were examined. Only one helminth species was found and identified as the digenean trematode Telorchis atenuatta (Goldberger, 1911), present in the 7.6% of the animals analysed. This is the first report of the parasite in sliders from Spain. Although conclusions are preliminary due to the limited sampling, our results suggest that the presence of red-eared turtles in new habitats may increase the risk of introducing new microorganisms and new diseases with them, altering the sanitary status of the autochthonous terrapins Mauremys leprosa (Schweigger, 1812) and Emys orbicularis (Linnaeus, 1758).

Morphological and molecular analysis of metacercariae of Diphtherostomum brusinae (Stossich, 1888) Stossich, 1903 from a new bivalve host Mytilus galloprovincialis

The digenean trematode Diphtherostomum brusinae (Stossich, 1888) Stossich, 1903 presents a complex life cycle that may involve more than one intermediate host. The present study represents the first description of the metacercariae from D. brusinae infecting the labial palps of a new intermediate host, Mytilus galloprovincialis, in the Aveiro estuary, Portugal. The morphology of this parasitic stage was studied by light (LM) and scanning electron microscopy (SEM) and some differences were reported (body and sucker sizes, and spine distribution and shape). In this work, the 18S partial region of the ribosomal DNA was sequenced from D. brusinae metacercariae isolated from M. galloprovincialis collected in different localities of the Aveiro estuary. In addition, sequences from the same region of the 18S rDNA were obtained from D. brusinae cercariae and metacercariae, hosted by Nassarius reticulatus and Cerastoderma edule, respectively. No intraspecific polymorphism was detected in the 18S partial region, since there was 100% homology among all the sequences analysed. The same comparison was made for the ITS1, and we observed intraspecific polymorphism in this region. To our knowledge, this is the first report of D. brusinae metacercariae infecting the mussel M. galloprovincialis with support from morphological and molecular data.