Cercariae of intertidal nassariid gastropods: Taxonomy, specificity and ecology

"The biodiversity and ecology of marine cercariae in Australia is poorly known. Although nassariid gastropods are widely distributed in Australia and are expected to have a diverse parasite fauna similar to that of other nassariids worldwide, their cercariae have not been characterised. Nassarius dorsatus, N. olivaceus and N. pullus were collected at three separate estuarine localities in central Queensland between August 2004 and August 2006 and their emergent cercariae characterised using morphological and molecular methods. Fifteen morphotypes of cercariae from six families that infect fishes and birds as definitive hosts were characterised. The highest richness was within the Acanthocolpidae (nine morphotypes) which comprised three cercariae interpreted as species of Stephanostomum and a complex of six unusual cercariae with a ventral keel. Molecular characterisation of these six cercariae supported their placement in the Acanthocolpidae and indicated that they comprised only three species, two of which displayed marked phenotypic plasticity. Molecular analysis supported the characterisation of the three species of Stephanostomum and suggested that one of the unusual cercariae was a species of Stephanostomoides. Dissection of fish from Sandy Point provided a single match; the sequence for a metacercaria in Valamugil buchanani, the blue-tail mullet, was identical to that of one of the species of Stephanostomum. There were two cercariae from the Echinostomatidae and one from each of the Lepocreadiidae, Microphallidae, Opecoelidae and Zoogonidae. Molecular analyses supported the placement of the cercariae from the Echinostomatidae, Lepocreadiidae, Opecoelidae and Zoogonidae within their respective families. The cercaria from the Lepocreadiidae was identified by comparison to known adult data and putatively corresponds to Prodistomum keyam, a parasite of Monodactylus argenteus, the silver moony or butter bream, common in the study region. Morphological analysis supported the characterisation of both of the Echinostomatidae as species of Himasthla. Although no other cercariae could be characterised to genus level, the opecoelid cercaria was determined to be in the subfamily Opecoelinae. Nassarius olivaceus was infected by three acanthocolpid cercariae and one cercaria from each of the other families. Nassarius dorsatus was infected by five acanthocolpid cercariae and one each from the Echinostomatidae and Lepocreadiidae. Nassarius pullus was infected by a single cercaria from the Echinostomatidae. Most cercariae were specific to a single species of gastropod; only four infected two species and none infected all three. Cercariae that infect fish as a definitive host were found at all three localities; but cercariae that infect birds were restricted to a single locality, Sandy Point at Corio Bay, important for domestic and migratory bird species. These results emphasise the relative significance of Sandy Point as an important bird habitat. In total, 126 infections were diagnosed (115 by cercarial emergence), with both the highest frequency of infection and number of infections in N. olivaceus (frequency 5.3%; n=102; 81% of infections), followed by N. dorsatus (1.1%; 20; 16%) and N. pullus (0.2%; 4; 3%). The highest number of infections occurred at Sandy Point (66; 52% of infections), followed by Cawarral Creek (38; 30%) and then Ross Creek (22; 17%). The highest number of infections was by the Acanthocolpidae (54; 43%). Diagnosis of infection by emergence was suitable for all cercariae except the opecoelid, which was underdiagnosed. Nassarius olivaceus had the longest survival in captivity (~4.5 y), followed by N. dorsatus (~0.8 y) and N. pullus (~0.3 y). Almost all infections remained patent for the life of the gastropod. Families commonly found in nassariids worldwide include Acanthocolpidae, Echinostomatidae, Lepocreadiidae and Zoogonidae. Nassariid gastropods are also the dominant marine first intermediate hosts for the Acanthocolpidae, Lepocreadiidae and Zoogonidae and one of the three most common first intermediate host families for species of marine Himasthla. Molecular relationships also indicate that the Acanthocolpidae and Brachycladiidae may actually be comprised of three discrete groups, the Stephanostomum + Monostephanostomum group, the Brachycladiidae, and a group of non-Stephanostomum species comprising Pleorchis, Tormopsolus and Stephanostomoides. This research adds information about the parasite assemblages of nassariid gastropods in Australia, adding 12 new cercarial species and 16 new host-parasite records. Cercarial reports for the Acanthocolpidae, Echinostomatidae, Lepocreadiidae, Opecoelidae and Zoogonidae have been comprehensively reviewed and patterns of association between nassariids and their common parasite families presented. As well, this research has provided a potential avenue for a review of the Acanthocolpidae, a family which is currently considered a ‘catch-all’ group.

Use of parasites as indicators of estuarine health and the presence of important host species

Trematode parasites in snail hosts have been proposed as potential bioindicators of estuarine ecosystem health. Both the definitive and snail host must be present in the system for successful transmission and the diversity and abundance of larval trematodes in snail first intermediate host populations directly reflects the diversity and abundance of definitive hosts in the ecosystem. In addition, trematodes in snail hosts are negatively influenced by environmental impacts and abundance and diversity are lower in impacted habitats. Estuarine snails proposed as suitable bioindicators include species from the Potamididae, Cerithiidae, Nassariidae, Batillariidae and Hydrobiidae. The northern hemisphere snail Nassarius obsoletus hosts nine species of parasite from eight families that infect fish, birds or reptiles. Very little is known about parasites of snails in Australia estuaries. A study of three estuarine locations in Capricornia compared the parasites of three species of Nassarius: N. dorsatus, N. olivaceus and N. pullus. Trematodes from six families that infect fish and birds were collected, including eight species from a single family of fish parasites. Fish parasites were present across all locations, but bird parasites were collected from a single location. This location is listed as an important bird habitat, both for endangered and migratory species. No reptile parasites were collected from these locations, although a reptile parasite was recently reported from N. dorsatus in Townsville. In addition to indicating ecosystem health, parasites in snail hosts may indicate the relative importance of selected locations for particular types of definitive host species; especially birds, reptiles or mammals. Using parasite information as an additional bioindicator for estuarine health may help to inform the presence of important definitive hosts

Host specificity and ecology of digenean parasites of nassariid gastropods in Central Queensland, Australia, with comments on host-parasite associations of the Nassariidae

Nassariid gastropods are well represented in Australia, and Nassarius dorsatus (Röding), Nassarius olivaceus (Bruguière), and Nassarius pullus (Linnaeus) are common in the intertidal mangroves of the Capricornia region in Central Queensland. A survey of the digenean trematode fauna of nassariids of the region revealed cercariae of 12 species from six trematode families associated with these gastropods. Six species of Acanthocolpidae were found, including three putative species of the genus Stephanostomum. Additionally, two species of Himasthlidae and one species each of Microphallidae, Opecoelidae, Zoogonidae, and Lepocreadiidae were identified. The majority of the trematode species identified were found in only one host species; only four infected more than one species of nassariid. Sixteen new hostparasite combinations were added to the fauna known to infect the Nassariidae. Broader examination of host-parasite associations shows that families using nassariids commonly as first intermediate host are the Acanthocolpidae, Lepocreadiidae, and Zoogonidae, as well as Himasthla spp. from the Himasthlidae. Nassariid gastropods are also the dominant marine first intermediate hosts for the Acanthocolpidae, Lepocreadiidae, and Zoogonidae and one of the three most common first intermediate host families for marine Himasthla spp. © 2018, Yale Journal of Biology and Medicine Inc. All rights reserved

Phenotypic plasticity of six unusual cercariae in nassariid gastropods and their relationships to the Acanthocolpidae and Brachycladiidae (Digenea)

Species-level relationships of a complex of six putative acanthocolpid cercariae with ventral keels from nassariid gastropods from Capricornia, Queensland (Cercaria capricornia I–VI) were explored using mitochondrial and nuclear DNA sequence data. Analysis of the large subunit of nuclear ribosomal DNA (rDNA), the internal transcribed spacer 2 rDNA and subunit 3 of the nicotinamide-adenine-dinucleotide dehydrogenase mitochondrial gene indicates that the six distinct morphotypes previously identified relate to only three discrete genotypes. Consequently, the six cercarial morphotypes that were inferred to represent six species and distinguished previously based on morphology and behaviour, are considered here to be comprised of three species (i.e. Cercaria capricornia I and II are considered the same species, C. capricornia types III to V are considered to belong to a second species and C. capricornia VI is considered to comprise the third species in this complex). This recognition of reduced diversity following molecular analysis runs contrary to the more frequently reported uncovering of cryptic diversity, especially when larval trematodes are examined. Phylogenetic analysis of these morphologically unusual cercariae supports their inclusion in the Acanthocolpidae. Bayesian inference analysis identifies three distinct clades: (a) Stephanostomum + Monostephanostomum; (b) the Brachycladiidae and Pleorchis; and (c) Tormopsolus and the present cercariae. Genus-level identification of these cercariae remains uncertain given the current lack of available comparative genetic data on nucleic acid databases. © 2017 Elsevier B.V

Vigilance to limit the bidirectional introduction and co-circulation of virus serotypes aims to reduce risk of severe dengue disease in North East India

Dengue is a serious re-emerging arboviral disease that presents a global human health challenge. Infection is caused by one of several recognised serotypes of dengue virus, a member of the mosquito-transmitted Flaviviridae family of human pathogens which also includes yellow fever, Zika and West Nile viruses. The greatest burden of dengue is borne by countries in the Asia Pacific region, in particular South East Asia and the Indian subcontinent where infection breaks out with increasing frequency and intensity. The World Health Organization South East Asia Region has a population of 1.3 billion people and includes 11 countries, of which Thailand, Myanmar and India have the highest reported incidence of dengue. Infection outcomes vary from the commonly asymptomatic or a self-limiting fever to more severe manifestations such as dengue haemorrhagic fever and dengue shock syndrome. Although primary infection with one serotype elicits lifelong immunity against homologous reinfection, severe dengue is linked to an antibody-related immunopathological response that is triggered by secondary infection with a heterologous serotype. The principal vectors of transmission are the day-biting mosquitoes Aedes aegypti and Ae. albopictus, the respective preference of which for urban and rural habitats has facilitated the rapid and continued spread of dengue across tropical and sub-tropical climatic zones. The narrow elevated corridor of North East India was considered historically to be non-endemic for dengue and as such separated areas of high endemicity in the Bengal Delta and the other zones of India from those in Thailand and Myanmar. In these regions the predominant serotype is usually distinct, so a breach of this geographical bottleneck would potentially enable the co-circulation of virus serotypes, thus posing an increased risk of severe disease upon secondary infection. This alarming scenario may now be realised following the notification of clinical cases in the states of Arunachal Pradesh, Assam and Manipur in recent years. While incidence is still sporadic in North East India limitation of local outbreaks is required through vigilant adherence to the integrated implementation of clinical management, diagnosis, surveillance and vector control in order to prevent the transcontinental spread of dengue

A complex of putative acanthocolpid cercariae (Digenea) from Nassarius olivaceus and N. dorsatus (Gastropoda: Nassariidae) in Central Queensland, Australia

Cercariae capricornia I–VI, six new cercariae putatively identified as belonging to the Acanthocolpidae, are described and named from prosobranch gastropods of the family Nassariidae collected from the intertidal zone in the Capricornia region, Central Queensland, Australia. Four species are reported from Nassarius olivaceus and two from N. dorsatus. The cercariae have a unique and complex three- dimensional body shape, including a keel, which differentiates them from previously described acanthocolpid cercariae. These are the first cercariae to be described from these gastropods

A review of the currently recognised zoogonid cercariae, including the identification and emergence ecology of Cercaria capricornia XI (Digenea: Zoogonidae) from Nassarius olivaceus (Gastropoda: Nassariidae) in Central Queensland, Australia : New zoogonid cercaria from Australia

In the present study, we review the known zoogonid cercariae, summarise their life-cycles and first intermediate host distributions, and present a new cercaria, Cercaria capricornia XI (Digenea: Zoogonidae), which was found in one of three nassariid gastropods, Nassarius olivaceus (Bruguière), surveyed in the intertidal zone in the Capricornia region of Central Queensland, Australia. Morphological data and molecular analysis of the ITS2 rDNA region support placement of this cercaria in the family Zoogonidae but do not allow any further resolution of its identity. There are now fifteen cercariae described as belonging to the Zoogonidae; thirteen of these, including the present species, infect neogastropods as first intermediate hosts and two use vetigastropods. This study reinforces the pattern that the Nassariidae is by far the most commonly reported family for the Zoogonidae. Given its richness we predict that the Nassariidae will prove to harbour many more zoogonid species

Two new Stephanostomum-like cercariae (Digenea: Acanthocolpidae) from Nassarius dorsatus and N. olivaceus (Gastropoda: Nassariidae) in Central Queensland, Australia

Two new Stephanostomum-like cercariae, Cercaria capricornia VII and Cercaria capricornia VIII (Digenea: Acanthocolpidae), are described from the nassariid gastropods Nassarius dorsatus and Nassarius olivaceus collected from the intertidal zone in the Capricornia region of Central Queensland, Australia. Morphological analysis of these new cercariae was augmented with DNA sequence data from the large subunit (LSU) ribosomal DNA region to aid in identification. Bayesian inference analysis of the LSU rDNA revealed that these putative acanthocolpid cercariae nested well within a clade containing species of Stephanostomum, which along with morphological data, suggests they are species of Stephanostomum. Comparative analysis of LSU rDNA sequences also indicates that these two cercariae are not S. adlardi, S. bicoronatum, S. tantabiddii or S. cf. uku, all species known from Australian fishes. The secondary structure of the internal transcribed spacer 2 (ITS2) rDNA region was inferred for these two cercariae using minimum free energy modelling algorithms. Both cercarial types displayed a four helix ITS2 secondary structure model and differed from each other by two compensatory base changes (CBCs) and nine hemi-CBCs

A review of the currently recognised opecoelid cercariae, including the identification and emergence ecology of Cercaria capricornia XII (Digenea: Opecoelidae) from Nassarius olivaceus (Gastropoda: Nassariidae) in Central Queensland, Australia

A new opecoelid cercaria, Cercaria capricornia XII, is reported from Nassarius olivaceus in Capricornia, Central Queensland, Australia. Combined molecular and morphological data indicate that this cercaria is a member of the subfamily Opecoelinae. Cercaria capricornia XII is the first known opecoeline cercaria reported from a nassariid gastropod. Cercaria capricornia XII can be distinguished fromother opecoelid cercariae by the combination of the presence of a 2-pointed stylet, body length and width, and the size of the tail. The emergence pattern for C. capricornia XII in captivitywas erratic; rapid emergences of thousands of cercariae were interspersed by periods that sometimes exceeded a month in which no emergence occurred. There was no detectable pattern to or stimulus of the emergence. The molluscan host range of opecoelids is analysed in detail. Gastropods from the Buccinoidea, Cerithioidea and Rissooidea are hosts to both opecoeline and plagioporine cercariae, but the dominant subfamily infecting the Cerithioidea and Rissooidea is the Plagioporinae. The dominant marine host gastropod superfamily for opecoeline cercariae is the Buccinoidea; the family Nassariidae is contained in the Buccinoidea. The range of gastropod superfamilies known as hosts of plagioporines is much broader than that for opecoelines, which may be explained by the relative size of the two opecoelid families and perhaps by indications that the Plagioporinae is polyphyletic

A quantitative PCR (TaqMan) assay for pathogenic Leptospira spp

Background: Leptospirosis is an emerging infectious disease. The differential diagnosis of leptospirosis is difficult due to the varied and often "flu like" symptoms which may result in a missed or delayed diagnosis. There are over 230 known serovars in the genus Leptospira. Confirmatory serological diagnosis of leptospirosis is usually made using the microscopic agglutination test (MAT) which relies on the use of live cultures as the source of antigen, often performed using a panel of antigens representative of local serovars. Other techniques, such as the enzyme linked immunosorbent assay (ELISA) and slide agglutination test (SAT), can detect different classes of antibody but may be subject to false positive reactions and require confirmation of these results by the MAT. Methods: The polymerase chain reaction (PCR) has been used to detect a large number of microorganisms, including those of clinical significance. The sensitivity of PCR often precludes the need for isolation and culture, thus making it ideal for the rapid detection of organisms involved in acute infections. We employed real-time (quantitative) PCR using TaqMan chemistry to detect leptospires in clinical and environmental samples. Results and Conclusions: The PCR assay can be applied to either blood or urine samples and does not rely on the isolation and culture of the organism. Capability exists for automation and high throughput testing in a clinical laboratory. It is specific for Leptospira and may discriminate pathogenic and non-pathogenic species. The limit of detection is as low as two cells