Euclinostomum heterostomum (Rudolphi, 1809) (Trematoda): life-cycle,growth and development of the metacercaria and adult

The life-history of Euclinostomum heterostomum (Rudolphi, 1809), a clinostomid trematode occurring in the oesophagus and buccal cavity of pond heron Ardeola grayii and cattle egret Bubulcus ibis at Visakhapatnam, India, has been traced from egg to adult stage, based on field and laboratory studies. Eggs released by gravid flukes hatched into miracidia in 10 days. The snail Indoplanorbis exustus served as the first intermediate host and the intramolluscan phase, involving the development of two redial generations and cercariae, took two months. Natural infections with the metacercaria were found in the kidneys and liver of the fish Channa punctatus and muscles of C. orientalis. Laboratory experiments demonstrated that more than 2 months are required for the development of the cercaria into the infective metacercarial stage. The morphology of all of the stages in the life-cycle has been studied in detail, as well as the growth characteristics of the metacercaria within the fish host. Two phases in the metacercarial growth were noted, a lag phase followed by an exponential phase. Allometric growth parameters of body proportions and organs of the metacercaria and the adult are compared. The validity of various species included in the genus Euclinostomum is discussed.     

Growth and development of Pleurogenoides orientalis (Srivastava, 1934) (Trematoda: Lecithodendriidae) in the intermediate and definitive hosts

Two phases with an intervening brief lag-phase are recognized during the growth of the metacercaria of Pleurogenoides orientalis in the intermediate host (nymphs of Tramea limbata), the first phase being characterized by rapid growth and organogeny and the second phase by limited growth and development. The phases of growth and development of the metacercaria are correlated with various events in the cyst-forming process. The growth of the adult fluke in the definitive host (Rana cyanophlyctis) is characterized by a short initial lag-phase followed by a rapid phase during which there is rapid growth. Comparison of allometric growth parameters of various organs of the metacercaria and of the adult revealed definite allometric shifts. The validity of various species included in the genus Pleurogenoides is examined in the light of observations made on intraspecific variation in P. orientalis.     

Morphological variation in the cosmopolitan fish parasite Neobenedenia girellae (Capsalidae: Monogenea)

Intra-species morphological variation presents a considerable problem for species identification and can result in taxonomic confusion. This is particularly pertinent for species of Neobenedenia which are harmful agents in captive fish populations and have historically been identified almost entirely based on morphological characters. This study aimed to understand how the morphology of Neobenedenia girellae varies with host fish species and the environment. Standard morphological features of genetically indistinct parasites from various host fish species were measured under controlled temperatures and salinities. An initial field-based investigation found that parasite morphology significantly differed between genetically indistinct parasites infecting various host fish species. The majority of the morphological variation observed (60%) was attributed to features that assist in parasite attachment to the host (i.e. the posterior and anterior attachment organs and their accessory hooks) which are important characters in monogenean taxonomy. We then experimentally examined the effects of the interaction between host fish species and environmental factors (temperature and salinity) on the morphology of isogenic parasites derived from a single, isolated hermaphroditic N. girellae infecting barramundi, Lates calcarifer. Experimental infection of L. calcarifer and cobia, Rachycentron canadum, under controlled laboratory conditions did not confer host-mediated phenotypic plasticity in N. girellae, suggesting that measured morphological differences could be adaptive and only occur over multiple parasite generations. Subsequent experimental infection of a single host species, L. calcarifer, at various temperatures (22, 30 and 32?°C) and salinities (35 and 40‰) showed that in the cooler environments (22?°C) N. girellae body proportions were significantly smaller compared with warmer temperatures (30 and 32?°C; P?<?0.0001), whereas salinity had no effect. This is evidence that temperature can drive phenotypic plasticity in key taxonomic characters of N. girellae under certain environmental conditions.     

Hidden diversity and evolutionary trends in malacosporean parasites (Cnidaria: Myxozoa) identified using molecular phylogenetics

Malacosporeans represent a small fraction of myxozoan biodiversity with only two genera and three species described. They cycle between bryozoans and freshwater fish. In this study, we (i) microscopically examine and screen different freshwater/marine fish species from various geographic locations and habitats for the presence of malacosporeans using PCR; (ii) study the morphology, prevalence, host species/habitat preference and distribution of malacosporeans; (iii) perform small subunit/large subunit rDNA and Elongation factor 2 based phylogenetic analyses of newly gathered data, together with all available malacosporean data in GenBank; and (iv) investigate the evolutionary trends of malacosporeans by mapping the morphology of bryozoan-related stages, host species, habitat and geographic data on the small subunit rDNA-based phylogenetic tree. We reveal a high prevalence and diversity of malacosporeans in several fish hosts in European freshwater habitats by adding five new species of Buddenbrockia and Tetracapsuloides from cyprinid and perciform fishes. Comprehensive phylogenetic analyses revealed that, apart from Buddenbrockia and Tetracapsuloides clades, a novel malacosporean lineage (likely a new genus) exists. The fish host species spectrum was extended for Buddenbrockia plumatellae and Buddenbrockia sp. 2. Co-infections of up to three malacosporean species were found in individual fish. The significant increase in malacosporean species richness revealed in the present study points to a hidden biodiversity in this parasite group. This is most probably due to the cryptic nature of malacosporean sporogonic and presporogonic stages and mostly asymptomatic infections in the fish hosts. The potential existence of malacosporean life cycles in the marine environment as well as the evolution of worm- and sac-like morphology is discussed. This study improves the understanding of the biodiversity, prevalence, distribution, habitat and host preference of malacosporeans and unveils their evolutionary trends.     

Phylogenetic relationships among monogenean gill parasites (Dactylogyridea, Ancyrocephalidae) infesting tilapiine hosts (Cichlidae): systematic and evolutionary implications

We studied the systematics of 14 species of monogenean (Ancyrocephalidae) gill parasites from West African tilapiine hosts (Cichlidae) using both morphological and genetic data. With these tools, we were able to: (i) confirm the validity of the previously described morphological parasite species and of the genus Scutogyrus; (ii) propose that some stenoxenous species (i.e., parasite species with more than one host) may be composed of sister species (e.g., Cichlidogyrus tilapiae); (iii) state that the use of the morphology of the haptoral sclerites is more suitable to infer phylogenetic relationships than the morphology of the genitalia (which seems to be more useful to resolve species-level identifications, presumably because of its faster rate of change). These results imply that: (i) the specificity of these monogenean parasites is greater than initially supposed (what were thought to be stenoxenous species may be assemblages of oïoxenous sister species); (ii) related species groups (i.e., “tilapiae,” “halli,” and “tiberianus”) have to be, as genus Scutogyrus, validated within the 54 ancyrocephalid species described from 18 species of tilapiine hosts in West Africa, (iii) the group “tilapiae,” due to its morphology and host range, have to be considered as being the most primitive; (iv) the occurrence of lateral transfers and parallel speciation processes are necessary to describe the repartition of the newly described parasite groups on the three host genera studied (Tilapia, Oreochromis, and Sarotherodon).     

Brachylaima asakawai sp. nov. (Trematoda: Brachylaimidae), a rodent intestinal fluke in Hokkaido,Japan, with a finding of the first and second intermediate hosts

In the 1970’s and 1980’s, an unknown species of the genus Brachylaima (Trematoda: Brachylaimidae) had been recorded from the intestines of Rattus norvegicus and Apodemus speciosus in Hokkaido, Japan. The rodent fluke was characteristic in extending a bilateral vitellarium till the level of posterior margin of anterior testis and in keeping almost the same-sized spherical ovary and testes. In this study, the rodent fluke was rediscovered from A. speciosus, Apodemus argenteus, and Myodes rufocanus in Hokkaido. The resultant parasite collection enabled us to make a mitochondrial DNA (mtDNA) barcode for tracking its intermediate hosts. The metacercaria of the rodent fluke was detected frequently from the kidney of three species of land snails (Discus pauper, Succinea lauta, and Ainohelix editha). However, its sporocyst with cercariae was found only from the hepatopancreas of D. pauper, a fairly small snail. The wide-spectrum of the second intermediate host seems to increase the chance of transmitting the parasite to various mammals and birds. The use of indigenous land snails as the first and second intermediate hosts, the distinctiveness of the mtDNA sequence, and the characteristic morphology of all the developmental stages prompted us to propose Brachylaima asakawai sp. nov. for the rodent intestinal fluke in Hokkaido. The present field survey suggests that the life cycle of the new species is primarily dependent on a predator-prey relationship between rodents and D. pauper.     

Novel and known myxobolids (Cnidaria,Myxozoa) infecting Chondrostoma angorense (Cypriniformes: Leuciscidae) in Turkey

Turkey has more than 200 endemic freshwater fish species, one of which is the Ankara nase, Chondrostoma angorense Elvira, 1987 (Cypriniformes: Leuciscidae), a food fish in northern Turkey. Like most endemic fish species in Turkey, its myxosporean parasite fauna (Cnidaria: Myxosporea) are not yet described. We surveyed twenty C. angorense from Lâdik Lake in northern Turkey, and identified two myxosporean parasites from gills of these fish: Myxobolus arrabonensis Cech, Borzák, Molnár, Székely, 2015, and a co-infection of a novel species, Myxobolus polati sp. nov. We characterized both infections based on myxospore morphology, morphometry, tissue tropism, small subunit ribosomal DNA sequence and phylogenetic analysis. Plasmodia of both species were observed in gills, but had distinct tropism: M. arrabonensis is an intrafilamental vascular type, and M. polati sp. nov. is an intralamellar vascular type. We identified M. arrabonensis on the basis of myxospore characters and 100% similarity to the type DNA sequence from the closely-related host C. nasus. The small subunit ribosomal DNA sequence of M. polati sp. nov. (1946 base pairs; GenBank Accession number MH392318) had a maximum similarity of 98% with any Myxobolus sp. from other Eurasian cypriniforms. Phylogenetic analysis revealed that M. polati sp. nov. is most closely related to gill-infecting Myxobolus diversicapsularis from Rutilus rutilus (L.). The present study is the first record of myxosporean species infecting C. angorense comprising a novel species, M. polati sp. nov. and a known species M. arrabonensis.     

Two novel myxosporean parasite species of Ceratomyxa Thélohan, 1892 from the banded cusk-eel Raneya brasiliensis (Kaup) (Ophidiiformes: Ophidiidae) off Patagonia,Argentina

We described two novel myxozoan parasite species Ceratomyxa argentina n. sp. and Ceratomyxa raneyae n. sp. from the gall bladder of Raneya brasiliensis (Kaup) from the Patagonian coast of Argentina. Both species can be distinguished from other ceratomyxids by myxospore and polar capsule (nematocyst) morphology and morphometry, fish host and geographic locality. Phylogenetic reconstruction using ssrDNA gene sequences showed that the two new species are placed in a long-branching ceratomyxid clade which also include Ceratomyxa appendiculata Thélohan, 1892, Ceratomyxa anko Freeman, Yokoyama and Ogawa, 2008, Ceratomyxa pantherini Gunter, Burger and Adlard, 2010 and Pseudoalataspora kovalevae Kalavati, MacKenzie, Collins, Hemmingsen and Brickle, 2013. This study documents additional biodiversity of marine myxozoans in the South Atlantic, a region still largely unexplored for this group of parasitic cnidarians.     

范尼道佛吸虫的生活史研究

范尼道佛吸虫的生活史包括毛蚴、胞蚴、尾蚴、囊蚴和成虫5个阶段。它的第一中间宿主为淡水壳菜,第二中间宿主为多种小型鲤形目和鲇形目鱼类,终末宿主是鳜。文中对这种吸虫的胞蚴、尾蚴、囊蚴和成虫的形态特征以及尾蚴的行为特征进行了观察和描述。在用人工方法成功地将尾蚴感染青后,对不同发育阶段的囊蚴在鱼体内的形态变化和器官发育进行了观察。       

Observations on the morphology and life-cycle of Procerovum varium (Onji & Nishio, 1916) (Trematoda: Heterophyidae)

The life-cycle of Procerovum varium (Digenea: Haplorchiinae) was studied experimentally and the morphology of stages in the life-cycle has been described and illustrated. Infections with adult flukes were found in the pond heron Ardeola grayii and heavy infections with metacercariae were found, attached to the liver of the fish Oryzias melastigma (Oryziatidae) occurring in a freshwater stream situated in Visakhapatnam, India. The cercariae developing in the snail Thiara tuberculatapossessed typical haplorchiine features and were characterised by the presence of numerous cystogenous glands. Early stages of metacercarial development occurred free in the muscles of the fish intermediate host. The larvae reached the liver at 5 days post-infection, encystment commenced 2 days later and 15-day-old metacercariae were found to be infective to chicks, ducks and mice that served as suitable experimental hosts. The adult flukes obtained from natural and experimental infections showed many intraspecific variations, especially in the size and shape of the expulsor which depend on the quantity of sperm it contains. The validity of various species described in the genus and differentiated on the basis of differences in the size of the expulsor has been examined. It is concluded that only three species of the genus, namely P. varium, P. cheni and P. calderoni, are valid. "P. sisoni" of Chen (1949) is confirmed as a synonym of P. varium. P. varium is reported for the first time from India.     

Morphological description of two bucephalid trematodes collected from freshwater fishes in the Uji River, Kyoto, Japan

The morphology of two species of bucephalids (Bucephalidae; Digenea; Trematoda), which since 1999 has caused a fish disease at the Uji River, Kyoto Prefecture, Japan, is described. Parabucephalopsis parasiluri Wang, 1985 was first recorded in the Uji River in 2000, and Prosorhynchoides ozakii (Nagaty, 1937) in 2005. The definitive host of both species is the Lake Biwa catfish (Silurus biwaensis), and the second intermediate hosts include many fish species from several families. P. parasiluri is an introduced parasite that invaded with its first intermediate host, golden mussels (Limnoperna fortunei), from the Asian continent. P. ozakii may also be an introduced species, although its first intermediate host has not been identified.     

The Acanthocolpidae (Digenea) of fishes from the north-east Atlantic: the status of Neophasis Stafford, 1904 (Digenea) and a study of North Atlantic forms

The genus Neophasis is defined. The taxonomic status of the genus is discussed in the light of its life-cycle, hosts and cercarial and adult morphology. It is considered to be closest to the family Acanthocolpidae due to the utilisation of a fish second intermediate host, the form of the cercarial excretory system and, in the adult, the presence of a uterine seminal receptacle and the absence of an external seminal vesicle. The taxonomic value of morphological features are discussed and a key to the north Atlantic species given. The following species are described: Neophasis oculatus (Levinsen) from Myoxocephalus scorpius off West Greenland (type-material), Norway and Denmark, Lycodes esmarkii off NW Scotland and the Faroes, and L. vahli off Newfoundland; N. burti n. sp. (distinguished from N. oculatus by sucker-ratio and testicular configuration) from Myoxocephalus octodecemspinosus off Nova Scotia (type-locality) and New Brunswick, also? immature in Gadus morhua from the Gulf of St. Lawrence; N. anarrhichae (Nicoll) from Anarhichas lupus in the North Sea and off the Faroes; and N. pusilla Stafford from A. lupus off ‘eastern Canada’ (type-material) and Nova Scotia. The only other species in the genus is N. symmetrorchis Machida from the NW Pacific Ocean.     

Evidence that a lineage of teleost-infecting blood flukes (Aporocotylidae) infects bivalves as intermediate hosts

The family Aporocotylidae is recognized as having the widest intermediate host usage in the Digenea. Currently, intermediate host groups are clearly correlated with definitive host groups; all known life cycles of marine teleost-infecting aporocotylids involve polychaetes, those of freshwater teleost-infecting aporocotylids involve gastropods, and those of chondrichthyan-infecting aporocotylids involve bivalves. Here we report the life cycle for a marine elopomorph-infecting species, Elopicola bristowi Orélis-Ribeiro & Bullard in Orélis-Ribeiro, Halanych, Dang, Bakenhaster, Arias & Bullard, 2017, as infecting a bivalve, Anadara trapezia (Deshayes) (Arcidae), as the intermediate host in Moreton Bay, Queensland, Australia. The cercaria of E. bristowi has a prominent finfold, distinct anterior and posterior widenings of the oesophagus, a tail with symmetrical furcae with finfolds, and develops in elongate to oval sporocysts. We also report molecular data for an unmatched aporocotylid cercaria from another bivalve, Megapitaria squalida (G. B. Sowerby I) (Veneridae), from the Gulf of California, Mexico, and six unmatched cercariae from a gastropod, Posticobia brazieri (E. A. Smith) (Tateidae), from freshwater systems of south-east Queensland, Australia. Phylogenetic analyses demonstrate the presence of six strongly-supported lineages within the Aporocotylidae, including one of elopomorph-infecting genera, Elopicola Bullard, 2014 and Paracardicoloides Martin, 1974, now shown to use both gastropods and bivalves as intermediate hosts. Of a likely 14 aporocotylid species reported from bivalves, six are now genetically characterised. The cercarial morphology of these six species demonstrates a clear distinction between those that infect chondrichthyans and those that infect elopomorphs; chondrichthyan-infecting aporocotylids have cercariae with asymmetrical furcae that lack finfolds and develop in spherical sporocysts whereas those of elopomorph-infecting aporocotylids have symmetrical furcae with finfolds and develop in elongate sporocysts. This morphological correlation allows predictions of the host-based lineage to which the unsequenced species belong. The Aporocotylidae is proving exceptional in is propensity for major switches in intermediate host use, with the most parsimonious interpretation of intermediate host distribution implying a minimum of three host switches within the family.     

Haplorchis pumilio and H. taichui in Vietnam discriminated using ITS-2 DNA sequence data from adults and larvae

Samples of Haplorchis taichui and Haplorchis pumilio of different life-stages (cercariae, metacercariae and adults) and from different host species (snail, fish, dog, cat and human) were collected in Nghe An and Nam Dinh Provinces in Vietnam. Samples from Thailand were available for comparison. All adults and metacercariae were initially identified using morphological criteria. Polymerase chain reaction (PCR) assays were developed for discriminating between the species. The complete sequence for the nuclear ribosomal internal transcribed spacer-2 (ITS-2) was obtained from one adult and one metacercaria of H. taichui and three adults and three metacercariae of H. pumilio from Vietnam. Sequences from cercariae from three different snails clustered with those of H. pumilio. Intra-individual variation in the ITS-2 region was detected by sequencing of cloned PCR products. These are the first sequences from Vietnamese Haplorchis spp. to be reported and demonstrate that H. taichui and H. pumilio can be identified unambiguously from any life-stage, including the cercarial stage that is difficult to identify using morphology. Discrepancies in the literature are discussed and examples of apparent misidentification highlighted. The data provide a resource to assist in taxonomic studies on heterophyids, in the design of probes for diagnosis and for field surveys to identify infection in snails.     

Short communication. First record of Torticaecum sp. (Trematoda: Didymozoidae) in the chaetognath Serratosagitta serratodentata (Krohn, 1853) from Caribbean waters

From samples collected in the Caribbean Sea during February, March, May and August 1991, a total of 22 508 holoplanktonic chaetognaths were caught. During the analyses of the samples, one non-cysted specimen of didymozoid metacercaria was found in the coelom of the chaetognath Serratosagitta serratodentata (prevalence 0.004%). Owing to the lack of an authentic stomach and the morphology of the intestine, this trematode seems to belong to the Torticaecum larval type. This is the first report of both the chaetognath species as a host and the geographical locality for this parasite.      

Molecular identification of developmental stages in Opecoelidae (Digenea)

Nuclear ribosomal DNA sequences represent a useful tool for distinction of poorly differentiated developmental stages, such as trematode cercariae or metacercariae. Here, the complete internal transcribed spacer region of the ribosomal DNA (ITS 1 + 5.8S + ITS 2) was sequenced for 29 specimens of the digenean family Opecoelidae, including 16 adult specimens and 13 undescribed larval stages (nine cercariae and four metacercariae) occurring in various marine host organisms. Six cercariae and three metacercariae were found to match their corresponding adult form. This work also revealed that cercariae of the same species are able to infect more than one gastropod host species, suggesting that the specificity for the first intermediate host within the Digenea may be lower than previously thought.     

Levisunguis subaequalis n. g., n. sp., a tongue worm (Pentastomida: Porocephalida: Sebekidae) infecting softshell turtles, Apalone spp. (Testudines: Trionychidae) in the southeastern United States

A new tongue worm (Pentastomida) belonging to the Sebekidae Sambon, 1922 (Porocephaloidea Sambon, 1922) is described based on exemplars collected from softshell terrapins Apalone spinifera aspera (Agassiz) and Apalone ferox (Schneider) in the southeastern United States; a new genus is erected to accommodate the new species. The new species belongs in the Sebekidae because adults possess four simple hooks arranged in a trapezoid pattern on the ventral surface of the cephalothorax, a mouth opening between the anterior and posterior pairs of hooks, a terminal anus, an elongated uterus with preanal uterine pore, and a Y-shaped seminal vesicle. Nymphs possess geminate hooks, and the new species has an aquatic life-cycle in which nymphs become encapsulated in the body cavity of a freshwater fish and mature in the lungs of a terrapin. The new genus is distinct from other genera in the Sebekidae primarily by differences in hook morphology and the fact that representatives use a terrapin as a definitive host. Nymphs infecting fish and presumed to be the new species matured as postlarval juveniles conspecific with the new species when they were fed to the eastern mud turtle, Kinosternon subrubrum (Lacépède). Nymphs of the new species are anatomically similar to but larger than nymphs of Sebekia mississippiensis Overstreet, Self & Vliet, 1985 found in the mesentery of fishes captured in Florida, U.S.A. Adults of the new species differ from those of S. mississippiensis based on hook features, chloride cell pore pattern on annuli, body size, and use of a turtle rather than crocodilian definitive host. The new species is the third North American member of the Sebekidae.