Ithyoclinostomum yamagutii n. sp. (Digenea: Clinostomidae) in the great blue heron Ardea herodias L. (Aves: Ardeidae) from Mississippi,USA

With only six recognised genera, the family Clinostomidae Lühe, 1901 remains a global research interest of parasitologists and ecologists. Recent efforts have focused on providing molecular data to investigate species diversity, elucidate life-cycles, and make inferences on the group’s evolutionary history. Of the clinostomid genera, the monotypic Ithyoclinostomum Witenberg, 1926 has remained more enigmatic compared to the commonly encountered Clinostomum Leidy, 1856. Recent morphological and molecular evidence from metacercariae suggests a second Ithyoclinostomum species may exist in freshwater cichlids in Central America and Mexico. In a recent survey of great blue herons Ardea herodias L. from commercial catfish production farms in Mississippi, USA, two specimens of an abnormally large (> 20 mm) clinostomid were encountered in the oesophagus of a single bird. These specimens were identified as an Ithyoclinostomum sp. morphologically distinct from the only nominal species Ithyoclinostomum dimorphum (Diesing, 1850). Using morphological and molecular data these adult specimens were confirmed as conspecific with the larval metacercariae previously described from Central America and Mexico and represent the novel species, Ithyoclinostomum yamagutii n. sp.

     

A large‐scale molecular survey of Clinostomum (Digenea,Clinostomidae)

Members of the genus Clinostomum Leidy, 1856 are parasites that mature in birds, with occasional reports in humans. Because morphological characters for reliable discrimination of species are lacking, the number of species considered valid has varied by an order of magnitude. In this study, sequences from the DNA barcode region of cytochrome c oxidase I (CO1) and/or internal transcribed spacer (ITS) from specimens from Mexico, Bolivia, Peru, Brazil, Kenya, China and Thailand were analysed together with published sequences from Europe, Africa, Indonesia and North America. Although ITS and CO1 distances among specimens were strongly correlated, distance‐based analysis of each marker yielded different groups. Putative species indicated by CO1 distances were consistent with available morphological identifications, while those indicated by ITS conflicted with morphological identifications in three cases. There was little overlap in sequence variation within and between species, particularly for CO1. Although ITS and CO1 distances tended to increase in specimens that were further apart geographically, this did not impair distance‐based species delineation. Phylogenetic analysis suggests a deep division between clades of Clinostomum inhabiting the New World and Old World, which parallels the distribution of their principal definitive hosts, the Ardeidae.     

Morphological and molecular differentiation of Clinostomum complanatum and Clinostomum marginatum (Digenea: Clinostomidae) metacercariae and adults

The separation of Clinostomum complanatum Rudolphi, 1814 and Clinostomum marginatum Rudolphi, 1819 has long been unclear. Recent data confirm the validity of the junior species, C. marginatum , by ～ 1% differences in its 18S rDNA sequences. We collected adults and metacercariae of C. complanatum and C. marginatum and found reliable morphological differences in the genital complex at both developmental stages. In addition, we identified basic morphometrics (distance between suckers, body width) in metacercariae that may be useful for discriminating the species. The morphological differences were supported by the comparison of sequences of internal transcribed spacers of ribosomal DNA and of the mitochondrial gene cytochrome c oxidase I (COI) from 39 specimens. In 36 specimens, the average divergence between the species was 7.3% in ITS and 19.4% in COI sequences. Two specimens from North America and 1 from Europe had sequences that did not allow them to be clearly allied with either species.     

Clinostomum complanatum and Clinostomum marginatum (Rudolphi, 1819) (Digenea: Clinostomidae) are separate species based on differences in ribosomal DNA

Infections by metacercariae of Clinostomum (Leidy, 1856) species adversely affect aquacultured fish and are potentially transmissible to humans. Molecular methodologies are efficient tools, which enable diagnosis of all life-history stages of trematodes in their diverse hosts. The small subunit of ribosomal DNA genes of adults of the Old World Clinostomum complanatum (Rudolphi, 1819) and the New World Clinostomum marginatum (Rudolphi, 1819), obtained from a little egret Egretta garzetta (Linnaeus, 1766) and the great blue heron Ardea herodias (Linnaeus, 1758), respectively, were amplified, sequenced, and aligned. The resulting alignment was used to develop a genetic assay to differentiate between these species.     

Microsatellite loci for great white herons and great blue herons (Aves,Ardeidae, Ardea herodias)

We used an enrichment technique to isolate 60 microsatellite loci in Ardea herodias. We developed primers for 17 loci, screened for variation in A. herodias and attempted to amplify these loci in three closely related species (A. alba, A. cinerea and A. cocoi). Fifteen loci were polymorphic in A. herodias. Observed heterozygosities ranged from 0.10 to 0.81. Two loci appeared to be monomorphic in A. herodias, but exhibited variation in product size among species within the genus. Our ability to amplify polymorphic products in closely related species suggests that these markers may be useful in other herons.     

Digenetic trematodes of the genus Clinostomum Leidy, 1856 (Digenea: Clinostomidae) from birds of Queensland, Australia, including C. wilsoni n. sp. from Egretta intermedia

Two species of Clinostomum previously described from Australia, C. hornum from Botaurus poiciloptilus (Australian bittern) and Nycticorax caledonicus (Nankeen night heron) and C. australiense from Pelecanus conspicillatus (Australian pelican), which have previously been synonymised with C. complanatum, are redescribed and recognised as valid species. In addition, C. complanatum is recorded from Egretta alba (large egret), E. garzetta (little egret), E. intermedia (plumed egret), N. caledonicus and Ardea novaehollandiae (white-faced heron). C. wilsoni n. sp. is described from E. intermedia from Queensland. C. wilsoni differs from the other three species in size and shape of the body and in the oral collar, oral sucker, intestinal caeca, caecal diverticula and position of testes. Taxonomic problems within the genus Clinostomum are discussed.     

<Emphasis Type=Italic>Ardea sytchevskayae</Emphasis> sp. nov., a new heron species (Aves: Ardeidae) from the Middle Miocene of Mongolia

A new heron species, Ardea sytchevskayae sp. nov., from the Middle Miocene Sharga locality is described. The new species is represented by an almost complete coracoid and a few partial coracoids, partial scapula, and caprometacarpus. A. sytchevskayae was a relatively large heron, of the same size as A. alba, and intermediate in morphology between A. alba and other species traditionally included in this genus. In addition, the Sharga locality has yielded a large bittern and a medium-sized heron of uncertain taxonomic position.     

Clinostomum attenuatum (Digenea) from the eye of Bufo marinus

Two specimens of the cane toad, Bufo marinus, were found with single worm infestations of the eye. One worm was excysted in the eye chamber, the other still encysted in the choroid layer; both worms were identified as Clinostomum attenuatum. This finding is both a new host record and a new site of development for this amphibian parasite.     

Bacteria associated with tegument of Clinostomum marginatum (Digenea)

Adults of Clinostomum marginatum freshly collected from a heron, Ardea herodias, were examined using transmission electron microscopy. Specimens from the mouth of the bird were encrusted with bacteria that were not removed by washing unless the saline contained antibiotics. There was no evidence that the attached bacteria were damaging to the trematode tegument. Three species of Gram-negative bacteria were isolated from the worm surfaces and identified; Achromobacter sp. was present in pure culture on 4 of 6 original cultures and in mixed culture with Edwardsiella tarda and Enterobacter agglomerans in 2 cultures. These species and 3 unidentified species of bacteria were isolated from the oral epithelium of the heron. Microorganisms were not seen attached to the surfaces of worms recovered from the esophagus. Because E. tarda and E. agglomerans were the only species isolated from the heron esophagus, the intimate bacterial-worm association in the heron mouth may be due specifically to Achromobacter sp.     

Pygidiopsis ardeae n. sp. (Digenea: Heterophyidae: Pygidiopsinae) in the grey heron Ardea cinerea L. from Denmark

A new species, Pygidiopsis ardeae (Heterophyidae, Pygidiopsinae), is described, using both light and stereoscan electron microscopy, from the grey heron Ardea cinerea L. (Ardeidae) from Denmark. The species has previously (as P. genata) been recorded in dogs fed on plaice and flounders from Danish waters. It developed experimentally in domestic chickens and pigeons. No species of Pygidiopsis Looss, 1907 has previously been described from birds in northern Europe. P. ardeae is very similar to the type-species, P. genata Looss, 1907, from Egypt, but the two species have different life-cycles. P. ardeae differs from P. piclaumoreli Dollfus & Capron, 1958 from Senegal in its size and the disposition of vitelline follicles. P. plana (Linton, 1928) Price, 1933, from a green heron from the eastern USA, is larger than P. ardeae and has a wide, sac-like prepharynx.     

Megalophallus deblocki n. sp. (Digenea: Microphallidae) from Rostrhamus sociabilis (Vieillot) (Aves: Accipitridae) in Paraguay

A new species in the little-known digenean fauna of Paraguayan birds is described in detail, especially in relation to its terminal genitalia. The new form, Megalophallus deblocki n. sp., is a microphallid from the intestine of the snail kite Rostrhamus sociabilis (Vieillot). It is differentiated in detail from its congeners, but differs mainly in having a substantially larger phallus (e.g. in relation to the ventral sucker), a smaller ventral sucker and smaller eggs. Comments are included on the functional morphology of the terminal genitalia.     

Megalophallus deblocki n. sp. (Digenea: Microphallidae) from Rostrhamus sociabilis (Vieillot) (Aves: Accipitridae) in Paraguay

A new species in the little-known digenean fauna of Paraguayan birds is described in detail, especially in relation to its terminal genitalia. The new form, Megalophallus deblocki n. sp., is a microphallid from the intestine of the snail kite Rostrhamus sociabilis (Vieillot). It is differentiated in detail from its congeners, but differs mainly in having a substantially larger phallus (e.g. in relation to the ventral sucker), a smaller ventral sucker and smaller eggs. Comments are included on the functional morphology of the terminal genitalia.     

Helminth communities of herons (Aves: Ardeidae) in southern Italy

The helminth communities of nine species of herons from southern Italy were studied and compared. Of 24 taxa found including seven digeneans, seven nematodes, six cestodes and four acanthocephalans, only five taxa were found in more than one heron species, and five of the 21 taxa that could be identified to species level were classified as ‘heron specialists’. The total number of helminth species per heron species ranged from 1 in Botaurus stellaris to 9 in Ixobrychus minutus with infection levels generally low. A statistical comparison was carried out for herons with a sample size > 5. At the infracommunity level, only I. minutus clearly differed from other heron species. Diversity parameters of heminth infracommunities did not significantly differ among heron species. Species richness ranged from just 0.3 to 2.3 helminth taxa per individual host, and the Brillouin index, from 0 to 0.3. Total helminth abundance did not exceed 40 worms per host except in a single case of Ardeola ralloides. Infracommunities clearly were dominated by single helminth species. The present study confirms a depauperate helminth community in herons from southern Italy. Comparison with data from Spain and the Czech Republic showed strong quantitative similarities with values obtained in the present study. Results also suggest that the composition of local helminth communities are strongly variable depending on geographical location as is demonstrated by comparison with data from other European areas. However, whether herons in Europe naturally host depauperate helminth communities or these communities are depauperate because of other factors is unknown.     

The effects of age, sex and breed of chicken on susceptibility to Clinostomum complanatum (Digenea: Clinostomatidae)

Susceptibility and development of Clinostomum complanatum in the chicken (Gallus gallus domesticus) have been studied. The progenetic metacercariae of C. complanatum developed into ovigerous worms in 3 days in the buccal cavity of the chicken and survived only 6-7 days in this experimental host. The susceptibility is influenced by the age, sex and breed/type of the host. Both percent susceptibility and recovery of the worms decreased with increase in the age of the host. The egg laying chickens were found to be least susceptible to the infection. Mature male chickens were more susceptible than the females. The single comb white Leghorn (SCWL) was found to be susceptible while the local type (LT) and the cross breed (SCWLxLT) were not susceptible to C. complanatum. The possible reasons for these variations have been discussed.     

In vitro Excystation of Metacercariae of Clinostomum complanatum (Rudolphi, 1819) (Digenea: Clinostomatidae)

In vitro excystation of metacercariae of Clinostomum complanatum (Rudolphi, 1819) was studied. Excystation was an active process, initiated by a rise in temperature. The metacercariae excysted in enzymic as well as non enzymic medium, but the rate of excystation was affected by the temperature and/or pH of the medium. The optimal excystation was obtained in pepsin-Tyrode at pH 2.3-3.5 and a temperature of 42°C. Trypsin treatment was not required.     

A bittern (Aves: Ardeidae) from the early Miocene of New Zealand

Herons (Aves: Ardeidae) are rare in the fossil record globally. Fossil taxa referred to Ardeinae and Nycticoracini are known from as early as the early Oligocene and ardeids undetermined to subfamily include some as old as the early Eocene. In Australasia, the pre-Pliocene record is restricted to one species from the early Miocene of New Zealand. On the basis of a tarsometatarsus and a coracoid we describe a new species of bittern (Ardeidae: Botaurinae) from the St Bathans Fauna, of early Miocene age, from Otago, New Zealand. This is only the third and the oldest pre-Quaternary record for Botaurinae globally.     

Oswaldotrema nacinovici gen. nov. sp. nov. (Digenea: Philophthalmidae) from Numenius phaeopus (Aves: Scolopacidae) in Brazil

A new genus, Oswaldotrema gen. nov. is proposed. Oswaldotrema nacinovici sp. nov. is descibed from Numenius phaeopus Latham, 1790. Differentiation from the other related genera, namely Philophthalmus, Pygorchis, Proctobium, Parorchis, Echinostephila, Cloacitrema, Pittacium, Ophthalmotrema, Skrjabinovermis and Paratrema, was based on morphological characters, mainly on those referring to the body surface, body shape, head, esophagus, pharynx, acetabulum, vitellaria, vitelline reservoir and seminal vesicle.     

The helminth infracommunities in three species of herons (Aves: Ardeidae) from Ukraine

This survey analysed the helminth communities associated with three Ukrainian heron species (Ardea cinerea, Ardea purpurea, Nycticorax nycticorax) collected throughout 1947–2019. The sample analysed included 128 specimens, and 55 helminth species were identified. Of these, the most diverse taxonomic group was trematodes with 29 species, followed by cestodes with 14 species, nematodes with nine, and acanthocephalans with three species. The helminth infracommunities were found to be similar across all three host species. Furthermore, the distance to the coastal regions impacts both diversity and species richness in infracommunities.     

The life-cycle of Helicometra gibsoni n. sp. (Digenea: Opecoelidae)

The life-cycle of Helicometra gibsoni n. sp., an opecoelid trematode of marine fishes, is elucidated and the life-history stages are described. Natural infections with the cercariae, which are typically opecoelid in many respects but characterised by having very long, extensile tails, were found in the intertidal snail Anachis terpsichore collected from the Lawson's Bay Coast, Bay of Bengal. Metacercariae were found in the thoracic muscles of the shooting shrimp Alphaeus malabaricus as prominent, reddish-brown, oval cysts. Laboratory experiments demonstrated that 14 days are required for the development of cercaria into the infective metacercarial stage in shrimps. Natural infections with the adult fluke were found in Scorpaenopsis cirrhosus. Therapon jarbua served as an experimental host. Fourteen-day-old metacercariae fed to T. jarbua developed into egg-producing adults in 40–43 days. Intra-specific variations noted in the flukes obtained from natural and experimental infections are presented. H. fasciata of Madhavi (1975) is regarded as a synonym of H. gibsoni.