First morphological and molecular characterization of cystacanths of Corynosoma evae Zdzitowiecki, 1984 (Acanthocephala: Polymorphidae) from Antarctic teleost fishes.

Cystacanths of the polymorphid acanthocephalan Corynosoma evae Zdzitowiecki, 1984 were examined and redescribed based on newly collected material from teleost fishes from coastal waters of the Galindez Island (Argentine Islands, West Antarctica). Detailed morphological data, measurements and photomicrographs, including scanning electron microscopy images, are presented. Our morphological and morphometrical analyses confirmed the validity of C. evae; however, three key characteristics of taxonomic importance (i.e., the number of rows of hooks on the proboscis, and the number and arrangement of genital spines in males) showed significant morphological variability. In addition, a genital spine in the posterior body end of a female is reported for the first time. This study provides the first sequences of the small and large subunits nuclear ribosomal RNA genes (SSU and LSU) and the mitochondrial cytochrome c oxidase subunit 1 (cox1) for C. evae. Maximum likelihood and Bayesian inference analyses of the SSU + LSU + cox1 and the cox1 datasets placed C. evae as a sister lineage to a clade formed by C. validum Van Cleave, 1953 and C. villosum Van Cleave, 1953, although with low support. In contrast, the position of C. evae in the phylogenetic analysis of the SSU + LSU dataset remained unresolved. Finally, C. arctocephali Zdzitowiecki, 1984 from pinnipeds from the subantarctic and Antarctic regions is considered as a valid species.

Trematodes from Antarctic teleost fishes off Argentine Islands, West Antarctica: molecular and morphological data.

In 2014-2015 and 2019-2021, teleost fishes off Galindez Island (Antarctic Peninsula) were examined for trematodes. Combined morphological and molecular analyses revealed the presence of eight trematode species of four families (Hemiuridae, Lecithasteridae, Opecoelidae, Lepidapedidae) from five fish species. Only adult trematodes were found and all of them are Antarctic endemics with their congeners occurring on other continents. The hemiuroids, Elytrophalloides oatesi (Leiper & Atkinson, 1914), Genolinea bowersi (Leiper & Atkinson, 1914), and Lecithaster macrocotyle Szidat & Graefe, 1967 belong to the most common Antarctic species and together with Lepidapedon garrardi (Leiper & Atkinson, 1914) and Neolebouria georgiensis Gibson, 1976 they were recorded as the least host-specific parasites. The originally sub-Antarctic Neolepidapedon macquariensis Zdzitowiecki, 1993 is a new record for the Antarctic Peninsula and Parachaenichthys charcoti (Vaillant), is a new host record. Neolebouria terranovaensis Zdzitowiecki, Pisano & Vacchi, 1993 is considered a synonym of N. georgiensis because of identical morphology and dimensions. The currently known phylogenetic relationships within the studied families are supported, including the polyphyly of Macvicaria Gibson & Bray, 1982 with the future need to accommodate its Antarctic species in a new genus. The validity of M. georgiana (Kovaleva & Gaevskaja, 1974) and M. magellanica Laskowski, Jezewski & Zdzitowiecki, 2013 needs to be confirmed by further analyses. Genetic sequence data are still scarce from Antarctica, and more studies applying integrative taxonomic approaches and large-scale parasitological examinations of benthic invertebrates are needed to match sequences of larval stages to those of well-characterised adults and to elucidate trematode life-cycles.

Molecular Study of Echinococcus granulosus Cestodes in Ukraine and the First Genetic Identification of Echinococcus granulosus Sensu Stricto (G1 Genotype) in the Country.

INTRODUCTION: Cystic echinococcosis is a globally distributed zoonotic disease of great medical and veterinary importance, which is caused by the cestode Echinococcus granulosus sensu lato. In Ukraine, two areas of the prominent circulation of the parasite are established, the southern steppe zone with sheep as the main transmitter, and the northern forest-steppe zone and Polissia, where pigs are mainly responsible for maintaining the E. granulosus transmission. METHODS: Given that only a few studies have so far addressed the genetic diversity of the parasite in Ukraine, we have sequenced partial mitochondrial genes of cytochrome c oxidase 1 (789 bp), NADH dehydrogenase 1 (602 bp) and 12S rRNA (333-334 bp) in pig metacestodes from the Sumy region (farms close to Sumy, northeastern Ukraine) and the Kyiv region (a farm in Bila Tserkva, central Ukraine). RESULTS: Four isolates from four pigs in the Sumy region were identified as E. canadensis (G7 genotype), the major E. granulosus s.l. species circulating in Eastern Europe, including the three microvariants (G7A, G7B, G7C). Three isolates from the two pigs in the Kyiv region were classified as E. granulosus s.s. (G1 genotype), including one microvariant (G1A). CONCLUSION: To our knowledge, this is the first genetic record of E. granulosus s.s. with the presumed highest infectivity and virulence among the E. granulosus s.l. species in Ukraine. The finding has implications for public health as local control programmes should take into consideration different development rate of this parasite in dogs and the greater risk of the species for human infection.

Helminth Assemblages of the Antarctic Black Rockcod, Notothenia coriiceps (Actinopterygii: Nototheniidae) in Coastal Waters near Galindez Island (Argentine Islands, West Antarctic): Temporal Changes in the Endoparasite Community.

PURPOSE: Analysis and comparison of the helminth assemblages in Antarctic rockcod Notothenia coriiceps collected near the UAS "Akademik Vernadsky" (Argentine Islands, West Antarctica) in 2002 and 2014-2015 were performed to characterise the parasite community and investigate the temporal changes in helminth assemblages and infection parameters. METHODS: All specimens of N. coriiceps (n = 194) were caught at depths of 10-30 m. Parasites (22,856 helminth specimens and 15,057 cysts) were collected manually and identified based on their morphology. Statistical analysis of the quantitative data was performed using the Quantitative Parasitology 3.0 (QP 3.0), Paleontological Statistics (PAST v. 3.1), and PRIMER 6 software. RESULTS: Twenty-seven species of four taxonomic groups were recorded: trematodes (8 species), cestodes (4), nematodes (5), and acanthocephalans (10). Helminth samples collected in 2002 and 2014-2015 showed a rather high similarity in species composition. The species richness was higher in the sample collected in 2014-2015, while the evenness and diversity in the two samples were similar. The dissimilarity between helminth infracommunities in the two samples appeared to be statistically significant. Larval cestodes Diphyllobotrium sp., the acanthocephalan Metacanthocephalus rennicki, and the trematode Neoleoburia antarctica were found to make the most significant impact on the dissimilarity. CONCLUSIONS: The analysis of the composition and structure of helminth community in N. coriiceps revealed the changes that have happened during the last decade. At least some of the changes are attributed to the changes in marine ecosystems in Western Antarctica.

Diphyllobothrium sprakeri n. sp. (Cestoda: Diphyllobothriidae): a hidden broad tapeworm from sea lions off North and South America.

BACKGROUND: The systematic of several marine diphyllobothriid tapeworms of pinnipeds has been revised in recent years. However, 20 species of Diphyllobothrium from phocids and otariids are still recognized as incertae sedis. We describe a new species of Diphyllobothrium from the intestine of California sea lions Zalophus californianus (Lesson) (type-host) and South American sea lions Otaria flavescens (Shaw). METHODS: Zalophus californianus from the Pacific coast of the USA and O. flavescens from Peru and Argentina were screened for parasites. Partial fragments of the large ribosomal subunit gene (lsrDNA) and the cytochrome c oxidase subunit 1 (cox1) mitochondrial gene were amplified for 22 isolates. Properly fixed material from California sea lions was examined using light and scanning electron microscopy. RESULTS: A total of four lsrDNA and 21 cox1 sequences were generated and aligned with published sequences of other diphyllobothriid taxa. Based on cox1 sequences, four diphyllobothriid tapeworms from O. flavescens in Peru were found to be conspecific with Adenocephalus pacificus Nybelin, 1931. The other newly generated sequences fall into a well-supported clade with sequences of a putative new species previously identified as Diphyllobothrium sp. 1. from Z. californianus and O. flavescens. A new species, Diphyllobothrium sprakeri n. sp., is proposed for tapeworms of this clade. CONCLUSIONS: Diphyllobothrium sprakeri n. sp. is the first diphyllobothriid species described from Z. californianus from the Pacific coast of North America, but O. flavescens from Argentina, Chile and Peru was confirmed as an additional host. The present study molecularly confirmed the first coinfection of two diphyllobothriid species in sea lions from the Southern Hemisphere.

Analysis of nad2 and nad5 enables reliable identification of genotypes G6 and G7 within the species complex Echinococcus granulosus sensu lato.

The larval stages of tapeworms in the species complex Echinococcus granulosus sensu lato cause a zoonotic disease known as cystic echinococcosis (CE). Within this species complex, genotypes G6 and G7 are among the most common genotypes associated with human CE cases worldwide. However, our understanding of ecology, biology and epidemiology of G6 and G7 is still limited. An essential first step towards this goal is correct genotype identification, but distinguishing genotypes G6 and G7 has been challenging. A recent analysis based on complete mitogenome data revealed that the conventional sequencing of the cox1 (366 bp) gene fragment mistakenly classified a subset of G7 samples as G6. On the other hand, sequencing complete mitogenomes is not practical if only genotype or haplogroup identification is needed. Therefore, a simpler and less costly method is required to distinguish genotypes G6 and G7. We compared 93 complete mitogenomes of G6 and G7 from a wide geographical range and demonstrate that a combination of nad2 (714 bp) and nad5 (680 bp) gene fragments would be the best option to distinguish G6 and G7. Moreover, this method allows assignment of G7 samples into haplogroups G7a and G7b. However, due to very high genetic variability of G6 and G7, we suggest to construct a phylogenetic network based on the nad2 and nad5 sequences in order to be absolutely sure in genotype assignment. For this we provide a reference dataset of 93 concatenated nad2 and nad5 sequences (1394 bp in total) containing representatives of G6 and G7 (and haplogroups G7a and G7b), which can be used for the reconstruction of phylogenetic networks.

Multifocal Necrotizing Myopathy in Northern Elephant Seal ( Mirounga angustirostris) Pups, San Miguel Island, California.

A field study addressing causes of mortality in freshly dead northern elephant seals ( Mirounga angustirostris, Gill, 1866) was conducted on San Miguel Island, California, in February 2015. Necropsies were performed on 18 pups ranging in age from stillbirths to approximately 7 to 8 weeks. The primary gross diagnoses in these pups included trauma, myopathy, starvation/emaciation, infections, congenital anomalies, and perinatal mortality. However, 6 (33%) had a previously unrecognized myopathy characterized by multiple white streaks that were most obvious within the inner layer of the abdominal wall and the small innermost ventral intercostal muscles. Following histological examination, 2 more pups from San Miguel Island and 6 pups from The Marine Mammal Center (Sausalito, California) were found to have similar lesions. Histologically, the lesions within the skeletal muscles were characterized by a multifocal polyphasic, mild to severe, acute to subacute necrotizing myopathy with mineralization. Acute necrosis and degeneration characterized by pyknotic nuclei, eosinophilic cytoplasm and cytoplasmic vacuolization were found in smooth muscle myocytes within the urinary bladder and digestive system. Degeneration of myocytes was present in the tunica media of a few small- to medium-sized vessels and was characterized by a vacuolar degeneration and occasionally necrosis. This condition has been termed multifocal necrotizing myopathy. A cause of this myopathy was not identified.

The benefits of analysing complete mitochondrial genomes: Deep insights into the phylogeny and population structure of Echinococcus granulosus sensu lato genotypes G6 and G7.

Cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of the species complex Echinococcus granulosus sensu lato. Within this complex, genotypes G6 and G7 have been frequently associated with human CE worldwide. Previous studies exploring the genetic variability and phylogeography of genotypes G6 and G7 have been based on relatively short mtDNA sequences, and the resolution of these studies has often been low. Moreover, using short sequences, the distinction between G6 and G7 has in some cases remained challenging. The aim here was to sequence complete mitochondrial genomes (mitogenomes) to obtain deeper insight into the genetic diversity, phylogeny and population structure of genotypes G6 and G7. We sequenced complete mitogenomes of 94 samples collected from 15 different countries worldwide. The results demonstrated that (i) genotypes G6 and G7 can be clearly distinguished when mitogenome sequences are used; (ii) G7 is represented by two major haplogroups, G7a and G7b, the latter being specific to islands of Corsica and Sardinia; (iii) intensive animal trade, but also geographical isolation, have likely had the largest impact on shaping the genetic structure and distribution of genotypes G6 and G7. In addition, we found phylogenetically highly divergent haplotype from Mongolia (Gmon), which had a higher affinity to G6.

The cestode community in northern fur seals (Callorhinus ursinus) on St. Paul Island, Alaska.

The diversity and ecology of cestodes from the northern fur seals, Callorhinus ursinus (NFS), were examined using newly collected material from 756 humanely harvested subadult males between 2011 and 2014. NFSs were collected from five different haul-outs on St. Paul Island, Alaska. A total of 14,660 tapeworms were collected with a prevalence of 98.5% and intensity up to 107 cestodes per host (mean intensity 19.7 ± 16.5 SD). Three species of tapeworms were found: Adenocephalus pacificus (Diphyllobothriidea) was the most prevalent (prevalence 97.4%), followed by Diplogonoporus tetrapterus (49.7%), and 5 immature specimens of Anophryocephalus cf. ochotensis (Tetrabothriidea) (0.5%). Most of the cestodes found in the NFS were immature (69.7%). However, only 0.9% of cestodes were in larval (plerocercoid) stages. The species composition, prevalence and intensity of cestodes from these NFSs were not statistically different between the five separate haul-outs. Significant increases in the intensity of NFS infections were observed during the study period.

High morphological plasticity and global geographical distribution of the Pacific broad tapeworm Adenocephalus pacificus (syn. Diphyllobothrium pacificum): molecular and morphological survey.

The most important causative agent of human diphyllobothriosis in South America, Diphyllobothrium pacificum, is transferred to the original genus Adenocephalus Nybelin, 1931; revised and redescribed on the basis of the evaluation of an extensive material collected mainly from northern fur seal, Callorhinus ursinus, from St. Paul Island, Alaska. Detailed analysis of morphological and morphometrical data shows a high variability in most of the characteristics traditionally used in diagnosis of diphyllobothriid tapeworms. Phylogenetic analyses based on newly characterised sequences of mitochondrial cytochrome c oxidase subunit 1 and nuclear large subunit ribosomal RNA genes consistently reveal Adenocephalus pacificus as a sister lineage to the clade formed of the remaining Diphyllobothrium species and other genera (Digramma, Diplogonoporus, Ligula). Despite the generally similar morphology, A. pacificus can be differentiated from the closely related taxa in the presence of transverse papilla-like tegumental protuberances distributed anteriorly, separated by narrow semicircular grooves on the ventral surface of proglottids between their anterior margin and the anterior edge of the male gonopore, and relatively small eggs. A. pacificus displays a relatively low host specificity (found in 9 of 16 otariids, and in accidental hosts such as man, dog and jackal, the latter representing a new host) and a uniquely wide geographical distribution on both hemispheres. In addition, suitability of morphological criteria used in diagnostics of diphyllobothriid cestodes is discussed.