Cox1 Sequences Research Articles

Global analysis of cytochrome c oxidase subunit 1 (cox1) gene variation in Dibothriocephalus nihonkaiensis (Cestoda: Diphyllobothriidae)

The cestode Dibothriocephalus nihonkaiensis (syns. Diphyllobothrium nihonkaiense and Diphyllobothrium klebanovskii), the broad fish tapeworm, is a parasitic agent of intestinal infection acquired by consumption of raw or undercooked Pacific salmon, Onchorhynchus spp. Sequencing studies conducted about a decade ago revealed the presence of two major lineages (A and B) in the broad fish tapeworm population within Asian coastal areas. However, in spite of the accumulation of sequence data on GenBank recently, no further genetic analyses of D. nihonkaiensis have been attempted. The present study assessed for the first time the global cox1 variation in D. nihonkaiensis. Novel partial cox1 sequences of 14 isolates of D. nihonkaiensis from 12 patients were generated, and a global genetic analysis was performed using the 14 novel and 79 previously published sequences for isolates from definitive and second intermediate hosts of this species was performed. A total of 48 haplotypes of three haplotype groups (Types A, B and C) were identified, and co-infections with genetically different D. nihonkaiensis were highlighted in humans and Pacific salmon.

Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), a new entomopathogenic nematode from Poland.

A new species of entomopathogenic nematodes, Steinernema sandneri n. sp., was recovered by baiting from Poland. Its morphological traits indicate that the new species is a member of the feltiae-kraussei group. A body length of 843 (708–965) μm, a more anterior position of excretory pore (56 μm), and the lower D% value (40 vs > 46) discriminate this species from most of the other group members. The first-generation males of S. sandneri n. sp. can be distinguished from the other clade members by a 60 μm long spicule, a relatively long gubernaculum (GS% = 79), and the position of the excretory pore (80 μm). Phylogenetic analysis of the ITS rDNA, D2D3 of 28 S rDNA, and cox1 sequences confirmed that S. sandneri n. sp. is a new species of the feltiae-kraussei group, closely related to S. kraussei and S. silvaticum.

Molecular and Morphological Data Confirmed First Record of Abbreviata kazakhstanica Markov and Paraskiv, 1956 (Spirurida: Physalopteridea) in Iran.

Background:The genus Abbreviata (Spirurida: Physalopteridea) currently contains 47 species. Physalopteridae nematodes infect a large number of vertebrates, including mammals, birds, reptiles and amphibians. The current study is a report of the first morphological and molecular identification of A. kazakhstanica (Spirurida: Physalopteridea) in Pseudopus apodus in Iran.Methods:Eleven road-killed P. apodus, were collected from, Iran during 2016–2018. The nematodes were isolated from stomach. After morphological study, the genomic DNA of the parasites was extracted using CTAB method. The DNA was used for PCR amplification of cytochrome c oxidase subunit I (cox1). The PCR products were sequenced, the sequence data were analyzed and multiple alignments were conducted using the Clustal Omega.Results:After detailed microscopic examination, the A. kazakhstanica was identified. The cox1 sequences confirmed the species of helminth. The new sequences of A. kazakhstanica were submitted to GenBank under the accession number MK578751-2.Conclusion:Regarding the limited data on parasitological status of Iranian reptiles, more specific and comprehensive investigations are needed to identify the parasitic fauna.

Dictyota cyanoloma (Dictyotales, Phaeophyceae), a Newly Introduced Brown Algal Species in California.

Here, we report for the first time the presence of Dictyota cyanoloma in southern California. Dictyota cyanoloma is conspicuous in harbors and bays by its distinctive bright blue-iridescent margins. This species was originally described from Europe, but subsequent studies have revealed that it represented an introduction from Australia. The current distribution of D. cyanoloma comprises southern Australia and the North East Atlantic, including the Mediterranean Sea and the Macaronesian islands. The presence of D. cyanoloma in southern California is supported by molecular cox1 and psbA gene sequences. A reconstruction of the invasive history based on nine polymorphic microsatellite markers reveals a close affinity of the Californian specimens with European populations. Dictyota cyanoloma in the United States appears to be (so far) restricted to the Californian coast from San Diego Bay in the south to Santa Catalina Island and Long Beach Harbor in the north. A correlative species distribution model suggests gradually declining habitat suitability north of the Southern Californian Bight and high suitability in Baja California, including the Gulf of California. Finally, its widespread abundance in bays and harbors suggests shipping is a likely transport mechanism.

Occurrence of Plasmopara destructor causing Downy Mildew on Impatiens walleriana in Brazil.

Impatiens walleriana (Balsaminaceae), popularly known as Impatiens, is an African succulent and a popular ornamental plant worldwide (GBIF, 2019). In Brazil it is broadly grown indoors and outdoors, including in public parks of Curitiba, State of Paraná (Viezzer et al. 2018). In September 2018, I. walleriana plants showing typical downy mildew symptoms were observed in wastelands and gardens in Curitiba. The symptoms included adaxial chlorotic leaf spots with abundant white sporulation on abaxial side (Supplementary figure 1). The disease led to severe defoliation of the plants and the incidence of the plant disease varied from 20 to 80% of plants in an area ranging from 400 to 40,000 m2. A representative sample was deposited in herbarium of the Museu Botânico Municipal de Curitiba (MBM 331601). The following morphology was observed: Sporangiophores (n = 30), hyaline, thin walled, emerging through stomata, 407.3 to 551.1 μm long, slightly swollen base, first branch at 165.8 to 324.7 μm from base, end branches 5.1 to 13.1 μm long, sporangia (n = 50) hyaline, thin-walled subglobose to ovoid, from 12.8 to 21.9 μm x 12.5 to 17.9 μm, slightly papillate. Due to morphological and genetic variations within the species Plasmopara obducens, Görg et al. (2017) proposed the new species P. velutina and P. destructor. The morphology of the Curitiba specimen was equivalent to that described for P. destructor (Görg et al. 2017). DNA was extracted from LEMIDPRTf-19-02 isolate and the ITS1 and cox2 regions were PCR amplified as described in Görg et al. (2017). The resulting sequences were deposited in GenBank (ITS1, MT680628; cox2, MT952335). A BLASTn analysis of the sequences revealed 100% homology with ITS (MF372742) and cox2 (MF372728) sequences of type strain of P. destructor (GLM-F107554). A Bayesian phylogenetic analysis was performed to compare the sequences from this study with reference sequences for P. obducens, P. destructor and P. velutina (Görg et al. 2017; Salgado-Salazar et al. 2018). The oomycete from Curitiba grouped in a reliable clade with P. destructor (Supplementary figure 2). Pathogenicity was carried out by ex vivo and in vivo tests. For ex vivo, stems with approximately four healthy leaves of I. walleriana (n = 10) were embedded in aluminum grid inside of gerbox with the stem bases immersed in distilled water. The inoculation of five stems was carried out by spraying a suspension with 6 x 104 sporangia mL-1 on the abaxial side of the leaves. Five stems with leaves inoculated with sterile water were used as controls. They were incubated in a growth chamber in the dark for 48 h at 20 °C and another 12 days in a 12 h light photoperiod. The confirmation of pathogenicity in plants (in vivo) was obtained with the inoculation of I. walleriana seedlings (one-month old) grown in 2 dm3 aluminum pots. The inoculation methodology and number of plants were the same as the stems test. After the inoculation, plants were incubated in a growth chamber for 48 h in the dark at 20 °C with 100% RH with nebulization, and another 10 days at a photoperiod of 12 hours of light. For both tests, abundant sporulation was observedwith morphology equivalent to Plasmopara destructor described by Görg et al. (2017). No disease developed on control plants. To our knowledge, this is the first report of P. destructor on I. walleriana in Brazil (Farr and Rossman 2019, Silva et al. 2019) representing a potential loss to flower production and a reduction in flowering period in public gardens and parks.

Anisakis simplex (s.s.) larvae (Nematoda: Anisakidae) hidden in the mantle of European flying squid Todarodes sagittatus (Cephalopoda: Ommastrephidae) in NE Atlantic Ocean: Food safety implications

Few reports exist upon the occurrence and localization of zoonotic anisakid nematodes in T. sagittatus, especially in the mantle of the squid. The occurrence and site of infection of larval anisakids in 98 T. sagittatus caught West off St. Kilda, NE Atlantic Ocean, were investigated. Squids were examined for anisakids using the UV-Press method. In total, 689 nematodes were detected in the viscera and mantle. According to morphology, all the larvae (L3) were assigned to genus Anisakis. Diagnostic allozymes and mtDNA cox2 sequence analysis permitted to genetically identify all larvae as Anisakis simplex (s.s.) (N = 100). Overall prevalence (P = 81%) and mean intensity (mI = 8.6) of infection with A. simplex are provided. Most of the larvae present in the mantle cavity were embedded in the stomach wall or attached in the outer layer of the stomach and caecum (49%). Over a third of squids (37%) hosted A. simplex (s.s.) larvae in the mantle. A novel schematized representation of larvae distribution in the mantle is provided, showing where they were mostly located. According to the results obtained, the risk of anisakiasis associated with consumption of raw or undercooked T. sagittatus should be considered.

Morphological and Molecular Identification of Spirometra Tapeworms (Cestoda: Diphyllobothriidae) from Carnivorous Mammals in the Serengeti and Selous Ecosystems of Tanzania.

Spirometra tapeworms (Cestoda: Diphyllobothriidae) collected from carnivorous mammals in Tanzania were identified by the DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and internal transcribed spacer 1 (ITS1), and by morphological characteristics. A total of 15 adult worms were collected from stool samples and carcasses of Panthera leo, Panthera pardus, and Crocuta crocuta in the Serengeti and Selous ecosystems of Tanzania. Three Spirometra species: S. theileri, S. ranarum and S. erinaceieuropaei were identified based on morphological features. Partial cox1 sequences (400 bp) of 10 specimens were revealed. Eight specimens showed 99.5% similarity with Spirometra theileri (MK955901), 1 specimen showed 99.5% similarity with the Korean S. erinaceieuropaei and 1 specimen had 99.5% similarity with Myanmar S. ranarum. Sequence homology estimates for the ITS1 region of S. theileri were 89.8% with S. erinaceieuropaei, 82.5% with S. decipiens, and 78.3% with S. ranarum; and 94.4% homology was observed between S. decipiens and S. ranarum. Phylogenetic analyses were performed with 4 species of Spirometra and 2 species of Dibothriocephalus (=Diphyllobothrium). By both ML and BI methods, cox1 and ITS1 gave well supported, congruent trees topology of S. erinaceieuropaei and S. theileri with S. decipiens and S. ranarum forming a clade. The Dibothriocephalus species were sisters of each other and collectively forming successive outgroups. Our findings confirmed that 3 Spirometra species (S. theileri, S. ranarum, and S. erinaceieuropaei) are distributed in the Serengeti and Selous ecosystems of Tanzania.

Prevalence and Genetic Characterization of Two Mitochondrial Gene Sequences of Strobilocercus Fasciolaris in the Livers of Brown Rats (Rattus norvegicus) in Heilongjiang Province in Northeastern China.

Rodents constitute the largest and most successful group of mammals worldwide. Brown rats (Rattus norvegicus) are one of the most common rodent species, and they serve as intermediate hosts of Hydatigera taeniaeformis. Although there have been a few studies reporting on the presence of the larval form of H. taeniaeformis (strobilocercus fasciolaris) in brown rats worldwide, little information is available on the genetic characterization of this parasite, with no molecular data from China. Therefore, from April 2014 to March 2016, this study was carried out to understand the prevalence and genetic characters of strobilocercus fasciolaris in brown rats captured in Heilongjiang Province in northeastern China. The livers of brown rats were collected and examined for the presence of cysts. Each cyst was identified based on morphological observation: the larvae with the naked eye and the scolexes under a microscope. The results were confirmed by polymerase chain reaction (PCR) and sequencing of the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 4 (nad4) genes. At the investigated sites, 11.8% (13/110) of the brown rats were infected with strobilocercus fasciolaris. Based on sequence analysis, there were 10 and six haplotypes regarding the cox1 and the nad4 loci, with 24 and 42 polymorphic sites, respectively (degree of intraspecific variation: 0.3%–4.4% and 0.6%–4.7%, respectively). Twelve nucleotide sequences (six of the 10 at the cox1 locus and all six at the nad4 locus) have not previously been described. Base differences in three of the six novel cox1 gene sequences and five of the six novel nad4 gene sequences caused amino acid changes. Phylogenetic analyses of the cox1 and nad4 gene sequences based on neighbor-joining and Bayesian inference trees indicated that all the strobilocercus fasciolaris isolates belonged to Hydatigera taeniaeformis sensu stricto (s.s.). This is the first report on the genetic characterization of strobilocercus fasciolaris in brown rats in China. The findings of novel cox1 and nad4 nucleotide and amino acid sequences may reflect the region-specific genetic characterization of the parasite. The data will be useful to explore the biological and epidemiological significance of the intraspecific variation within H. taeniaeformis s.s.

First report on the phylogenetic relationship, genetic variation of Echinococcus shiquicus isolates in Tibet Autonomous Region, China

BackgroundCystic or alveolar echinococcosis caused by the larval stages of Echinococcus spp. is a very severe zoonotic helminth infection. Echinococcus shiquicus is a newly discovered species that has only been reported in the Qinghai and Sichuan provinces of the Qinghai-Tibet plateau, China where, to date, it has only been confirmed in Tibetan foxes and wild small mammal populations of the Tibetan plateau. Information on its genetic and evolutionary diversity is scanty. The aim of this study was to investigate the prevalence of E. shiquicus in plateau pikas (Ochotona curzoniae), a known intermediate host, and to determine the genetic variation and phylogenetic relationship of the E. shiquicus population in the Tibet region of China based on mitochondrial DNA.MethodsEchinococcus shiquicus samples were collected from Damxung and Nyêmo counties (located in Tibet Autonomous Region, China). The mitochondrial cox1 and nad1 gene sequences were analyzed, and the genetic diversity and epidemiology of E. shiquicus in the region were discussed based on the results.ResultsThe prevalence of E. shiquicus in pikas in Damxung and Nyêmo counties was 3.95% (6/152) and 6.98% (9/129), respectively. In combination with previous public sequence data, the haplotype analysis revealed 12 haplotypes (H) characterized by two distinct clusters (I and II), and a sequence distance of 99.1–99.9% from the reference haplotype (H1). The diversity and neutrality indices for the entire E. shiquicus populations were: haplotype diversity (Hd) ± standard deviation (SD) 0.862 ± 0.035; nucleotide diversity (Hd ± SD) 0.0056 ± 0.0003; Tajima's D 0.876 (P > 0.05); and Fu’s F 6.000 (P > 0.05).ConclusionsThis was the first analysis of the newly discovered E. shiquicus in plateau pikas in the Tibet Autonomous Region of China. The neutrality indices suggest a deficiency of alleles, indicative of a recent population bottleneck.Graphical

New records of Chondracanthus saundersii and Schottera koreana (Gigartinales, Rhodophyta) from Japan based on molecular and morphological analyses

SUMMARYWe report two red algal species, Chondracanthus saundersii C.W. Schneider & C.E. Lane (Gigartinaceae, Gigartinales) and Schottera koreana M.S. Calderon, T.H. Seo & S.M. Boo (Phyllophoraceae, Gigartinales), from Japan for the first time based on molecular and morphological analyses. A C. saundersii specimen was collected at a depth of 35 m offshore of Mageshima Island, Kagoshima Prefecture, Japan. The rbcL and COI‐5P (cox1) sequences of C. saundersii form a clade with the type specimen from Bermuda and additional specimens from Cuba and Brazil. Morphologically, C. saundersii is characterized by the presence of secondary attachments, compressed to flattened axes up to 1 mm in width, and a looser medulla. Two specimens of S. koreana were collected in lower intertidal to subtidal waters at Ohara, Chiba Prefecture, and Matsuyama, Ehime Prefecture, Japan. The rbcL sequences of S. koreana were closely related to reported sequences of specimens from South Korea including the type specimens. Morphologically, S. koreana is characterized by the presence of unicellular colorless hair developed from cortical cells.

Morphology, genetic characterization and molecular phylogeny of pinworm Skrjabinema longicaudatum n. sp. (Oxyurida: Oxyuridae) from the endangered Tibetan antelope Pantholops hodgsonii (Abel) (Artiodactyla: Bovidae)

BackgroundThe Tibetan antelope Pantholops hodgsonii (Abel) (Artiodactyla: Bovidae) is an endangered species of mammal endemic to the Qinghai-Tibetan Plateau. Parasites and parasitic diseases are considered to be important threats in the conservation of the Tibetan antelope. However, our present knowledge of the composition of the parasites of the Tibetan antelope remains limited.MethodsLarge numbers of nematode parasites were collected from a dead Tibetan antelope. The morphology of these nematode specimens was observed using light and scanning electron microscopy. The nuclear and mitochondrial DNA sequences, i.e. small subunit ribosomal DNA (18S), large subunit ribosomal DNA (28S), internal transcribed spacer (ITS) and cytochrome c oxidase subunit 1 (cox1), were amplified and sequenced for molecular identification. Moreover, phylogenetic analyses were performed using maximum likelihood (ML) inference based on 28S and 18S + 28S + cox1 sequence data, respectively, in order to clarify the systematic status of these nematodes.ResultsIntegrated morphological and genetic evidence reveals these nematode specimens to be a new species of pinworm Skrjabinema longicaudatum (Oxyurida: Oxyuridae). There was no intraspecific nucleotide variation between different individuals of S. longicaudatum n. sp. in the partial 18S, 28S, ITS and cox1 sequences. However, a high level of nucleotide divergence was revealed between the new species and its congeners in 28S (8.36%) and ITS (20.3–23.7%) regions, respectively. Molecular phylogenetic results suggest that the genus Skrjabinema should belong to the subfamily Oxyurinae (Oxyuroidea: Oxyuridae), instead of the subfamily Syphaciidae or Skrjabinemiinae in the traditional classification, as it formed a sister relationship to the genus Oxyuris.ConclusionsA new species of pinworm Skrjabinema longicaudatum n. sp. (Oxyurida: Oxyuridae) is described. Skrjabinema longicaudatum n. sp. represents the first species of Oxyurida (pinworm) and the fourth nematode species reported from the Tibetan antelope. Our results contribute to the knowledge of the species diversity of parasites from the Tibetan antelope, and clarify the systematic position of the genus Skrjabinema.

Seasonal dynamics and molecular differentiation of three natural Anopheles species (Diptera: Culicidae) of the Maculatus group (Neocellia series) in malaria hotspot villages of Thailand

BackgroundAnopheles sawadwongporni Rattanarithikul & Green, Anopheles maculatus Theobald and Anopheles pseudowillmori (Theobald) of the Anopheles maculatus group (Diptera: Culicidae) are recognized as potential malaria vectors in many countries from the Indian subcontinent through Southeast Asia to Taiwan. A number of malaria vectors in malaria hotspot areas along the Thai-Myanmar border belong to this complex. However, the species distribution and dynamic trends remain understudied in this malaria endemic region.MethodsMosquitoes of the Maculatus group were collected using CDC light traps every other week from four villages in Tha Song Yang District, Tak Province, Thailand from January to December 2015. Adult female mosquitoes were morphologically identified on site using taxonomic keys. Molecular species identification was performed by multiplex PCR based on the internal transcribed spacer 2 (ITS2) region of ribosomal DNA (rDNA) and sequencing of the cox1 gene at a DNA barcoding region in a subset of 29 specimens.ResultsA total of 1328 An. maculatus (sensu lato) female mosquitoes were captured with An. maculatus, An. sawadwongporni and An. pseudowilmori accounting for 75.2, 22.1 and 2.7% respectively. The field captured mosquitoes of the Maculatus group were most abundant in the wet season and had a preferred distribution in villages at higher elevations. The phylogenetic relationships of 29 cox1 sequences showed a clear-cut separation of the three member species of the Maculatus group, with the An. pseudowillmori cluster being separated from An. sawadwongporni and An. maculatus.ConclusionsThis study provides updated information for the species composition, seasonal dynamics and microgeographical distribution of the Maculatus group in malaria-endemic areas of western Thailand. This information can be used to guide the planning and implementation of mosquito control measures in the pursuance of malaria transmission.Graphical

Molecular data reveal unexpected species diversity of tapeworms of Australasian reptiles: revision of Kapsulotaenia (Cestoda: Proteocephalidae).

Species diversity and interrelationships of tapeworms of the genus Kapsulotaenia Freze, 1963 (Proteocephalidae: Acanthotaeniinae), parasites of lizards, especially monitors (Varanus spp.) in the Australasian region, were re-assessed using an interdisciplinary approach. Molecular phylogenetic analyses of newly characterized lsrDNA and cox1 sequences confirmed monophyly of the genus, which is typified by the presence of eggs in capsules, and also indicated a strict (oioxenous) level of host specificity of its species thus revealing unexpected species diversity. Diagnoses of insufficiently described species were amended based on a study of the types and freshly collected specimens and, in addition, three new species were described. A list of ten species of the genus recognized as valid is provided, including illustrations of taxonomically important structures of poorly known taxa. Kapsulotaenia beveridgei n. sp. from V. rosenbergi in Australia differs from all other species of Kapsulotaenia but K. frezei and K. saccifera by having a lower number of testes and an absence of banana-shaped clusters of eggs. Kapsulotaenia cannoni n. sp. from V. gouldii can be distinguished from all species but K. chisholmae by a smaller scolex diameter and from all remaining species by its bigger cirrus-sac ratio and a bigger Mehlis' gland/proglottid width ratio. Kapsulotaenia cannoni n. sp. differs from K. chisholmae, by the presence of an armed cirrus and a lower number of eggs in cluster (3-7 versus 8-13). Kapsulotaenia nybelini n. sp., which also occurs in V. gouldii, differs from K. tidswelli, K. frezei and K. beveridgei by having a greater number of testes, and it differs from K. varia by having a smaller relative size of the ovary. It differs from K. saccifera by the absence of banana-shaped cluster, and from K. pythonis by the number of eggs in clusters. Identification keys for all species of Kapsulotaenia and genera of the Acanthotaeniinae are also provided, together with SEM micrographs of three species, including two newly described species.

Molecular identification of Stylops advarians (Strepsiptera: Stylopidae) in western Canada.

Strepsiptera are an enigmatic order of insects with extreme sexual dimorphism which makes it difficult to "match-up" free-living adult males with parasitic conspecific females of the Stylopidia, and free-living females of the Mengenillidae using morphological characters. Species identification is further complicated for the Stylopidia because adult females are endoparasitic and neotenic. Therefore, we used DNA sequencing of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) to confirm the species identity of adult strepsipterans that were morphologically identified as Stylops advarians. These specimens, collected from Saskatoon (Saskatchewan, Canada), included one adult male, and eight females, the latter of which had been collected from solitary bees (Andrena milwaukeensis). Also included in the analyses were three pools of first-instar larvae that had emerged from three of the females. The results of the molecular analyses revealed that all specimens had an identical cox1 sequence, and belonged to a clade, with total statistical support (bootstrap value of 100%), that contained specimens of S. advarians from New York and Maine (USA). Hence, the results were consistent with the morphological identification of S. advarians. This study demonstrates the usefulness of a molecular approach for the identification of endoparasitic adult female and larval strepsipterans, life cycle stages that lack significant morphological characters for species identification.

Molecular Identification of Zoonotic Parasites of the Genus Anisakis (Nematoda: Anisakidae) from Fish of the Southeastern Pacific Ocean (Off Peru Coast).

The study aims to perform, for the first time, the molecular identification of anisakid larvae in commercial fish from the Southeastern Pacific Ocean off the Peru coast, and to provide data on their infection level by fishing ground, fish host, and site of infection. Fish specimens (N = 348) from the northern and the central coast of Peru were examined for parasites. The fish fillets were examined by the UV-press method. Anisakis spp. larvae (N = 305) were identified by mtDNA cox2 sequences analysis and by the ARMS-PCR of the locus nas10 nDNA. Two hundred and eighty-eight Anisakis Type I larvae corresponded to Anisakis pegreffii, whereas 17 Anisakis Type II larvae clustered in a phylogenetic lineage distinct from Anisakis physeteris deposited in GenBank, and corresponding to a phylogenetic lineage indicated as Anisakis sp. 2, previously detected in fish from both Pacific and Atlantic waters. Anisakis pegreffii was found to infect both the flesh and viscera, while Anisakis sp. 2 occurred only in the viscera. The average parasitic burden with A. pegreffii in the examined fish species from the two fishing grounds was significantly higher than that observed with Anisakis sp. 2. The results obtained contribute to improve the knowledge on the distribution and occurrence of Anisakis species in Southeastern Pacific waters and their implications in seafood safety for the local human populations.

<p><strong><em>S</em></strong><strong><em>arcopeltis gen. nov. </em></strong><strong>(Gigartinaceae, Rhodophyta), with <em>S</em>. <em>skottsbergii comb. nov. </em>from southern South America and<em> S. antarctica</em> <em>sp. nov.</em> from the Antarctic Peninsula</strong></p>

Gigartina skottsbergii is the currently accepted name for the large, peltate species of Gigartinaceae distributed in Argentina, Chile, and along the Antarctic Peninsula. However, phylogenetic analyses of DNA sequences for more than twenty years indicate that two species are passing under the name G. skottsbergii, and both DNA sequences and morpho-anatomy indicate that the species are assignable to an undescribed genus. To formally validate the new genus and species, we performed next generation sequencing (NGS) on the lectotype material of G. skottsbergii to document its genetic identity and obtained its complete mitogenome and partial plastid genome. Phylogenetic analysis of rbcL and cox1 sequences of the lectotype along with recent collections of G. skottsbergii from southern South America and Antarctica, confirmed the generic distinction and the presence of two morphologically similar taxa. We propose Sarcopeltis gen. nov., containing the generitype S. skottsbergii comb. nov. from southern South America, and S. antarctica sp. nov. from the Antarctic Peninsula. Sarcopeltis is characterized by the following suite of features: 1) peg-like secondary haptera that adhere to the substratum, 2) absence of an envelope surrounding each cystocarp, 3) presence of extensive, terminal, tubular gonimoblast filaments that fuse with surrounding vegetative cells, and 4) tetrasporangia formed from secondary filaments entirely in the medulla. This study demonstrates that genetic analysis of type material, together with recently obtained DNA sequences from field collected specimens, can provide clear and objective taxonomic conclusions.

Gayliella nanaoensis sp. nov. (Ceramiaceae, Rhodophyta) from China

ABSTRACT Because morphological variation in Gayliella species is extensive, a molecular-assisted taxonomic approach is essential for accurate species identification. In China, six Gayliella species have been recorded, but no molecular-assisted identification has been used. In this study, we used detailed morphological analyses combined with rbcL and cox1 sequence analyses to identify Gayliella specimens from Nan’ao Island, Shantou city, China. Morphological analyses revealed that our specimens can be distinguished from other species of Gayliella by a combination of morphological characters: branching at intervals 4(–5) on main axes, 4–6 on lateral axes; five periaxial cells; short basipetal corticating filaments only 1–2 cells long; ovoid gland cells developed from periaxial cells; and tetrasporangia whorled at a node. Molecular sequence analysis also revealed sufficient sequence divergence between our specimens and other Gayliella species to separate them. Therefore, based on combined morphological and molecular analyses, we describe the specimens from Nan’ao Island, Shantou city, China, as Gayliella nanaoensis sp. nov.

Parasites of an Arctic scavenger; the wolverine (Gulo gulo).

Parasites are fundamental components within all ecosystems, shaping interaction webs, host population dynamics and behaviour. Despite this, baseline data is lacking to understand the parasite ecology of many Arctic species, including the wolverine (Gulogulo), a top Arctic predator and scavenger. Here, we combined traditional count methods (i.e. adult helminth recovery, where taxonomy was confirmed by molecular identification) with 18S rRNA high-throughput sequencing to document the wolverine parasite community. Further, we investigated whether the abundance of parasites detected using traditional methods were associated with host metadata, latitude, and longitude (ranging from the northern limit of the boreal forest to the low Arctic and Arctic tundra in Nunavut, Canada). Adult parasites in intestinal contents were identified as Baylisascaris devosi in 72% (n = 39) of wolverines and Taenia spp. in 22% (n = 12), of which specimens from 2 wolverines were identified as T. twitchelli based on COX1 sequence. 18S rRNA high-throughput sequencing on DNA extracted from faeces detected additional parasites, including a pseudophyllid cestode (Diplogonoporus spp. or Diphyllobothrium spp.), two metastrongyloid lungworms (Angiostrongylus spp. or Aelurostrongylus spp., and Crenosoma spp.), an ascarid nematode (Ascaris spp. or Toxocara spp.), a Trichinella spp. nematode, and the protozoan Sarcocystis spp., though each at a prevalence less than 13% (n = 7). The abundance of B. devosi significantly decreased with latitude (slope = -0.68; R2 = 0.17; P = 0.004), suggesting a northerly limit in distribution. We describe B. devosi and T. twitchelli in Canadian wolverines for the first time since 1978, and extend the recorded geographic distribution of these parasites ca 2000 km to the East and into the tundra ecosystem. Our findings illustrate the value of molecular methods in support of traditional methods, encouraging additional work to improve the advancement of molecular screening for parasites.

Two species of Dioncopseudobenedenia Yamaguti, 1965 (Monogenea: Capsalidae) from Naso unicornis (Forsskål) (Acanthuridae) and Coryphaena hippurus Linnaeus (Coryphaenidae) in western Japan, with a description of D. elongata n. sp. from N. unicornis.

Three species of Dioncopseudobenedenia Yamaguti, 1965 (Monogenea: Capsalidae: Benedeniinae) have been described, all found parasitising Naso spp. (Acanthuridae) and Siganus lineatus (Siganidae), in Hawaii, the Great Barrier Reef, and New Caledonia in the Pacific. In this paper, D. elongata n. sp. is described, found parasitising the gills of N. unicornis (Forsskål) caught off Okinawa-jima Island. Dioncopseudobenedenia kala Yamaguti, 1965 is also reported, found parasitising the gills of N. unicornis, and those of a new host, the common dolphinfish Coryphaena hippurus Linnaeus (Coryphaenidae), off Okinawa and Kochi prefectures, Japan. The new species differed from known species in both nuclear (28S rDNA and ITS1-5.8S-ITS2) and mitochondrial DNA (cox1) sequences. Morphologically, the new species differed from other species of Dioncopseudobenedenia in possessing a longer accessory sclerite and a thick, short penis. No species of Dioncopseudobenedenia have been detected during past examinations of C. hippurus; this instance was probably an accidental infection. A key for identification of the species of Dioncopseudobenedenia is also provided herein.

Sarcocystis spp. infection in South American deer huemul (Hippocamelus bisulcus) and pudu (Pudu puda) from Patagonian National Parks, Argentina.

Sarcocystis spp. are intracellular protozoan parasites with heteroxenous life cycles. This study described Sarcocystis spp. infection in adult South American native deer huemul (Hippocamelus bisulcus) and pudu (Pudu puda). Heart, diaphragm, tongue, and skeletal muscle samples were collected from 5 huemuls and 2 pudus, found dead in National Parks. Direct microscopic examination, transmission electron microscopy, PCR, and sequencing were performed. Sarcocystis spp. microscopic thin-walled cysts were identified in 3 huemuls and 1 pudu. Several cysts from 1 huemul and 1 pudu were observed by TEM; ultrastructure was similar to previously reported as cyst wall type 17 and types 2 and 8, respectively. Fragments of the 18S rRNA and cytochrome c oxidase subunit I (cox1) genes were amplified and sequenced from 3 individual cysts from 2 huemuls and 2 cysts from the pudu. The sequences from huemuls showed a high identity among them (> 99%) at both amplified targets. The highest identities were > 99.7% at 18S rRNA and 93% at cox1 with S. tarandivulpes sequences. The 18S rRNA gene sequences from pudus showed an identity > 99.7% with Sarcocystis sp., S. taeniata, and S. linearis sequences, while the cox1 sequences were different, one showing 99.42% identity with S. venatoria and the other 98.22% with S. linearis. A single species, similar to S. tarandivulpes, was identified in all huemul samples while 2 molecularly different Sarcocystis spp. were found in 1 pudu with high similarities to either S. venatoria or to S. linearis, S. taeniata-like, and S. morae. Based on the cox1 sequence identities, at least the Sarcocystis sp. in huemuls might represent a new species, primarily occurring in this host. Additional sarcocyst isolates from both hosts need to be examined molecularly in order to firmly establish whether these species are indeed native to huemuls and/or pudus or are derived from introduced deer species.