SSU rDNA Sequences Research Articles

Additions to the genus Kirschsteiniothelia (Dothideomycetes); Three novel species and a new host record, based on morphology and phylogeny.

During a survey of microfungi associated with forest plants, four specimens related to Kirschsteiniothelia were collected from decaying wood in Guizhou, Hainan and Yunnan Provinces, China. Kirschsteiniothelia species have sexual and asexual forms. They are commonly found as saprophytes on decaying wood and have been reported as disease-causing pathogens in humans as well. In this study, we introduce three novel Kirschsteiniothelia species (K.bulbosapicalis, K.dendryphioides and K.longirostrata) and describe a new host record for K.atra, based on morphology and multi-gene phylogenetic analyses of a concatenated ITS, LSU and SSU rDNA sequence data. These taxa produced a dendryphiopsis- or sporidesmium-like asexual morph and detailed descriptions and micromorphological illustrations are provided. Furthermore, we provide a checklist for the accepted Kirschsteiniothelia species, including detailed host information, habitat preferences, molecular data, existing morphological type, country of origin and corresponding references.

Prasiolopsis wulf-kochii (Prasiolales, Trebouxiophyceae), a New Species Occurring in Hairs of the Sloth Bradypus tridactylus.

The monotypic genus Prasiolopsis has been known for a long time, but is often overlooked because of difficulties in identification and the morphological variability between uniseriate filaments and cell packages forming pseudoparenchymatic thalli depending on age. We investigated a strain (SAG 84.81) originally denoted as Trichophilus welckeri, which was isolated from the hairs of the sloth Bradypus tridactylus, and compared it with other available strains of Prasiolopsis and of the sister genus Pseudomarvania. Our investigations clearly showed that this strain differed in morphology, especially of the chloroplast, from those originally described for Trichophilus. Phylogenetic analyses of the SSU and ITS rDNA sequences revealed that the strain SAG 84.81 is sister to several strains of P. ramosa within the Prasiola clade (Trebouxiophyceae). Using the ITS-2/CBC approach, we clearly demonstrated that this strain represented a new species of Prasiolopsis, which we proposed here as P. wulf-kochii. In addition, we evaluated the ITS-2/CBC approach by comparing it with the two species of Pseudomarvania. All investigated strains showed CBCs and HCBCs, which support their species delimitation. The sequencing data of Trichophilus welckeri available in GenBank were phylogenetically re-evaluated by including all representatives of the Ulotrichales (Ulvophyceae). Our analyses showed that these sequences formed their own lineage within this order.

Molecular detection of Babesia and Hepatozoon species and morphological characteristics of Babesia species in Japanese wild boars

We investigated intraerythrocytic Babesia parasites in 21 Japanese wild boars, Sus scrofa leucomystax, captured in Wakayama Prefecture on the mainland from 2008 to 2009 and in 31 Japanese wild boars from 2011 to 2013 in Kochi Prefecture on Shikoku Island, Japan. We detected small subunit ribosomal RNA (18S rRNA) gene (SSUrDNA) fragments of a Babesia species in 17 boars from Wakayama and 18 boars from Kochi. The nearly full SSUrDNA sequence (1669 bps) of this species was determined. A FASTA search revealed that the SSUrDNA sequence of the Babesia sp. in Japanese wild boars was the most homologous to those of several Babesia isolates reported as Babesia gibsoni. Phylogenetic analysis showed that the Babesia sp. found in Japanese wild boars was the closest relative to B. gibsoni but made a different clade from B. gibsoni. The Babesia sp. in Japanese wild boars was completely different from Babesia sp. Suis found in a European domestic pig, Sus scrofa domesticus. By microscopic examination, ring-shaped, oval and pear-shaped small sized intraerythrocytic parasites were observed on blood smears of 12 of 18 Japanese wild boars whose blood smears could be examined in Wakayama. We also detected SSUrDNA fragments of a Hepatozoon species in 6 of the 21 wild boars from Wakayama. The nearly full SSUrDNA sequence (1774 bps) of the Hepatozoon sp. was shown to be identical to that of Hepatozoon apri.

Morphological and Molecular Characterization of Three Myxosporean Species of the Genera Myxobolus, Henneguya, and Myxidium (Cnidaria: Myxozoa) Infecting Freshwater Fish, Isolated for the First Time in Japan.

The majority of myxosporean species (Cnidaria: Myxozoa) of the genera Myxobolus (35 species), Henneguya (8 species), and Myxidium (9 species) from freshwater or brackish fish in Japan were recorded more than 30 years ago (accumulatively 81.1% [43/53]). The re-discovery and molecular-genetic characterization of these species is a current research priority. During our myxosporean survey in Japanese freshwater fish, we detected three species that had never been recorded in Japan, but in the Russian Far East (Sakhalin Island, and Maritime Province): Myxobolus tribolodonus sp. n., forming cysts in the gills of Tribolodon sachalinensis (syn. M. marinus sensu Aseeva, 2000; M. marinus sensu Sokolov et Frolova, 2015, recorded from the gills of Pseudaspius (syn. Tribolodon) spp.); Henneguya pungitii Achmerov, 1953, forming cysts in the subcutis of external skin and buccal submucosa of Pungitius sinensis; and Myxidium salvelini Konovalov et Shulman, 1966, in the urinary bladder of Oncorhynchus masou ishikawae. These new isolates were characterized by integrated taxonomic approaches, i.e., myxospore morphology and molecular-genetic characterization of the small subunit ribosomal RNA gene (SSU rDNA). These new isolates were phylogenetically differentiated from any species whose SSU rDNA sequences were deposited in the DNA databases, and concurrently compared with recorded species based on classical morphological criteria. All three species were differentiated from myxosporeans previously recorded in Japan, indicating new distribution records out of the Russian Far East. For reliable species identification, accumulation of at least SSU rDNA sequences of known species worldwide is critically important.

Mass mortality event of round sardinella Sardinella aurita Valenciennes associated with Glugea Thélohan, 1891 microsporidian infection off the southern Italian coast.

Intracellular parasites of the genus Glugea Thélohan, 1891 (Microsporidia) comprise about 34 putative species capable of causing high morbidity and mortality in freshwater and marine teleost fishes. In this study, we report on the first mass mortality event associated with Glugea sp. infecting free-ranging round sardinella Sardinella aurita in the southern Tyrrhenian Sea (Italy). Here, we describe the ultrastructure of mature spores of this microsporidian and characterize it molecularly, as well as report its phylogenetic position. Most of the affected fish showed an irregular swelling of its abdomen. At necropsy, a variable number of xenomas, spherical to ellipsoidal in shape, were found in the peritoneal cavity strongly attached to the viscera of all fish. Histological analysis revealed varying severity of chronic inflammation along with occasional necrosis in visceral organs associated with multiple xenoma proliferation. These pathological findings were considered the main cause of this mass mortality event. Morphologically, the present material was closely related to G. sardinellesis and G. thunni. The phylogenetically closest taxa to the newly SSU rDNA sequence were G. thunni and an erroneusly identified G. plecoglossi, which were very closely related to each other, also suggesting that all these sequences might belong to the same species.

Cryptosporidium occultus in disguise

As data accumulate in GenBank, the difficulties of delineating species of Cryptosporidium based on nuclear small subunit ribosomal RNA (ssu rRNA) gene information alone becomes increasingly evident. Here, we summarize currently available evidence suggesting that several ssu rDNA sequences primarily referred to as Cryptosporidium suis (some of them from non-suid hosts) should be considered Cryptosporidium occultus.

Synopsis of the species of OrtholineaShulman, 1962 (Cnidaria: Myxosporea: Ortholineidae)

A synopsis of Ortholinea Shulman, 1962 (Cnidaria: Myxosporea: Ortholineidae) is presented and identifies 26 nominal species presently allocated within this genus. Species morphological and morphometric features, tissue tropism, type-host, and type-locality are provided from original descriptions. Data from subsequent redescriptions and reports is also given. Accession numbers to sequences deposited in GenBank are indicated when available, and the myxospores were redrawn based on original descriptions. The information gathered shows that Ortholinea infect a wide taxonomic variety of freshwater and marine fish. Nonetheless, the broad host specificity reported for several species is not fully supported by morphological descriptions and requires molecular corroboration. The members of this genus are coelozoic and mainly parasitize the urinary system, with few species occurring in the gallbladder. Ortholinea visakhapatnamensis is the only exception, being histozoic in the visceral peritoneum. Molecular data of the small subunit ribosomal RNA gene (SSU rDNA) is available for about one third of Ortholinea species, with genetic interspecific variation ranging between 1.65% and 29.1%. Phylogenetic analyses reveal Ortholinea to be polyphyletic, with available SSU rDNA sequences clustering within the subclades of the highly heterogenous freshwater urinary clade of the oligochaete-infecting lineage. The life cycles of two Ortholinea species have been clarified based on molecular inferences and identify triactinomyxon actinospores as counterparts, and marine oligochaetes of the family Naididae as permissive hosts to this genus.

The first record of \(\textit{Kudoa yasunagai}\) (Hsieh & Chen, 1984) (Myxosporea: Multivalvulida) parasitizing the brain of barhead spinefoot \(\textit{Siganus virgatus}\) (Valenciennes) from Vietnam

A brain-infecting myxozoan, Kudoa yasunagai (Hsieh & Chen, 1984), was identified using both morphological and molecular methods in a marine fish from Van Phong Bay, Khanh Hoa province, Vietnam. Cysts containing spores were observed in the brain of one out of 10 Siganus virgatus (Valenciennes) individuals (10%). The myxospores were radially symmetrical, containing a majority of 7 equal shell valves and polar capsules. The spores measured 6.6 ± 0.1 (6.4‒6.7) µm in length, 7.6 ± 0.5 (7.0‒8.4) µm in width, and 7.0 ± 0.4 (6.5‒7.7) µm in thickness. Polar capsules measured 2.8 ± 0.2 (2.6‒3.0) µm in length and 1.4 ± 0.1 (1.2‒1.5) µm in width, containing a filament inside. The SSU rDNA sequence from the Vietnamese specimen showed > 99.94 % identity with K. yasunagai from Japan and Australia. This report marks a new geographical and host record for K. yasunagai.

Identification of new amoebae strains in rainbow trout (Oncorhynchus mykiss, Walbaum) farms affected by nodular gill disease (NGD) in Northeastern Italy.

Nodular gill disease (NGD) is an emerging condition associated with amoeba trophozoites in freshwater salmonid farms. However, unambiguous identification of the pathogens still must be achieved. This study aimed to identify the amoeba species involved in periodic NGD outbreaks in two rainbow trout (Oncorhynchus mykiss) farms in Northeastern Italy. During four episodes (February-April 2023), 88 fish were euthanized, and their gills were evaluated by macroscopic, microscopic and histopathological examination. The macroscopic and microscopic severity of the lesions and the degree of amoebae infestation were scored and statistically evaluated. One gill arch from each animal was put on non-nutrient agar (NNA) Petri dishes for amoeba isolation, cultivation and subsequent identification with SSU rDNA sequencing. Histopathology confirmed moderate to severe lesions consistent with NGD and mild to moderate amoeba infestation. The presence of amoebae was significantly correlated with lesion severity. Light microscopy of cultured amoebae strains and SSU rDNA analysis revealed the presence of a previously characterized amoeba Naegleria sp. strain GERK and several new strains: two strains from Hartmannelidae, three vannelid amoebae from the genus Ripella and cercozoan amoeba Rosculus. Despite the uncertainty in NGD etiopathogenesis and amoebae pathogenic role, identifying known and new amoebae leans towards a possible multi-aetiological origin.

Henneguya (Cnidaria: Myxobolidae) species infecting Oligosarcus jenynsii (Characiformes: Characidae) in a Neotropical shallow lake from Argentina: morphological and molecular characterisation.

Two previously undescribed myxozoan species, Henneguya sardellae sp. n. and H. margaritae sp. n., found infecting connective tissues of the Neotropical characid fish Oligosarcus jenynsii (Günther) from Argentina are morphologically and molecularly characterised. Mature spores of H. sardellae sp. n. are ellipsoid, with two, straight and visibly fused caudal appendages cleaved at its blunt terminal end; measuring 33.5 ± 1.2 (30.9-35.5) μm in total length, spore body 17.5 ± 0.6 (16.3-18.6) µm, 7.8 ± 0.4 (7.0-8.8) µm wide and 6.9 ± 0.2 (6.6-7.2) µm thick, with two elongated, unequally-sized polar capsules situated at anterior end, and 11-13 turns of polar tubules. Mature spores of H. margaritae sp. n. are pyriform, with two caudal appendages visible fused together and much longer than spore body, with unequal endings; measuring 35.9 ± 2.8 (29.2-40.7) µm in total length, spore body 11.5 ± 0.9 (9.2-13.0) µm long, 5.8 ± 0.4 (5.1-6.7) µm wide and 5.5 ± 0.2 (5.1-5.8) µm thick, with two polar capsules similar in size, pyriform polar capsules containing polar tubules with 4-5 coils. Both species showed a membraneous sheath surrounding the spore body and caudal appendages; in H. sardellae sp. n. this feature can deploy laterally. Phylogenetic analyses based on SSU rDNA sequences showed that H. sardellae sp. n. and H. margaritae sp. n. clustered with other myxobolids parasitising Characiformes in Brazil, Cichliformes in Mexico and Cyprinodontiformes in Mexico and the United States. The description of these two new species of Henneguya as the first described species of the genus that parasitise freshwater fish in Argentina highlights the importance of further research on the diversity and distribution of myxozoans in this region.

Morphological and Molecular Characterization of Two New Species of Ceratomyxa Thélohan, 1892 (Cnidaria: Myxosporea) from the Marine Ornamental Fish Zanclus cornutus (Linnaeus, 1758) off Lakshadweep Islands, Arabian Sea.

The present study provides the complete morphological and molecular description of two new species of myxosporeans, Ceratomyxa zancli n. sp. and Ceratomyxa cornuti n. sp. infecting the gallbladder of Zanclus cornutus from the Lakshadweep Islands, Arabian Sea. Zanclus cornutus were screened for the presence of myxosporeans, and the recovered myxospores were morphologically characterized using Nomarski Differential Interference Contrast (DIC) optics. The sequences of SSU rDNA were employed for molecular and phylogenetic studies. Both the parasites exhibited a prevalence of 21% each. C. zancli n. sp. is characterized by broadly cresentic myxospores with convex anterior and slightly concave to straight posterior margins and rounded ends. Spore valves two, unequal, measured 9.6 ± 0.7μm × 25.2 ± 1.3μm. Polar capsules two, unequal, spherical, measured 4 ± 0.6μm × 3.5 ± 0.6μm. Polar filament exceptionally long and arranged irregularly. Myxospores of C. cornuti n. sp. are elongated with convex anterior and slightly concave to straight posterior margins. Spore valves two, unequal, measured 7.00 ± 0.4μm × 26.56 ± 1.8μm. Polar capsules spherical, unequal, measured 3.52 ± 0.2 × 3.36 ± 0.35. Molecular analysis of C. zancli n. sp. (ON818297) and C. cornuti n. sp. (ON818298) resulted in 1469 and 1491bp long SSU rDNA sequences, respectively. Molecularly C. zancli n. sp. is close to C. diplodae and C. barnesi with 91.39% similarity, while C. cornuti n. sp. appears closer to C. robertsthomsoni with 97.46% similarity. In phylogenetic analyses, C. zancli n. sp. branched separately within the Ceratomyxa clade while C. cornuti n. sp. clustered with C. robertsthomsoni and C. thalassomae. Based on the differences in morphological, morphometric, molecular, and phylogenetic characteristics, as well as differences in the host and geographic location, the above two species of myxosporeans are considered novel. The study forms the first report of a species of Ceratomyxa from Z. cornutus.

Solving the Myxidium rhodei (Myxozoa) puzzle: insights into its phylogeny and host specificity in Cypriniformes.

Myxidium rhodei Léger, 1905 (Cnidaria: Myxozoa) is a kidney-infecting myxosporean that was originally described from the European bitterling Rhodeus amarus. Subsequently, it has been documented based on spore morphology in more than 40 other cypriniform species, with the roach Rutilus rutilus being the most commonly reported host. This study introduces the first comprehensive data assessment of M. rhodei, conducted through morphological, ecological and molecular methods. The morphological and phylogenetic analyses of SSU rDNA sequences of Myxidium isolates obtained from European bitterling and roach did not support parasite conspecificity from these fish. In fact, the roach-infecting isolates represent three distinct parasite species. The first two, M. rutili n. sp. and M. rutilusi n. sp., are closely related cryptic species clustering with other myxosporeans in the freshwater urinary clade, sharing the same tissue tropism. The third one, M. batuevae n. sp., previously assigned to M. cf. rhodei, clustered in the hepatic biliary clade sister to bitterling-infecting M. rhodei. Our examination of diverse cypriniform fishes, coupled with molecular and morphological analyses, allowed us to untangle the cryptic species nature of M. rhodei and discover the existence of novel species. This underscores the largely undiscovered range of myxozoan diversity and highlights the need to incorporate sequence data in diagnosing novel species.

First molecular identification of the trematode Pulmonicola cochleotrema (Platyhelminthes: Opisthotrematidae) in West Indian manatees (Trichechus manatus, Sirenia: Trichechidae) from Puerto Rico and Florida.

West Indian manatees (Trichechus manatus) harbor a variety of endoparasites, including the nasal trematodePulmonicola cochleotrema, which infects the respiratory tract, especially the nasal passages. Previous studies have described and identified this digenean using morphological data only. This study presents the first molecular identification ofP. cochleotremain West Indian manatees from Puerto Rico and Florida. Samples of the trematode were collected from seven manatees found stranded dead at both locations. The small subunit ribosomal DNA (18S rDNA) was amplified from each sample using universal primers for different regions of the gene, resulting in a consensus sequence of 1871 base pairs. The phylogenetic reconstruction was carried out using DNA sequences of other species of digenean parasites from other hosts, including a trematode of the same taxonomic family from another sirenian species. Specimens collected from both locations show the same molecular identity using SSU rDNA sequence data. The identity ofP. cochleotremawas confirmed using the Basic Local Alignment Search Tool from the National Center for Biotechnology Information database, yielding a high similarity of 98.8 % withOpisthotrema dujonisand 98.2 % withLankatrema mannarenselocated in the same clade in our analysis. The latter two digeneans belong to the Opisthotrematidae as doesP. cochleotrema and previous studies reported them infecting the Eustachian tubes, esophagus, and digestive tract in dugongs (Dugong dugon). These findings evidence that the nasal trematode of manatees in Florida and Puerto Rico and the dugong, all inhabiting and feeding in marine environments, will have a marine mollusk as an intermediate host, probably a gastropod. The question remains, which species of nasal trematode are harbored by lotic-dwelling manatees in other parts of their distribution like South America.

Post-mortem ‘soft flesh’ in three commercial fish species from off Atlantic Morocco associated with the myxosporean parasites Kudoa thyrsites and K. encrasicoli (Myxozoa)

Small pelagic fishes represent one of the most important food resources off the Northwest coast of Africa. Despite their economic significance, little is known about the infections with flesh invading myxosporean parasites of genus Kudoa (Cnidaria, Myxozoa). Heavy infections in the flesh may be associated with post-mortem myoliquefaction, commonly known as ‘soft flesh’. This condition may reduce the quality and marketability of the fish fillet, resulting in both economic losses to the fishing industry and loss of consumer confidence. In this study, we investigated Kudoa-induced ‘soft flesh’ occurrence in European anchovy Engraulis encrasicolus, European pilchard Sardina pilchardus, and Atlantic chub mackerel Scomber colias caught in 2019 off the Moroccan Atlantic coast. Five hundred specimens of each fish species were examined for ‘soft flesh’ by texture testing and visual inspection 48 h post-catch. ‘Soft flesh’ occurred in 0.2 % of the European anchovies, 1.4 % of the European pilchard, and in 4.4 % of the Atlantic chub mackerel. Microscopic examination of muscle samples revealed that ‘soft flesh’-affected fish were infected with myxospores of K. thyrsites-like morphotype. Analysis of the kudoid SSU rDNA sequence obtained from European pilchard and the Atlantic chub mackerel identified these as K. thyrsites (100 % identity), whereas analysis of the sequence from European anchovy identified the presence of K. encrasicoli (100 % identity). Even if there are no known human health consequences associated with the ingestion of these Kudoa species, the unsightly appearance of some infected fillets is a food quality issue, that can eventually lead to reduced marketability and value.

Morphogenesis of an anaerobic ciliate Heterometopus palaeformis (Kahl, 1927) Foissner, 2016 (Ciliophora, Armophorea) with notes on its morphological and molecular characterization

The morphology, morphogenesis, and molecular phylogeny of Heterometopus palaeformis (Kahl, 1927) Foissner, 2016 were studied using microscopical observations on live and protargol-stained specimens as well SSU rRNA gene sequencing. The morphogenetic data for the genus are presented for the first time. Compared to other metopids, the morphogenesis of H. palaeformis is distinct since its (1) perizonal stripe rows 4 and 5 are involved in the formation of the opisthe’s adoral polykinetids; (2) perizonal stripe rows 3–5 and two adjacent preoral dome kineties contribute to most of the opisthe’s paroral membrane while perizonal stripe rows 1 and 2 contribute very little; (3) four kinety rows are formed to the left of the opisthe’s adoral zone of polykinetids. The Chinese population resembles the original and neotype populations well in terms of general morphology — characterized by a life size of 55–120 × 10–20 μm, an elongate ellipsoidal body with a hardly spiralized flat preoral dome, about 18 somatic kineties and 20 adoral polykinetids. The SSU rDNA sequence of the present population exhibits a disparity of 1.33%–2.22% divergence from sequences of other populations. Nevertheless, phylogenetic analysis reveals that populations of H. palaeformis form a separate, stable cluster within the paraphyletic Metopidae clade.

Description of Holostichides (Extraholostichides) eastensis tianjinensis subgen. nov. subspec. nov. (Ciliophora, Hypotricha) from northern China

The morphology and morphogenesis of a new urostylid, Holostichides (Extraholostichides) eastensis tianjinensis subgen. nov. subspec. nov. were analyzed. The new subspecies differs from the nominotypical subspecies H. (Extraholostichides) eastensis eastensis Wang et al., 2022 by the relatively long frontoterminal row (about 60% vs. 30% of body length), colorless cortical granules (vs. dark brown), two (vs. one) parabuccal cirri, and usually an extra cirrus behind the first midventral pair (vs. lacking). Based on the difference in the frontal ciliature, we split Holostichides into two subgenera: H. (Extraholostichides) subgen. nov. (type species Holostichides eastensis Wang et al., 2022; with a short cirral row behind the middle frontal cirrus) and H. (Holostichides) Foissner, 1987 (type species Holostichides chardezi Foissner, 1987; lacking this short row). The main morphogenetic characters of the new subspecies are very similar to those of H. (Extraholostichides) eastensis eastensis except for some minor differences. Phylogenetic analyses based on SSU rDNA sequences indicate that H. (Extraholostichides) subgen. nov. is monophyletic and nested within the monophyletic genus Holostichides, which is sister to Eschaneustyla lugeri.

Phylogenetic analysis of the Cladophora coelothrix complex, including the description of the new genus Leliaertia (Cladophorales, Ulvophyceae)

ABSTRACT Molecular phylogenetic studies have revealed the polyphyletic nature of Cladophora as traditionally circumscribed, which is separated into four main clades: Cladophoraceae, Siphonocladus, Pithophoraceae and Pseudocladophoraceae. Several taxonomic proposals have recently been made to resolve its polyphyletic nature. The present work focuses on the cryptic diversity of Cladophora coelothrix, within the Siphonocladus clade. Based on new collections from northeastern Brazil and phylogenetic analyses of partial SSU and LSU rDNA sequence alignments, we propose the new genus and species Leliaertia repens to accommodate one of the cryptic species hitherto included in C. coelothrix, occurring in the tropical Atlantic, Indian and Pacific Oceans. Leliaertia repens was recovered in the Siphonocladus clade and is phylogenetic distinct from the genuine Cladophora species in the Cladophoraceae. The new species occurs in marine and estuarine environments and forms conspicuous mats of semi-prostrate filaments. It represents a cryptic species and genus since morphological features, including the diameter and length:diameter ratio of the apical cell, show high levels of overlap with other species, including C. socialis. Our phylogenetic data indicate that L. repens, C. coelothrix and C. socialis represent a complex of cryptic species with at least four lineages, some of which co-occur in tropical seas.

A full-length SSU rRNA-based workflow for high-resolution monitoring of nematode communities reveals direct and indirect responses to plant-based manipulations

Agricultural intensification has resulted in a decline in soil biodiversity, and concerns about the deterioration of the biological condition of soils prompted the development of measures to restore soil life. Due to the overwhelming biodiversity of soils, evaluation of such measures is not straightforward, and proxies are used to assess soil health. Because of their trophic diversity, high abundance, and relatively well-characterized ecologies, nematodes are often used as soil health indicators. However, the scarcity of informative morphological characters hampers the upscaling of this proxy. Here we present a community analysis approach that uses nanopore sequencing to generate full-length sequences of small subunit ribosomal DNAs (SSU rDNA). Cover cropping is a common agricultural practice that stimulates soil life, and we mapped the effects of ten cover crop treatments on nematode communities in a field experiment. These analyses included the monitoring of a high-impact plant-parasite, Meloidogyne chitwoodi. In total, 132 nematode samples were analyzed, and 65 nematode taxa were detected, mostly at the species level, including representatives of all trophic groups. As a validation, all samples were analyzed microscopically for M. chitwoodi, and a comparison of count and DNA read data revealed highly similar results. Treatments did not only affect plant-parasitic nematodes but also free-living nematodes in a cover crop-specific manner. Free-living nematodes from the same trophic group, and even congeneric species, responded differentially to plant-mediated manipulations of the soil microbiome. Hence, nanopore-based SSU rDNA sequencing could facilitate a substantial refinement of the use of nematodes as indicators for soil health.

Description and development of Auerbachia ignobili n. sp. (Cnidaria: Myxosporea: Bivalvulida) from the giant trevally, Caranx ignobilis (Forsskål, 1775) from Indian waters.

The present study describes a new species of myxosporean, Auerbachia ignobili n. sp., infecting the hepatic bile ducts of Caranx ignobilis (Forsskål, 1775). Myxospores are club-shaped with a broad anterior region and a narrow, slightly curved and blunt caudal extension, measuring 17.4 ± 1.5 μm in length and 7.5 ± 7.4 μm in width. Shell valves asymmetrical, with a faint suture line, and enclosed a single, elongate-elliptical polar capsule with a ribbon-like polar filament, arranged in 5-6 coils. Developmental stages included early and late presporogonic stages, pansporoblast, and sporogonic stages with monosporic and disporic plasmodia. A. ignobili n. sp. differs from the other described species of Auerbachia in the shape and dimensions of the myxospores and polar capsules. The molecular analysis generated ~1400 bp long SSU rDNA sequences and the present species exhibited a maximum similarity 94.04-94.91% with A. chakravartyi. Genetic distance analysis indicated the lowest interspecies divergence of 4.4% with A. chakravartyi. In phylogenetic analysis, A. ignobili n. sp. was positioned independently with a high bootstrap value (1/100) and appeared as sister to A. maamouni and A. chakravartyi. Fluorescent in situ hybridization and histology indicates that the parasite develops within the hepatic bile ducts. Histological studies did not reveal any pathological changes. Considering the morphological, morphometric, molecular, and phylogenetic differences coupled with the differences in host and geographic locations, the present myxosporean is treated as a new species and named A. ignobili n. sp.

Morphological, histological and phylogenetic identification of three species of Myxobolus (Myxosporea: Myxobolidae) infecting different carp lineages in China.

The global cultivation of common carp Cyprinus carpio is developed primarily for either food or recreational purposes and myxosporean infections causing significant economic losses are regularly troublesome for carp farmers. However, most myxosporean species are poorly characterized, making it difficult to correctly elaborate on their parasitism mode and determine pathogenicity. Here, based on an integrative approach, we supplemented fundamental information on three Myxobolus species infecting different carp lineages in China. Myxobolus elliptoides Wu et Chen, 1987; Nine new species of Myxosporida from freshwater fish of Wuhu lake, Hubei, China. Acta Hydrobiologica Sinica, 11, 161 formed yellowish pseudocysts in the anal fin of common carp and were observed with ellipsoidal myxospores and two polar capsules of distinct dimensions. The dermal tissue within the fin ray was regarded as the type site for growing plasmodia of M. elliptoides. Myxobolus basilamellaris Lom et Molnar, 1983; Myxobolus basilamellaris sp. n. (myxozoa, Myxosporea), a parasite of the gills of common carp (Cyprinus carpio L.). Folia Parasitologica, 30, 1 aggregated ellipsoidal myxospores at the base of the gill filaments of koi carp juvenile, causing significant tissue damage. Myxobolus artus Achmerov, 1960, enwrapped in diffuse pseudocysts, was observed throughout the trunk muscle of mirror carp. Its oblate myxospores were asymmetrical and contained two unequal-sized polar capsules. According to the localization of large plasmodia, inter- and intramuscular preferences were displayed by M. artus. For precise species identification, we provided the SSU rDNA sequences for each species of Myxobolus. Among them, M. elliptoides was molecularly characterized for the first time, showing the highest 94.21% identity to Thelohanellus sinensis (KY469292). Phylogenetically, the affinity of both M. artus and M. basilamellaris to their conspecific species derived from different carp lineages was highly supported. Moreover, the intermixed cluster of Myxobolus species, including M. elliptoides, with those of Thelohanellus species provided evidence for querying the monophyletic evolution history of these taxa. This work updates the appreciation of the concerned species and enhances our understanding of the parasite fauna of common carp.