Effects of lipid extraction on stable isotope ratios of carbon and nitrogen in muscles of freshwater fish.

The extraction of lipids by the Folch method from the muscles of all the fish studied led to statistically significant differences in the values of δ15N. At the same time, lipid extraction led to a statistically significant increase in δ13C in pike and roach, and to a statistically insignificant decrease in δ13C in perch and bream. Thus, lipid extraction cannot serve as a universal method of sample preparation for the analysis of the isotopic composition of carbon (13C/12C) and nitrogen (15N/14N) in fish muscles. The differences between the δ13C values in the samples before and after lipid extraction were statistically investigated by different models. It is shown that mathematical correction method models can be used, but the results are depending on the fish types.

Invasive Alien Slug Arion vulgaris Moquin-Tandon, 1855 (Gastropoda: Pulmonata: Arionidae) in Moscow Parks and Its Co-introduced Parasite Alloionema appendiculatum Schneider, 1859.

PURPOSE: The present study investigates the origin of Arion vulgaris slugs in the parks of Moscow city and their parasites. METHODS: Snails and slugs inhabiting green areas of Moscow city were collected in the summer season of 2020 and examined on the presence of gastropod-associated nematodes and trematodes using morphological and molecular methods. RESULTS: The presence of the alien slug species, Arion vulgaris, was recorded in several locations, and the mitochondrial gene-based analysis has shown that slug populations inhabited Moscow parks originated from West and Central Europe. Out of a total of 15 gastropod species examined, A. vulgaris was the only species infected by the nematode Alloionema appendiculatum Schneider, 1859, a larval parasite of molluscs. It is the first record of this nematode from the territory of the Russian Federation. COX1 mtDNA sequences of A. appendiculatum obtained from 3 populations of infected slugs were identical with those from Western and Central Europe similarly to their gastropod hosts thus indicating that the nematodes travelled with their hosts. No parasites dangerous for humans or animals were found. CONCLUSION: The complex life cycle of A. appendiculatum includes a free-living stage in soil which offers a source of infection for other potentially susceptible gastropod species but the capacity of A. appendiculatum to change hosts in local conditions needs to be further investigated. The particular susceptibility and tolerance of A. vulgaris to nematodes in our study was in concordance with earlier data while in contradiction with the enemy release hypothesis.

Mercury levels in sediment, fish and macroinvertebrates of the Boroo River, northern Mongolia, under the legacy of gold mining.

Gold mining is currently one of the main anthropogenic sources of mercury in the environment. In this study, the total mercury content was measured in bottom sediments, benthic macroinvertebrates (mayfly larvae), and fish (Siberian dace) along the Boroo River in northern Mongolia. There was a gold recovery plant in the middle reaches of the river until the mid-twentieth century; an accident there in the 1950s caused a mercury spill. We found an increased content of mercury in measured ecosystem components near the plant compared to the upper reaches of the river. The mercury content in sediments varied from trace amounts in the upper Boroo to 2200 ng/g dry weight (dw) in the vicinity of the plant ruins. The mercury content in mayfly larvae ranged from 50 to 2940 ng/g dw and had a spatial pattern as sediments, with the highest concentrations near the plant. The mercury content in sediments was lower at the mouth of the Boroo River than near the plant, reflecting the lower boundary of the mercury spill. Maximum values of mercury content in fish muscle were found at the river's mouth and were several times higher than in other rivers of northern Mongolia. Median mercury content in muscles of dace from the lower Boroo in 2016 has doubled since studies in 2010-2012, which may be the result of current mercury releases from gold mining.

The re-description of Synoecnemahirsutum Timm, 1959 (Synoecneminae, Ungellidae, Drilonematoidea) from a pheretimoid earthworm in Vietnam with the analysis of its phylogenetic relationships.

Synoecnemahirsutum Timm, 1959 (Ungellidae, Drilonematoidea), found in the body cavity of the pheretimoid earthworm at the border of Laos and Vietnam, was re-described and illustrated. The mitochondrial genome of S.hirsutum obtained with Illumina HiSeq sequencing is the first annotated mitochondrial genome as a representative of the superfamily Drilonematoidea. The phylogeny inferred from the analysis of 12 mitochondrial genes has shown some similarity of S.hirsutum with a cephalobid Acrobeloidesvarius.

Structure, development, and evolutive patterns of spermatozoa in rhabditid nematodes (Nematoda: Rhabditida).

Spermatogenesis of five rhabditid nematodes was studied using transmission electron microscopy and is described herein. Structure and development of nematode sperm in all available representatives of the extensive order Rhabditida have been analysed and the main characteristics of each infraorder are discussed. The ancestral sperm of the order Rhabditida was reconstructed using maximum likelihood and Bayesian methods based on 44 ultrastructural sperm characters. The hypothetical ancestral spermatogenesis of the order Rhabditida agrees with the previously suggested "rhabditid" pattern and appears to be conserved throughout the order Rhabditida. Despite the enormous variation of rhabditid nematodes, few groups deviate from the ancestral pattern. This conserved pattern can be informative within the phylum Nematoda at order level, but poses limitations when used in taxonomic and phylogenetic analysis within Rhabditida.

Angiostoma norvegicum n. sp. (Nematoda: Angiostomatidae) a parasite of arionid slugs in Norway.

Angiostoma norvegicum n. sp. (Angiostomatidae) is described from the oesophagus, crop and the buccal mass of five species of slugs of the family Arionidae, Arion vulgaris (Moquin-Tandon), Arion ater (L.), Arion fasciatus (Nilsson), Arion fuscus (Müller) and Arion rufus/Arion ater hybrid), collected throughout Norway. Angiostoma norvegicum n. sp. was found parasitising arionids at seven of the 30 sample sites examined (23.3%), and 9.9% of all Arion spp. were infected with this nematode. The new species is characterised by its large size (4.0-8.6 mm long) and in having: lateral alae; 6 + 6 papillae at the cephalic end; a large circular mouth aperture; a spacious stoma; a pharyngeal basal bulb without valvular apparatus; an excretory pore near the base of bulb; a distal part of posterior ovary always outstretched; an anterior ovary distally nearly always outstretched; a vulva situated anterior to mid-body; long, nearly straight spicules and a small gubernaculum; three circumcloacal papillae and caudal genital papillae (GP) arranged in a pattern 1+2/3+3 with GP 5 and GP 8 opened on dorsal side of narrow bursa not reaching tail tip; short conical tails in both sexes with tips supplied by 4 short, unequal denticles. Morphologically, A. norvegicum n. sp. is similar to Angiostoma limacis Dujardin, 1845, which diagnostic characteristics are given based on examination of specimens from Norway and the UK. Conversely, the phylogenetic analyses based on D2D3 large subunit (LSU) rRNA gene sequences performed in the present study did not support the morphological affinity of these two species. Phylogenetic analyses demonstrated that although Angiostoma spp. cluster together, A. norvegicum n. sp. forms a tight monophyletic clade with the milacid nematode parasites Angiostoma margaretae Ross, Malan & Ivanova, 2011 and Angiostoma milacis Ivanova & Wilson, 2009.

Synoecnema watinagii (Drilonematoidea: Ungellidae: Synoecneminae), a new nematode species parasitic in earthworms from the Philippines with the first molecular and SEM data for the genus.

A new species of the genus Synoecnema-S. watinagii sp. n.-is described and illustrated. The species is characterized by its small size, lack of sexual dimorphism apart from sexual characters, males with pericloacal disk, females with anterior vulva position and lacking an anus, and similar caudal organs in both sexes in the shape of long, deep slits on the surface of the posterior half of body. Sequences of D2-D3 LSU and SSU rDNA and SEM images were obtained for the first time for the genus Synoecnema. Phylogenetic analysis of these sequences supported the validity of the genus and elucidated its relationships within Synoecneminae and Drilonematoidea. The earthworm host was characterized by its CoxI mt DNA sequence.

A new genus and two new nematode species (Drilonematoidea: Ungellidae: Synoecneminae) parasitic in two morphs of Drawida ghilarovi Gates, endemic earthworm from the Russian Far East.

Drasico n. g. is erected to accommodate two new species of nematode, Drasico nemoralis n. sp. and D. paludigenus n. sp., recovered from coelomic cavities of Drawida ghilarovi Gates, endemic earthworms of the Russian Far East. The new genus is characterised by the following unique for the Synoecneminae characters: apical portion of the head attenuated, cephalic hooks displaced to the base of attenuated portion, amphids displaced posterior to cephalic hooks, excretory duct short and weak, males possessing several genital papilliform sensilla. The new species are differentiated by the size, number and disposition of the male genital sensilla (larger and more numerous in D. nemoralis n. sp.); the body shape of females (with thinner neck and wider mid-body in D. paludigenus n. sp.) and the ovarian tube arranged in transversal folds in D. paludigenus (vs longitudinal folds in D. nemoralis n. sp.). Nucleotide sequences of D2-D3 expansion segment of 28S rDNA for the two new species differed at 13 positions. Phylogenetic analysis revealed close relationships of Drasico n. g. with species of Siconema Timm, 1966. The host species was represented by two morphs (blue-grey forest and tar-black meadow-swamp morph) with intraspecific divergence of 16-17% for cytochrome c oxidase subunit 1 (COI) gene, and each host morph was found infected by a different nematode species. A co-infection with the plectid nematode Creagrocercus drawidae Ivanova & Spiridonov, 2011 was recorded together with D. nemoralis n. sp. in the blue-grey forest morph.

Morphological and molecular characterisation of Steinernema costaricense Uribe-Lorío, Mora & Stock, 2007 (Panagrolaimorpha: Steinernematidae) isolate from Bush Augusta State Park, Missouri, USA.

A new isolate of Steinernema costaricense Uribe-Lorío, Mora & Stock, 2007 (Panagrolaimorpha: Steinernematidae) was discovered from the Bush Augusta State Park, Missouri, USA, and its morphological and molecular examination was carried out. Morphologically, adults of the Missouri isolate are very close to S. costaricense from Costa Rica. Infective juveniles (IJs) from Missouri are characterised by mean body length of 843 µm, mean body width of 33 µm, mean tail length of 80 µm, mean pharynx length of 143 µm, excretory pore at 62 µm from anterior extremity, high cephalic papillae, and lateral field formula 2-6-2 with two central ridges less prominent. They resemble IJs of S. costaricense from Costa Rica in having high cephalic papillae, similar lateral field pattern and pharynx length, but are twice shorter. Based on the nearly complete identity between the sequence of D2-D3 domains of LSU rDNA of the Missouri isolate and those of S. costaricense retrieved from GenBank and the high similarity of their bacterial symbionts, the new isolate was identified as S. costaricense. The phylogenetic affinities among S. costaricense and the species of the "bicornutum" group proposed by the authors of its original description based on LSU rDNA analysis, is debated. In the present study, all three methods of analysis for the ITS region showed that four species of Steinernema from the Americas (S. rarum de Doucet, 1986, S. scarabaei Stock & Koppenhöfer, 2003, S. unicornum Edgington, Buddie, Tymo, France, Merino & Hunt, 2009 and S. costaricense Missouri isolate) formed a weakly supported clade although bootstrap support for the sister-group relationship between the new isolate and S. scarabaei was always high. In phylogenies inferred from D2-D3 LSU rDNA S. costaricense never formed a clade with S. rarum and S. unicornum.

Angiostoma margaretae n. sp (Nematoda: Angiostomatidae), a parasite of the milacid slug Milax gagates Draparnaud collected near Caledon, South Africa.

Angiostoma margaretae n. sp. (Angiostomatidae) is described from the oesophagus of the slug Milax gagates Draparnaud collected near Caledon in the Western Cape Province of South Africa. The new species closely resembles another parasite of a milacid slug, A. milacis Ivanova & Wilson, 2009, with a similar head, stoma and spicule shape, the presence of distally outstretched ovaries, coiled oviducts, the same number of caudal papillae and enlarged rectal glands. However, A. margaretae differs from the latter by having: a shorter, wider tail with a rounded vs pointed tip; the distal parts of both ovaries with a particular hook-like shape due to an expansion closely following the short initial zone; ovoviparous females; and a different arrangement of male papillae. A. margaretae is comparable with A. limacis Dujardin, 1845, A. asamati (Spiridonov, 1985), A. coloaense (Pham Van Luc, Spiridonov & Wilson, 2005) and A. stammeri (Mengert, 1953), which have a similar stoma shape and size, but can be readily differentiated by the presence of distally outstretched vs reflexed ovaries and the presence vs lack of enlarged rectal glands. The new species has a similar arrangement of the ovaries to A. kimmeriense Korol & Spiridonov, 1991 and A. zonitidis Ivanova & Wilson, 2009, but is clearly differentiated by the lack of an off-set lip region and presence of a large bowl-shaped vs tubular stoma and less numerous male caudal papillae (seven pairs vs nine in A. kimmeriense and 10 in A. zonitidis).

Molecular phylogeny of slug-parasitic nematodes inferred from 18S rRNA gene sequences.

Terrestrial molluscs are diverse and are infected by many nematodes. We propose a phylogeny of slug-parasitic nematodes using 18S rRNA gene sequences from nematodes isolated from slugs collected from six countries. Eight species, representing six families of nematodes were identified and trees inferred placed them within four (I, III, IV and V) out of the five clades of Nematoda, indicating multiple origins of slug parasitism. Five species representing three families formed a monophyletic group in clade V. Although these species are closely related, their morphology has changed greatly, suggesting adaptive radiation to fill different niches within the host.

Two new species of Angiostoma Dujardin, 1845 (Nematoda: Angiostomatidae) from British terrestrial molluscs.

Two new species of Angiostoma Dujardin, 1845 taken from the intestines of terrestrial molluscs (Gastropoda: Stylommatophora) collected in the UK are described and illustrated. Angiostoma milacis n. sp. is parasitic in milacid and agriolimacid slugs and is closely related to A. asamati Spiridonov, 1985. Angiostoma zonitidis n. sp. was found in a zonitid snail and is most closely related to A. kimmeriense Korol & Spiridonov, 1992 (emend.), a species also from zonitid snails. An amended generic diagnosis is proposed. The distribution of A. limacis and A. milacis sp. n. in hosts is discussed and a key to the species of Angiostoma is presented.