Morphological and molecular characterisation of Paralongidorus francolambertii sp. n. (Nematoda: Longidoridae) from Serbia

Nematology ◽  
2017 ◽  
Vol 19 (6) ◽  
pp. 681-695 ◽  
Author(s):  
László Barsi ◽  
Francesca De Luca
Keyword(s):  
Phylogenetic Analyses ◽  
The Body ◽  
Likelihood Method ◽  
Rrna Gene ◽  
28S Rrna ◽  
Sister Relationship ◽  
Large Size ◽  
Tilia Tomentosa ◽  
Amphidial Fovea ◽  
Cuticular Layer

Paralongidorus francolambertii sp. n., a bisexual species found in the rhizosphere of silver lime (Tilia tomentosa) and common juniper (Juniperus communis), is described. The species is characterised by its medium to large size (L = 5.86-8.29 mm) and slender body (a = 143-197), a lip region flattened with rounded profile, clearly offset from the body by a deep constriction, a narrow neck between the head and body, a shoulder-like body posterior to the neck, a stirrup-shaped amphidial fovea, with conspicuous slit-like aperture, lying on the lateral cuticular collar, a moderately long odontostyle ca 140 μm long, a guide ring located at ca 28 μm from anterior end, a tail terminus with thickened outer cuticular layer in both sexes, and males with spicules ca 52 μm long. The D2-D3 expansion domains of the 28S rRNA gene and the ITS-containing region of P. francolambertii sp. n. were amplified and sequenced. Phylogenetic analyses by using the Maximum Likelihood method showed that P. francolambertii sp. n. had a sister relationship with P. rex and that all Paralongidorus species formed a well-supported group.

Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

Zootaxa ◽  
2020 ◽  
Vol 4819 (2) ◽  
pp. 295-315
Author(s):  
HIROSHI KAJIHARA
Keyword(s):  
16S Rrna ◽  
Vascular System ◽  
Phylogenetic Analyses ◽  
Far East ◽  
Muscle Layer ◽  
The Body ◽  
Rrna Gene ◽  
28S Rrna ◽  
Anterior Portion ◽  
Longitudinal Muscle Layer

The heteronemertean Cerebratulus orochi sp. nov. is described based on material collected intertidally at a muddy beach in Akkeshi, northern Japan. For the last 80 years, the species has been confused with Cerebratulus marginatus Renier, 1804; the latter was originally described from the Adriatic and once believed to occur in many places in the northern hemisphere including Japan. Cerebratulus orochi sp. nov. is morphologically different from all the congeners including C. marginatus by the following combination of characters: several layers of diagonal-muscle meshwork coated with connective tissue, proximo-distally distributed in cross section from the distal portion of the body-wall outer longitudinal muscle layer to the cutis-gland zone throughout the anterior portion of the body from the precerebral to the foregut regions; the cephalic vascular system consisting of lateral and mid-dorsal vessels; and the sub-rhynchocoelic vessel possessing a pair of antero-lateral diverticula before the former forks posteriorly into a pair of lower lateral vessels in the post-cerebral, pre-oral region. Previous records of C. marginatus from Japanese waters are no longer considered to be substantiated. Multi-locus phylogenetic analyses based on the mitochondrial 16S rRNA and cytochrome c oxidase subunit I (COI), as well as the nuclear 18S rRNA, 28S rRNA, and histone H3 genes among heteronemerteans comprising the “Cerebratulus clade” indicated that C. orochi sp. nov. was closely related to C. cf. marginatus from the US Pacific coast. A MegaBLAST search at the NCBI website with the 16S rRNA gene sequence from C. orochi sp. nov. followed by a couple of species delimitation analyses suggests that larvae of the species are also distributed in Vostok Bay, Far East Russia.

Description and molecular characterisation of Paralongidorus litoralis sp. n. and P. paramaximus Heyns, 1965 (Nematoda: Longidoridae) from Spain

Nematology ◽  
2008 ◽  
Vol 10 (1) ◽  
pp. 87-101 ◽  
Author(s):  
Juan E. Palomares-Rius ◽  
Sergei A. Subbotin ◽  
Blanca B. Landa ◽  
Nicola Vovlas ◽  
Pablo Castillo
Keyword(s):  
Maximum Likelihood ◽  
Phylogenetic Analyses ◽  
Large Body ◽  
Original Description ◽  
The Body ◽  
Southern Spain ◽  
Rrna Gene ◽  
28S Rrna ◽  
Gene Sequences ◽  
Data Set

Abstract Paralongidorus litoralis sp. n., a new bisexual species of the genus, is described and illustrated by light microscopy, scanning electron microscopy and molecular studies from specimens collected in a coastal sand dune soil around roots of lentisc (Pistacia lentiscus L.) from Zahara de los Atunes (Cadiz), southern Spain. Paralongidorus litoralis sp. n. is characterised by the large body size (7.5-10.0 mm), a rounded lip region, clearly offset from the body by a collar-like constriction, and bearing a very large stirrup-shaped, amphidial fovea, with conspicuous slit-like aperture, a very long and flexible odontostyle ca 190 μm long, guiding ring located at 35 μm from anterior end, and males with spicules ca 70 μm long. In addition, identification data of a Spanish population of P. paramaximus Heyns, 1965 recovered from sandy soil of a commercial citrus orchard at Alcala de Guadaira (Seville), southern Spain, agree very well with the original description of the species from South Africa. The 18S rRNA and D2 and D3 expansion regions of 28S rRNA gene sequences were obtained for P. litoralis sp. n. and P. paramaximus. Phylogenetic analyses of P. litoralis sp. n. and P. paramaximus rRNA gene sequences and of Longidoridae sequences published in GenBank were done using maximum likelihood and Bayesian inference. In trees generated from the 18S data set Paralongidorus clustered as an external clade from Longidorus, and in trees generated from D2-D3 of 28S dataset Paralongidorus was monophyletic and nested within Longidorus. Maximum likelihood test supported the hypothesis of validity of the Paralongidorus genus.

scholarly journals Morphological and molecular characterisation of Paralongidorus plesioepimikis n. sp. (Nematoda: Longidoridae) from southern Spain

Nematology ◽  
2013 ◽  
Vol 15 (3) ◽  
pp. 363-378 ◽  
Author(s):  
Juan E. Palomares-Rius ◽  
Carolina Cantalapiedra-Navarrete ◽  
Carlos Gutiérrez-Gutiérrez ◽  
Gracia Liébanas ◽  
Pablo Castillo
Keyword(s):  
Anterior Part ◽  
Phylogenetic Analyses ◽  
Southern Spain ◽  
Rrna Gene ◽  
28S Rrna ◽  
Data Set ◽  
Cephalic Lobe ◽  
Pharyngeal Gland ◽  
Amphidial Fovea ◽  
Partial 18S

Paralongidorus plesioepimikis n. sp. is described and illustrated by light microscopy, scanning electron microscopy and molecular studies from specimens collected in a sandy soil in the rhizosphere of stone pine (Pinus pinea) in Bonares (Huelva Province), southern Spain. The new species is characterised by a very long body (9.71-14.11 mm), an expanded and rounded lip region, with a clear constriction followed by a depression posterior to the amphidial aperture, a very long and flexible odontostyle (202-227 μm), guiding ring located at 36.5-44.0 μm from anterior end, dorsal pharyngeal gland nucleus in anterior part of bulb, one subventral pair of nuclei near middle of bulb, vulva at 33-38%, and a dorsally convex tail with rounded terminus (32-39 μm long). SEM observations showed a very large amphidial fovea with conspicuous aperture ca three-fourths as wide as lip region and cephalic papillae appearing as small apertures, each located just anterior to a distinct cephalic lobe. The sequences of the D2-D3 expansion segments of 28S rRNA, partial 18S rRNA and ITS rRNA gene for P. plesioepimikis n. sp. were obtained. Phylogenetic analyses of P. plesioepimikis n. sp. rRNA gene sequences and of Paralongidorus spp. and Longidorus spp. sequences published in GenBank were done using Maximum Likelihood and Bayesian inference. Paralongidorus species (including P. plesioepimikis n. sp.) clustered together, except for P. bikanerensis which clustered within Longidorus spp. and was clearly separated from all other Paralongidorus spp. in trees generated from the D2-D3 expansion segments of 28S and partial 18S data set. ML analysis using SH-test for the validity of Paralongidorus was performed and showed the validity of the genus using the D2-D3 expansion segment of 28S and partial 18S.

Molecular and morphological characterisation of Paralongidorus iranicus n. sp. and P. bikanerensis (Lal & Mathur, 1987) Siddiqi, Baujard & Mounport, 1993 (Nematoda: Longidoridae) from Iran

Nematology ◽  
2012 ◽  
Vol 14 (4) ◽  
pp. 427-443 ◽  
Author(s):  
Majid Pedram ◽  
Ebrahim Pourjam ◽  
Somayeh Namjou ◽  
Mohammad Reza Atighi ◽  
Carolina Cantalapiedra-Navarrete ◽  
...  
Keyword(s):  
Phylogenetic Analyses ◽  
Large Body ◽  
Original Description ◽  
Phoenix Dactylifera ◽  
Rrna Gene ◽  
28S Rrna ◽  
Northern Iran ◽  
Data Set ◽  
Eastern Iran ◽  
Amphidial Fovea

Paralongidorus iranicus n. sp., a new bisexual species of the genus, is described and illustrated by light microscopy, scanning electron microscopy and molecular studies from specimens collected in the rhizosphere of Scots pine (Pinus sylvestris) from the Caspian (Khazar) seashore, Nour, northern Iran. Paralongidorus iranicus n. sp. is characterised by the large body size (7.8-11.4 mm), a rounded lip region, clearly set off by a collar-like constriction at level of, or slightly posterior to, the amphidial aperture, and bearing a very large, stirrup-shaped, amphidial fovea, with conspicuous slit-like aperture, a very long and flexible odontostyle ca 170 μm long, guiding ring located at 34 μm from anterior end and males with spicules ca 80 μm long. In addition, data from an Iranian population of P. bikanerensis recovered from the rhizosphere of palm (Phoenix dactylifera L.) in Bam, Kerman province, south-eastern Iran, agree very well and are very close to the original description of the species from India. The D2 and D3 expansion regions of 28S rRNA gene, ITS1, and 18S rRNA sequences were obtained for P. iranicus n. sp. and P. bikanerensis. Phylogenetic analyses of P. iranicus n. sp. and P. bikanerensis rRNA gene sequences and of Longidorus spp. sequences published in GenBank were done using Maximum Likelihood and Bayesian inference. Paralongidorus species (including P. iranicus n. sp.) clustered together; however, P. bikanerensis clustered within Longidorus spp. and was clearly separated from all other Paralongidorus spp. in trees generated from the D2-D3 expansion segments of 28S and partial 18S data set, respectively.

Steinernema borjomiense n. sp. (Rhabditida: Steinernematidae), a new entomopathogenic nematode from Georgia

Nematology ◽  
2018 ◽  
Vol 20 (7) ◽  
pp. 653-669 ◽  
Author(s):  
Oleg Gorgadze ◽  
Elena Fanelli ◽  
Manana Lortkhipanidze ◽  
Alberto Troccoli ◽  
Medea Burjanadze ◽  
...  
Keyword(s):  
New Species ◽  
18S Rrna ◽  
Entomopathogenic Nematode ◽  
Phylogenetic Analyses ◽  
Tail Length ◽  
Morphological Characters ◽  
18S Rrna Gene ◽  
The Body ◽  
Rrna Gene ◽  
A New Species

Summary A new species of entomopathogenic nematode, Steinernema borjomiense n. sp., was isolated from the body of the host insect, Oryctes nasicornis (Coleoptera: Scarabaeidae), in Georgia, in the territory of Borjomi-Kharagauli. Morphological characters indicate that the new species is closely related to species of the feltiae-group. The infective juveniles are characterised by the following morphological characters: body length of 879 (777-989) μm, distance between the head and excretory pore = 72 (62-80) μm, pharynx length = 132 (122-142) μm, tail length = 70 (60-80) μm, ratio a = 26.3 (23.0-29.3), H% = 45 (40-51), D% = 54 (47-59), E% = 102 (95-115), and lateral fields consisting of seven ridges (eight incisures) at mid-body. Steinernema borjomiense n. sp. was molecularly characterised by sequencing three ribosomal regions (the ITS, the D2-D3 expansion domains and the 18S rRNA gene) and the mitochondrial COI gene. Phylogenetic analyses revealed that S. borjomiense n. sp. differs from all other known species of Steinernema and is a member of the monticolum-group.

scholarly journals First Report of Ditylenchus dipsaci on Garlic in Minnesota

Plant Disease ◽  
2012 ◽  
Vol 96 (11) ◽  
pp. 1707-1707 ◽  
Author(s):  
D. S. Mollov ◽  
S. A. Subbotin ◽  
Carl Rosen
Keyword(s):  
The Body ◽  
Parasitic Nematodes ◽  
Rrna Gene ◽  
28S Rrna ◽  
Gene Sequences ◽  
Surrounding Water ◽  
Pharyngeal Bulb ◽  
First Report ◽  
Plant Parts ◽  
Ditylenchus Dipsaci

In the summer of 2011, two independent garlic samples from Morrison and Dakota counties and in 2012 one garlic sample from Carver county in Minnesota were submitted by commercial growers to the University of Minnesota Plant Disease Clinic for disease analyses. Symptoms of the above-ground plant parts were stunting and chlorosis. Symptoms of bulbs were necrosis, underdevelopment, and distortion. Upon microscopic examination, phytonematodes exuded into the surrounding water droplet. Nematodes were present in the protective leaves, abscission zone, and cloves in all submitted bulbs (n = 18) for analyses. Morphometric examination of females, males, and juveniles determined that they were Ditylenchus dipsaci. Nematodes extracted from garlic cloves were fixed in TAF (97 ml formalin [40%], 2 ml triethanolamine, and 91 ml dH2O). Morphological observations and measurements were made under an Olympus BX51 microscope equipped with a Nomarski differential interference contrast. Female (n = 6) measurements were: L = 1.411 to 1.636 mm, a = 38 to 44, b = 5.8 to 8.0, c = 14 to 17, stylet = 11.5 to 12.3 μm, V = 79 to 81%, and tail = 95 to 105 μm. The body was almost straight, when heat relaxed, lip region flattened, median bulb oval, and isthmus elongate and slender. The basal pharyngeal bulb overlapped the intestine. The post-vulval uterine branch was about half of vulva-anus distance. The tail was conoid with a pointed terminus. Male (n = 9) measurements were: L = 1.372 to 1.558 mm, a = 40 to 50, b = 6.5 to 7.0, c = 14 to 16, stylet = 11.5 to 12.3 μm, spicules = 22 to 27 μm, and gubernaculum = 9 to 10 μm. The bursa was leptoderan and spicules were curved with simple gubernaculum. Morphology and morphometrics of females and males of D. dipsaci from Minnesota generally fit the descriptions provided for the type and other populations by Hopper (1) and other authors. Several specimens were also taken for molecular identification. DNA extraction, PCR, and sequencing protocols were as described by Subbotin et al. (2). The TW81 and AB28 primers were used for amplification of ITS-rRNA region and the D2A and D3B primers were used for amplification of the D2-D3 expansion segments of 28S rRNA gene. Comparison of the ITS and D2-D3 of 28 rRNA gene sequences showed 100 and 99% identity with corresponding gene sequences of D. dipsaci published in the GenBank (2). The sequences were submitted in the GenBank under accession numbers JX123258 and X123259. This nematode problem has not been known to occur in either of these locations previously. The most likely source of introduction of D. dipsaci are imported garlic seed bulbs. To our knowledge, this is the first report of D. dipsaci affecting garlic or any other crops in Minnesota. The garlic produced in these locations was considered unmarketable and complete loss to the farmers. The presence of D. dipsaci could have a significant economic impact in the emerging multi-million dollar garlic industry in Minnesota. References: (1) D. J. Hooper. Ditylenchus dipsaci. CIH Descriptions of Plant-Parasitic Nematodes Set 1, No. 14, 1972. (2) S. A. Subbotin et al. Phytopathology 95:1308, 2005.

Morphological and molecular characterisation of Heterodera koreana (Vovlas, Lamberti & Choo, 1992) Mundo-Ocampo, Troccoli, Subbotin, Del Cid, Baldwin & Inserra, 2008 (Nematoda: Heteroderidae) from bamboo in Japan

Nematology ◽  
2017 ◽  
Vol 19 (3) ◽  
pp. 333-350 ◽  
Author(s):  
Shigeyuki Sekimoto ◽  
Taketo Uehara ◽  
Takayuki Mizukubo
Keyword(s):  
Phylogenetic Analyses ◽  
Original Description ◽  
Cyst Nematode ◽  
Soil Samples ◽  
Rrna Gene ◽  
28S Rrna ◽  
Gene Sequences ◽  
Phyllostachys Edulis ◽  
Relationship Of ◽  
First Time

The Korean cyst nematode, Heterodera koreana, was recorded for the first time from Japan and characterised morphologically, morphometrically and molecularly. In total, 41 populations were detected from soil samples collected from the rhizosphere of four bamboo species in Japan: 31 populations from moso bamboo (Phyllostachys edulis), seven from madake (P. bambusoides), two from henon bamboo (P. nigra var. henonis) and one from fish pole bamboo (P. aurea). The morphology and morphometrics of the Japanese population were in agreement with those of the original description of H. koreana from South Korea and other subsequent descriptions from China and Iran, with the exception of some minor differences. The results of the phylogenetic analyses of the D2-D3 expansion segments of 28S rRNA gene and ITS rRNA gene sequences confirmed the species identification and phylogenetic relationship of H. koreana with other Heterodera species. The COI mtDNA gene sequences were obtained for the first time for H. koreana. Three COI haplotypes found in Japanese H. koreana populations showed a characteristic geographical distribution in Japan.

Mononchoides macrospiculum n. sp. (Nematoda: Neodiplogastridae) and Teratorhabditis synpapillata Sudhaus, 1985 (Nematoda: Rhabditidae): nematode associates of Rhynchophorus ferrugineus (Oliver) (Coleoptera: Curculionidae) in Italy

Nematology ◽  
2015 ◽  
Vol 17 (8) ◽  
pp. 953-966 ◽  
Author(s):  
Alberto Troccoli ◽  
Monica Oreste ◽  
Eustachio Tarasco ◽  
Elena Fanelli ◽  
Francesca De Luca
Keyword(s):  
Molecular Level ◽  
Phylogenetic Analyses ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
28S Rrna ◽  
Italian Population ◽  
Rhynchophorus Ferrugineus ◽  
Red Palm Weevil ◽  
Palm Weevil ◽  
Mitochondrial Cytochrome Oxidase

Several juvenile and adult nematodes were isolated after dissection of pupae and adults of the red palm weevil,Rhynchophorus ferrugineus, recovered from an infestedPhoenix canariensisChabaud exemplar in Bari, Italy. Two species of nematodes were recovered,Teratorhabditis synpapillataandMononchoides macrospiculumn. sp. which is described herein. The mitochondrial cytochrome oxidase I (COI), the ITS-containing region, the 18S rRNA gene (SSU) and the D2-D3 expansion domains of the 28S rRNA gene (LSU) were amplified and sequenced. The new species,M. macrospiculumn. sp., is described at morphological and molecular level. Phylogenetic analyses using SSU and LSU sequences placedM. macrospiculumn. sp. together withM. composticolaandM. striatus. The sequences of the Italian population ofT. synpapillataare identical to those ofT. synpapillatafrom Japan. This is the first report on the association ofM. macrospiculumn. sp. andT. synpapillatawith the red palm weevil in Europe.

scholarly journals First Report of Powdery Mildew Caused by Podosphaera xanthii on Cucurbita argyrosperma in Mexico

Plant Disease ◽  
2021 ◽  
Author(s):  
José Francisco Díaz-Nájera ◽  
Sergio Ayvar-Serna ◽  
Antonio Mena-Bahena ◽  
Guadalupe Arlene Mora-Romero ◽  
Karla Yeriana Leyva-Madrigal ◽  
...  
Keyword(s):  
Powdery Mildew ◽  
Microscopic Examination ◽  
Disease Incidence ◽  
Phylogenetic Analyses ◽  
Its Region ◽  
Likelihood Method ◽  
28S Rrna ◽  
Podosphaera Xanthii ◽  
Voucher Specimen ◽  
First Report

Cucurbita argyrosperma, commonly named as winter or cushaw squash, is highly sought for its seeds, which have important uses in culinary arts. During the autumn 2021, powdery mildew-like signs and symptoms were observed on cushaw squash in several commercial fields located in Cocula, Guerrero, Mexico. Signs were initially appeared as whitish powdery patches on both sides of leaves and then covering entire leaves and causing premature senescence. The disease incidence was estimated to be 80% in about 1000 plants in two fields. The mycelium was amphigenous, persistent, white in color, and occurred in dense patches. A voucher specimen was deposited in the Herbarium of the Colegio Superior Agropecuario del Estado de Guerrero under the accession number CSAEG22. For the morphological characterization by light microscopy, fungal structures were mounted in a drop of lactic acid on a glass slide. Microscopic examination showed nipple-shaped hyphal appressoria. Conidiophores (n = 30) were straight, 100 to 190 × 10 to 12 μm and produced 2 to 6 conidia in chains. Foot-cells were cylindrical, 41 to 78 μm long, followed by 1 to 2 shorter cells. Conidia (n = 100) were ellipsoid-ovoid to barrel-shaped, 29.5 to 39.1 × 19.4 to 22.7 μm, and contained conspicuous fibrosin bodies. Germ tubes were produced from a lateral position on conidia. Chasmothecia were not observed during the growing season. The morphological characters were consistent with those of the anamorphic state of Podosphaera xanthii (Braun and Cook 2012). For further confirmation, total DNA was extracted from conidia and mycelia following the CTAB method (Doyle and Doyle 1990), and the internal transcribed spacer (ITS) region and part of the 28S gene were amplified by PCR, and sequenced. The ITS region of rDNA was amplified using the primers ITS5/ITS4 (White et al. 1990). For amplification of the 28S rRNA partial gene, a nested PCR was performed using the primer sets PM3 (Takamatsu and Kano 2001)/TW14 (Mori et al. 2000) and NL1/TW14 (Mori et al. 2000) for the first and second reactions, respectively. Phylogenetic analyses using the Maximum Likelihood method, including ITS and 28S sequences of isolates of Podosphaera spp. were performed and confirmed the results obtained in the morphological analysis. The isolate CSAEG22 grouped in a clade with isolates of Podosphaera xanthii. The ITS and 28S sequences were deposited in GenBank under accession numbers OL423329 and OL423343, respectively. Pathogenicity was confirmed by gently dusting conidia from infected leaves onto ten leaves of healthy C. argyrosperma plants. Five non-inoculated leaves served as controls. The plants were maintained in a greenhouse at 25 to 35 ºC, and relative humidity of 60 to 70%. All inoculated leaves developed similar signs to the original observation after 10 days, whereas control leaves remained symptomless. Microscopic examination of the fungus on inoculated leaves showed that it was morphologically identical to that originally observed on diseased plants, fulfilling Koch’s postulates. Podosphaera xanthii has been previously reported on C. maxima, C. moschata, and C. pepo in Mexico (Yañez-Morales et al. 2009; Farr and Rossman 2021). To our knowledge, this is the first report of P. xanthii causing powdery mildew on C. argyrosperma in Mexico. This pathogen is a serious threat to C. argyrosperma production in Mexico and disease management strategies should be developed.

scholarly journals Молекулярно-генетическая изменчивость генов 18S-rRNA и 28S-rRNA у цестод рода Bothriocephalus Rud., 1808 (Cestoda: Bothriocephalidea) из рыб Чёрного моря

2020 ◽  
Author(s):  
Yu.V. Slynko ◽  
T.A. Polyakova ◽  
E.E. Slynko
Keyword(s):  
Black Sea ◽  
18S Rrna ◽  
Molecular Genetic Analysis ◽  
The Body ◽  
The Black Sea ◽  
Rrna Gene ◽  
28S Rrna ◽  
Crimean Peninsula ◽  
Nucleotide Variability ◽  
Scorpaena Porcus

Проведён молекулярно-генетический анализ фрагментов двух генов 18S-rRNA (длиной 568 п.н.) и 28S-rRNA (длиной 312 п.н.) цестод рода Bothriocephalus Rud., 1808, паразитирующих у скорпены Scorpaena porcus (Linnaeus, 1758) и у черноморской камбалы Scophthalmus maeoticus (Pallas, 1814), обитающих в Чёрном море. Материал был собран в северной части Чёрного моря возле побережья Крымского полуострова. Пробы тела паразитов фиксировали в 96 этаноле. В результате, как по каждому гену в отдельности, так и при их объединении установлено, что образец, извлечённый из скорпены (деп. в NCBI MH011407 18S-rRNA и MH000375 28S-rRNA), надёжно идентифицируется, как относящийся к кладе, содержащей B. timii, B. scorpii и B. australis, р-расстояние между нашим образцом и другими видами этой группы не превышает 1,6. Остальные три образца MH011408, MH011409, MH011410 (для гена 18s-rRNA) и MH000376 (для гена 28s-rRNA) сформировали отдельную кладу, состоящую из двух субклад: одна включает образцы МН011409 и МН011410, другая образец МН011408 (указаны только регистрационные номера для гена 18s-rRNA). Следует также отметить, что виды Bothriocephalus timii и Bothriocephalus scorpii дистанцированы всего лишь на 0,5, а Bothriocephalus timii и Bothriocephalus australis на 0,6. Гаплотипы вида Bothriocephalus claviceps составили внешнюю группу, р-расстояние от которого всех, как наших образцов, так и рассматриваемых видов комплекса scorpio , не опускалось ниже 26,3. Вместе с тем, дистанцированность объединённых гаплотипов цестод из черноморской камбалы на уровне 45 р-расстояний, а также значения бутстрепа позволяют полагать их близнецовыми видами (или подвидами) в пределах рода, по аналогии с генами мтДНК. В результате анализа нуклеотидной изменчивости данных фрагментов генов подтверждена принадлежность рассматриваемых экземпляров к роду Bothriocephalus, и они идентифицированы как виды, входящие в комплекс видов Bothriocephalus scorpii .Molecular genetic analysis of fragments of the two genes 18S-rRNA (568 bps long) and 28S-rRNA (312 bps long) cestodes of the genus Bothriocephalus Rud., 1808, parasitizing in Scorpaena porcus (Linnaeus, 1758) and at the Black Sea flounder Scophthalmus maeoticus (Pallas, 1814) living in the Black Sea. Material was collected in the northern Black Sea off the coast of the Crimean Peninsula. Samples of the body of parasites were fixed in 96 ethanol. As a result it was established both for each gene individually and when combining them that the sample extracted from scorpion ( in NCBI MH011407 18S-rRNA and MH000375 28S-rRNA) is reliably identified as referring to clade containing B. timii, B. scorpii and B. australis, the p-distance between our sample and other species of this group does not exceed 1.6. The remaining three samples MH011408, MH011409, MH011410 (for the 18s-rRNA gene) and MH000376 (for the 28s-rRNA gene) formed a separate clade which consists of two subclades: one includes samples MH011409 and MH011410 the other sample MH011408 (only registration numbers are indicated for MH011408 gene 18s-rRNA). It should also be noted that the species Bothriocephalus timii and Bothriocephalus scorpii are only 0.5 apart while Bothriocephalus timii and Bothriocephalus australis are 0.6 apart. Haplotypes of the species Bothriocephalus claviceps constituted an external group, the p-distance from which of all both our samples and the species of the scorpio complex under consideration did not fall below 26.3. At the same time the distance between the combined haplotypes of cestodes from the Black Sea flounder at the level of 45 p-distances as well as bootstrap values allow us to consider them to be twin species (or subspecies) within the genus by analogy with mtDNA genes. An analysis of the nucleotide variability of these gene fragments confirmed the affiliation of the examined species to the genus Bothriocephalus and they were identified as species forming part of the complex of species Bothriocephalus scorpii.

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