Evidence for a new species of <Emphasis Type="Italic">Anisakis</Emphasis> Dujardin, 1845: morphological description and genetic relationships between congeners (Nematoda: Anisakidae)

In the present study, a new biological species of Anisakis Dujardin, 1845, was detected in Kogia breviceps and K. sima from West Atlantic waters (coast of Florida) on the basis of 19 (nuclear) structural genes studied by multilocus allozyme electrophoresis. Fixed allele differences at 11 enzyme loci were found between specimens of both adults and larvae of the new species and the other Anisakis spp. tested. Reproductive isolation from A. brevispiculata Dollfus, 1968 was demonstrated by the lack of hybrid or recombinant genotypes in mixed infections in K. breviceps. Genetic distance of the new species from its closest relative, A. brevispiculata, was D(Nei)=0.79. The new species is morphologically different from the other species which have been genetically characterised and from the other Anisakis retained by Davey (1971) as valid or as species inquirendae: the name of Anisakis paggiae n. sp. is proposed for the new taxon. Anisakis Type II larvae (sensu Berland, 1961) from the European hake Merluccius merluccius in the northeastern Atlantic Ocean (Galician coast) and from the scabbard fish Aphanopus carbo in Central Atlantic waters (off Madeira), were identified as A. paggiae n. sp. Its genetic relationships with respect to the seven species previously characterised (A. simplex (Rudolphi, 1809) sensu stricto), A. pegreffii Campana-Rouget & Biocca, 1955, A. simplex, (A. typica (Diesing, 1860), A. ziphidarum Paggi et al., 1998, A. physeteris Baylis, 1923 and A. brevispiculata) were also inferred. Overall, a low genetic identity was detected at allozyme level between the eight Anisakis species. Interspecific genetic identity ranged from I(Nei)=0.68, between the sibling species of the A. simplex complex, to I(Nei)=0.00 (no alleles shared at the considered loci) when A. physeteris, A. brevispiculata and the new species were compared with the other species of the genus. Concordant topologies were obtained using both UPGMA and NJ tree analyses for the considered species. In both analyses, A. paggiae n. sp. clustered with A. brevispiculata. They also indicated two main clades, the first including A. physeteris, A. brevispiculata and A. paggiae n. sp., the second containing all of the remaining species (i.e. A. simplex (s.s.), A. pegreffii, A. simplex, A. typica and A. ziphidarum). A deep separation between these two main Anisakis clades, also supported by high bootstrap values at the major nodes, was apparent. This is also supported by differences in adult and larval morphology, as well as with respect to their main definitive hosts. A morphological key for distinguishing adult A. paggiae n. sp., A. physeteris and A. brevispiculata is presented. Allozyme markers for the identification of any life-history stage of the Anisakis spp. so far studied, as well as ecological data on their definitive host preferences and geographical distribution, are updated.     

<Emphasis Type="Italic">Longidorus</Emphasis><Emphasis Type="Italic">kheirii</Emphasis> n. sp. (Nematoda: Longidoridae) from Iran

Longidorus kheirii n. sp., a parthenogenetic species, was found in soil samples collected from the rhizosphere of Rosa sp. growing in a natural mountainous region close to Maragheh city, northwestern Iran. It is characterised by having a long body (6.7-9 mm), a 19.5-23 mum wide head continuous with the body contour, a truncate and slightly concave lip region with convex sides between the anterior end and the guide-ring, an odontostyle 113-130 mum long, an odontophore 69-97.5 mum long, a body width of 90.5-117.5 mum at the mid-body, a long, wide oesophageal bulb (149.5-193.5 x 39.5-48 mum), a tail length of 47-72 mum, a male with 11 ventromedian supplements and spicules of 85 mum in length, and four juvenile stages. The ribosomal 18S rDNA gene of L. kheirii n. sp., L. leptocephalus Hooper, 1961, L. profundorum Hooper, 1966 L. euonymus Mali & Hooper, 1973 and two unidentified species listed as Longidorus sp. 1 and Longidorus sp. 2, all recovered from northwestern Iran in the same survey, and the ITS1 of L. kheirii n. sp. and Longidorus sp. 1 were sequenced in order to investigate the phylogenetic relationships with other previously sequenced Longidorus species.     

<Emphasis Type="Italic">Aulacnema monodelphis</Emphasis> n. g., n. sp. and <Emphasis Type="Italic">Angiostoma coloaense</Emphasis> n. sp. (Nematoda: Rhabditida: Angiostomatidae) from terrestrial molluscs of Vietnam

Two species of nematodes, Angiostoma coloaense n. sp. and Aulacnema monodelphis n. g., n. sp. (Angiostomatidae: Rhabditida) are described from terrestrial molluscs of Vietnam. Both species are characterised by a long bowl-shaped buccal cavity and typically angiostomatid male bursal alae, spicules and gubernaculum. The new genus, Aulacnema n. g., is erected for the first reported monodelphic angiostomatid. Angiostoma coloaense n. sp. can be distinguished from other species of Angiostoma Dujardin, 1845 by having a buccal cavity length greater than its width and the absence of tail spikes on both males and females.     

<Emphasis Type="Italic">Steinernema boemarei</Emphasis> n. sp. (Nematoda: Steinernematidae), a new entomopathogenic nematode from southern France

A new entomopathogenic nematode, Steinernema boemarei n. sp., is described from southern France. Morphological, molecular (28S and ITS rDNA sequence data) and cross-hybridisation studies were used for diagnostics and identification purposes. Both molecular and morphological data indicate that the new species belongs to the ‘glaseri-group’ of Steinernema spp. Key morphological diagnostic traits for S. boemarei n. sp. include the presence of prominent deirids (cervical papillae) on adult males, the morphology of the spicules and gubernaculum, and the arrangement of the 23 genital papillae of the first generation males. Additionally, morphometric traits of the third-stage infective juvenile, including total body length (mean 1,103 μm), tail length (mean 86 μm), location of the excretory pore (mean 91 μm), and D% (mean 63), E% (mean 106) and H% (mean 41) values are definitive. In addition to these morphological characters, analysis of both 28S and ITS rDNA sequences depict this Steinernema species as a distinct and unique entity.     

Taxonomic characterization of <Emphasis Type="Italic">Haloferax</Emphasis> sp. ("<Emphasis Type="Italic">H. alicantei</Emphasis>") strain Aa 2.2: description of <Emphasis Type="Italic">Haloferax lucentensis</Emphasis> sp. nov.

An extremely halophilic archaeon, previously named as Haloferax sp. strain Aa 2.2 or "Haloferax alicantei" that has been extensively used for genetic studies with halobacteria, was taxonomically characterized by using phenotypic tests (including morphological, physiological, biochemical and nutritional features), DNA-DNA hybridization and 16S rRNA sequence phylogenetic analysis. This organism was isolated in 1986 by Torreblanca et al. from a pond of a Spanish saltern located in Alicante. The cells were pleomorphic, Gram negative and grew optimally at 25% NaCl. The polar lipid composition was similar to that of species of the genus Haloferax. The DNA G+C content of this strain was 64.5 mol%. Phylogenetic analysis based on 16S rRNA sequence comparison confirmed that this archaeon is a member of the genus Haloferax and was most closely related to Haloferax volcanii. DNA-DNA hybridization between strain Aa 2.2 and the type strain of all named species of the genus Haloferax revealed low levels of relatedness (25-2%), supporting the placement of this organism in a new species. On the basis of the phenotypic characteristics, molecular data and phylogenetic analysis we propose to name strain Aa 2.2 as a new species, Haloferax lucentensis sp. nov. The type strain is Aa 2.2 (=JCM 9276=NCIMB 13854=CIP 107410=DSM 14919=CECT 5871=CCM 7023).     

Monocotylids (Monogenea: Monopisthocotylea) from <Emphasis Type="Italic">Aetobatus</Emphasis> cf. <Emphasis Type="Italic">narinari</Emphasis> off New Caledonia,with a description of <Emphasis Type="Italic">Decacotyle elpora</Emphasis> n. sp.

Four species of monocotylid monogeneans were found on a spotted eagle ray Aetobatus cf. narinari caught off Noumea, New Caledonia, South Pacific. The host is illustrated for future possible identification, because it belongs to a currently unresolved species complex. Decacotyle octona (Young, 1967) Chisholm & Whittington, 1998, from the secondary gill lamellae, is figured and previously unreported details of the reproductive system are described. D. elpora n. sp. is described from numerous specimens found between the secondary gill lamellae. The new species, 780-1650 microm in body length, is smaller than D. octona, but has a longer sclerotised male copulatory organ. It is characterised by two dorsal haptoral accessory structures, each with three bumps, and a 9-shaped sclerotised male copulatory organ 240-290 microm in length. D. octona and Clemacotyle australis Young, 1967, both from the branchial cavity, and Thaumatocotyle pseudodasybatis Hargis, 1955, from the nasal tissue, are new geographical records for New Caledonia. It is suggested that differences in the monocotylid fauna between the Atlantic and Indo-Pacific forms of Aetobatus cf. narinari could be of interest for the systematic status of the ray, but the sporadic nature of the sampling probably casts doubt on the usefulness of the currently available data.     

Multilocus phylogeny of <Emphasis Type="Italic">Clonostachys</Emphasis> subgenus <Emphasis Type="Italic">Bionectria</Emphasis> from Brazil and description of <Emphasis Type="Italic">Clonostachys chloroleuca</Emphasis> sp. nov.

Phylogenetic analyses based on protein-encoding gene exons and introns of ATP citrate lyase (ACL1), beta tubulin (TUB), the largest subunit of RNA polymerase II (RPB1), and translation elongation factor 1-α (TEF1) are used for inferring the existence of a new Clonostachys species from the Cerrado biome in Brazil, described here as C. chloroleuca. The species produces dimorphic, primary, and secondary conidiophores that form consistently greenish conidial masses on artificial media. It resembles therefore C. rosea f. catenulata although it differs from this species by less adpressed branches in the secondary conidiophores. The new species is also phylogenetically related to C. byssicola and C. rhizophaga. Our inventory suggests that C. byssicola, C. chloroleuca, C. pseudochroleuca, C. rhizophaga, C. rogersoniana, and C. rosea commonly occur in native and agriculturally used soils of the Cerrado and Amazon Forest. Using sequences available from two genome-sequenced strains employed as biological control agents, we confirm the identity of the European strain IK726 as C. rosea and identify strain 67-1 from China as C. chloroleuca.     

<Emphasis Type="Italic">Contracaecum bioccai</Emphasis> n. sp. from the brown pelican <Emphasis Type="Italic">Pelecanus occidentalis</Emphasis> (L.) in Colombia (Nematoda: Anisakidae): morphology,molecular evidence and its genetic relationship with congeners from fish-eating birds

Contracaecum bioccai n. sp. is described from the brown pelican Pelecanus occidentalis (L.) in northern Colombia (Totumo Marsh) based on 20 enzyme loci studied using multilocus allozyme electrophoresis. Moreover, genetic relationships between the new taxon and related congeners are presented based on allozyme data-sets and sequence analyses (519 bp) of the mtDNA-cox2 gene. Fixed allele differences were found at some of the allozyme loci analysed in comparison with other Contracaecum spp. from pelicans and cormorants [i.e. the sibling species of the C. rudolphii Hartwich, 1964 complex, C. septentrionale Kreis, 1955, C. micropapillatum (Stossich, 1890), C. microcephalum (Rudolphi, 1809) and C. pelagicum Johnston & Mawson, 1942]. The genetic distance, at the allozyme level, between C. bioccai n. sp. and its congeners ranged from D ( Nei ) = 0.80 versus C. septentrionale to D ( Nei ) = 1.40 versus C. micropapillatum. The genetic distance at the mtDNA cox-2 level ranged, on average, from K-2P = 0.12 versus the C. rudolphii species complex to K-2P = 0.15 versus C. micropapillatum. An overall concordant tree topology, obtained from UPGMA and NJ tree analyses inferred from allozyme data, as well as from MP, UPGMA and NJ inferred from mtDNA-cox2 sequence analysis, showed C. bioccai n. sp. as a separated lineage to the other Contracaecum spp. A concordant result was also obtained by PCA analysis based on both the allozyme and mtDNA cox-2 data-sets. All of the tree topologies, derived from the phylogenetic analysis inferred from both allozymes and mtDNA data-sets, were in substantial agreement and depicted C. bioccai as closely related to the sibling species of the C. rudolphii complex (C. rudolphii A and C. rudolphii B) and C. septentrionale. Morphological analysis and a differential diagnosis based on male specimens of C. bioccai, which had been genetically characterised by both allozyme markers and mtDNA sequences analysis with respect to morphologically related congeners, enabled the detection of differences in a numbers of characters, including spicule length, the morphology of the distal end of the spicule and the distribution patterns of the distal caudal papillae.     

Mercury Tolerance of Thermophilic <Emphasis Type="Italic">Bacillus</Emphasis> sp. and <Emphasis Type="Italic">Ureibacillus</Emphasis> sp

Although resistance of microorganisms to Hg(II) salts has been widely investigated and resistant strains have been reported from many eubacterial genera, there are few reports of mercuric ion resistance in extremophilic microorganisms. Moderately thermophilic mercury resistant bacteria were selected by growth at 62 °C on Luria agar containing HgCl₂. Sequence analysis of 16S rRNA genes of two isolates showed the closest matches to be with Bacillus pallidus and Ureibacillus thermosphaericus. Minimum inhibitory concentration (MIC) values for HgCl₂ were 80 μg/ml and 30 μg/ml for these isolates, respectively, compared to 10 μg/ml for B. pallidus H12 DSM3670, a mercury-sensitive control. The best-characterised mercury-resistant Bacillus strain, B. cereus RC607, had an MIC of 60 μg/ml. The new isolates had negligible mercuric reductase activity but removed Hg from the medium by the formation of a black precipitate, identified as HgS by X-ray powder diffraction analysis. No volatile H₂S was detected in the headspace of cultures in the absence or presence of Hg²⁺, and it is suggested that a new mechanism of Hg tolerance, based on the production of non-volatile thiol species, may have potential for decontamination of solutions containing Hg²⁺ without production of toxic volatile H₂S.     

A new species of water mite <Emphasis Type="Italic">Atractides</Emphasis> (<Emphasis Type="Italic">Atractides</Emphasis>) <Emphasis Type="Italic">turcicus</Emphasis> sp. n. (Acari: Hydrachnidia: Hygrobatidae) from Turkey

In this study, the structural characteristics, unique features, various organ measurements of males and females of the water mite Atractides (Atractides) turcicus sp. n. from Turkey are described. In addition, the study compares their characteristics with related species.     

<Emphasis Type="Italic">Hexamermis popilliae</Emphasis> n. sp. (Nematoda: Mermithidae) parasitizing the Japanese beetle <Emphasis Type="Italic">Popillia japonica</Emphasis> Newman (Coleoptera: Scarabaeidae) in Italy

A new species of mermithid nematode, Hexamermis popilliae n. sp. (Nematoda: Mermithidae) is described from the Japanese beetle Popillia japonica Newman in Italy, an area of new introduction for this invasive pest. The combination of the following characters separates H. popilliae from other members of the genus Hexamermis Steiner, 1924: adult head obtuse; amphidial pouches slightly posterior to lateral head papillae in female but adjacent to lateral head papillae in males; amphidial openings large, well developed; amphidial pouches elliptical in females and oblong in males; cuticular vulvar cone well developed, vulvar lips greatly reduced or lacking, vagina curved at tip where meeting uteri, without reverse bend (not S-shaped), spicules slightly curved, with a slight bend in the basal portion, approximately equal to body width at cloaca. This is the first record of a species of Hexamermis parasitizing the Japanese beetle Popillia japonica. The only previous mention of mermithid nematodes from P. japonica was an undescribed species of Psammomermis in North America. Hexamermis popilliae will be evaluated as a potential biological control agent in an integrated control program of the Japanese beetle in Italy.     

<Emphasis Type="Italic">Pleuroxus</Emphasis>-like chydorids (Crustacea: Anomopoda) from South Africa,with the description of <Emphasis Type="Italic">Dumontiellus africanus</Emphasis> gen. n., sp. n.

Five species of Pleuroxus-like chydorids are found in new material from South Africa or are known from the literature. One species is here described as Dumontiellus africanus gen. n., sp. n., endemic to South Africa. Handling editor: K. Martens     

<Emphasis Type="Italic">Dracunculus mulbus</Emphasis> n. sp. (Nematoda: Spirurida) from the water python <Emphasis Type="Italic">Liasis fuscus</Emphasis> (Serpentes: Boidae) in northern Australia

A new species of Dracunculus Reichard, 1759 (Nematoda: Spirurida) is described from the tissues surrounding organs in the body-cavity of the water python Liasis fuscus Peters in northern Australia. One to 14 worms were recovered from 22% (27/120) of pythons examined. Males were located principally around the lungs, liver and heart of the hosts, and females were recovered from peritoneal tissue surrounding the intestines and lining the body-cavity. This species differs from previously described species of Dracunculus in the position of the papillae at the posterior end in males, and in the possession of thick, narrow caudal alae. Submedian cephalic papillae are single in both sexes. Dorsal and ventral anterior cephalic papillae are absent in males. This is the first report of a species of Dracunculus from the Australian region.     

<Emphasis Type="Italic">Nibribacter flagellatus</Emphasis> sp. nov., isolated from rhizosphere of <Emphasis Type="Italic">Hibiscus syriacus</Emphasis> and emended description of the genus <Emphasis Type="Italic">Nibribacter</Emphasis>

A Gram-stain negative, aerobic, motile by flagella, rod-shaped strain (THG-T16^T) was isolated from rhizosphere of Hibiscus syriacus. Growth occurred at 10–40 °C (optimum 28–30 °C), at pH 6.0–8.0 (optimum 7.0) and at 0–1.0% NaCl (optimum 0%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-T16^T were identified as Nibribacter koreensis KACC 16450^T (98.6%), Rufibacter roseus KCTC 42217^T (94.7%), Rufibacter immobilis CCTCC AB 2013351^T (94.5%) and Rufibacter tibetensis CCTCC AB 208084^T (94.4%). The DNA G+C content of strain THG-T16^T was determined to be 46.7 mol%. DNA–DNA hybridization values between strain THG-T16^T and N. koreensis KACC 16450^T, R. roseus KCTC 42217^T, R. immobilis CCTCC AB 2013351^T, R.tibetensis CCTCC AB 208084^T were 33.5?±?0.5% (31.7?±?0.7% reciprocal analysis), 28.1?±?0.2% (25.2?±?0.2%), 17.1?±?0.9% (10.2?±?0.6%) and 8.1?±?0.3% (5.2?±?0.1%). The polar lipids were identified as phosphatidylethanolamine, two unidentified aminophospholipids, an unidentified aminolipid and three unidentified lipids. The quinone was identified as MK-7 and the polyamine as sym-homospermidine. The major fatty acids were identified as C_16:1 ω5c, C_17:1 ω6c, iso-C_15:0, summed feature 3 (C_16:1 ω7c and/or C_16:1 ω6c) and summed feature 4 (iso-C_17:1 I and/or anteiso-C_17:1 B). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics, and DNA–DNA hybridization data, strain THG-T16^T represents a novel species of the genus Nibribacter, for which the name Nibribacter flagellatus sp. nov. is proposed. The type strain is THG-T16^T(=?KACC 19188^T?=?CCTCC AB 2016246^T).     

Free-living nematodes of the genus <Emphasis Type="Italic">Alaimella</Emphasis> Cobb 1920 (Nematoda: Leptolaimidae): A description of <Emphasis Type="Italic">A. macramphis</Emphasis> sp. n. from the White Sea and a revision of the genus

Alaimella cincta Cobb 1920 and Alaimella macramphis sp. n. are described and illustrated. Both the species were first recorded for the White Sea in northern Russia. A. cincta occurs in the shells of the agglutinated foraminiferan Reophax curtus, as well as freely in bottom sediments. A. macramphis sp. n. is described from a single male sampled from bottom sediments. A. macramphis sp. n. is distinguished from two previously known species of Alaimella (A. cincta Cobb 1920 and A. truncate Cobb 1920) by having a longer body, longer cephalic setae, and a wide amphid equal to the respective body diameter. The new species also differs from A. truncate by the distinct striation of the cuticular annulations. The Alaimella species are additionally characterized by having a posterior glandular widening of the esophagus. An emended diagnosis of the genus Alaimella Cobb 1920 and a key for species identification are provided.     

Molecular diversity and genetic relationships in <Emphasis Type="Italic">Secale</Emphasis>

The objective of this study was to quantify the molecular diversity and to determine the genetic relationships among Secale spp. and among cultivars of Secale cereale using RAPDs, ISSRs and sequence analysis of six exons of ScMATE1 gene. Thirteen ryes (cultivated and wild) were genotyped using 21 RAPD and 16 ISSR primers. A total of 435 markers (242 RAPDs and 193 ISSRs) were obtained, with 293 being polymorphic (146 RAPDs and 147 ISSRs). Two RAPD and nine ISSR primers generated more than 80% of polymorphism. The ISSR markers were more polymorphic and informative than RAPDs. Further, 69% of the ISSR primers selected achieved at least 70% of DNA polymorphism. The study of six exons of the ScMATE1 gene also demonstrated a high genetic variability that subsists in Secale genus. One difference observed in exon 1 sequences from S. vavilovii seems to be correlated with Al sensitivity in this species. The genetic relationships obtained using RAPDs, ISSRs and exons of ScMATE1 gene were similar. S. ancestrale, S. kuprijanovii and S. cereale were grouped in the same cluster and S. segetale was in another cluster. S. vavilovii showed evidences of not being clearly an isolate species and having great intraspecific differences.