Ecological Study of Nematode Parasitism in Ips Beetles from California and Idaho

Nematodes found in Ips paraconfusus from ponderosa pine in California were an undescribed species of Parasitaphelenchus, Contortylenchus elongatus, C. reversus, and C. brevicomi. C. elongatus, the most commonly found contortylenchid, was present in 98.2% of the contortylenchid-parasitized beetles. Only one nematode parasite of the gut, a Parasitorhabditis sp., was isolated. Although significant differences in parasitism were observed, they were by collection sites, rather than by elevation or bole sources (slash or standing). Significant changes in parasitism between fall and spring collections were observed but not at every site. Nematode parasitism in the F₁ generation of I. paraconfusus by Parasitaphelenchus, Contortylenchus, or Parasitorhabditis increased or decreased from the parent generation depending upon the experiment.Nematode parasites from I. pini included an undescribed Parasitaphelenchus sp., two undescribed Contortylenchus spp., C. reversus and Parasitylenchus (= Neoparasitylenchus) ovarius from the hemocel, and Parasitorhabditis ipini from the gut. Parasitaphelenchus sp. was found in 99% and 45.3% of the beetles from Idaho and California, respectively. Of the 1,000 I. pini from Idaho and California, 157 were parasitized by the contortylenchid species or P. ovarius.     

First report of parthenogenesis in the genus Bursaphelenchus Fuchs, 1937: a description of Bursaphelenchus okinawaensis sp. nov. isolated from Monochamus maruokai (Coleoptera: Cerambycidae)

An evolutionarily interesting nematode, Bursaphelenchus okinawaensis sp. nov., is described and illustrated. The new species has several characteristic morphological traits, i.e., four lateral lines in both sexes, lack of a vulval flap in females, and a triangular spicule shape and stout P4 caudal papillae in males, and characteristic biological traits, including phoretic association with Monochamus maruokai, a species of longhorn beetle, parthenogenetic reproduction, and a high frequency of dauer production. Bursaphelenchus okinawaensis sp. nov. shares several important traits with various phylogenetic groups within the genus. The new species shares its spicule shape with B. hellenicus and B. hofmanni. It shares four lateral lines, P4 caudal papillar structure (size and position), and female vulval shape with the 'hunti' group, although it was molecularly inferred to be phylogenetically closer to the 'xylophilus' group and B. africanus. The autapomorphic traits of B. okinawaensis sp. nov. are parthenogenetic reproduction and high frequency of dauer production. All other nominal Bursaphelenchus nematodes have bisexual reproduction and tightly synchronized dauer production. The unique morphological and biological traits of B. okinawaensis sp. nov. suggest genetic flexibility within the genus. The importance of the morphology and arrangement of the caudal papillae is discussed relative to the phylogeny of the genus.     

Morphology and Description of Bursaphelenchus platzeri n. sp. (Nematoda: Parasitaphelenchidae), an Associate of Nitidulid Beetles

Bursaphelenchus platzeri n. sp., an associate of nitidulid beetles in southern California, is described and illustrated. Adult males and females of B. platzeri n. sp. were examined by scanning electron microscopy for ultrastructural comparisons with other members of the genus. Bursaphelenchus cocophilus (red ring nematode) appears to be the closest related taxon to B. platzeri n. sp. based upon shared morphological features of the fused spicules, female tail shape, phoresy with non-scolytid beetles, and molecular analysis of the near full-length small subunit (SSU) rDNA. Unfortunately, sequence data from the D2D3 expansion segments of the large subunit (LSU) rDNA and partial mitochondrial DNA COI did not help resolve the relationship of nearest relative. In addition to significant molecular sequence differences in SSU, LSU, and COI, B. platzeri n. sp., which is an obligate fungal feeder, can be differentiated from B. cocophilus because it is an obligate parasite of palms. Bursaphelenchus platzeri n. sp. can be differentiated from all other species of Bursaphelenchus by the length and shape of the female tail and spicule morphology. The spicules are fused along the ventral midline and possess unfused cucullae; the fused unit appears to function as a conduit for sperm. Population growth of B. platzeri n. sp. was measured in a time-course experiment at 25°C in the laboratory on cultures of the fungus Monilinia fructicola grown on 5% glycerol-supplemented potato dextrose agar (GPDA). Nematode population densities rapidly increased from 25 to approximately 200,000/culture within 14 d and then plateaued for up to 28 d.     

Bursaphelenchus anatolius n. sp. (Nematoda: Parasitaphelenchidae), an Associate of Bees in the Genus Halictus

Bursaphelenchus anatolius n. sp., a phoretic associate of Halictus bees from Ankara, Turkey, is described and illustrated. Bursaphelenchus anatolius n. sp. is closest to B. kevini, which is phoretically associated with Halictus bees from the Pacific Northwest. Bursaphelenchus anatolius n. sp. and B. kevini appear to be sister taxa based upon several shared morphological features, similar life histories involving phoresy with soil-dwelling Halictus bees, and molecular analysis of the near-full-length small subunit rDNA, D2D3 expansion segments of the large subunit rDNA, and partial mitochondrial DNA COI. Bursaphelenchus anatolius n. sp. can be differentiated from all other species of Bursaphelenchus based upon spicule morphology. The paired spicules are uniquely shaped and ventrally recurved, and both B. anatolius n. sp. and B. kevini possess extending flaps that open when the spicules are protracted beyond the cloaca. Population growth of B. anatolius n. sp. was measured at 23 °C in the laboratory on cultures of the fungus Monilinia fructicola grown on lactic acid-treated, 5% glycerol-supplemented potato dextrose agar. Nematode population densities rapidly increased from 110 to about 110,000/9-cm-diam. dish within 21 days.     

Two New Species of Aphelenchoididae (Nemata) from Declining Red Pines in Maryland

Aphelenchoides resinosi n. sp. and Ektaphelenchus joyceae n. sp. are described and illustrated from red pines of the Allegheny plateau of Maryland, USA. The new species were found in trees infested with Bursaphelenchus xylophilus. Primary diagnostic characters of A. resinosi females are constriction of the head, basal stylet knobs, tong postuterine sac, two incisures in the lateral field, and conical tail four to five anal body widths long with a simple terminal mucro. Diagnostic characters of the males are two pairs of subventral caudal papillae and spicule shape: Primary diagnostic characters of E. joyceae females are a slight constriction of the head, six similar lips, conical tail, and short postuterine sac. Diagnostic characters of the males are spicule size and shape, a single row of spermatocytes, and one pair of caudal papillae. Within-tree distributions of A. resinosi and E. joyceae are presented. A total of 70% of both red-needled and chlorotic-needled trees in the study were positive for A. resinosi and E. joyceae. Branch hierarchy was related to the percentage of samples positive for A. resinosi.     

Sound production in Scolytidae: Female sonic stimulus of male pheromone release in two Dendroctonus beetles

The hypothesis that female sonic stimulus may evoke male pheromone release in a behavioural interaction analogous to the known male sonic stimulus of female pheromone release, was confirmed in Dendroctonus pseudotsugae, and also in D. brevicomis. In both species known male-produced substances collected from males stimulated by recorded female stridulation were identified by coupled gas chromatography/mass spectrometry. In a second test with D. pseudotsugae, male pheromone release during recorded female stridulation was evident in the change of male stridulation from the simple attractant chirp to the interrupted chirp, which is known to result from a medium concentration of 3,2-MCH. Also, the D. pseudotsugae male attractant chirp was synthesised with an electronic pulse generator and used to evoke pheromone release. It is concluded that the antiaggregative pheromone of this species is released by each sex at the sonic stimulus of the other sex.     

Species of Bursaphelenchus Fuchs, 1937 (Nematoda: Parasitaphelenchidae) and other nematode genera associated with insects from Pinus pinaster in Portugal

Insects associated with maritime pine, Pinus pinaster, in Portugal were collected and screened for the presence of Bursaphelenchus species. Nematodes were identified using Internal Transcribed Spacers‐Restriction Fragment Length Polymorphism (ITS‐RFLP) analysis of dauer juveniles and morphological identification of adults that developed from dauer juveniles on fungal cultures or on cultures in pine wood segments at 26°C. Several associations are described: Bursaphelenchus teratospicularis and Bursaphelenchus sexdentati are associated with Orthotomicus erosus; Bursaphelenchus tusciae, B. sexdentati and/or Bursaphelenchus pinophilus with Hylurgus ligniperda and Bursaphelenchus hellenicus with Tomicus piniperda, Ips sexdentatus and H. ligniperda. An unidentified Bursaphelenchus species is vectored by Hylobius sp. The previously reported association of Bursaphelenchus xylophilus with Monochamus galloprovincialis was confirmed. The association of Bursaphelenchus leoni with Pityogenes sp. is not definitively established and needs further studies for clarification. Other nematode genera besides Bursaphelenchus were found to be associated with the insects sampled, including two different species of Ektaphelenchus, Parasitorhabditis sp., Parasitaphelenchus sp., Contortylenchus sp. and other unidentified nematodes. The Ektaphelenchus species found in O. erosus is morphologically similar to B. teratospicularis found in the same insect; adults of both the species are found in cocoon‐like structures under the elytra of the insects.     

Redescription and Life History of Contortylenchus brevicomi,a Parasite of the Southern Pine Beetle Dendroctonus frontalis

Larval and adult life stages are described for Contortylenchus brevicomi (Massey) Rühm parasitizing a Mississippi population of Dendroctonus frontalis, the southern pine beetle. Fourth-stage larvae and free-living adult females of this species are identified and described for the first time. The life cycle of C. brevicomi can be reconstructed from this study. The adult female nematode lays eggs in a mature beetle. Larval development progresses within the hemocoel until fourth-stage larvae exit the host. Mating occurs in beetle galleries and only females enter an immature beetle host.     

Gastromermis kolleonis n. sp. (Nematoda: Mermithidae), a Parasite of Midges (Chironomus sp. Chironomidae) from Argentina

Gastromermis kolleonis n. sp. (Nematoda: Mermithidae) is described from the Arroyo Saldan River in Córdoba, Argentina. This species parasitizes midges of the genus Chironomus (Chironomidae: Diptera). It is distinguished from other members of the genus by the presence of six longitudinal chords, vulval flaps, degree of ventral displacement of the mouth, and size and shape of the spicule and amphids.     

<Emphasis Type="Italic">Capillaria appendigera</Emphasis> n. sp. (Nematoda: Capillariidae) from the goldbanded jobfish <Emphasis Type="Italic">Pristipomoides multidens</Emphasis> (Day) (Lutjanidae) and new records of other intestinal capillariids from marine perciform fishes off Australia

Based on light and scanning electron microscopical studies, a new nematode parasite, Capillaria appendigera n. sp. (Capillariidae), is described from the intestine of the goldbanded jobfish Pristipomoides multidens (Day) (Perciformes, Lutjanidae) from the Arafura Sea West, off the northern coast of Australia. The new species, belonging to the subgenus Procapillaria Moravec, 1987, differs from other congeneric species from fishes mainly in the length (0.92–1.13 mm), shape and structure of the spicule, obtuse spines on the spicule sheath and the structure of eggs. It is characterised, in the male, by the presence of two well-developed dorsolateral caudal lobes, a pair of lateral papillae, a heavily sclerotised spicule with many rough transverse grooves covering almost the entice spicule surface (except for spicule ends), a spinose spicule sheath, and in the female, by a subterminal anus, mostly the presence of a large vulval appendage and by eggs (size 54–69 × 27–33 µm) encapsulated by a conspicuous light-coloured superficial layer. Capillaria appendigera n. sp. is the 12th nominal species of capillariids recorded from fishes in Australian waters and the second known capillariid species parasitising fishes of the perciform family Lutjanidae. In addition, four unidentifiable, morphologically different types of capillariid females, probably representing undescribed species, were recorded from the intestines of marine fishes off the northern coast of Australia: Capillariidae gen. sp. 1 and Capillariidae gen. sp. 2 from Lutjanus johnii (Bloch) and L. malabaricus (Bloch & Schneider), respectively (both Lutjanidae), Capillariidae gen. sp. 3 from Protonibea diacanthus (Lacépède) (Sciaenidae) and Capillariidae gen. sp. 4 from Rachycentron canadum (Linnaeus) (Rachycentridae).     

Morphological and genetic analysis of the Coolia monotis species complex with the introduction of two new species,Coolia santacroce sp. nov. and Coolia palmyrensis sp. nov. (Dinophyceae)

The dinoflagellate genus Coolia Meunier is an important epi-benthic organism that is commonly found in association with other dinoflagellates known to cause ciguatera. Two closely related taxa, Coolia monotis and Coolia malayensis, make up the C. monotis species complex. In this study we introduce two new toxic species that should be included in that complex, Coolia palmyrensis Karafas, Tomas, York sp. nov. and Coolia santacroce Karafas, Tomas, York sp. nov., collected from the Palmyra Atoll in the Pacific Ocean and Saint Croix, US Virgin Islands, respectively. These two species can be distinguished morphologically by size, pore shape, pore density, and the relative size of the apical pore complex. The ITS1/5.8S/ITS2 and the D1/D2 regions of the LSU rDNA were used to provide molecular support of morphological observations using maximum likelihood and Bayesian analyses. Furthermore, C. palmyrensis and C. santacroce both showed cytotoxic effects on human derived cells in vitro.     

Morphological,Molecular, and Differential-Host Characterization of Meloidogyne floridensis n. sp. (Nematoda: Meloidogynidae), a Root-Knot Nematode Parasitizing Peach in Florida

A root-knot nematode, Meloidogyne floridensis n. sp., is described and illustrated from peach originally collected from Gainesville, Florida. This new species resembles M. incognita, M. christiei, M. graminicola, and M. hispanica, but with LM and SEM observations it differs from these species either by the body length, shape of head, tail and tail terminus of second-stage juveniles, body length and shape of spicules in males, and its distinctive female perineal pattern. This pattern has a high to narrowly rounded arch with coarsely broken and network-like striae in and around anal area, faint lateral lines interrupting transverse striae, a sunken vulva and anus, and large distinct phasmids. Molecular data from ribosomal IGS illustrate that M. floridensis n. sp. is different from the mitotic species M. arenaria, M. incognita, and M. javanica. Data from RAPDs confirm it and suggest that this new species lies in an intermediate phylogenetic position between the previous species and the meiotic species M. hapla, M. fallax, and M. chitwoodi. Differential host tests based on annual crops and on Prunus accessions are reported.     

Diplogasteroides asiaticus n. sp. is Associated with Monochamus alternatus in Japan

Diplogasteroides asiaticus n. sp. is described and illustrated, and its molecular profile and phylogenetic status within the family Diplogastridae are inferred. Morphologically, the new species is characterized by its stomatal structure, a tube-like stoma with three small, rod-like dorsal teeth and two subventral ridges; a spicule clearly ventrally bent at 1/3 from the anterior end; a gubernaculum with a rounded anterior end and sharply pointed distal end in lateral view; nine pairs of genital papillae with an arrangement of <v1, (v2, v3d), C, v4, ad, ph, (v5, v6, v7), pd>; a short tail spike in males; and a well-developed receptaculum seminis, i.e., the antiparallel blind sacs of the uteri beyond the vulva region and elongated conical tail in females. This new species is morphologically similar to D. haslacheri, but it can be distinguished by the morphology of the somewhat shorter tail in females. D. asiaticus n. sp. shares high sequence conservation with D. andrassyi as there is only one base pair difference in the nearly full-length 18S rDNA and seven base pair differences in the D2-D3 expansion segments of the 28S rDNA. Despite this sequence conservation, the species status of D. asiaticus n. sp. was confirmed using the biological species concept, as D. asiaticus n. sp. and D. andrassyi failed to generate viable F₂ progeny in hybridization tests.     

A survey on coccidian infection of Lake Biwa fishes in Japan,with the description of four new species of Goussia Labbé, 1896 (Apicomplexa)

Thirty-two specimens of 11 freshwater fish species from Lake Biwa, Japan were surveyed for coccidian infections. Four fish species proved to be infected with apicomplexans belonging to the genus Goussia. Altogether, seven Goussia species were found. Oöcysts of four species inhabiting the intestinal epithelium were shed at the sporulated stage. Two other intestinal species left the fish unsporulated. A few relatively large and aged oöcysts of a further species were found in the renal tubules. The sporulated intestinal species were described as Goussia biwaensis n. sp., G. grygieri n. sp., G. nipponica n. sp. and G. wakabayashii n. sp.     

Phylogeny of an Albugo sp. infecting Barbarea vulgaris in Denmark and its frequency of symptom development in natural populations of two evolutionary divergent plant types

The oomycete Albugo candida has long been considered a broad spectrum generalist pathogen, but recent studies suggest that it is diverged into several more specialized species in addition to the generalist Albugo candida sensu stricto. Whereas these species cause the disease white blister rust in many crucifer plants, asymptomatic endophytic infections may be important in the epidemiology of others. One of the plant species attacked by Albugo sp. is the wild crucifer Barbarea vulgaris ssp. arcuata, which is diverged into two phytochemically and genetically different types with different geographical distributions in Europe. These were previously shown to differ strongly in propensity to develop white rust upon controlled infections in the greenhouse. Here, we analyse the phylogenetic relatedness of this local Albugo sp. field isolate to other species and lines of Albugo spp., including others collected on B. vulgaris. We further ask whether the difference in incidence of white rust between the two types of B. vulgaris are also expressed in natural populations.     

Molecular epidemiological survey of Babesia bovis,Babesia bigemina,and Babesia sp. Mymensingh infections in Mongolian cattle

Bovine babesiosis caused by Babesia species is an economically significant disease of cattle. Severe clinical babesiosis in cattle is caused by Babesia bovis, B. bigemina, and the recently discovered Babesia sp. Mymensingh. Mongolia is an agricultural country with a large cattle inventory. Although previous studies have detected active infections of B. bovis and B. bigemina in Mongolian cattle, only a few provinces were surveyed. Additionally, the endemicity of Babesia sp. Mymensingh in Mongolia remains unknown. We screened blood DNA samples from 725 cattle reared in 16 of the 21 Mongolian provinces using B. bovis-, B. bigemina-, and Babesia. sp. Mymensingh-specific PCR assays. The overall positive rates of B. bovis, B. bigemina, and Babesia sp. Mymensingh were 27.9% (n = 202), 23.6% (n = 171), and 5.4% (n = 39), respectively. B. bovis and B. bigemina were detected in cattle in all surveyed provinces; whereas Babesia sp. Mymensingh was detected in 11 of the 16 surveyed provinces. On a per province basis, the B. bovis- B. bigemina-, and Babesia sp. Mymensingh-positive rates were 5.9–52.0%, 9.1–76.3%, and 0–35.7%, respectively. In conclusion, this is the first report of Babesia sp. Mymensingh in Mongolia. In addition, we found that species of Babesia that are capable of causing bovine clinical babesiosis, including B. bovis, B. bigemina, and Babesia sp. Mymensingh, are widespread throughout the country.     

Capsalid monogeneans of fishes from the Seto Inland Sea,Japan: Description of Benedenia kobudai n. sp. parasitic on Semicossyphus reticulatus (Perciformes: Labridae)

The Seto Inland Sea, the largest inland sea in Japan, is a rich fishery with high biodiversity and productivity. Monogeneans have been studied for more than 120 years, and 58 nominal species have been recorded in the Seto inland Sea. This study provided DNA information on five species of Benedenia sensu lato (Capsalidae) from marine fishes from the sea, and one of them, Benedenia kobudai n. sp., is described from Semicossyphus reticulatus (Perciformes: Labridae). This new species differs from the other congeners by the hooded appearance between the anterior attachment organs, the morphology of the penis, the absence of the lobe near the vaginal pore and the common genital pore, the position of the vaginal pore, the germarium lying near the slightly hexagonal testes, the morphology of the haptor, and the shape and position of the hamuli. Phylogenetic analysis showed B. kobudai n. sp. in a separate clade from the other Benedenia species, B. epinepheli, B. hoshinai, B. sekii, and B. seriolae collected from the sea. Each of the newly provided DNA sequences (28S rDNA, ITS1‐5.8S-ITS2, cox1) of the above four species are based on specimens obtained from the type hosts and/or type localities and are considered important for future taxonomic re-examination and confirmation of the reliability of the registered sequences. Furthermore, these four species are important causes of fish diseases in aquaculture, and it is expected that information on the distribution, host range, and occurrence of fish diseases for each capsalid secured by molecular identification will be accumulated.The Life Science Identifier (LSID): urn:lsid:zoobank.org:pub:26C15D17-CFD1-450D-9FCE-EFFF692D2133.     

Sex Attraction and Mating in Bursaphelenchus okinawaensis and B. xylophilus

The fungal feeding, hermaphroditic Bursaphelenchus okinawaensis is a laboratory model to understand the biology of Bursaphelenchus. The extent to which B. okinawaensis can be used to model Bursaphelenchus xylophilus mating was investigated. A chemotaxis assay was conducted to examine whether B. xylophilus and B. okinawaensis produce and respond to volatile sex attractants. Unmated B. xylophilus females were found to attract B. xylophilus males. Similarly, old (sperm depleted) but not young (sperm repleted) B. okinawaensis hermaphrodites attract B. okinawaensis males. Thus, in both species, sperm status corresponds to its ability to attract males. B. xylophilus males also produce a volatile pheromone that attracts both mated and unmated females. A second assay, in which the behavior of males on petri plates in the presence of different females or hermaphrodites of Bursaphelenchus was observed, revealed that B. xylophilus unmated females attract B. okinawaensis males, and B. okinawaensis old hermaphrodites attract B. xylophilus males. These observations suggested that the pheromones of Bursaphelenchus work to some extent across species. Mating behavior through spicule insertion occurs across species, suggesting that postcopulatory mechanisms prevent production of interspecific progeny. The hermaphroditic B. okinawaensis will be a useful model to conduct genetic studies for the understanding of the molecular mechanisms underlying mating behavior in Bursaphelenchus nematodes.     

木质包装材料中线虫一新种(线虫门,滑刃科)记述

记述了伞滑刃属线虫1新种,即弯尾伞滑刃线虫Bursaphelenchus curvicaudatus sp.nov..线虫样本自中国连云港口岸截获木质包装材料中分离获得.新种体长相对较长(雌虫767～960 μm;雄虫663～831 μm),口针较短(雌虫13.9～17.4 μm;雄虫13.9～16.5 μm),口针基部略微加厚,交合刺小(16.5～21.6 μm),末端盘状结构不明显,雌虫尾明显向腹面弯曲.另外,新种雄虫有6个尾乳突,与以前报道有7个尾乳突不同.交合刺形状、雌虫尾的形状及体长等特征能将新种与B.hofmanni、B.abietinus、B.fun-givorus、B.hellenicus、B. hylobianum、B.rainufi、B.eggersi以及B.corneolus区分开来.新种特有的限制性酶切图谱(PCR-ITS-RFLP图谱)是该分离物为1新种的分子证据.     

New species of oribatid mites (Acari: Oribatida) of the genera Belbodamaeus (Damaeidae), Malaconothrus (Malaconothridae) and Nothrus (Nothridae) from India

Three new oribatid mite species, Belbodamaeus indicus sp. n. (Damaeidae), Malaconothrus macrofoveolatus sp. n. (Malaconothridae) and Nothrus phylliformis sp. n. (Nothridae), are described from Indian soils. Belbodamaeus indicus sp. n. is clearly distinguishable from all species of Belbodamaeus by the absence of discidia, very long sensilli and morphology of parastigmatic tubercles Sa. Malaconothrus macrofoveolatus sp. n. is very similar morphologically to Malaconothrus yinae Yamamoto, Aoki, Wang & Hu, 1993 from China, however it differs from the latter by the morphology of notogastral and genital setae, size of body foveolae, epimeral formula, and number of genital setae. Nothrus phylliformis sp. n. is very similar morphologically to Nothrus mystax Mahunka, 1986 from Tanzania, however it differs from the latter by the smaller body size, length of interlamellar setae and the position of notogastral setae d ₁. An identification key to known species of Belbodamaeus is presented.