New record of Steinernema arenarium (Artyukhovsky) (Rhabditida: Steinernematidae) from Ukraine and a note on its distribution.

During a survey of the biodiversity of entomopathogenic nematodes in Ukraine, a population of Steinernema arenarium, strain Ch, was recovered in the sensitive Chornobyl Exclusion Zone. In the present work, this strain was morphologically and molecularly characterised using light microscopy and the sequences of the ITS and D2-D3 region of the 28S rDNA. In addition, we sequenced the ITS and D2-D3 regions of four populations of S. arenarium from a laboratory collection. Phylogenetic analyses were performed and the phylogenetic structure and geographic distribution of S. arenarium are discussed.

Alloionema californicum n. sp. (Nematoda: Alloionematidae): a new alloionematid from USA.

A new species of the family Alloionematidae was isolated from a rotten winged gourd at White Crane Garden, San Francisco, USA, sampled by Christopher Nelson in November 2010, and a live culture is deposited in Félix Lab Strain Database (http://www.justbio.com/worms/index.php), IBENS, Paris, France. Specimens from the culture have been examined. Both morphologically and molecularly, the nematode described herein as Alloionema californicum n. sp. differs from the other alloionematid species, A. appendiculatum and Neoalloionema tricaudatum. It is characterised by having a narrow stoma, 2.5-3.5 or 4 times longer than broad in adults or dauer juveniles respectively. Lateral fields are not present in adults but occur as one prominent ridge in dauers. Males have no bursa, six pairs of genital papillae and one single papilla. Dauers have large apparent phasmids in the middle of the tail. The ecology of the newly described species is unknown but probably it is a saprobic bacteriophagous nematode preferring rotting organic material.

Phasmarhabditis bonaquaense n. sp. (Nematoda: Rhabditidae), a new slug-parasitic nematode from the Czech Republic.

Phasmarhabditis bonaquaense n. sp. is described and illustrated from the body of Malacolimax tenellus, from the locality of Ceské Svýcary near the village of Dobrá Voda, the Czech Republic. Females are characterized by a body length of 2349 (1878-2626) µm and a cupola shaped tail with a long hyaline hair-like tail tip. Extremely prominent papilla-like phasmids present. Males 1829 (1414-2121) µm long. Peloderan bursa with nine pairs of rays (papillae), 1/1/1/2/1/3. One non-paired apparent papilla-like structure located near the ventral appendage anterior to the cloaca. Prominent papilla-like phasmids located close to the tail tip. Small subunit (18S), ITS, and D2-D3 expansion segments of the large subunit of ribosomal DNA were used to analyze the phylogenetic relationships of sequenced species in the genus Phasmarhabditis and other closely related species. Phasmarhabditis bonaquaense n. sp. varied from other related nematodes both in morphological characterizations and phylogenetic analysis. The life cycle of the newly described species is not well known but it is probably a facultative, mollusc-parasitic nematode able to survive permanently in the saprobic phase on decaying organic matter.

Steinernema poinari sp. n. (Nematoda: Steinernematidae) a new entomopathogenic nematode from the Czech Republic.

A new steinernematid nematode, herein described as Steinernema poinari sp. n., was recovered by baiting soil samples with the last instar wax moth larvae Galleria mellonella in three localities of southwest Bohemia, Czech Republic. Morphological and molecular data confirmed this nematode as a new species which belongs to the affine/intermedium group. S. poinari sp. n. was characterized by male, female and infective juvenile morphological observations. Male spicules are robust with a pointed tip, SW% = 109 (98-123) and GS% = 70 (58-87). The tail terminus of the first generation have a papillated mucron, whereas in the second generation a short filamentous mucron sometimes develops. Females of the first generation have a moderately protruding vulva, postanal swelling absent or slightly developed. Second generation females have a long filamentous mucron on the tail tip. Infective juvenile have a body length of 768 µm, lateral field formula 2, 5, 6, 5, 2, 1, ratio D% of 46 (40-55), E% of 84 (76-95) and H% of 50 (43-56). The new species was characterized by sequences of ITS and D2D3 regions of ribosomal DNA. S. poinari sp. n. can also be separated from S. affine and S. intermedium by cross-breeding tests.

Osteoplastic decompressive craniotomy--an alternative to decompressive craniectomy.

BACKGROUND: In spite of various degrees of brain expansion, decompressive surgery is usually carried out using decompressive craniectomy (DC). After craniectomy it is necessary to perform cranioplasty, which prolongs hospitalization and is not always without complications. Hence, in situations when cranial decompression is indicated, but DC would be too radical, we do not remove the bone flap, and we perform so-called osteoplastic decompressive craniotomy (ODC). The technique is detailed. OBJECTIVE: To demonstrate the effectiveness of ODC. METHODS: Twenty patients underwent ODC for brain edema under various pathological conditions. The diagnoses were as follows: 13 subdural hematomas, 3 cerebral contusions, 2 middle cerebral artery infarcts, 1 epidural hematoma and 1 arteriovenous malformation. The effect of ODC was assessed using postoperative ICP monitoring and the midline shift on CT. The ICP threshold for the additional removal of the bone flap was 25 mmHg. Clinical outcome was evaluated 6 months after surgery using the Glasgow Outcome Scale (GOS). RESULTS: Postoperative ICP was up to 25 mmHg in 18 patients and exceeded 25 mmHg in 2 cases. The mean midline shift on CT was 10 mm preoperatively and 3 mm postoperatively. The decompression during ODC was sufficient in 18 patients and insufficient in 2 in whom an additional removal of the bone flap was performed. Eight survivals had a favorable outcome (GOS 4-5); 12 patients had an unfavorable outcome (GOS 1-3), and of these, 4 died. CONCLUSION: Our limited study shows that ODC is effective in the treatment of intracranial hypertension in the selected subgroup of patients in whom DC would be too radical. The main advantage of this method is the elimination of further cranioplasty.

Does scavenging extend the host range of entomopathogenic nematodes (Nematoda: Steinernematidae)?

Living and freeze-killed natural and laboratory hosts, with different susceptibility to entomopathogenic nematodes, were exposed to the larvae of Steinernema affine and Steinernema kraussei in two different experimental arenas (Eppendorf tubes, Petri dishes), and the success of the colonisation and eventual progeny production were observed. Both nematodes were able to colonise both living and dead larvae of Galleria mellonella (Lepidoptera) and adult Blatella germanica (Blattodea) even though the progeny production in dead hosts was lower on average. Living carabid beetles, Poecilus cupreus, and elaterid larvae (Coleoptera) were resistant to the infection, however, both nematodes were able to colonise and multiply in several dead P. cupreus and in a majority of dead elaterid larvae. By scavenging, EPNs can utilise cadavers of insects that are naturally resistant to EPN infection, and so broaden their host range.

Mixed infection of Galleria mellonella with two entomopathogenic nematode (Nematoda: Rhabditida) species: Steinernema affine benefits from the presence of Steinernema kraussei.

The interactions of two sympatric entomopathogenic nematodes Steinernema affine and Steinernema. kraussei were studied in a series of laboratory experiments. Single species, simultaneous and sequential infections of Galleria mellonella were performed in Eppendorf tubes and the invasion rate, nematode progeny production and the number of hosts producing nematode progeny were observed. The invasion rate of S. affine was not affected by the mixed infection whereas the invasion of the latter species was strongly reduced. S. affine out-competed S. kraussei in all treatments and the progeny production of the latter species occurred only sporadically. In comparison to single species infections, per-host progeny production of S. affine was affected only in the treatments with a low dose of S. affine, where it was 30-50% lowered. Generally, in the presence of the latter species, S. affine was able to infect and multiply in a higher number of hosts in comparison to single species infection, especially at a low inoculation rate. S. affine invaded and multiplied also in hosts already infected and even killed by S. kraussei producing a normal amount of progeny. Generally the results suggest that the interactions between steinernematid species can be more complex, including a positive effect of one species on another.

Steinernema xueshanense n. sp. (Rhabditida, Steinernematidae), a new species of entomopathogenic nematode from the province of Yunnan, southeast Tibetan Mts., China.

During a random EPN survey in the northern part of the Yunnan province (southeastern Tibet, Dequen district, town of Dequen) in 2005, soil samples containing an unknown EPN species were collected. The new species is described herein as Steinernema xueshannense n. sp. named after the Xue Shan Mts. a mountain range between Yunnan and Tibet where the nematode was collected. The isolate is a new species belonging to the Steinernema feltiae /kraussei group. S. xueshanense n. sp. is characterized by male, female of both generations and infective juveniles (IJ). IJ lateral field with eight ridges, submarginal pair less distinct, formula 2, 7, 8, 7, 6, 4, 2, Hyaline portion occupies approximately one half of tail length. Second-generation males with distinct mucron and moderately curved spicules. Females wuth a characteristic cone on the tail. Infective juveniles of S. xueshanense n. sp. differ from S. akhursti, S. cholashanense S. kraussei, S. oregonense by different number of ridges in lateral fields. Species which have same number of eight lateral ridges, such as S. silvaticum, S. thanhi, S. weiseri differs from S. xueshanense n. sp. by less prominent sublateral pair whereas those species have all ridges equally spaced and prominent. Only S. feltiae possess the same pattern of ridges as S. xueshanense n. sp., but this species differ by shape of spicules with oblongate manubrium. The description of S. sangi gives only a few characters to compare this species with S. xueshanense n. sp. The exception is the excretory pore position of IJs which is at 40% of pharynx length whereas in other species of this group, including S. xueshanense n. sp., it is situated approximately at 50%. Cross-breedings, sequences of ITS and D2/D3 regions of the ribosomal DNA confirmed the new species identity.

Steinernema ichnusae sp. n. (Nematoda: Steinernematidae) a new entomopathogenic nematode from Sardinia Island (Italy).

A new Steinernema species was isolated from three different sandy soil samples along the Platamona Beach, in the north-west coast of Sardinia Island (Italy). This new species is characterized by the following morphological characters: infective third-stage juvenile with a body length of 866+/-61 (767-969)microm, distance from head to excretory pore of 63+/-2.7 (59-68)microm, tail length of 81+/-3.2 (76-89)microm, ratio E (%) 77+/-3.4 (68-83); male tail with a mucron only in the second generation, spicule length of 66+/-1.4 (64-67)microm and gubernaculum length of 44+/-1.4 (43-46)microm in the first generation male; female of first generation with a slight vulval protrusion and ratio D (%) of 53+/-4.0 (47-63). The new species differs distinctly from the related species (S. feltiae, S. kraussei, S. litorale, S. oregonense and S. cholashanense) in some morphometric values such as percentage of hyaline portion, ratios of gubernaculum/spicule length, spicule head length/width. The DNA analyses of the internal transcribed spacers and D2D3 regions show that the studied nematode isolates are a new species. Cross hybridisation tests with S. feltiae, S. kraussei, S. litorale, S. weiseri and S. oregonense showed that these species were reproductively isolated.

Steinernema cholashanense n. sp. (Rhabditida, Steinernematidae) a new species of entomopathogenic nematode from the province of Sichuan, Chola Shan Mountains, China.

During a random survey of entomopathogenic nematodes in the provinces of Sichuan and Gansu (eastern Tibet) in 2004, soil samples from several sites were collected and tested for the incidence of entomopathogenic nematodes. A new species was collected in this survey and it is described herein as Steinernema cholashanense n. sp. Steinernema cholashanense n. sp. is characterized by morphology and morphometry of the IJ and male. For the IJ, the new species can be recognized by the average body length 843 microm, esophagus length 125 microm, H%=39% and E%=81%. The lateral field pattern is 2, 5, 7, 4, 2. The male of the first generation is characterized by spicule shape and length and especially with prominent velum and the presence of a mucron on both generations. The average body length of the IJ of S. cholashanense n. sp. (843 microm) is shorter than that of S. oregonense (980 microm), S. kraussei (951 microm) and S. litorale (909 microm), similar to that of S. feltiae (849 microm), but longer than that of S. weiseri (740 microm), S. jollietti (711 microm) and S. hebeiense (658 microm). Esophagus length of the new species (125 microm) is closer to that of S. jollieti (123 microm) but longer than that of S. weiseri (113 microm) and shorter than that of S. oregonense (132 microm), S. kraussei (134 microm) and S. feltiae (136 microm). E% of the new species (81) is similar to that of S. kraussei (80), but smaller than that of S. jollieti (88), S. weiseri (95), S. oregonense (100) and S. feltiae (119). Spicule head length of the new species is almost the same as its width, this character is similar to that of S. kraussei but it is different from this species by its prominent velum. The new species can be recognized further by characteristics of sequences of ITS and D2D3 regions and cross hybridization with closely related species, S. feltiae, S. kraussei and S. oregonense.

Natural population dynamics of entomopathogenic nematode Steinernema affine (Steinernematidae) under dry conditions: Possible nematode persistence within host cadavers?

The effect of dry conditions on the population dynamics of the entomopathogenic nematode (EPN) Steinernema affine was studied for one month in the exceptionally dry period in the summer of 2003 in the oak wood in the vicinity of Ceské Budejovice, Czech Republic. Soil moisture, soil temperature, and the abundance of suitable insect hosts were monitored. The abundance of infective juveniles (IJs) was correlated with soil moisture and both these values were gradually decreasing during the study period and finally rapidly increased at the end of the investigation. During this period there was a decline in the number of insects suitable as hosts for S. affine, but not in numbers of unsuitable insects. We hypothesise that the observed decrease in IJ numbers was probably caused by the persistence of IJs in host cadavers due to low ambient moisture.

Steinernema sichuanense n. sp. (Rhabditida, Steinernematidae), a new species of entomopathogenic nematode from the province of Sichuan, east Tibetan Mts., China.

Steinernema sichuanense n. sp. is characterized by male, female and IJ. For male, the spicules are robust with prominent rostrum; gubernaculum has blunt anterior end; cuneus is arrow-shaped, pointed posteriorly. Second-generation male has a prominent mucron. For female, tail usually has one to four papillae-like projections on tail tip; post anal swelling is absent. For IJ, body length is about 710 microm; lateral field has six ridges; the formula of lateral field is 2, 5, 6, 4, 2 with two prominent submarginal ridges; tail usually has a dorsal depression. In Steinernema affine/intermedium group, the IJ of S. sichuanense n. sp. differs from S. affine by its absence of the internal tail spine; differs from Steinernema beddingi by its six ridges in lateral field compared to 4 for S. beddingi. For male mucron is absent in both generations of S. affine, S. intermedium and S. beddingi, whereas it is present in the second-generation of S. sichuanense sp. n. Morphology and morphometrics of spicules and gubernacula of the four species in S. affine/intermedium group are quite different based on SEM photographs. For female, the postanal swelling is absent in the first-generation of S. sichuanense n. sp. whereas S. affine and S. intermedium have slight swelling and S. beddingi has conspicuous swelling. The new species is further recognized by characterization of sequences of ITS and D2/D3 regions of the ribosomal DNA. The symbiotic bacterium associated to S. sichuanense belongs to the species Xenorhabdus bovienii.

Seasonal dynamics of entomopathogenic nematodes of the genera Steinernema and Heterorhabditis as a response to abiotic factors and abundance of insect hosts.

The seasonal dynamics of entomopathogenic nematodes (EPNs) of the genus Steinernema and Heterorhabditis were studied during one season in meadow and oak wood habitats, in the vicinity of Ceské Budejovice, Czech Republic. The influences of soil temperature, moisture, and abundance of suitable hosts on EPN dynamics were investigated. The host range of these nematodes, in both habitats was also observed. A total of four EPN species were found in both habitats. Steinernema affine was the dominant species both in oak wood and in meadow. Additionally, the oak wood habitat was inhabited by S. kraussei and S. weiseri; the meadow habitat by Heterorhabditis bacteriophora. The mean abundance of total EPN community was 28,000ind./m(2) in oak wood and 11,000ind./m(2) in meadow. The seasonal dynamics of entomopathogenic nematodes in both habitats were characterized by high nematode densities in the beginning of the season, followed by a rapid decrease, and then stabilization. EPN abundances did not show any apparent correlation with soil temperature and moisture, but they were negatively correlated with the abundance of suitable insect hosts. Inter- and intraspecific competition for limited nutrients (hosts) probably played a major role in EPN seasonal dynamics. Broad host range of entomopathogenic nematodes in both habitats was predominantly represented by dipteran and coleopteran larvae. Most common hosts belonged to the families Asilidae, Bibionidae, and Empididae (Diptera), as well as Carabidae and Curculionidae (Coleoptera).