Morphological, molecular and ecological characterization of a native isolate of Steinernema feltiae (Rhabditida: Steinernematidae) from southern Chile.

BACKGROUND: Steinernema feltiae is an entomopathogenic nematode used in biological control programs with a global distribution. Populations of this species show phenotypic plasticity derived from local adaptation and vary in different traits, such as location and host penetration. The aim of this work was to describe a Chilean isolate of this nematode species, using integrative approaches. METHODS: Nematode morphological and morphometric studies were conducted along with molecular analysis of nuclear genes. The symbiotic bacterium was also identified by sequencing the 16S rRNA gene. Some ecological characteristics were described, including the temperature requirements for the nematode life cycle and the effect of soil water content for optimal reproduction. RESULTS: Morphometric characterization revealed a large intra-specific variability. The isolate identity was also corroborated with the analysis of nuclear genes. Based on the 16S gene, its symbiont bacteria, Xenorhabdus bovienii, was identified. The lowest, optimal and highest temperatures found to limit the infestation and reproduction on Galleria mellonella were 10, 20 and 30 °C, respectively; the emergence from the host larvae occurred approximately 10 days after inoculation. Differences were observed in offspring, and 120 infective juveniles (IJ)/larva was the most prolific dose at 20 °C. The soil water content did not affect the number of IJ invaders, penetration efficacy and IJ emergence time or offspring per larva, but it caused a delay in achieving full mortality at the permanent wilting point with respect to saturation and field capacity. CONCLUSIONS: For the first time, a Chilean isolate of S. feltiae is described in detail considering morphological, molecular and ecological aspects. The isolate was shown to be efficient in soil containing water, with optimal temperatures ranging from 15 to 25 °C for host infestation and production of an abundant offspring; these characteristics would allow its potential use as control agents in a wide geographical area of the country.

First case of fatal equine meningoencephalitis caused by Halicephalobus gingivalis in Mexico.

Aberrant nematode larval migration in the CNS of horses is rare but frequently fatal; one of the main etiological agents involved in this illness is Halicephalobus gingivalis. This soil nematode has been associated with several fatal equine meningoencephalitis reports worldwide; however, it had never been diagnosed in horses of Mexico. A 10 year-old Andalusian horse presented dysphagia, fever, weakness, prostration and ataxia; the patient expired during the medical attention. Post mortem examination was performed and no gross alterations were found. Histopathology revealed meningoencephalitis, vasculitis and intralesional adult nematodes, larvae and eggs compatible with Halicephalobus spp. A polymerase chain reaction (PCR) for the nuclear large subunit ribosomal RNA gene (LSU rDNA) of nematodes was performed from formalin-fixed and paraffin wax-embedded sections of brain. Posterior nucleotide sequence analysis of the amplified fragment identified the agent as H. gingivalis. To our knowledge, this is the first confirmed report of Halicephalobiasis in Mexico.

A new species of Diomedenema (Nematoda, Rhabditida, Spiruromorpha) from Spheniscus magellanicus (Aves, Sphenisciformes) found on the southern coast of Brazil.

A new species of the genus Diomedenema, a spiruromorph nematode, collected from the lung of Spheniscus magellanicus (Sphenisciformes) found on the southern coast of the state of Rio Grande do Sul, Brazil, is described. The new species is differentiated from the only previously described species of the genus, D. diomedeae Johston & Mawson, 1952, by males possessing a set of caudal papillae with three pairs of precloacal, two pairs of adcloacal and one pair of postcloacal papillae; precloacal papillae with the papillae of the first two pairs being closer to each other than those of the third pair; a longer and pointed tail in males; and females with the vulva at mid-body. This is the first report of a nematode infecting the lung of a sphenisciforme host.

Morphological and molecular characterisation of an isolate of Steinernema diaprepesi Nguyen & Duncan, 2002 (Rhabditida: Steinernematidae) from Argentina and identification of its bacterial symbiont.

Entomopathogenic nematodes of the families Heterorhabditidae Poinar, 1976 and Steinernematidae Chitwood & Chitwood, 1937 are used for biological control of insect pests. An isolate of Steinernema diaprepesi Nguyen & Duncan, 2002 was recovered from a carrot field in the locality of Santa Rosa de Calchines (Santa Fe Province, Argentina). These nematodes were characterised based on morphological, morphometric and molecular studies. Their symbiotic bacterium was identified as Xenorhabdus doucetiae Tailliez, Pagès, Ginibre & Boemare, 2006 by sequencing the 16S rRNA gene. The isolate of S. diaprepesi studied exhibits some morphometric differences with the original description, especially in the first generation adults. This is the first description of the species in Argentina.

Steinernema goweni n. sp. (Rhabditida: Steinernematidae), a new entomopathogenic nematode from Zulia State, Venezuela.

A nematode from the genus Steinernema was isolated in Zulia state (North-western, Venezuela). Morphological, morphometric and molecular studies indicated that this nematode belongs to the "bicornutum" group and is described herein as Steinernema goweni n. sp. Steinernema goweni n. sp. is characterised by morphometrics of its infective juveniles, with body length 640 (531-719) µm, pharynx length of 119 (109-126) µm, tail of 67 (59-89) µm, c ratio = 9 (6-11) and E% = 77 (48-94). First generation male specimens can be recognised by the shape and size of spicules (55 (50-57) µm long) and gubernaculum (35 (30-40) µm long), and %D value 42 (28-59), which is at the lower limit within the "bicornutum" group. The number of genital papillae in males is also distinctive for S. goweni n. sp. presenting up to 27 (13 pairs + 1) papillae in 15% of specimens. The phasmids can be seen in scanning electron microscopy in all stages, characters not previously reported within the "bicornutum" group. Phylogenetic analyses of the "bicornutum" group based on both ITS and D2D3 regions showed a clear separation of S. goweni n. sp. from the other species. In both analyses S. goweni n. sp. formed a strongly supported group of American species.

Steinernema brazilense n. sp. (Rhabditida: Steinernematidae), a new entomopathogenic nematode from Mato Grosso, Brazil.

A new entomopathogenic nematode, Steinernema brazilense n. sp., was isolated from a single soil sample collected from a natural forest in Mato Grosso do Sul state, Brazil. S. brazilense n. sp. is characterized morphologically by features of infective juveniles (IJ), males and females. For the IJ, body length averaging 1157 (1023-1284)microm, distance from anterior end to excretory pore 95 (87-102)microm, from anterior end to end of esophagus 148 (139-153)microm, tail length 85 (80-104)microm, D% and E% values 63 (58-70) and 106 (95-118.0), respectively. Lateral field pattern variable; the formula for the arrangement of ridges from head to tail is: 2, 4, 6, 8, 6, 2. For the male, the diagnostic characters include spicule averaging 83 (75-89)microm; D% about 65; the ratio SW% about 192. The length of spicule head is greater than width. Lateral field with one narrow ridge. First generation females are characterized by the presence of a ventral postanal swelling. S. brazilense n. sp. is morphologically close to Steinernema diaprepesi. It can be differentiated from S.diaprepesi by its longer IJ body length (1157 vs 1002microm), longer distance from anterior end to excretory pore (110 vs 75microm), a longer tail length (103 vs 83microm); males of the new species with longer spicule (83 vs 79microm). The new species can be distinguished further from other members of Steinernema glaseri group by characteristics of rDNA of ITS and D2D3 regions.

Molecular and morphological characterization of heterorhabditid entomopathogenic nematodes from the tropical rainforest in Brazil.

Despite massive losses of primary forest, the Amazonian rainforest remains an extremely rich source of biodiversity. In recent years, entomopathogenic nematodes (EPNs) have been isolated from soil in various parts of the world and used successfully as biological control agents against numerous insect pests. Therefore, a sampling in the rainforest of Monte Negro, Rondônia, Brazil was conducted with the aim of discovering new strains and/or species of EPNs for future development as biological control agents. From 156 soil samples taken at nine collecting sites, 19 isolates were obtained, all of them belonging to the genus Heterorhabditis. Four strains were subjected to detailed morphological and molecular evaluation. Based on morphometrics and internal transcribed spacer (ITS) sequence data, the strains LPP1, LPP2 and LPP4 were identified as Heterorhabditis indica, whereas LPP7 was considered Heterorhabditis baujardi. Comparative analysis of the ITS1 sequence of H. indica and H. baujardi isolates showed a polymorphic site for the restriction enzyme Tth 111 that could be used to distinguish the two species. Consequently, strains LPP1, LPP2, LPP3, LPP4, and LPP9 were identified as H. indica, whereas LPP5, LPP7, LPP8 and LPP10 were identified as H. baujardi.

Molecular and morphological characterization of heterorhabditid entomopathogenic nematodes from the tropical rainforest in Brazil

Despite massive losses of primary forest, the Amazonian rainforest remains an extremely rich source of biodiversity. In recent years, entomopathogenic nematodes (EPNs) have been isolated from soil in various parts of the world and used successfully as biological control agents against numerous insect pests. Therefore, a sampling in the rainforest of Monte Negro, Rondônia, Brazil was conducted with the aim of discovering new strains and/or species of EPNs for future development as biological control agents. From 156 soil samples taken at nine collecting sites, 19 isolates were obtained, all of them belonging to the genus Heterorhabditis. Four strains were subjected to detailed morphological and molecular evaluation. Based on morphometrics and internal transcribed spacer (ITS) sequence data, the strains LPP1, LPP2 and LPP4 were identified as Heterorhabditis indica, whereas LPP7 was considered Heterorhabditis baujardi. Comparative analysis of the ITS1 sequence of H. indica and H. baujardi isolates showed a polymorphic site for the restriction enzyme Tth 111 that could be used to distinguish the two species. Consequently, strains LPP1, LPP2, LPP3, LPP4, and LPP9 were identified as H. indica, whereas LPP5, LPP7, LPP8 and LPP10 were identified as H. baujardi.

Steinernema costaricense n. sp. and S. puntauvense n. sp. (Rhabditida: Steinernematidae), two new entomopathogenic nematodes from Costa Rica.

Steinernema costaricense n. sp. and S. puntauvense n. sp. were recovered during a survey for native entomopathogenic nematodes in Costa Rica. Morphological data, molecular (28S rDNA sequence data) studies and cross-hybridisation tests were used for diagnostic and identification purposes. Additionally, 28S rDNA sequence data were used to assess the evolutionary relationships of the new species with other Steinernema spp. Morphological diagnostic features for S. costaricense n. sp. include: the body size of the infective juvenile (av. 1,696); the presence of protruding 'horn-like' cephalic papillae; the position of the excretory pore in the infective juvenile (av. 77 microm) and the first generation male (av. 117 microm); the D% value of the infective juvenile (av. 53) and the first generation male (av. 56); the E% value of the infective juvenile (av. 85); and the morphology of the spicules and gubernaculum of the first generation male. Diagnostic traits for S. puntauvense n. sp. are: the position of the excretory pore in the infective juveniles (av. 25 microm); the shape and size of the spicules and gubernaculum of the first generation male; and the shape of the tail of the first generation female. In addition to these traits, 28S rDNA sequences analysis and hybridisation tests showed that both new Steinernema species are distinct and unique entities.

Two new species of nematodes (Rhabditida: Diplogasteridae and Rhabditidae) parasites of Gryllodes Iaplatae (Orthoptera: Gryllidae) in Argentina

Cephalobium magdalensis n.sp. (Rhabditida: Diplogasteridae) found in Magdalena, Buenos Aires, and Cruznema lincolnensis n. sp (Rhabditida: Rhabditidae) found in Lincoln, Buenos Aires, parasitizing the cricket Gryllodes laplatae (Orthoptera: Gryllidae) are described and illustrated. C. magdalensis n. sp. is characterized by having the excretory pore between the pseudobulb and the basal bulb and seven pairs of postanal papillae in the male. C. lincolnensis n.sp. can be distinguished by having meanly two pairs of preanal papillae, one pair of adanal papillae and six pairs of postanal papillae in the male.

Two new species of nematodes (Rhabditida: Diplogasteridae and Rhabditidae) parasites of Gryllodes laplatae (Orthoptera: Gryllidae) in Argentina.

Cephalobium magdalensis n. sp. (Rhabditida: Diplogasteridae) found in Magdalena, Buenos Aires, and Cruznema lincolnensis n. sp. (Rhabditida: Rhabditidae) found in Lincoln, Buenos Aires, parasitizing the cricket Gryllodes laplatae (Orthoptera: Gryllidae) are described and illustrated. C. magdalensis n. sp. is characterized by having the excretory pore between the pseudobulb and the basal bulb and seven pairs of postanal papillae in the male. C. lincolnensis n. sp. can be distiguished by having meanly two pairs of preanal papillae, one pair of adanal papillae and six pairs of postanal papillae in the male.