Ultrastructural study of anther parasitism of Ficus laevigata by Ficophagus laevigatus (Aphelenchoididae).

Transmission electron microscopy (TEM) was used to compare the ultrastructural differences between healthy male florets (anthers) and one floret parasitized by Ficophagus laevigatus in late phase C syconia of Ficus laevigata from southern Florida. Previous light-microscopic examination of paraffin-sectioned material showed that F. laevigatus-infested anthers of F. laevigata manifested as malformed, often with aberrant pollen and hypertrophied epidermal cells closest to regions containing propagating nematodes. Female florets or fig wasp-parasitized female florets were not observed to be parasitized by nematodes. Considering that plant-feeding in the Aphelenchoididae is purportedly much less specialized than in certain groups of the Tylenchomorpha, where specialized hypertrophied feeder cells are produced in response to nematode feeding, we examined the putative induced response in this unusual aphelenchoidid system with the higher resolution afforded by TEM. TEM confirmed the expression of significant epidermal cell hypertrophy of the anther and anther filament in the presence of propagating nematodes, which was expressed as cell enlargement (2-5X), fractionation of large electron-dense stores into smaller aggregates, irregularly shaped nuclei enclosed by an elongated nuclear envelope, nucleolus enlargement, increased organelle production, and apparent metabolism with increased numbers of mitochondria, pro-plastids, and endoplasmic reticulum, as well as increased thickening of the cell walls. Pathological effects were observed in adjacent cells/tissue (e.g., anther and anther filament parenchymal cells, pollen tubes, pollen, and endothecium) with apparent diminishment as the distance from propagating nematodes increased (which was also probably affected by number of nematodes). Some TEM sections captured previously undocumented ultrastructural highlights of propagating individuals of F. laevigatus.

Morphological and Molecular Characterization of Labrys filiformis n. sp. (Rhabditida: Tylenchidae) from Iran.

Labrys filiformis n. sp., the second species of the rare genus Labrys , was recovered from natural forests of Gilan province and is described based upon morphological and molecular characters. The new species is characterized by its smooth cuticle under light microscopy, lateral field with two incisures forming a single plain band, lip region continuous with body contour, dorso-ventrally flattened and forming four poorly prominent lobes, having a dorso-ventrally narrower protuberant labial plate laterally extended to the amphidial margins, oral area (oral plate) dorso-ventrally elongated and embedded in the labial plate with six small labial sensilla surrounding the slightly prominent oral aperture, amphidial apertures as longitudinally lemniscatic slits bordered by the labial plate extensions which are overlapped at the middle length of amphids, stylet delicate, 6 to 7 µm long, elongate weakly developed fusiform median bulb with weak valve, wide excretory pore with long and heavily sclerotized duct, offset spermatheca filled with small spheroid sperm cells, 106 to 127 µm long elongate-conoid tail with filiform distal region and finely rounded tip. Molecular phylogenetic analyses were performed using a near-full length fragment of the 18S rDNA and the D2-D3 expansion segments of the 28S rDNA using Bayesian inference and maximum likelihood methods. In the inferred phylogenetic tree with 18S rDNA, the new species has a close affinity with several isolates of the type species, Labrys chinensis . The reconstructed phylogenetic tree using partial 28S rDNA, revealed the new species is nested inside the putative monophyletic group of several populations of L. chinensis .

Effects of Infection by Belonolaimus longicaudatus on Rooting Dynamics among St. Augustinegrass and Bermudagrass Genotypes.

Understanding rooting dynamics using the minirhizotron technique is useful for cultivar selection and to quantify nematode damage to roots. A 2-yr microplot study including five bermudagrass ('Tifway', Belonolaimus longicaudatus susceptible; two commercial cultivars [TifSport and Celebration] and two genotypes ['BA132' and 'PI 291590'], which have been reported to be tolerant to B. longicaudatus) and two St. Augustinegrass ('FX 313', susceptible, and 'Floratam' that was reported as tolerant to B. longicaudatus) genotypes in a 5 x 2 and 2 x 2 factorial design with four replications, respectively, was initiated in 2012. Two treatments included were uninoculated and B. longicaudatus inoculated. In situ root images were captured each month using a minirhizotron camera system from April to September of 2013 and 2014. Mixed models analysis and comparison of least squares means indicated significant differences in root parameters studied across the genotypes and soil depths of both grass species. 'Celebration', 'TifSport' and 'PI 291590' bermudagrass, and 'Floratam' St. Augustinegrass had significantly different root parameters compared to the corresponding susceptible genotypes (P ≤ 0.05). Only 'TifSport' had no significant root loss when infested with B. longicaudatus compared to non-infested. 'Celebration' and 'PI 291590' had significant root loss but retained significantly greater root densities than 'Tifway' in B. longicaudatus-infested conditions (P ≤ 0.05). Root lengths were greater at the 0 to 5 cm depth followed by 5 to 10 and 10 to 15 cm of vertical soil depth for both grass species (P ≤ 0.05). 'Celebration', 'TifSport', and 'PI 291590' had better root vigor against B. longicaudatus compared to Tifway.

Phylogenetic Status and Morphological Characters of Rhabditolaimus anoplophorae (Rhabditida: Diplogastridae).

Rhabditolaimus anoplophorae Kanzaki and Futai was re-isolated from its type host (carrier), the cerambycid beetle Anoplophora malasiaca, collected in an experimental field of the Forestry and Forest Products Research Institute, Tsukuba, Ibaraki, Japan. The nematode was cultured on nematode growth medium plates seeded with Escherichia coli OP50, and its morphological characters and molecular profile were examined to modernize the description. Scanning electron microscopic and light microscopy revealed the presence of four stomatal flaps, a very long gymnostom, a single ventral papilla in males, and a horizontal slit-like vulval opening in females. The positions of the deirids, hemizonids, phasmids, and rectal glands are additionally described, and the absence of a male bursa was confirmed. Phylogenetically, the genus forms a well-supported clade in the family Diplogastridae. Rhabditolaimus anoplophorae is a member of the monophyletic Rhabditolaimus clade and is closely related to R. leuckarti and several undescribed species.

Description of Schistonchus altissimus n. sp. (Nematoda: Aphelenchoididae), an associate of Ficus altissima in China.

A new nematode species was recovered from the syconia of Ficus altissima from the residential area of Huajingxincheng, Guangzhou, Guangdong Province, China during a survey of nematode diversity. Schistonchus altissimus n. sp. is characterised by having females with a short post-uterine sac, an ovoid spermatheca and a conoid tail with a mucron in the female, excretory pore located near the lip; and males with amoeboid sperm, a conoid tail without a mucron and three pairs of subventral papillae, no gubernaculum, and hook-shaped spicules with a cucullus and a thorn-shaped rostrum. Schistonchus altissimus n. sp. is typologically differentiated from all other described species in this genus, except for S. microcarpus, by having a spicule with cucullus on the male tail tip. Schistonchus altissimus n. sp. is easily differentiated from other sequenced species by the partial small subunit rRNA gene (SSU), D3 expansion segment of the large subunit rRNA gene (LSU) and mitochondrial DNA subunit I (mtCOI). Phylogenetic analysis with partial SSU sequences suggests that S. altissimus n. sp. is in a highly supported monophyletic clade with two Chinese species (S. microcarpus and S. centerae) and two neotropical species (S. aureus and Schistonchus sp. ex Ficus colubrinae Standl.). Based on inferences using LSU D3 sequence data, S. altissimus n. sp. has a closer relationship with four Chinese species (S. centerae, S. fistulosus, S. guangzhouensis and S. microcarpus) than with S. hirtus and S. superbus, also from China.

Nematodes from galls on Myrtaceae. IV. Fergusobia from flat leaf galls on Eucalyptus and Corymbia, with descriptions of two new species.

Two new species of Fergusobia are described. Both were collected from flat leaf galls from South Australia, one on Eucalyptus microcarpa and the other on E. porosa. Fergusobia microcarpae n. sp. Davies is characterised by the combination of a C-shaped parthenogenetic female with a short, broadly rounded conoid tail, an arcuate to open C-shaped infective female with an hemispherical tail tip, and arcuate to J-shaped males with angular spicules and short peloderan bursa. Fergusobia porosae n. sp. Davies is similar in having an arcuate to C-shaped parthenogenetic female with a small conoid tail, an almost straight to arcuate infective female with an hemispherical tail tip, and males that are almost straight to barely J-shaped with angular spicules and short peloderan bursa. They differ in that the bodies of parthenogenetic and infective females of F. microcarpae n. sp. are more curved than in F. porosae n. sp. Other known similar forms of Fergusobia/Fergusonina galls are outlined and the larval shield morphologies of their associated mutualistic Fergusonina fly species are discussed where known. An inventory of all known Fergusobia/Fergusonina associations from flat leaf galls from Corymbia spp. and Eucalyptus spp. is presented. Relationships of Fergusobia nematodes were inferred from analysis of sequences of 28S rDNA D2/D3 domains and a portion of mitochondrial DNA cytochrome oxidase subunit I (mtCOI). Nematodes from flat leaf galls appeared in two clades. 

Nematodes from galls on Myrtaceae. V. Fergusobia from large multilocular shoot bud galls from Angophora and Eucalyptus in Australia, with descriptions of six new species.

Six new species of Fergusobia, from large multilocular shoot bud galls on two species of Angophora and four species of Eucalyptus from both subgenera Eucalyptus and Symphyomyrtus, are described. Fergusobia cosmophyllae Davies n. sp. is characterized by the combination of a C-shaped parthenogenetic female with a short arcuate conoid tail, a broad (small a ratio) arcuate infective female with an hemispherical tail tip, and an arcuate to J-shaped male with broad, angular spicules and short bursa.  Fergusobia delegatensae Davies n. sp. has an open C-shaped parthenogenetic female with a broadly conoid tail, an infective female of variable shape with an hemispherical tail tip, and a male of open C-shape with a crenate bursa that arises 40-70% along the length of the body from the tail tip and terminates just anterior to the cloaca. Fergusobia diversifoliae Davies n. sp. has a C-shaped parthenogenetic female with a conoid tail, an arcuate infective female with a hemispherical tail tip, and an arcuate, C- or J-shaped male with angular spicule and a long peloderan bursa. Fergusobia floribundae Davies n. sp. has a C-shaped parthenogenetic female with a narrow, arcuate, conoid tail, an arcuate infective female with a hemispherical tail tip, and an arcuate or J-shaped male with an angular spicule and a short to mid-body length peloderan bursa. Fergusobia minimus Lisnawita n. sp. has a C-shaped parthenogenetic female with a conoid tail, an arcuate to open C-shaped infective female with a hemispherical tail tip, and an arcuate to open C-shaped male with an angular spicule and a peloderan bursa arising at about 10-30% of body length. Fergusobia pimpamensis Davies n. sp. has an open C to C-shaped parthenogenetic female with a narrow conoid tail, an arcuate to open C-shaped infective female with a hemispherical tail tip, and an arcuate to C-shaped male with an arcuate spicule and a long, crenate, peloderan bursa. An inventory of all known Fergusobia/Fergusonina associations from terminal shoot bud galls is presented. The larval shield morphology of the associated mutualistic Fergusonina species is discussed where known. Analyses of DNA sequences of D2/D3 and COI suggested that the six new species are distributed between three clades of Fergusobia.

The nature and frequency of chimeras in eukaryotic metagenetic samples.

Pyrosequencing of an artificially assembled nematode community of known nematode species at known densities allowed us to characterize the potential extent of chimera problems in multi-template eukaryotic samples. Chimeras were confirmed to be very common, making up to 17% of all high quality pyrosequencing reads and exceeding 40% of all OCTUs (operationally clustered taxonomic units). Typically, chimeric OCTUs were made up of single or double reads, but very well covered OCTUs were also present. As expected, the majority of chimeras were formed between two DNA molecules of nematode origin, but a small proportion involved a nematode and a fragment of another eukaryote origin. In addition, examples of a combination of three or even four different template origins were observed. All chimeras were associated with the presence of conserved regions with 80% of all recombinants following a conserved region of about 25bp. While there was a positive influence of species abundance on the overall number of chimeras, the influence of specific-species identity was less apparent. We also suggest that the problem is not nematode exclusive, but instead applies to other eukaryotes typically accompanying nematodes (e.g. fungi, rotifers, tardigrades). An analysis of real environmental samples revealed the presence of chimeras for all eukaryotic taxa in patterns similar to that observed in artificial nematode communities. This information warrants caution for biodiversity studies utilizing a step of PCR amplification of complex DNA samples. When unrecognized, generated abundant chimeric sequences falsely overestimate eukaryotic biodiversity.

Bursaphelenchus suri n. sp.: A second Bursaphelenchus syconial parasite of figs supports adaptive radiation among section Sycomorus figs.

The nematode genus Bursaphelenchus is a highly divergent group. This genus mainly consists of mycophagous entomophilic species, but some species have specialized as obligate or facultative plant parasites, facultative insect parasites, or exhibit feeding dimorphism (phenotypic plasticity) leading to mycophagous and predatory forms. In the present study, a new Bursaphelenchus species, B. suri n. sp. was isolated from fresh syconia (figs) of Ficus sur and is described and illustrated based on its typological characters and molecular phylogenetic status. The new species is characterized by its highly derived feeding structures found in obligate plant parasites, lip possessing a labial disc and a long and thick stylet with a long conus and extremely well-developed basal swellings. In addition, slender body of both sexes is characteristic of the species. The new species is phylogenetically and typologically closely related to B. sycophilus, i.e., these two species share the characteristic feeding structures and form a well-supported clade within the B. fungivorus group in the genus. Biologically, these two species are both isolated from fresh figs of the section Sycomorus. However, the new species differs from B. sycophilus by the length of the female post-uterine sac and the shape of the male spicule, i.e., the new species has a long post-uterine sac and spicule condylus without dorsal recurvature. Thus, the new species is the second obligate fig parasite of the genus, and the evolutionary relationship between the B. suri n. sp. and B. sycophilus clade and section Sycomorus figs is hypothesized as an example of adaptive radiation with more species to be discovered.

Field Responses of Bermudagrass and Seashore paspalum Cultivars to Sting and Spiral Nematodes.

Belonolaimus longicaudatus and Helicotylenchus spp. are damaging nematode species on bermudagrass (Cynodon spp.) and seashore paspalum (Paspalum vaginatum) in sandy soils of the southeastern United States. Eight bermudagrass and three seashore paspalum cultivars were tested for responses to both nematode species in field plots for two years in Florida. Soil samples were taken every three months and nematode population densities in soil were quantified. Turfgrass aboveground health was evaluated throughout the growing season. Results showed that all bermudagrass cultivars, except TifSport, were good hosts for B. longicaudatus, and all seashore paspalum cultivars were good hosts for H. pseudorobustus. Overall, bermudagrass was a better host for B. longicaudatus while seashore paspalum was a better host for H. pseudorobustus. TifSport bermudagrass and SeaDwarf seashore paspalum cultivars supported the lowest population densities of B. longicaudatus. Seashore paspalum had a higher percent green cover than bermudagrass in the nematode-infested field. Nematode intolerant cultivars were identified.

Molecular diversity and relationships of fig associated nematodes from South Africa.

Nematodes of figs and fig wasps have received limited attention in Africa since their discovery in 1973. Sixteen of the 25 species of native South African figs were sampled for nematode associates using molecular barcoding with three loci (SSU, LSU D2-D3 and mtCOI) and fourteen (93%) were positive for at least one nematode species. Thirty-three putative species of nematodes were identified and classified according to the loci that were amplified and successfully sequenced. Fourteen putative nematode species were classified as Aphelenchoididae, of which nine were identified as Ficophagus from four species of Ficus from the section Galoglychia (i.e., five ex F. burkei including one shared with F. natalensis, one ex F. glumosa, one ex F. lutea, and one ex F. stuhlmannii) and one species ex F. sur from the section Sycomorus. In addition, there were four nematode species classified as Schistonchus s.s. from section Galoglychia figs (i.e., one ex F. burkei, two ex F. trichopoda, and one ex F. glumosa). There was also one species of Bursaphelenchus nematode recovered from F. sur from the section Sycomorus. Sixteen putative nematode species were classified as Diplogastridae, of which eight occurred in two clades of what is currently called Parasitodiplogaster with one (P. salicifoliae) being recovered from two Ficus species in the section Urostigma (F. salicifolia and F. ingens) and seven diplogastrids being associated with six species of Ficus from the section Galoglychia (i.e., two ex F. burkei including P. sycophilon, one ex F. stuhlmannii, one ex F. burtt-davyi, one ex F. trichopoda, one ex F. abutilifolia and one ex F. sansibarica). Three Acrostichus spp., a Teratodiplogaster and a Pristionchus species were recovered from F. sur and two Teratodiplogaster spp. and Pristionchus sycomori were recovered from F. sycomorus with both Ficus species belonging to the subgenus and section Sycomorus. The identities of the previously described T. martini and Parasitodiplogaster doliostoma (= Pristionchus sp. 35) are discussed. Lastly, there was a panagrolaimid identified from F. petersii.

Nematode epibionts on skin of the Florida manatee, Trichechus manatus latirostris.

A survey for the presence of nematodes on the skin of the native Florida manatee, Trichechus manatus latirostris from Crystal River, Florida was conducted during annual manatee health assessments. A putative isolate of Cutidiplogaster manati (Diplogastridae) and two other nematodes belonging to the same family were recovered from mid-dorsal tail skin-scrapings from all sampled winter-collected healthy wild adult manatees during two successive years (2018-2019). Qualitative abundance estimates of these three species of diplogastrid nematodes suggest that an average wild Florida manatee adult might possess between 30,000 and 120,000 nematodes on its tail dorsum and that the entire body dorsum including the tail might possess 160,000-640,000 nematodes in roughly equal ratios. Attempts to culture these nematodes on a variety of different culture media were unsuccessful but examination of the mouth (stomatal) morphology suggests specialized feeding on microbes such as diatoms or predation on other nematodes. No skin lesions were observed during the 2018-2019 samplings suggesting that under normal conditions these nematodes are highly specialized free-living epibionts of the skin that are tightly bound to this niche and horizontally transferred between individual manatees in an analogous fashion to human skin mites (Demodex folliculorum and D. brevis). Molecular phylogenetic inferences using sequences of near full length SSU and D2-D3 expansion segments of LSU rRNA genes revealed a putative new morphospecies in Cutidiplogaster sister to C. manati that was monophyletic with several named Mononchoides species, and another putative new morphospecies that formed a clade with several undescribed species similar in appearance to Mononchoides as well as Tylopharyx, Eudiplogasterium, Paroigolaimella and Sachsia.

Ecometagenetics confirm high tropical rainforest nematode diversity.

The general patterns of increasing biodiversity from the poles to the equator have been well documented for large terrestrial organisms such as plants and vertebrates but are largely unknown for microbiota. In contrast to macrobiota, microbiota have long been assumed to exhibit cosmopolitan, random distributions and a lack of spatial patterns. To evaluate the assumption, we conducted a survey of nematode diversity within the soil, litter and canopy habitats of the humid lowland tropical rainforest of Costa Rica using an ultrasequencing ecometagenetic approach at a species-equivalent taxonomic level. Our data indicate that both richness and diversity of nematode communities in the tropical rainforests of Costa Rica are high and exceed observed values from temperate ecosystems. The majority of nematode species were unknown to science, providing evidence for the presence of highly endemic (not cosmopolitan) species of still completely undiscovered biodiversity. Most importantly, the greater taxonomic resolution used here allowed us to reveal predictable habitat associations for specific taxa and thus gain insights into their nonrandom distribution patterns.

Effects of formulation and host nematode density on the ability of in vitro-produced pasteuria endospores to control its host Belonolaimus longicaudatus.

The effect of nematode population density at the time of application and formulations of in vitro-produced Pasteuria spp. endospores on the final population density of Belonolaimus longicaudatus was studied in an 84-d-long pot bioassay. The experiment utilized a factorial design consisting of 30 or 300 B. longicaudatus /100 cm(3) of sandy soil and three formulations of in vitro-produced Pasteuria spp. endospores (nontreated, granular, or liquid). No differences were observed in percent endospore attachment between nematode inoculum levels during either trial. Granular and liquid formulations of in vitro-produced endospores suppressed nematode population densities by 22% and 59% in the first trial and 20% and 63% in the second, respectively compared with the nontreated control. The liquid formulation increased percent endospore attachment by 147% and 158%, respectively, compared with the granular formulation. The greatest root retention by the host plant was observed at the lower B. longicaudatus inoculation level following application of the liquid formulation. While both the granular and liquid formulations reduced B. longicaudatus population densities in the soil, the liquid spore suspension was most effective.

Teratodiplogaster fignewmani gen. nov., sp. nov. (Nematoda: Diplogastridae) from the syconia of Ficus racemose in Australia.

During the course of a survey of fig-associated nematodes in eastern Australia, we discovered an unusual diplogastrid nematode that we describe herein as Teratodiplogaster fignewmani gen. nov., sp. nov. This nematode was isolated as adults and juveniles from the syconia of Ficus racemose in Queensland and Western Australia. It is presumed to be associated with the agaonid fig wasp, Ceratosolen fusciceps, for dispersion to new phase-B sycones. Teratodiplogaster fignewmani gen. nov., sp. nov. was inferred to be the sister taxon to the genus Parasitodiplogaster based upon molecular phylogeny using nearly full-length sequences of the SSU and D2/D3 LSU ribosomal RNA genes. It is sufficiently distinct in morphology (with many autapomorphies) to justify a new monotypic genus with the hypothesis that further species of Teratodiplogaster gen. nov. will be discovered when sycones from other Ficus from tropical Australasia and possibly Africa are sampled.

The first report of a putative "entomoparasitic adult form" in Bursaphelenchus.

A putative "entomoparasitic adult form" of Bursaphelenchus luxuriosae was found in the tracheal system and body cavity of its cerambycid vector beetle, Acalolepta luxuriosa. Morphologically, this form is intermediate between the usual mycophagous adult and the phoretic dauer juvenile, i.e., it shares the primary and secondary reproductive features with the mycophagous phase but shares a degenerate digestive tract with JIV dauer juveniles. In addition, the "parasitic form" has specialized characters relative to the mycophagous phase analogous to dicyclic Hexatylina, e.g., vacuole-like dots (assumed to be sensory organs) at the anterior end, a very long vulval flap, conical female tail, and elongated male spicules. The presence of insect-parasitic juveniles has been reported in several Bursaphelenchus species, but this is the first confirmed report of a putative "entomoparasitic adult form" in the genus. Thus, the "parasitic adult form" of B. luxuriosae is hypothesized to be an autapomorphic character of this species in the genus. The physiological impact of the parasitism on the host beetle is assumed to be weak because no clear symptoms were observed in the infested beetles. Also, no nematode eggs or propagative juveniles were observed in the beetle host, suggesting that more research is needed to confirm the nature of the association (parasitic vs. endophoretic).

Feeding Dimorphism in a Mycophagous Nematode, Bursaphelenchus sinensis.

Phenotypic plasticity has been widely reported in animals and can drive investment in new biological characters that engender ecological adaptability. The nematode family Diplogastridae, especially Pristionchus pacificus with its dramatic stomatal (feeding) dimorphism, has become an important model system to analyze the evolutionary and developmental aspects of polyphenism. However, this plasticity has not been confirmed in other nematode groups. In the present study, we experimentally examined the feeding dimorphism of a fungal feeding free-living nematode, Bursaphelenchus sinensis. In a laboratory culturing experiment, the nematode expressed dimorphism, i.e., a small proportion of the population manifested as a predatory form. This form only occurred in females and was not clearly influenced by the presence of potential prey species. In addition, the ratio of the predatory form to the mycophagous form varied among different fungal food species grown in monoculture on different culture media. The predatory form of B. sinensis was typologically similar to the monomorphic (specialized) predators belonging to the same family. However, some essential morphological characters were slightly different from the specialized predators, and their behaviours were clearly disparate, suggesting that predation in B. sinensis is derived from a different phylogenetic origin than that of the specialized predators.

Nematodes from galls on Myrtaceae. XI. Descriptions of five new species of Fergusobia from Australia.

Five new species of Fergusobia are described. Fergusobia janetae Davies n. sp. is characterized by the combination of an arcuate to straight, spindle-shaped parthenogenetic female with a relatively small oesophageal gland, an extensile uterus and a conoid tail, an arcuate infective female with a bluntly rounded tail tip, and a C-shaped male with an arcuate to angular spicule and bursa arising at more than 90% of body length. Fergusobia robustae Davies n. sp. is characterized by the combination of a small, C-shaped parthenogenetic female with a short cylindroid tail with a broadly rounded tip; an arcuate to C-shaped, relatively broad, infective female with a tail tip that is almost hemispherical; and an arcuate male with a strong angular spicule and a bursa arising at 30-40% of body length. Fergusobia pruinosae Davies n. sp. is characterized by the combination of a medium sized, open C-shaped, parthenogenetic female in which the cuticle does not swell upon fixation, with a strongly sclerotised stylet, with a more or less narrowly conoid tail with a bluntly or broadly rounded tip; infective female that is arcuate to J-shaped with a notched tail tip; and open C-shaped males with stout, angular spicules and bursa arising near the secretory/excretory pore. Fergusobia pauciflorae Davies n. sp. is characterized by the combination of a medium sized, arcuate, parthenogenetic female with a small stylet, and a conoid tail with a bluntly rounded tip; a small, open C to J-shaped infective female with a broadly rounded to almost hemispherical tail tip; and straight to arcuate males with stout, angular spicules and bursa at ~70% of body length anterior to the cloaca. Fergusobia obliquae Davies n. sp. has a relatively broad, arcuate, parthenogenetic female having a small but strongly sclerotised stylet, and a short conoid tail with a bluntly rounded tip; an arcuate, infective female with an almost hemispherical tail tip; and C-shaped males with arcuate to angular (not heavily sclerotised) spicules and bursa arising 50-80% of body length anterior to tail. Sequencing of the 18S and 28S rDNA domains for F. janetae n. sp. and of mitochondrial DNA cytochrome oxidase subunit I (mtCOI) for F. pauciflorae n. sp. confirm that they are distinct species.

Morphological and molecular characteristics of Parasitodiplogaster religiosae n. sp. (Nematoda: Diplogastrina) associated with Ficus religiosa in China.

A new nematode species of the genus Parasitodiplogaster was recovered from syconia of Ficus religiosa at the Guangxiao Temple, Guangzhou, China. It is described herein as P. religiosae n. sp. and is characterised by possessing the longest and thinnest spicule of all currently described males in the genus, an elongated laterally "ε-shaped" and ventrally rhomboid-like gubernaculum, a stoma without teeth, consisting of a ring-like cheilostom with indistinct anteriolateral projections, a tube-like gymnostom and a funnel-like stegostom, monodelphic with a mean vulval position of 66%. There are three pre-cloacal and six post-cloacal male genital papillae with the arrangement P1, P2, P3, (C, P4), P5, P6d, P7, P8, P9d, Ph. This new species was easily differentiated from other members of the genus by DNA sequences of partial small subunit rRNA gene (SSU) and the D2-D3 expansion segments of the large subunit rRNA gene (LSU). Phylogenetic analysis also corroborated its reasonable placement within a well-supported monophyletic clade with other Parasitodiplogaster species and within the australis-group that includes P. australis and P. salicifoliae that are all associates of fig wasp pollinators (Platyscapa sp.) of figs of the subsection Urostigma.