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1.  Phylogenetic Status and Morphological Characters of Rhabditolaimus anoplophorae (Rhabditida: Diplogastridae) 
Journal of Nematology  2014;46(1):44-49.
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.
PMCID: PMC3957571  PMID: 24644370
molecular profiles; morphology; re-isolation; Rhabditolaimus anoplophorae; taxonomy
2.  Molecular Characterization and Development of Real-Time PCR Assay for Pine-Wood Nematode Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae) 
PLoS ONE  2013;8(11):e78804.
Bursaphelenchus xylophilus, the pine-wood nematode (PWN), is the causal agent of pine wilt disease, one of the most damaging emerging pest problems to forests around the world. It is native to North America where it causes relatively minor damage to native conifers but is labeled an EPPO-A-2 pest and a quarantine nematode for many countries outside of the United States because of its potential for destruction to their native conifers. Exports of wood logs and commodities involving softwood packaging materials now require a lab test for the presence/absence of this regulated nematode species. We characterized the DNA sequences on the ribosomal DNA small subunit, large subunit D2/D3, internal transcribed spacer (ITS) and mitochondrial DNA cytochrome oxidase subunit one on the aphelenchid species and described the development of a real-time-PCR method for rapid and accurate identification of PWN targeting the ITS-1. A total of 97 nematode populations were used to evaluate the specificity and sensitivity of this assay, including 45 populations of B. xylophilus and 36 populations of 21 other species of Bursaphelenchus which belong to the abietinus, cocophilus, eggersi, fungivorus, hofmanni, kevini, leoni, sexdentati, and xylophilus groups and one unassigned group from a total of 13 groups in the genus Bursaphelenchus; 15 populations of Aphelenchoides besseyi, A. fragariae, Aphelenchoides species and Aphelenchus avenae; and one population of mixed nematode species from a soil sample. This assay proved to be specific to B. xylophilus only and was sensitive to a single nematode specimen regardless of the life stages present. This approach provides rapid species identification necessary to comply with the zero-tolerance export regulations.
doi:10.1371/journal.pone.0078804
PMCID: PMC3823978  PMID: 24244367
3.  Nematode Spatial and Ecological Patterns from Tropical and Temperate Rainforests 
PLoS ONE  2012;7(9):e44641.
Large scale diversity patterns are well established for terrestrial macrobiota (e.g. plants and vertebrates), but not for microscopic organisms (e.g. nematodes). Due to small size, high abundance, and extensive dispersal, microbiota are assumed to exhibit cosmopolitan distributions with no biogeographical patterns. This assumption has been extrapolated from local spatial scale studies of a few taxonomic groups utilizing morphological approaches. Recent molecularly-based studies, however, suggest something quite opposite. Nematodes are the most abundant metazoans on earth, but their diversity patterns are largely unknown. We conducted a survey of nematode diversity within three vertical strata (soil, litter, and canopy) of rainforests at two contrasting latitudes in the North American meridian (temperate: the Olympic National Forest, WA, U.S.A and tropical: La Selva Biological Station, Costa Rica) using standardized sampling designs and sample processing protocols. To describe nematode diversity, we applied an ecometagenetic approach using 454 pyrosequencing. We observed that: 1) nematode communities were unique without even a single common species between the two rainforests, 2) nematode communities were unique among habitats in both rainforests, 3) total species richness was 300% more in the tropical than in the temperate rainforest, 4) 80% of the species in the temperate rainforest resided in the soil, whereas only 20% in the tropics, 5) more than 90% of identified species were novel. Overall, our data provided no support for cosmopolitanism at both local (habitats) and large (rainforests) spatial scales. In addition, our data indicated that biogeographical patterns typical of macrobiota also exist for microbiota.
doi:10.1371/journal.pone.0044641
PMCID: PMC3439412  PMID: 22984536
4.  Reverse Taxonomy for Elucidating Diversity of Insect-Associated Nematodes: A Case Study with Termites 
PLoS ONE  2012;7(8):e43865.
Background
The molecular operational taxonomic unit (MOTU) has recently been applied to microbial and microscopic animal biodiversity surveys. However, in many cases, some of the MOTUs cannot be definitively tied to any of the taxonomic groups in current databases. To surmount these limitations, the concept of “reverse taxonomy” has been proposed, i.e. to primarily list the MOTUs with morphological information, and then identify and/or describe them at genus/species level using subsamples or by re-isolating the target organisms. Nevertheless, the application of “reverse taxonomy” has not been sufficiently evaluated. Therefore, the practical applicability of “reverse taxonomy” is tested using termite-associated nematodes as a model system for phoretic/parasitic organisms which have high habitat specificity and a potential handle (their termite host species) for re-isolation attempts.
Methodology
Forty-eight species (from 298 colonies) of termites collected from the American tropics and subtropics were examined for their nematode associates using the reverse taxonomy method and culturing attempts (morphological identification and further sequencing efforts). The survey yielded 51 sequence types ( =  MOTUs) belonging to 19 tentatively identified genera. Within these, four were identified based on molecular data with preliminary morphological observation, and an additional seven were identified or characterized from successful culturing, leaving eight genera unidentified.
Conclusions
That 1/3 of the genera were not successfully identified suggests deficiencies in the depth of available sequences in the database and biological characters, i.e. usually isolated as phoretic/parasitic stages which are not available for morphological identification, and too many undiscovered lineages of nematodes. Although there still is the issue of culturability of nematodes, culturing attempts could help to make reverse taxonomy methods more effective. However, expansion of the database, i.e., production of more DNA barcodes tied to biological information by finding and characterizing additional new and known lineages, is necessary for analyzing functional diversity.
doi:10.1371/journal.pone.0043865
PMCID: PMC3429485  PMID: 22952792
5.  The Nature and Frequency of Chimeras in Eukaryotic Metagenetic Samples. 
Journal of Nematology  2012;44(1):18-25.
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.
PMCID: PMC3593260  PMID: 23482827
chimera; biodiversity; metagenetics; multi-template PCR; nematode; ultrasequencing
6.  Field Responses of Bermudagrass and Seashore paspalum Cultivars to Sting and Spiral Nematodes 
Journal of Nematology  2011;43(3-4):201-208.
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.
PMCID: PMC3547344  PMID: 23430148
Belonolaimus longicaudatus; bermudagrass; Cynodon spp.; field; Helicotylenchus spp.; Paspalum vaginatum; resistance; seashore paspalum; spiral nematodes; sting nematodes
7.  Evaluation of Econem™, a Formulated Pasteuria sp. Bionematicide, for Management of Belonolaimus longicaudatus on Golf Course Turf1 
Journal of Nematology  2011;43(2):101-109.
In 2010, a turfgrass bionematicide containing in vitro produced Pasteuria sp. for management of Belonolaimus longicaudatus was launched under the tradename Econem™. Greenhouse pot studies and field trials on golf course fairways and tee boxes evaluated Econem at varied rates and application frequencies. Trials on putting greens compared efficacy of three applications of Econem at 98 kg/ha to untreated controls and 1,3-dichloropropene at 53 kg a.i/ha. Further putting green trials evaluated the ability of three applications of Econem at 98 kg/ha to prevent resurgence of population densities of B. longicaudatus following treatment with 1,3-dichloropropene at 53 kg a.i./ha. None of the Econem treatments in pot studies were effective at reducing B. longicaudatus numbers (P ≤ 0.05). Econem was associated with reduction in population densities of B. longicaudatus (P ≤ 0.1) on only a single sampling date in one of the eight field trials and did not improve turf health in any of the trials (P > 0.1). These results did not indicate that Econem is an effective treatment for management of B. longicaudatus on golf course turf.
PMCID: PMC3380457  PMID: 22791919
Belonolaimus longicaudatus; bermudagrass; biological control; biopesticide; Cynodon spp.; nematode management; Pasteuria sp.; sting nematode; turfgrass
8.  Effects of Formulation and Host Nematode Density on the Ability of In Vitro-Produced Pasteuria Endospores to Control its Host Belonolaimus longicaudatus 
Journal of Nematology  2010;42(2):87-90.
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 cm3 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.
PMCID: PMC3380477  PMID: 22736843
Belonolaimus longicaudatus; biological control; formulation; management; Pasteuria spp.; sting nematode; suppression; turfgrass
9.  Stomatal Ultrastructure, Molecular Phylogeny, and Description of Parasitodiplogaster laevigata n. sp. (Nematoda: Diplogastridae), a Parasite of Fig Wasps 
Journal of nematology  2006;38(1):137-149.
Parasitodiplogaster comprises a potentially large radiation of nematode species that appear to be parasitically bound to their Agaonid fig wasp hosts, which are mutualistically associated in the syconia (figs) of the diverse plant genus Ficus. Parasitodiplogaster laevigata n. sp. is described and illustrated as an associate of the fig wasp, Pegoscapus sp. from Ficus laevigata from southern Florida. It is the first species of Parasitodiplogaster reported from North America and is closest to P. trigonema from F. trigonata from Panama. Parasitodiplogaster laevigata n. sp. can be differentiated from all described species of Parasitodiplogaster based on stomatal morphology (presence of a large dorsal and a right subventral tooth) in the adults of both sexes, molecular comparisons of two expansion segments (D2,D3) of the large subunit (LSU) rRNAgene, and fig-fig wasp host affinities. The ultrastructure of P. laevigata n. sp. was elucidated using TEM and SEM for comparisons with other species of Parasitodiplogaster. The stoma of P. laevigata n. sp. possesses a nonsegmented cheilostomal ring that connects to the longitudinal body musculature per- and interradially, a claw-like dorsal tooth, a right subventral tooth, and telostegostomatal apodemes arising from the dorsal side of each subventral sector. The unification of the pro-, meso-, and metastegostom with the gymnostom in P. laevigata n. sp. and further simplification in other described species may be due to derived adaptations associated with the internal parasitism of fig wasps.
PMCID: PMC2586439  PMID: 19259439
Agaonidae; buccal capsule; Ficus, fig wasp; Hymenoptera; LSU rRNA; molecular phylogeny; morphology; nematode; parasitism; Diplogastridae; Parasitodiplogaster laevigata n. sp.; Pegoscapus sp.; stoma; taxonomy
10.  Morphology and Description of Bursaphelenchus platzeri n. sp. (Nematoda: Parasitaphelenchidae), an Associate of Nitidulid Beetles 
Journal of nematology  2006;38(1):150-157.
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.
PMCID: PMC2586433  PMID: 19259440
Bursaphelenchus platzeri n. sp.; Carpophilus humeralis, Coleoptera; morphology; mycophagy; nematode; Nitidulidae; Parasitaphelenchidae; phylogeny; scanning electron microscopy; systematics; taxonomy
11.  Bursaphelenchus anatolius n. sp. (Nematoda: Parasitaphelenchidae), an Associate of Bees in the Genus Halictus 
Journal of Nematology  2005;37(3):336-342.
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.
PMCID: PMC2620966  PMID: 19262882
Bursaphelenchus anatolius n. sp.; cytochrome oxidase subunit I; Halictidae; Halictus (Argalictus); Hymenoptera; large subunit rRNA; molecular phylogeny; morphology; mycophagy; nematode; Parasitaphelenchidae; phoresy; small subunit rrna; taxonomy
12.  Schistonchus aureus n. sp. and S. laevigatus n. sp. (Aphelenchoididae): Associates of Native Floridian Ficus spp. and Their Pegoscapus Pollinators (Agaonidae) 
Journal of Nematology  2001;33(2-3):91-103.
Two new species of Schistonchus were recovered from the hemocoel of adult female fig wasps, Pegoscapus spp. (Agaonidae), and the syconia of Ficus spp. native to Florida. They are described here as Schistonchus aureus n. sp., associated with Pegoscapus mexicanus and Ficus aurea, and Schistonchus laevigatus n. sp., associated with Pegoscapus sp. and Ficus laevigata. The Florida species of Schistonchus are differentiated from each other by host plant and fig wasp associates, number of incisures in the lateral field, male spicule shape, and mucronate male tail of S. aureus n. sp. Schistonchus aureus n. sp. and S. laevigatus n. sp. are differentiated from other members of the genus by the absence of a lip sector disc as observed with scanning electron microscopy for comparison with S. caprifici and S. macrophylla. Excretory pore position for both Florida species, S. altermacrophylla, and S. africanus is less than 50% (one half) the distance between the top of the head and the top of the metacorpus, as opposed to other species (S. caprifici, S. hispida, S. macrophylla, and S. racemosa), where it is located near or below the level of the metacorpus. Males of both species from Florida can be distinguished from S. altermacrophylla and S. africanus by the more posteriad positioning of the three pairs of caudal papillae; the anteriormost pair of papillae in the Florida species are adcloacal vs. precloacal in the other two species.
PMCID: PMC2638131  PMID: 19266003
Agaonidae; Aphelenchoididae; Ficus; fig; nematode; new species; parasitism; Pegoscapus; phoresy; scanning electron microscopy; Schistonchus aureus n. sp.; Schistonchus laevigatus n. sp.; taxonomy
13.  Histological Comparisons of Parasitism by Schistonchus spp. (Nemata: Aphelenchoididae) in Neotropical Ficus spp. 
Journal of Nematology  1999;31(4):393-406.
Syconia (enclosed infructescences) infested with host-specific species of Schistonchus (Aphelenchoididae) were collected from six species of Ficus (Moraceae) native to Florida or Panama. They were sectioned and histologically examined to assess the effects of parasitism. Parasitism by Schistonchus spp. was associated with hypertrophied cells, tissue necrosis, and the presence of an exudate in all species. Occasional hypertrophy of the outer epidermal cells occurred on seed florets, wasp florets, and on the endothecial cells of male florets in F. aurea (subgenus Urostigma) from Florida. Aberrations of the inner mesocarp occurred under the hypertrophied cells on seed florets. In F. laevigata (subgenus Urostigma) from Florida, Schistonchus sp. infested immature male florets and was associated with hypertrophy of endothecial cells, epidermal cells of the anther filaments, and anthers. Schistonchus sp. also caused aberrations of the anther filament, anthers, and pollen. Ficus poponoei (subgenus Urostigma) and F. glabrata (subgenus Pharmacosycea), both from Panama, had hypertrophied outer epidermal cells on seed florets. Ficus poponoei also had Schistonchus sp. within the pedicel of an aborted floret, with hypertrophy of the cortical parenchyma. Ficus trigonata (subgenus Urostigma) from Panama had hypertrophy of the outer epidermis of seed florets. When the outer epidermis on these florets was missing, the inner mesocarp was hypertrophied. Ficus maxima (subgenus Pharmacosycea) from Panama had hypertrophy on the outer epidermis of seed and aborted florets. Schistonchus spp. were not found in wasp larvae or pupae in any of the Ficus spp. examined. Hypertrophy was never observed in the absence of Schistonchus spp.
PMCID: PMC2620391  PMID: 19270912
Aphelenchoididae; Ficus aurea; Ficus glabrata; Ficus laevigata; Ficus maxima; Ficus poponoei; Ficus trigonata; fig; floret wall morphology; histopathology; life history; nematode; parasitism; Schistonchus spp.
14.  Nematodes Associated with Fig Wasps, Pegoscapus spp. (Agaonidae), and Syconia of Native Floridian Figs (Ficus spp.) 
Journal of Nematology  1995;27(1):1-14.
Syconia in successive developmental phases from Ficus laevigata Vahl (F. citrifolia Miller sensu DeWolf 1960) (Moraceae) and successive life stages of its fig wasp pollinator, Pegoscapus sp. (P. assuetus (Grandi) sensu Wiebes 1983) (Agaonidae) were dissected to elucidate their association with two undescribed species of nematodes. Parasitodiplogazter sp. (Diplogasteridae) are transported by female Pegoscapus sp. into the cavity of a phase B syconium as third-stage juveniles (J3), where they molt to the J4 stage and greatly increase in size in the hemocoel of the fig wasp after it begins to pollinate and oviposit in female florets. The J4 exit the wasp cadaver in a phase B or early phase C syconium, and molt to adults that mate and lay eggs. New J3 infect the next generation of female or male wasps as they emerge from their galls in phase D figs. Mated entomogenous females of Schistonchus sp. (Aphelenchoididae) are transported in the hemocoel of female wasps to the fig cavity of a phase B syconium. Female Schistonchus sp. exit the wasp and parasitize immature male florets causing an exudate, the development of hypertrophied epidermal cells of the anther filaments and anthers, and aberrations of the anther filament, anthers, and pollen. At least one generation of Schistonchus sp. occurs in the male florets. Entomogenous females appear at about the time that fig wasps molt to adults in their galls in late phase C syconia. Another Schistonchus sp. was recovered from females of P. mexicanus (Ashmead) (P. jimenezi (Grandi) sensu Wiebes 1983) and from the syconia of F. aurea Nuttall and appears to have a life cycle similar to that described for the Schistonchus sp. from F. laevigata.
PMCID: PMC2619580  PMID: 19277255
Agaonidae; Aphelenchoididae; Diplogasteridae; Ficus aurea; Ficus laevigata; fig; life history; nematode; parasitism; Parasitodiplogaster sp.; Pegoscapus spp.; Schistonchus spp.
15.  Host Status of Different Bermudagrasses (Cynodon spp.) for the Sting Nematode, Belonolaimus longicaudatus 
Journal of Nematology  1992;24(4S):749-756.
Thirty-seven warm-season bermudagrass (Cynodon spp.) accessions, two cool-season grasses (Lolium perenne and Festuca arundinacea), 'Transvala' digitgrass (Digitaria decumbens), and Sorghum bicolor were evaluated to determine host suitability and susceptibility to the sting nematode, B. longicaudatus, in a 140-day microcell bioassay. All seven of the evaluated commercial cultivars of Cynodon were suitable hosts for B. longicaudatus but varied in their tolerance to the nematode. 'Midiron,' 'Tifdwarf,' 'Tifgreen,' 'Tifgreen II,' 'Tifway II,' and 'Tufcote' were sensitive, with reductions in root weight of >24%, whereas 'Tifway' appeared to be relatively tolerant with only a 4% reduction in root dry weight. Twenty other Cynodon accessions showed decreases (P ≤ 0.05) in root dry weight relative to uninoculated plants of the same germplasm and (or) > 11% root reductions. In addition to 'Tifway,' 10 other Cynodon accessions and L. perenne, F. arundinacea, D. decumbens, and S. bicolor appeared to be relatively more tolerant of B. longicaudatus than the other accessions evaluated.
PMCID: PMC2629856  PMID: 19283057
Belonolaimus longicaudatus; bermudagrass; Cynodon spp.; digitgrass; grass; host-plant resistance; nematode; perennial ryegrass; sorghum; sting nematode; tall rescue
16.  Host suitability of Ixora spp. for the Root-knot Nematodes Meloidogyne incognita Race 1 and M. javanica 
Journal of Nematology  1992;24(4S):722-728.
Eight commonly cultivated Ixora species or cultivars were tested for their suitability as hosts and their level of tolerance to Meloidogyne incognita race 1 and M. javanica in a greenhouse study. Twenty weeks postinoculation with 5,000 eggs per pot, M. incognita race 1 and M. javanica produced galls and formed egg masses on roots of all eight Ixora species or cultivars tested. However, only M. javanica-infected 'Petite Yellow' and 'Maui' had decreases (P ≤ 0.05) in root wet weights, suggesting that the other cultivars were more tolerant to these root-knot nematode species. Differential host suitability to each Meloidogyne species was based on the relative number of galls, galls per gram root weight, egg masses, and second-stage juveniles produced per plant. 'Bonnie Lynn,' 'Maui,' and 'Petite Red' were good to excellent hosts for both Meloidogyne spp. Ixora coccinea was a good host for M. incognita race 1 but less suitable for M. javanica. 'Singapore' and 'Petite Yellow' were poor hosts for M. incognita race 1 but excellent hosts for M. javanica. 'Nora Grant' and I. casei 'Super King' were poor hosts for both species of root-knot nematodes.
PMCID: PMC2629859  PMID: 19283052
host-parasite relationship; Ixora spp.; Meloidogyne incognita race 1; M. javanica; nematode; ornamental; root-knot nematode; woody ornamental
17.  Dynamics of Belonolaimus longicaudatus Parasitism on a Susceptible St. Augustinegrass Host 
Journal of Nematology  1992;24(3):432-437.
St. Augustinegrass (Stenotaphrum secundatum) cv FX-313 was used as a model laboratory host for monitoring population growth of the sting nematode, Belonolaimus longicaudatus, and for quantifying the effects of sting nematode parasitism on host performance in two samples of autoclaved native Margate fine sand with contrasting amounts of organic matter (OM = 7.9% and 3.8%). Following inoculation with 50 Belonolaimus longicaudatus per pot, nematodes peaked at a mean of 2,139 nematodes per pot 84 days after inoculation, remained stable through 168 days at 2,064 nematodes per pot, and declined at 210 days. The relative numbers of juveniles and adults demonstrated senescence after 84 days. Root dry weight of nematode-inoculated plants increased briefly to an apparent equilibrium 84 days after inoculation, whereas root weights of uninoculated controls continued to increase, exceeding those of inoculated plants from 84 to 210 days (P < 0.01). At 210 days, uninoculated plants had 227% the root dry weight of inoculated plants. Transpiration of FX-313 was reduced by nematodes (P < 0.0001) at 84 and 126 days after inoculation; reduction was first observed at 42 days and last observed 168 days after inoculation (P < 0.05). OM content affected all plant performance variables at multiple dates, and generally there were no inoculation x OM content interactions. OM content had no effect on nematode numbers per pot, although there was a slight (P < 0.05) increase in the number of nematodes per gram root dry weight in the low-OM soil compared with the high-OM soil.
PMCID: PMC2619280  PMID: 19283019
Belonolairnus longicaudatus; nematode; population dynamics; resistance screening; soil organic matter; St. Augustinegrass; Stenotaphrum secundatum; sting nematode; turfgrass
18.  Susceptibility of Diploid St. Augustinegrasses to Belonolaimus longicaudatus 
Journal of Nematology  1991;23(4S):604-610.
A fine-textured, dwarf St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze) genotype, FX-313, was severely damaged in plots in the third year of evaluation in sandy soil in southern Florida. Damage was associated with numerous (> 40/100-cm³ soil) sting nematodes, Belonolaimus longicaudatus Rau. Damage was ameliorated (P < 0.05) by fenamiphos applied broadcast at 2.2 g a.i./m², and B. longicaudatus numbers were reduced (P < 0.01), compared with untreated plots. Root dry weights of four diploid (2n = 18) St. Augustinegrasses--FX-261, FX-299, FX-313, and Seville--were reduced (P < 0.001) by B. longicaudatus in a temperature- and light-controlled experiment. Estimated daily transpiration, an indicator of plant health, was reduced (P < 0.001) after 112 days to 3.32 g/pot for inoculated plants, compared with 5.10 g /pot for uninoculated plants. Genotypes did not differ in nematode number per pot (mean 551/215 cm² soil) 128 days after inoculation, but differed (P < 0.05) in nematode numbers on a root dry weight basis, with FX-313 and Seville representing the extremes, 12,300 and 4,000 B. longicaudatus/g root dry weight, respectively. The diploid St. Augustinegrasses evaluated were good hosts for B. longicaudatus, but field data and controlled inoculation demonstrate genetic variation in susceptibility.
PMCID: PMC2619225  PMID: 19283170
Belonolaimus longicaudatus; breeding; fenamiphos; nematode; resistance; St. Augustinegrass; Stenotaphrum secundatum; sting nematode; turfgrass
19.  Isolates of the Pasteuria penetrans Group from Phytoparasitic Nematodes in Bermudagrass Turf 
Journal of Nematology  1990;22(4S):750-762.
A survey was conducted between 1985 and 1989 of isolates of the Pasteuria penetrans group on phytoparasitic nematodes in bermudagrass (Cynodon spp.) turf in southern Florida. Six different isolates of the P. penetrans group were observed from five different species of phytoparasitic nematode hosts. Five of the bacterial isolates were different (P ≤ 0.01) in sporangium diameter, endospore width, and ratio of sporangium diameter to endospore width. All locations surveyed had one or more isolates present, suggesting that the Pasteuria penetrans group is widespread in its distribution in southern Florida. Three survey sites had high densities of Belonolaimus longicaudatus, with more than 60% of the host population encumbered with a large-spored isolate of Pasteuria (mean sporangium diameter = 6.10 μm). One of these sites was monitored for 16 months during which the proportion of nematodes encumbered with this Pasteuria isolate remained constant. Soil infested with this isolate was not suppressive to Pasteuria-free populations of B. longicaudatus grown on bermudagrass for 6 months after controlled soil inoculation. However, the proportion of spore-encumbered and parasitized B. longicaudatus after 6 months was 73%, which was similar to the 74% level observed at the field site. The uhrastructure of mature sporangia of the large-spored isolates of Pasteuria from B. longicaudatus and Hoplolaimus galeatus is described and compared with ultrastructural descriptions of P. penetrans sensu strictu and P. thornei from the literature. These B. longicaudatus and H. galeatus isolates of Pasteuria appear to be distinct from the known species and may warrant new species status.
PMCID: PMC2619098  PMID: 19287792
bacterial parasite; Belonolaimus longicaudatus; bermudagrass; biological control; Helicotylenchus microlobus; Hoplolaimus galeatus; Meloidogyne spp.; Pasteuria penetrans group; Tylenchorhynchus annulatus; ultrastructure
20.  Teratorhabditis palmarum n. sp. (Nemata: Rhabditidae): An Associate of Rhynchophorus palmarum and R. cruentatus 
Journal of Nematology  1990;22(3):337-347.
Teratorhabditis palmarum n. sp., an associate of the palm weevils Rhynchophorus palmarum and R. cruentatus is described. Teratorhabditis palmarum was isolated from newly emerged adults and cocoons of R. palmarum from red-ring diseased coconut palms, Cocos nucifera, in Trinidad and Ecuador, and from red-ring diseased oil palms, Elaeis guineensis, in Colombia. Teratorhabditis palmarum was also associated internally with newly emerged adults of R. cruentatus from mature transplanted cabbage palmettos, Sabal palmetto, in Florida. Dauer juveniles of T. palmarum infested the genital capsule and body cavity of newly emerged adult female and male palm weevils. Adult nematodes isolated from cocoons and dauer juveniles from newly emerged palm weevils were cuhurable on bacterial lawns on several solid media. Females of T. palmarum have a single anteriorly directed ovary; vulva at 93-96% of the body length; short, hemispherical spicate tail; three or four teeth in the metastom; cuticle with distinct transverse punctations that change abruptly at the level of the procorpus to indentations of alternating size and arrangement; and eggs with cuticular sculpturing. Males have a crenate, peloderan bursa with a 2 + 5 + 3 pattern of bursal rays (7 extend to the margin of bursa); spicules linear, completely fused at the distal tip and dorsally for 50% of the total spicule length.
PMCID: PMC2619040  PMID: 19287730
coconut palm; Cocos nucifera; Colombia; Ecuador; Elaeis guineensis; entomophilic nematode; Florida; new species; oil palm; red ring disease; Rhadinaphelenchus cocophilus; Sabal palmetto; Teratorhabditis palmarum n. sp.; Tobago; Trinidad
21.  Association of the Red Ring Nematode and Other Nematode Species with the Palm Weevil, Rhynchophorus palmarum 
Journal of Nematology  1990;22(2):143-149.
The palm weevil, Rhynchophorus palmarum (L.), was collected in cocoons from red ring-diseased coconut palms (Cocos nucifera L.) in Trinidad and Tobago. Juveniles of five species of nematodes were extracted from the genitalia and macerated bodies of newly emerged adults of the palm weevil: Rhadinaphelenchus cocophilus (Cobb) Goodey (the red ring nematode), Teratorhabditis sp., Diplogasteritus sp., Mononchoides sp., and Bursaphelenchus sp. Over 90% of newly emerged weevil females and males were infested internally with red ring nematode juveniles, and over 47% of the weevils contained more than 1,000 red ring nematodes each. There was no significant correlation between weevil body length and the number of red ring nematodes carried internally by each weevil. Teratorhabditis sp. and Diplogasteritus sp. were extracted from over 50% of the palm weevils, and Monochoides sp. and Bursaphelenchus sp. were found in a small proportion of the weevils. Field-collected adult weevils were also internally and externally infested with a Rhabditis sp., which was not observed in or on weevils allowed to emerge from field-collected cocoons.
PMCID: PMC2619033  PMID: 19287703
Bursaphelenchus sp.; Cocos nucifera; Diplogasteritus sp.; entomophilic nematode; Mononchoides sp.; palm weevil; red ring nematode; Rhabditis sp.; Rhadinaphelenchus cocophilus; Rhynchophorus palmarum; Teratorhabditis sp.; Trinidad
22.  Commensal Nematodes in the Glands, Genitalia, and Brood Cells of Bees (Apoidea) 
Journal of Nematology  1990;22(2):150-161.
Seven species of bees from the eastern United States, representing four families in the Apoidea, were dissected and examined for nematode associates. Dufour's glands in females of Halictus ligatus, Augochlora pura mosieri, and Augochlorella gratiosa (Halictidae) from Florida were infested with dauer juveniles of Aduncospiculum halicti (Diplogasteridae). The Dufour's glands of Colletes thoracicus (Colletidae) females from Maryland were infested with dauer juveniles of a new species of Koerneria sp. (Diplogasteridae), and abdominal glands of females of Andrena alleghaniensis (Andrenidae) from New York were infested with dauer juveniles of another new species of Koerneria. The lateral and median oviducts, Dufour's glands, and poison sacs in females of Anthophora abrupta (Anthophoridae) from Maryland and Alabama were infested with dauer juveniles of a new species of Bursaphelenchus sp. (Aphelenchoididae). Cross sections of the nematode-infested poison sacs of A. abrupta revealed two types of humoral host defense reactions.
PMCID: PMC2619032  PMID: 19287704
Andrenidae; Aduncospiculum halicti; Anthophoridae; bees; Bursaphelenchus sp.; Colletidae; commensal; Diplogasteridae; Halictidae; Koerneria spp.
23.  Evaluation of Three Nematicides for the Control of Phytoparasitic Nematodes in 'Tifgreen II' Bermudagrass 
Journal of Nematology  1988;20(Annals 2):46-49.
Three nematicides were evaluated for control of Belonolaimus longicaudatus, Hoplolaimus galeatus, Criconemella spp., and Meloidogyne spp. in 'Tifgreen II' bermudagrass mowed at golf course fairway height (1.3 cm) in Fort Lauderdale, Florida. Bermudagrass plots were treated with fenamiphos (13.5 kg a.i./ha), oxamyl (13.5 kg a.i./ha), or 30% formaldehyde (6.4 liter a.i./ha). The plots treated with fenamiphos or formaldehyde were split 14 days later and one-half of each plot received two biweekly applications of formaldehyde. Forty-two days after the treatments were applied, the turfgrass vigor ratings and dry root weights in plots treated with fenamiphos were higher (P < 0.05) than the control, oxamyl, or formaldehyde treatments. The population levels of B. longicaudatus were suppressed (P < 0.05) in the fenamiphos, fenamiphos plus formaldehyde, and oxamyl treatments.
PMCID: PMC2618884  PMID: 19290302
Belonolaimus longicaudatus; bermudagrass; chemical control; Criconemella spp.; fenamiphos; formaldehyde; lance nematode; Hoplolaimus galeatus; Meloidogyne spp.; oxamyl; ring nematode; root-knot nematode; sting nematode; turfgrass
24.  Solarization for Nematode Disinfestation of Small Volumes of Soil 
Journal of Nematology  1988;20(Annals 2):41-45.
Several polyethylene plastics were evaluated as potential materials for disinfesting small volumes of soil containing nematodes. Bursaphelenchus seani, cultured on the fungus Monilinia fructicola in petri dishes, was used to bioassay the survival and reproductive capability of nematodes buried 7.5 cm deep in Margate fine sand (soil moisture = 4.9%). The soil was exposed to sunlight for 6 days in May 1987. The highest mean temperatures recorded at 7.5 cm deep were 38 ± 1 C, 43 ± 1 C, 43 ± 1 C, and 50 ± 1 C for the no plastic, clear plastic, black plastic, and clear + black plastic treatments, respectively. The temperature in the clear + black plastic treatment exceeded 47 C for more than 2 hours on clear days. Nematode survival averaged 98 ± 3%, 78 ± 22%, 38 ± 38%, and 0 ± 0%, whereas the reproductive success of B. seani following treatment was 100, 100, 75, and 0% for the no plastic, clear plastic, black plastic, and clear + black plastic treatments, respectively. Bursaphelenchus seani in petri dishes and Belonolaimus longicaudatus and Hoplolaimus galeatus in soil died when exposed to 48 ± 2 C for 2 hours.
PMCID: PMC2618878  PMID: 19290301
Belonolaimus longicaudatus; Bursaphelenchus seani; Hoplolaimus galeatus; lance nematode; physical control; potted plant; preplant control; solarization; sting nematode

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