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In recent years, some golf course superintendents in Florida have reported that the turf health is no longer as great, and nematode responses to fluopyram have decreased. The objective of this research was to determine if the mechanism of the reported reduced efficacy was attributable to either: i ) enhanced degradation accelerating its breakdown in the soil, or ii ) reduced sensitivity to the nematicide in the nematode populations. In a field experiment, soil and nematodes were collected from small plots that had been treated multiple times over four years, for only one year, or never treated. Soil and nematodes were additionally collected from commercial turf sites where either multiple applications of fluopyram had been made for numerous years, or it had never been used. Bioassay experiments found no evidence of enhanced degradation. However, M. graminis collected from small field plots and commercial sites with long-term use of fluopyram were less sensitive to fluopyram in-vitro than those from small plots and commercial sites where fluopyram had not been used. These results indicate that nematicide resistance is a likely cause of reduced fluopyram efficacy on golf-course turf in Florida., (© 2023 Christian L. Kammerer et al., published by Sciendo.)

Specimens of a tylenchid nematode were recovered in 2019 from soil samples collected from a corn field, located in Pickens County, South Carolina, USA. A moderate number of Tylenchus sp. adults (females and males) were recovered. Extracted nematodes were examined morphologically and molecularly for species identification, which indicated that the specimens of the tylenchid adults were a new species, described herein as Tylenchus zeae n. sp. Morphological examination and the morphometric details of the specimens were very close to the original descriptions of Tylenchus sherianus and T. rex . However, females of the new species can be differentiated from these species by body shape and length, shape of excretory duct, distance between anterior end and esophageal intestinal valve, and a few other characteristics given in the diagnosis. Males of the new species can be differentiated from the two closely related species by tail, spicules, and gubernaculum length. Cryo-scanning electron microscopy confirmed head bearing five or six annules; four to six cephalic sensilla represented by small pits at the rounded corners of the labial plate; a small, round oral plate; and a large, pit-like amphidial opening confined to the labial plate and extending three to four annules beyond it. Phylogenetic analysis of 18S rRNA gene sequences placed Tylenchus zeae n. sp. in a clade with Tylenchus arcuatus and several Filenchus spp., and the mitochondrial cytochrome oxidase c subunit 1 ( COI ) gene region separated the new species from T. arcuatus and other tylenchid species. In the 28S tree, T. zeae n. sp. showed a high level of sequence divergence and was positioned outside of the main Tylenchus-Filenchus clade., (© 2023 Kantor et al., published by Sciendo.)

Nematodes of the genus Auanema are interesting models for studying sex determination mechanisms because their populations consist of three sexual morphs (males, females, and hermaphrodites) and produce skewed sex ratios. Here, we introduce a new undescribed species of this genus, Auanema melissensis n. sp., together with its draft nuclear genome. This species is also trioecious and does not cross with the other described species A. rhodensis or A. freiburgensis . Similar to A. freiburgensis , A. melissensis ' maternal environment influences the hermaphrodite versus female sex determination of the offspring. The genome of A. melissensis is ~60 Mb, containing 11,040 protein-coding genes and 8.07% of repeat sequences. Using the estimated ancestral chromosomal gene content (Nigon elements), it was possible to identify putative X chromosome scaffolds., (© 2022 Tandonnet et al., published by Sciendo.)

Mitochondrial genomes have widely been used as molecular markers in understanding the patterns and processes of nematode evolution. The species in genus Cruznema are free-living bacterivores as well as parasites of crickets and mollusks. The complete mitochondrial genome of C. tripartitum was determined through high-throughput sequencing as the first sequenced representative of the genus Cruznema . The genome is comprised of 14,067 bp nucleotides, and includes 12 protein-coding, two rRNA, and 22 tRNA genes. Phylogenetic analyses based on amino acid data support C. tripartitum as a sister to the clade containing Caenorhabditis elegans and Oscheius chongmingensis . The analysis of gene arrangement suggested that C. tripartitum shares the same gene order with O. chongmingensis , Litoditis marina , Diplocapter coronatus , genus Caenorhabditis , and Pristionchus pacificus . Thus, the mitochondrial gene arrangement is highly conserved in the family Rhabditidae as well as some species in Diplogasteridae., (© 2022 Du et al., published by Sciendo.)

During a recent inventory survey of the nematofauna of tea plantation at Zhejiang Province, China, Discocriconemella parasinensis n. sp. was detected in the rhizosphere of Camellia sinensis . The new species can be characterized by having the uninterrupted rounded labial disc, en face view showing rectangular-rounded labial plate without submedian lobes, R = 82.6 (80-86), Rex = 22 (21-23), stylet length of 68.3 (59-76) μm, excretory pore located 1-2 annuli posterior to the esophageal bulb, vulva open, postvulval body elongated conoid, and tail conoid with bilobed terminus. Morphologically, the species shares the same lip-type with D. discolabia , D. mauritiensis , D. mineira , D. perseae , and D. sinensis . Phylogenetic relationships of the new species based on D2-D3 expansion segments of 28S, ITS, and 18S rRNA genes revealed that D. parasinensis n. sp. formed a separated clade from other criconematid species, thereby supporting its status as a new species of the genus. The new species showed close phylogenetic relationships with Criconemoides geraerti ., (© 2022 Junxia Li et al., published by Sciendo.)

Fumigant use in perennial crops can be reduced through prolonging the life of existing orchards. The longer an orchard remains healthy and productive, the less often it will be terminated, fumigated, and replanted. Two trials were conducted to determine the effectiveness of DiTera, a toxin produced by the fungus ( Myrothecium verrucaria) and Nema-Q, an extract of the soap bark tree ( Quillaja saponaria ) for management of root-lesion ( Pratylenchus vulnus ) and ring ( Mesocriconema xenopla) nematodes on walnuts. In the first trial, spring and fall treatments of DiTera were applied each year for four years to variety 'Chandler' scion on seedling 'Paradox' rootstock, and to own-rooted 'Chandler' trees. On 'Paradox' rootstock, both DiTera and Nema-Q increased walnut yields ( P  ≤ 0.05) and produced more vigorous trees ( P  ≤ 0.05) without significant reductions in nematode populations ( P  ≤ 0.05). A second trial was conducted with three rates of DiTera and four rates of Nema-Q, combinations of the two products, and an untreated control on 'Chandler' scion on 'Paradox' rootstock. The highest rate of Nema-Q ( P  ≤ 0.05), and a combination treatment of DiTera plus Nema-Q ( P  ≤ 0.05) increased trunk circumference. Several treatments showed reductions in root-lesion and ring nematodes ( P  ≤ 0.05). Bionematicides can improve yield, growth, and vigor in walnut orchards infested with plant-parasitic nematodes., (© 2021 Authors.)

Epidorylaimus procerus sp. n., collected from a natural habitat in Vietnam, is described and illustrated. It is distinguishable by its 2.16 to 2.46-mm-long body, lip region offset by depression and 15 to 17-µm broad, odontostyle 32 to 35-µm long, neck 415 to 461-µm long, pharyngeal expansion occupying 47 to 52% of the total neck length, uterus 76 to 130-µm long or 1.0 to 1.5-body diameters, vulva transverse ( V  = 40-43), caudal region conical elongate (157-186 µm, c  =12.1-14.4, c'  = 4.4-5.5) with blister-like bodies, and hyaline portion occupying one-fourth its length, and males absent. Molecular analysis shows a close relationship of the new species and E. lugdunensis , supporting monophyly of the genus Epidorylaimus ., (© 2020 Authors.)

Two populations of Paratylenchus lepidus and P. minor were detected in the rhizosphere of Elaeocarpus sp. and Chinese red pine from Taizhou and Hangzhou, Zhejiang Province, China. Previously, P. lepidus has been reported from China whereas P. minor was originally described from India decades ago in the rhizosphere of peach but was never reported thereafter. In this study, both species were characterized morphologically and molecularly coupled with SEM observations. Morphologically, both species have four incisures in the lateral field, vulval present (SEM observations), stylet less than 30 μm long and cephalic region without submedian lobes. Phylogenetically, both species grouped with paratylenchid species having short stylets. Both species can be differentiated from each other by the shape of lip region (rounded in P. lepidus and narrow truncated in P. minor ) and tail terminus (pointed in P. lepidus and a broadly rounded in P. minor ) and several morphomemtrical values.The study provided an updated description of P. lepidus and P. minor and a first record of P. minor from China. In addition, both species are the first paratylenchid species recorded from Elaeocarpus sp. and Pinus tabuliformis , respectively., (© 2019 Authors.)

The United States Department of Agriculture Nematode Collection (USDANC) is one of the largest and most valuable in existence and includes millions of specimens housed in over 39,800 permanent slides and 9,300 vials. This collection preserves type specimens of nematodes to serve as a reference for identifications and future taxonomic revisions. Also, the collection provides useful information on nematode hosts, occurrence, and distribution. The present list includes type specimens added to the USDANC since 1998. Since that time, the collection has expanded, with 474 type species mounted and preserved on 2,564 glass slides and 180 vials. We encourage nematologists throughout the world to deposit type specimens in the USDANC for use by future generations.

Three populations of neotylenchid nematodes were isolated in Ningbo, P. R. China, from white pine lumber (Pinus monticola) imported from the USA. The nematodes were morphologically intermediate between Hexatylus and Deladenus. The nematodes were molecularly characterized based on sequences of the rDNA small subunit 18S, large subunit 28S D2/D3, and internal transcribed spacer sequences. The phylogenetic inferences placed the nematodes with other neotylenchid nematodes, i.e., Fergusobia and Rubzovinema. Based on the morphology and phylogenetic analysis, this nematode is described herein as Delatylus andersoni n. gen., n. sp. The new genus/species is characterized by the female body habitus ranging from nonobese to semiobese and from straight to dorsally curved when heat relaxed, cephalic framework with six unequal sized lip sectors, lateral fields having 10 to 12 lines, 4 to 5 guide rings on the stylet, excretory pore posterior to the nerve ring, spermatheca diminished or absent, vulvar opening large, and relative proximity of vulva to the anus. Detailed morphological and molecular characterization of the new genus/species is presented along with the comparison of the related genera.

Xiphinema parachambersi n. sp. recovered in Ningbo, China, from the rhizosphere of ornamental plants ( Gardenia jasminoides and Euonymus hamiltonianus ) imported from Japan is described. The new species is characterized by a long female body 1,830 to 2,109  μ m long, odontostyle 105 to 116  μ m long, reproductive system mono-opisthodelphic, vulva located anteriorly at 25.2 to 27.7% of total body, long ovary 119 to 292  μ m with simple uterus and Z-organ absent, female tail elongated conoid with a rounded terminus. Only two juvenile stages were available to study and no male was found. The polytomous identification codes for this new species are A1, B4, C2, D23, E1, F2, G2, H2, I2, J2, K?, L1 and it belongs to the morphospecies group 1. Phylogenetic analysis based on the 18S, ITS1 and 28S D2/D3 sequences of the new species showed close relationships with X. chambersi . Morphologically, the new species is similar to X. chambersi , X. hangzhouense , and X. winotoi but can be differentiated by morphological characters and DNA sequences. To help identify the species, a diagnostic key to the group 1 species is presented., (© The Society of Nematologists.)

Deladenus posteroporus n. sp. isolated from packaging wood originated from Canada and from white pine lumber from the United States, both intercepted in the port of Ningbo, China, is described and illustrated. Both mycetophagous and infective forms were recovered from the Canadian sample, whereas only the mycetophagous form was found in the U.S., Sample: The new species is characterized by the posteriorly positioned excretory pore relative to the hemizonid in both mycetophagous and infective forms, by a broadly rounded tail end in mycetophagous females and lateral fields with 11 to 12 lines midbody in both mycetophagous females and males. The partial 18S, complete internal transcribed spacer, and partial 28S D2/D3 rRNA genes were amplified and sequenced. Phylogenetic analyses of the genes grouped the new species with previously sequenced species of Deladenus in a fully supported clade. This is the first report of Deladenus species with a known infective stage to have the excretory pore positioned posterior to the hemizonid.

Although marigold (Tagetes patula) is known to produce allelopathic compounds toxic to plant-parasitic nematodes, suppression of Meloidogyne incognita can be inconsistent. Two greenhouse experiments were conducted to test whether marigold is more effective in suppressing Meloidogyne spp. when it is active rather than dormant. Soils infested with Meloidogyne spp. were collected and conditioned in the greenhouse either by 1) keeping the soil dry (DRY), 2) irrigating with water (IRR), or 3) drenching with cucumber (Cucumis sativus) leachate (CL) for 5 wk. These soils were then either planted with cucumber, marigold or remained bare for 10 wk. Suppression of nematode by marigold was then assayed using cucumber. DRY conditioning resulted in the highest number of inactive nematodes, whereas CL and IRR had higher numbers of active nematodes than DRY. At the end of the cucumber bioassay, marigold suppressed the numbers of Meloidogyne females in cucumber roots if the soil was conditioned in IRR or CL, but not in DRY. However, in separate laboratory assays, marigold root leachate slightly reduced M. incognita J2 activity but did not reduce egg hatch (P > 0.05). These finding suggest that marigold can only suppress Meloidogyne spp. when marigold is actively growing. This further suggests that marigold will more efficiently suppress Meloidogyne spp. if planted when these nematodes are in active stage.

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.

The effects of soil type, irrigation, and population density of Rotylenchulus reniformis on cotton were evaluated in a two-year microplot experiment. Six soil types, Fuquay sand, Norfolk sandy loam, Portsmouth loamy sand, Muck, Cecil sandy loam, and Cecil sandy clay, were arranged in randomized complete blocks with five replications. Each block had numerous plots previously inoculated with R. reniformis and two or more noninoculated microplots per soil type, one half of which were irrigated in each replicate for a total of 240 plots. Greatest cotton lint yields were achieved in the Muck, Norfolk sandy loam, and Portsmouth loamy sand soils. Cotton yield in the Portsmouth loamy sand did not differ from the Muck soil which averaged the greatest lint yield per plot of all soil types. Cotton yield was negatively related to R. reniformis PI (initial population density) in all soil types except for the Cecil sandy clay which had the highest clay content. Supplemental irrigation increased yields in the higher yielding Muck, Norfolk sandy loam, and Portsmouth loamy sand soils compared to the lower yielding Cecil sandy clay, Cecil sandy loam, and Fuquay sand soils. The Portsmouth sandy loam was among the highest yielding soils, and also supported the greatest R. reniformis population density. Cotton lint yield was affected more by R. reniformis Pi with irrigation in the Portsmouth loamy sand soil with a greater influence of Pi on lint yield in irrigated plots than other soils. A significant first degree PI × irrigation interaction for this soil type confirms this observation.

The D2-D3 expansion segments of the 28S ribosomal RNA (rRNA) were sequenced and compared to predict secondary structures for Hoplolaiminae species based on free energy minimization and comparative sequence analysis. The free energy based prediction method provides putative stem regions within primary structure and these base pairings in stems were confirmed manually by compensatory base changes among closely and distantly related species. Sequence differences ranged from identical between Hoplolaimus columbus and H. seinhorsti to 20.8% between Scutellonema brachyurum and H. concaudajuvencus. The comparative sequence analysis and energy minimization method yielded 9 stems in the D2 and 6 stems in the D3 which showed complete or partial compensatory base changes. At least 75% of nucleotides in the D2 and 68% of nucleotides in the D3 were related with formation of base pairings to maintain secondary structure. GC contents in stems ranged from 61 to 73% for the D2 and from 64 to 71% for the D3 region. These ranges are higher than G-C contents in loops which ranged from 37 to 48% in the D2 and 33-45% in the D3. In stems, G-C/C-G base pairings were the most common in the D2 and the D3 and also non-canonical base pairs including A•A and U•U, C•U/U•C, and G•A/A•G occurred in stems. The predicted secondary model and new sequence alignment based on predicted secondary structures for the D2 and D3 expansion segments provide useful information to assign positional nucleotide homology and reconstruction of more reliable phylogenetic trees.

Rotylenchulus reniformis is one of the major nematode pests capable of reducing cotton yields by more than 60%, causing estimated losses that may exceed millions of dollars U.S. Therefore, early detection of nematode numbers is necessary to reduce these losses. This study investigates the feasibility of using remotely sensed hyperspectral data (reflectances) of cotton plants affected with different nematode population numbers with self-organizing maps (SOM) in correlating and classifying nematode population numbers extant in a plant's rhizosphere. The hyperspectral reflectances were classified into three classes based on R. renifomis population numbers present in plant's rhizosphere. Hyperspectral data (350-2500 nm) were also sub-divided into Visible, Red Edge + Near Infrared (NIR) and Mid-IR region to determine the sub-region most effective in spectrally classifying the nematode population numbers. Various combinations of different feature extraction and dimensionality reduction methods were applied in different regions to extract reduced sets of features. These features were then classified using a supervised-SOM classification method. Our results suggest that the overall classification accuracies, in general, for most methods in most regions (except visible region) varied from 60% to 80%, thereby, indicating a positive correlation between the nematode numbers present in plant's rhizosphere and the corresponding plant's hyperspectral signatures. Results showed that classification accuracies in the Mid-IR region were comparable to the accuracies obtained in other sub-regions. Finally, based on our findings, the use of remotely-sensed hyperspectral data with SOM could prove to be extremely time efficient in detecting nematode numbers present in the soil.

A new Parasitorhabditis species with males and females was discovered from the southern pine beetle Dendroctonus frontalis and its galleries in loblolly pine, Pinus taeda, growing in Mississippi. Females of the new species have a cupola-shaped tail with a small spike; males possess a 2 + (3+2) + 3 ray pattern on the tail fan with ray 10 reaching the margin, and a distinctive stomatal tooth. Parasitorhabditis frontali n. sp. has some similarities to P. hylurgi Massey, 1974 from Hylurgops pinifex in New York, USA, P. terebranus Massey, 1974 from D. terebrans (Olivier, 1795) in Texas USA, P. ligniperdae Fuchs, 1915 from Hylergops ligniperda (Fabricius, 1787) and P. dendroctoni Rühm, 1956 from D. micans (Kugelann, 1794) in Europe, P. ateri Fuchs, 1915 isolated from the beetle Hylastes ater (Paykull, 1800) in Germany, and P. malii Devdariani and Kakulia,1970 from Scolytus mali (Bechstein, 1805) within the republic of Georgia. Morphometrics for 44 species of Parasitorhabditis are provided to update older keys. Parasitorhabditis frontali n. sp. was initially grown on Malt Extract (ME) agar with its own microbial contaminants that included a bacterium and fungus. The nematode also grew and reproduced after slices of ME agar with nematodes and microbial contaminants were transferred to water agar. It was killed by E. coli on NGM agar plates commonly used to raise other Rhabditida. Drawings of diagnostic anatomy and low-temperature SEM images of bodies, heads, and tails are provided for cultured specimens from pine beetle frass.

Hp-FAR-1 is a major, secreted antigen of the parasitic nematode Heligmosomoides polygyrus, a laboratory mouse model frequently used to study the cellular mechanisms of chronic helminth infections. The DNA encoding Hp-FAR-1 was recovered by screening a fourth larval (L₄) H. polygyrus cDNA expression library using antibodies raised against L₄ stage excretory/secretory (E/S) proteins. Predictions of secondary structure based on the Hp-FAR-1 amino acid sequence indicated that an alpha-helix predominates in Hp-FAR-1, possibly with some coiled-coil conformation, with no beta-structure. Fluorescence-based ligand binding analysis confirmed that the recombinant Hp-FAR-1 (rHp-FAR-1) binds the fluorescent fatty acid analog 11-((5-[dimethylaminoaphthalene-1-sulfonyl)amino)undecanoic acid (DAUDA), and by competition oleic acid. RT-PCR amplification of the hp-far-1 gene indicated that the gene is transcribed in all parasitic stages of the organism's life cycle. The presence of a secreted FAR protein in the well-defined laboratory model of H. polygyrus provides an excellent model for the further study and analysis of the in vivo role of secreted FAR proteins in parasitism, and supports the mounting evidence that secreted FAR proteins play a major role in nematode parasitism.

Hoplolaimus columbus is an important nematode pest which causes economic loss of crops including corn, cotton, and soybean in the Southeastern United States. DNA sequences of the ITS1-5.8S-ITS2 region of ribosomal DNA from H. columbus were aligned and analyzed to characterize intraspecific genetic variation between eleven populations collected from Georgia, Louisiana, North Carolina, and South Carolina. In comparative sequence analysis with clones from either one or two individuals obtained from the eleven populations, we found variability existed among clones from an individual and that clonal diversity observed from within individuals was verified by PCR-RFLP. PCR-RFLP analysis with Rsa I and Msp I restriction enzymes yielded several fragments on 3.0% agarose gel that corresponded to different haplotypes in all populations and the sum of digested products exceeded the length of undigested PCR products, which revealed that ITS heterogeneity existed in a genome of H. columbus. This indicates that heterogeneity may play a role in the evolution of this parthenogenetic species.

Three new species of the genus BrevitobriusTsalolikhin, 1981 are described. Brevitobrilus glandulatus n. sp. is characterized by conspicuous sphincter between pars dilatata and uterus; two pairs of vaginal glands; spicules having elliptical capitula with small proximal stiffening piece; proximally-arcuate gubernaculum; S3 and S4 smaller than other supplements; S6 out of spicular range and 57-60 micropapillae. Brevitobrilus dimorphicus n. sp. is diagnosed by sexual dimorphism in labial sensilla and amphids; thick-walled rectum with a diverticulum protruding into intestinal lumen and males with boat-shaped spicules and S6 occasionally slightly smaller than other supplements. Brevitobrilus allahabadensis n. sp. possesses large amphids of 28-33% of corresponding labial diameter in both sexes; vagina and uterus with muscular, plicate walls; well developed sphincter between vas deferens and ejaculatory duct; capitulate spicules with sloping ventral and angular dorsal walls; S3, S4 and S6 smaller than other supplements, S6 close to cloaca and 28-37 micropapillae. The relationships of the species of genus Brevitobrilus have been assessed using morphological characters subjected to parsimony and a non cladistic key to identification of species is given.

Rotylenchulus reniformis is the predominant parasitic nematode of cotton in the Mid South area of the United States. Although variable levels of infection and morphological differences have been reported for this nematode, genetic variability has been more elusive. We developed microsatellite-enriched libraries for R. reniformis, produced 1152 clones, assembled 694 contigs, detected 783 simple sequence repeats (SSR) and designed 192 SSR-markers. The markers were tested on six R. reniformis cultures from four states, Texas, Louisiana, Mississippi and Georgia, in the USA. Based on performance we selected 156 SSR markers for R. reniformis from which 88 were polymorphic across the six reniform nematode populations, showing as the most frequent motif the dinucleotide AG. The polymorphic information content of the markers ranged from 0.00 to 0.82, and the percentage of multiallelic loci of the isolates was between 40.9 and 45.1%. An interesting finding in this study was the genetic variability detected among the three Mississippi isolates, for which 22 SSR markers were polymorphic. We also tested the level of infection of these isolates on six cotton genotypes, where significant differences were found between the Texas and Georgia isolates. Coincidentally, 62 polymorphic markers were able to distinguish these two populations. Further studies will be necessary to establish possible connections, if any, between markers and level of pathogenicity of the nematode. The SSR markers developed here will be useful in the assessment of the genetic diversity of this nematode, could assist in management practices for control of reniform nematode, be used in breeding programs for crop resistance, and help in detecting the origin and spread of this nematode in the United States.

In October 1985 during a survey of fauna of the Great Smoky Mountains National Park, Ernest Bernard recovered a limited number of specimens of a non-described species of Xiphinema (Nematoda: Longidoridae) and sent them to the senior author. The species is distinct from other species by its large size and having Z-organs in the genital tract. During July 2006, Dr. Bernard's survey crew took samples in the area where the species was first found and was successful in finding it again. Without Dr. Bernard's efforts, this species could not have been described and thus the new species is named X. bernardi n. sp. in his honor. Several female and juvenile specimens of the new species were recovered in a sample from a mixed forest of maple, hemlock, and silverbell. It is distinct from all others in Xiphinema group 4 species (with Z-organs) by having a longer total stylet length, 259.8 to 284.2 μm vs < 253 μm for all other species in this group. Xiphinema bernardi n. sp. is distinctive because of its long body length (4.45 to 6.00 mm), tail shape, and c' ratio. Of the group 4 species, it most closely resembles X. phoenicis. Second, third and fourth stage juvenile descriptions and morphometrics are included. The polytomous key code for X. bernardi n. sp. is A4-B1-C6-D56-E56-F(4)5-G4-H2-I34-J5-K?-L1. Molecular approaches using the internal transcribed spacer 1 sequences of nuclear ribosomal DNA suggested that X. bakeri and X. diversicaudatum are the most closely related species from the species examined.

DNA sequences of the D2-D3 expansion segments of the 28S gene of ribosomal DNA from 23 taxa of the subfamily Hoplolaiminae were obtained and aligned to infer phylogenetic relationships. The D2 and D3 expansion regions are G-C rich (59.2%), with up to 20.7% genetic divergence between Scutellonema brachyurum and Hoplolaimus concaudajuvencus. Molecular phylogenetic analysis using maximum likelihood and maximum parsimony was conducted using the D2-D3 sequence data. Of 558 characters, 254 characters (45.5%) were variable and 198 characters (35.4%) were parsimony informative. All phylogenetic methods produced a similar topology with two distinct clades: One clade consists of all Hoplolaimus species while the other clade consists of the rest of the studied Hoplolaiminae genera. This result suggests that Hoplolaimus is monophyletic. Another clade consisted of Aorolaimus, Helicotylenchus, Rotylenchus, and Scutellonema species. Phylogenetic analysis using the outgroup species Globodera rostocheinsis suggests that Hoplolaiminae is paraphyletic. In this study, the D2-D3 region had levels of DNA sequence divergence sufficient for phylogenetic analysis and delimiting species of Hoplolaiminae.

DNA sequence analyses of the nuclear ribosomal ITS1 region of the ribosomal DNA and D1-D3 expansion segments of the 28S gene were conducted to characterize the genetic variation of six amphimictic Hoplolaimus species, including H. magnistylus, H. concaudajuvencus, H. galeatus, Hoplolaimus sp. 1, Hoplolaimus sp. 2 and Hoplolaimus sp. 3, and two closely related parthenogenetic species, H. columbus and H. seinhorsti. PCR amplifications of the combined D1-D3 expansion segments and the ITS1 region each yielded one distinct amplicon. In the D1-D3 region, there was no nucleotide sequence variation between populations of H. columbus, H. magnistylus, Hoplolaimus sp. 2 and Hoplolaimus sp. 3, whereas the ITS1 sequences had nucleotide variation among species. We detected conserved ITS1 regions located at the 3' and 5' end of ITS1 and also in the middle of the ITS1 among Hoplolaimus species. These regions were compared with sequences of distantly related Heterodera and Globedera. PCR-RFLP and sequence analysis of ITS1 and 28S PCR products revealed that several haplotypes existed in the same genome of H. columbus, H. magnistylus, H. seinhorsti, H. concaudajuvencus and Hoplolaimus sp. 1. Maximum likelihood and maximum parsimony analysis using the combined ITS1 and D1-D3 expansion segment sequences always produced trees with similar topology; H. columbus and H. seinhorsti grouped in one clade and the other six species (H. galeatus, H. concaudajuvencus, H. magnistylus, Hoplolaimus sp. 1, Hoplolaimus sp. 2, Hoplolaimus sp. 3) grouped in another. Molecular analysis supports morphological schemes for this genus to be divided into two groups based on several phenotypic traits derived from morphological evolution.

Damage to cotton by Rotylenchulus reniformis below plow depth was evaluated in a sandy clay loam soil at Weslaco, Texas. In December 1999, 14 holes on 51-cm centers were dug 91 cm deep along the planting bed and adjacent furrow and 2 ml of 1,3-dichloropropene was placed 91, 61, and 30 cm deep as each hole was refilled and packed. This technique eliminated 96%, 81%, and 74% of R. reniformis down to 107 cm at distances 0, 25, and 51 cm laterally from the point of application (P

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 degrees 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.

The possible impact of Rotylenchulus reniformis below plow depth was evaluated by measuring the vertical distribution of R. reniformis and soil texture in 20 symptomatic fields on 17 farms across six states. The mean nematode population density per field, 0 to 122 cm deep, ranged from 0.4 to 63 nematodes/g soil, and in 15 fields more than half of the R. reniformis present were below 30.5 cm, which is the greatest depth usually plowed by farmers or sampled by consultants. In 11 fields measured, root density was greatest in the top 15 cm of soil; however, roots consistently penetrated 92 to 122 cm deep by midseason, and in five fields in Texas and Louisiana the ratio of nematodes to root-length density within soil increased with depth. Repeated sampling during the year in Texas indicated that up to 20% of the nematodes in soil below 60 cm in the fall survived the winter. Differences between Baermann funnel and sugar flotation extraction methods were not important when compared with field-to-field differences in nematode populations and field-specific vertical distribution patterns. The results support the interpretation that R. reniformis below plow depth can significantly impact diagnosis and treatment of cotton fields infested with R. reniformis.

The most effective management program for soybean cyst nematode, Heterodera glycines, is a crop rotation that uses nonhost crops and resistant soybean cultivars. However, little is known about the effects of rotation crops and overwintering on H. glycines biology. These experiments were initiated to determine the effects of seven alternative crops on H. glycines' ability to infect and mature on subsequent soybean crops, and to assess the viability of eggs during the overwintering months. Rotation studies were conducted for 2 years in each of two naturally infested fields, and overwintering tests were conducted in three consecutive growing seasons in one naturally infested field. Rotation crop and fallow treatments did not have a consistent effect on the ability of H. glycines to infect soybean or mature. Soybean yields were often higher following fallow or a nonhost crop than following soybean, although not usually significantly so. Heterodera glycines egg viability did not differ (P < 0.05) between overwintering months at 0-to-10 or 10-to-20-cm soil depths. These results suggest that H. glycines' ability to infect a subsequent soybean crop and develop to maturity is not diminished by nonhost crops or during the winter months.

To enable investigation of nematode mitochondrial DNA evolution, methodology has been developed to amplify intact nematode mitochondrial genomes in preparative yields using a rolling circle replication strategy. Successful reactions were generated from whole cell template DNA prepared by alkaline lysis of the rhabditid nematode Caenorhabditis elegans and a mermithid nematode, Thaumamermis cosgrovei. These taxa, representing the two major nematode classes Chromodorea and Enoplea, maintain mitochondrial genomes of 13.8 kb and 20.0 kb, respectively. Efficient amplifications were conducted on template DNA isolated from individual or pooled nematodes that were alive or stored at -80 degrees C. Unexpectedly, these experiments revealed that multiple T. cosgrovei mitochondrial DNA haplotypes are maintained in our local population. Rolling circle amplification products can be used as templates for standard PCR reactions with specific primers that target mitochondrial genes or for direct DNA sequencing.

FOUR NATIVE PLANT COMMUNITY TYPES (IN DECREASING ELEVATION: montane bog, rain forest, wet mesic forest, drier forest) on Molokai were sampled for nematodes. Six samples of 10 cores each were gathered from each community. Nematodes were extracted from 200 cm(3) soil by elutriation. All extracted nematodes were counted and identified to species-level taxa. Sixty-seven species were identified among the four plant communities; only eight species occurred in all four communities. Species diversity and evenness were greater in the rain forest and mesic forest than in the bog and the drier forest, but the drier forest and mesic forest had similar species communities. The bog nematode community was not similar to the other three sites. In a presence/absence cluster analysis, all six bog sample assemblages clustered together. The rain forest samples also clustered together but were associated with the mesic forest sample closest to the rain forest edge. Of 11 nematode orders collected, Tylenchida accounted for 40% to 73% of all individuals, followed by Dorylaimida (5% to 17%). Diplogasterida were absent. No plant-parasitic nematodes of known Hawaiian agricultural importance or occurrence were collected in these native plant communities. Calculated nematode densities (76,000 to 321,300/m(2)) were comparable to those reported for some other Pacific tropical forests.

Endospores of Pasteuria penetrans were evaluated for their vertical distribution in field soil and their downward movement through soil in the laboratory. In the field trial, the number of endospores attached to second-stage juveniles (J2) of Meloidogyne arenaria race 1 varied greatly in different soil depths. There were higher percentages of J2 with endospores attached in former weed fallow plots during the first 3 years of growing peanut than in former bahiagrass and rhizomal peanut plots (P 0.05). In laboratory trials, P. penetrans endospores were observed to move throughout the soil through the percolation of water. After one application of water, some endospores were detected 25 to 37.5 cm deep. Endospores were present at the greatest depth, 37.5 to 50 cm, after the third application of water. These results indicate that rain or water applications by irrigation are likely to move endospores to deeper levels of the soil, but the majority of endospores remain in the upper 0-to-30-cm depth.

An unusual population of Meloidogyne hapla, earlier thought to be an undescribed species, was found causing large galls, without adventitious roots, and substantial damage to coffee in Maui, Hawaii. Only in Brazil had similar damage to coffee been reported by this species. Unlike M. exigua from South and Central America, this population reproduced well on coffee cv. Mokka and M. incognita-susceptible tomato but poorly on tomato with the Mi resistance gene. Characterization included SEM images, esterase isozymes, and five DNA sequences: i) the D3 segment of the large subunit (LSU-D3 or 28S) rDNA, ii) internal transcribed spacer (ITS-1) rDNA, iii) intergenic spacer (IGS) rDNA, iv) the mitochondrial interval from cytochrome oxidase (CO II) to 16S mtDNA, and v) the nuclear gene Hsp90. Sequences for ITS-1, IGS, and COII were similar to other M. hapla populations, but within species ITS-1 variability was not less than among species. One LSU-D3 haplotype was similar to a previously analyzed population with two minor haplotypes. Hsp90 exhibited some variation between Maryland and Hawaiian populations distinct from other species. Females were narrow with wide vulval slits, large interphasmidial distances, and more posterior excretory pores; 20% of perineal patterns had atypical perivulval lines. Males had a low b ratio (<12 microm). Juveniles had a short distance between stylet and dorsal gland orifice. Juvenile body length was short (<355 microm) and was different between summer and winter populations.

Thripinema fuscum is a natural enemy of Frankliniella fusca in peanut. Laboratory experiments were conducted to determine the reproductive biology of T. fuscum as affected by gender and stage of development of the host and to determine the effects of parasitism on host longevity, fecundity, and mortality. The adult females of F. fusca were the most readily parasitized (P < 0.001) in the laboratory experiments followed by the second instars, the first instars, and the adult males. One generation of T. fuscum developed within the parasitized larvae and adults, with the males and females emerging only during the adult stage of the host. Parasitism did not cause mortality of the host. Parasitism affected male longevity (P < 0.001) but not female longevity. The adult female thrips that were parasitized as first or second instars did not lay eggs, and the adult females stopped laying eggs within 3 days of being parasitized. The female-to-male sex ratio of T. fuscum emerging from parasitized male and female F. fusca was 22 and 18 to 1, respectively. More T. fuscum emerged from female hosts than from male hosts (P < 0.001). More emerged from hosts parasitized as larvae compared with hosts parasitized as adults (P < 0.05). The intrinsic capacity of increase of T. fuscum ranged between 0.29 and 0.37 when parasitizing the adult males and females and between 0.18 and 0.21 when parasitizing the larval males and females. Percent parasitism of F. fusca was estimated in peanut fields. The flowers were the primary site for aggregation of the adults of F. fusca and for the free-living females of T. fuscum to parasitize new hosts. As under laboratory conditions, field parasitism of adult males was less than parasitism of adult females in 2001 and 2002 (P < 0.01 and 0.001, respectively). Our study indicates that T. fuscum is a potential biological control agent capable of suppressing F. fusca populations in peanut.

The long-term persistence and suppressiveness of Pasteuria penetrans against Meloidogyne arenaria race 1 were investigated in a formerly root-knot nematode suppressive site following 9 years of continuous cultivation of three treatments and 4 years of continuous peanut. The three treatments were two M. arenaria race 1 nonhost crops, bahiagrass (Paspalum notatum cv. Pensacola var. Tifton 9), rhizomal peanut (Arachis glabrata cv. Florigraze), and weed fallow. Two root-knot nematode susceptible weeds commonly observed in weed fallow plots were hairy indigo (Indigofera hirsuta) and alyce clover (Alysicarpus vaginalis). The percentage of J2 with endospores attached reached the highest level of 87% in 2000 in weed fallow, and 63% and 53% in 2002 in bahiagrass and rhizomal peanut, respectively. The percentage of endospore-filled females extracted from peanut roots grown in weed fallow plots increased from nondetectable in 1999 to 56% in 2002, whereas the percentages in bahiagrass and rhizomal peanut plots were 41% and 16%, respectively. Over 4 years, however, there was no strong evidence that endospores densities reached suppressive levels because peanut roots, pods, and pegs were heavily galled, and yields were suppressed. This might be attributed to the discovery of M. javanica infecting peanut in this field in early autumn 2001. A laboratory test confirmed that although the P. penetrans isolate specific to M. arenaria attached to M. javanica J2, no development occurred. In summary, P. penetrans increased on M. arenaria over a 4-year period, but apparently because of infection of M. javanica on peanut at the field site root-knot disease was not suppressed. This was confirmed by a suppressive soil test that showed a higher level of soil suppressiveness than occurred in the field (P

The effects of the application of poultry litter at 0.0, 6.7, 13.4, and 20.1 tons/ha on population changes during the growing season on nematode communities were evaluated in two cotton production fields in North Carolina. Numbers of bactivorous nematodes increased at midseason in response to the rate at which litter was applied but decreased with increasing litter application rates at cotton harvest. Numbers of fungivores at cotton harvest were related positively to the rate of litter applied, and this affected a positive increase in the fungivore-to-bacterivore ratio at this sampling date. The rate at which poultry litter was applied resulted in an increase in the bacterivore to plant-parasite ratio, and this corresponded with increased cotton lint yield. Trophic diversity was increased by litter application rate at cotton harvest at one location but not at another. The plant-parasite maturity index was greater consistently at one site than at a second site where the Hoplolaimus columbus population density was above the damage threshold for cotton. The population density of H. columbus was suppressed with increasing rates of poultry litter application, but other plant-parasitic nematodes were affected marginally.

The discovery of Meloidogyne mayaguensis is confirmed in Florida; this is the first report for the continental United States. Meloidogyne mayaguensis is a virulent species that can reproduce on host cultivars bred for nematode resistance. The perineal patterns of M. mayaguensis isolates from Florida show morphological variability and often are similar to M. incognita. Useful morphological characters for the separation of M. mayaguensis from M. incognita from Florida are the male stylet length values (smaller for M. mayaguensis than M. incognita) and J2 tail length values (greater for M. mayaguensis than M. incognita). Meloidogyne mayaguensis values for these characters overlap with those of M. arenaria and M. javanica from Florida. Enzyme analyses of Florida M. mayaguensis isolates show two major bands (VS1-S1 phenotype) of esterase activity, and one strong malate dehydrogenase band (Rm 1.4) plus two additional weak bands that migrated close together. Their detection requires larger amounts of homogenates from several females. Amplification of two separate regions of mitochondrial DNA resulted in products of a unique size. PCR primers embedded in the COII and 16S genes produced a product size of 705 bp, and amplification of the 63-bp repeat region resulted in a single product of 322 bp. Nucleotide sequence comparison of these mitochondrial products together with sequence from 18S rDNA and ITS1 from the nuclear genome were nearly identical with the corresponding regions from a M. mayaguensis isolate from Mayaguez, Puerto Rico, the type locality of the species. Meloidogyne mayaguensis reproduced on cotton, pepper, tobacco, and watermelon but not on peanut. Preliminary results indicate the M. mayaguensis isolates from Florida can reproduce on tomato containing the Mi gene. Molecular techniques for the identification of M. mayaguensis will be particularly useful in cases of M. mayaguensis populations mixed with M. arenaria, M. incognita, and M. javanica, which are the most economically important root-knot nematode species in Florida, and especially when low (<25) numbers of specimens of these species are recovered from the soil.

The putative mutualism between different host-specific Fergusobia nematodes and Fergusonina flies is manifested in a variety of gall types involving shoot or inflorescence buds, individual flower buds, stems, or young leaves in the plant family Myrtaceae. Different types of galls in the early-to-middle stages of development, with host-specific species of Fergusobia/Fergusonina, were collected from Australian members of the subfamily Leptospermoideae (six species of Eucalyptus, two species of Corymbia, and seven species of broad-leaved Melaleuca). Galls were sectioned and histologically examined to assess morphological changes induced by nematode/fly mutualism. The different gall forms were characterized into four broad categories: (i) individual flower bud, (ii) terminal and axial bud, (iii) 'basal rosette' stem, and (iv) flat leaf. Gall morphology in all four types appeared to result from species-specific selection of the oviposition site and timing and number of eggs deposited in a particular plant host. In all cases, early parasitism by Fergusobia/Fergusonina involved several layers of uninucleate, hypertrophied cells lining the lumen of each locule (gall chamber where each fly larva and accompanying nematodes develop). Hypertrophied cells in galls were larger than normal epidermal cells, and each had an enlarged nucleus, nucleolus, and granular cytoplasm that resembled shoot bud gall cells induced by nematodes in the Anguinidae.

In a study of relationships among selected cyst-forming and noncyst-forming species of Heteroderoidea, combined sequences comprised of DNA from part of the conserved 18S ribosomal RNA gene (rDNA) plus the complete ITS rDNA segment were more similar to analyses based on the ITS data alone than to analyses based on the 18S data alone. One of the two noncyst-forming species, Ekphymatodera thomasoni, grouped with cyst-forming species of Heteroderoidea. Bilobodera flexa, also a noncyst-forming species, was separated from all the other taxa by a long branch. Afenestrata koreana, with a weakly sclerotized cyst, grouped closely with H. bifenestra. These observations suggest that phylogenetic analyses using molecular data may aid in our understanding of the evolution of cyst formation in nematodes, including the possibility of secondary loss. The usefulness of molecular phylogenetic analyses in nematodes may depend more on the particular selection of taxa than on mere addition of data from additional genes.

The effect of Steinernema riobrave and Heterorhabditis bacteriophora on population density of Mesocriconema xenoplax in peach was studied in the greenhouse. Twenty-one days after adding 112 M. xenoplax adults and juveniles/1,500 cm(3) soil to the soil surface of each pot, 50 infective juveniles/cm(2) soil surface of either S. riobrave or H. bacteriophora were applied. Another entomopathogenic nematode application of the same density was administered 3 months later. The experiment was repeated once. Mesocriconema xenoplax populations were not suppressed (P

The impact of Meloidogyne graminicola on growth and yield of lowland rainfed rice was assessed with and without carbofuran in a rice-wheat rotation area of northwestern Bangladesh. The experiment was conducted on farmer fields and at a research station, with experimental plots arranged in a randomized complete block design. Prior to transplanting, rice seedling height and dry weight were greater (P

Two experiments were conducted to assess nutrient partitioning in coffee (Coffea arabica cv. Typica land race Guatemala) infected with Meloidogyne konaensis. Nutrient levels were quantified from soil, roots, and leaves. In the first experiment, 500-cm3 aliquants of a Kealakekua Andisol were infested with four initial population densities of M. konaensis ranging from 0 to 1,500 freshly hatched second-stage juveniles. Coffee plants (~3 months old) were transplanted into the soil and grown for 25 weeks. Plants responded to nematode infection with decreases (P < 0.05) in concentrations of Ca, Mg, P, and B and increases (P < 0.05) in concentrations of Mn, Cu, Zn, and Ca/B in the roots. Mn and Cu uptake by roots was decreased (P < 0.05) by nematode infection even though concentrations of Mn and Cu increased (P < 0.05) in the roots. Concentrations of Ca and Mg also decreased (P < 0.05) in the leaves, whereas the concentration of Zn increased (P < 0.05). In the second experiment, the soil was amended with Zn at 0 or 5 mg/kg soil and infested with M. konaensis at 0, 100, 1,000 or 10,000 eggs/1,200 cm(3) soil. Three-month-old coffee seedlings of similar height were weighed and transplanted into pots and then placed in a greenhouse and grown under 50% shade for 23 weeks. Concentrations of P, K, Ca, Mg, Mn, B, and Zn increased in roots of nematode-free plants growing in Zn-amended soil. The beneficial effects due to the Zn amendment were not apparent in nematode-infected plants. Mn, B, and Zn uptake by coffee roots and P and B concentrations in coffee leaves responded similarly. Management of M. konaensis is necessary to achieve optimal nutrient management in coffee.

Alternatives to reduce or modify nematicide use for minimizing groundwater contamination in Easter lily were explored in two field trials. Alternatives to standard 1,3-dichloropropene (1,3-D) plus phorate injection in the first trial were: (i) delaying applications until after winter rains, (ii) removing roots from planting stock, (iii) 1,3-D via drip irrigation, (iv) a chitin-urea soil amendment, (v) the registered insecticide disulfoton, and (vi) several nonregistered nematicides. None of the treatments equaled the standard treatment. In the second trial, potential benefits of adding a systemic nematicide, oxamyl (OX), or a fungicide, metalaxyl (MX), to the standard treatment were explored. Preplant drip irrigation applications of metam sodium (MS), sodium tetrathiocarbonate (ST), and emulsifiable 1,3-D were evaluated alone and in combination with postplant applications of OX and MX. Several drip-applied treatments performed comparably to the standard treatment with respect to the most important criteria of crop quality, bulb circumference. Metam-sodium in combination with either or both OX and MX, 1,3-D plus OX and MX, and ST plus OX and MX provided the best results.

A mixed population of Meloidogyne arenaria race 1 and M. javanica race 3 is reported on peanut from a field in Levy County, Florida. Confirmation of M. javanica on peanut is based on esterase and malate dehydrogenase isozyme patterns resolved on polyacrylamide slab gels following electrophoresis, and perineal patterns. Up to 29% of 290 individual females collected from peanut roots in the field in autumn 2002 showed a typical esterase J3 phenotype for M. javanica. This is the third report of M. javanica infecting peanut in the United States.

The synthesis and localization of an endospore surface epitope associated with the development of Pasteuria penetrans was determined using a monoclonal antibody (MAb) as a probe. Nematodes, uninfected or infected with P. penetrans, were harvested at 12, 16, 24, and 38 days after inoculation (DAI) and then examined to determine the developmental stage of the bacterium. Vegetative growth of P. penetrans was observed only in infected nematodes harvested at 12 and 16 DAI, whereas cells at different stages of sporulation and mature endospores were observed at 24 and 38 DAI. ELISA and immunoblot analysis revealed that the adhesin-associated epitope was first detected at 24 DAI, and increased in the later stages of sporogenesis. These results indicate that the synthesis of adhesin-related proteins occurred at a certain developmental stage relative to the sporulation process, and was associated with endospore maturation. Immunofluorescence microscopy indicated that the distribution of the epitope is nearly uniform on the periphery of each spore, as defined by parasporal fibers. Immunocytochemistry at the ultrastructural level indicated a distribution of the epitope over the parasporal fibers. The epitope also was detected over other structures such as sporangium and exosporium during the sporogenesis process, but it was not observed over the cortex, inner-spore coat, outer-spore coat, or protoplasm. The appearance of the adhesin epitope first at stage III of sporogenesis and its presence on the parasporal fibers are consistent with an adhesin-related role in the attachment of the mature endospore to the cuticle of the nematode host.

Belonolaimus longicaudatus is a serious problem on bermudagrass, a common warm-season turfgrass, in Florida. The cancellation of organophosphate nematicides necessitates that new management tools be identified for use on sports turf. Postplant application of 1,3-dichloropropene (1,3-D) on bermudagrass was evaluated for management of B. longicaudatus on golf course fairways and driving ranges. A series of 10 experiments were conducted to evaluate the effectiveness of 1,3-D in reducing population densities of B. longicaudatus and enhancing bermudagrass recovery from nematode damage. In 5 of 10 experiments, 1,3-D injected at 46.8 liters/ha was effective in reducing population densities of B. longicaudatus (P < 0.05) compared to untreated plots 2 to 4 weeks after treatment. One month after treatment, population densities of B. longicaudatus ranged from 59% to 97% of those in untreated plots. Nematode suppression generally lasted 2 months or less. Turf visual performance was improved following injection with 1,3-D (P < 0.05) over untreated plots when other factors were not limiting. Turf root development also was enhanced following injection with 1,3-D. Postplant injection of 1,3-D could be a useful nematode management tool for certain sports turf applications.

Intracellular bacteria of the genus Wolbachia are among the most abundant endosymbionts on the planet, occurring in at least two major phyla-the Arthropoda and Nematoda. Current surveys of Wolbachia distribution have found contrasting patterns within these groups. Whereas Wolbachia are widespread and occur in all three major subphyla of arthropods, with estimates placing them in at least several million arthropod species, the presence of nematodes carrying Wolbachia is currently confined to the filariids, in which they occur at appreciable frequencies. It has been hypothesized that Wolbachia entered the ancestor of modern-day filariids in a single acquisition event, and subsequently cospeciated with their filariid hosts. To further investigate this hypothesis, we examined the broader distribution of Wolbachia in nematodes using a polymerase chain reaction (PCR) assay in a diverse set of nonfilariid species. The assay consisted of three different types of PCR screens on adults of 20 secernentean nematode species (14 rhabditids, 2 strongylid parasites of vertebrates; 1 diplogasterid; 3 cephalobid relatives, 1 myolaim, and 1 filariid) and two adenophorean species (plectids). Two PCR screens were specific to the 16S rDNA and ftsZ protein coding gene of Wolbachia; and the third screen was specific to the 18S rDNA of the nematodes. Based upon our results, we conclude that Wolbachia are absent in all 21 non-filariid species encompassing all the major groups of the Secernentea and two species of Adenophorea, from which the Secernentea derived. The absence of Wolbachia in these non-filariids is consistent with the hypothesis that Wolbachia entered the nematode phylum once, in an ancestral lineage of extant filariids.

Pasteuria spp. include endospore-forming bacterial pathogens of cladoceran crustaceans and plant-parasitic nematodes. Propagation of these nematode pathogens requires attachment of soilborne endospores to nematode hosts, infection, growth, sporulation, and release of endospores to repeat the cycle of infection and propagation. The ability of these bacteria to suppress the levels of plant-parasitic nematodes in the field has made them particularly promising candidates for biocontrol of nematode diseases of plants. Genes encoding 16S ribosomal RNA have been sequenced for the cladoceran (water flea) parasite and type species, Pasteuria ramosa, and for Pasteuria spp. isolated from root-knot (Meloidogyne arenaria race 1 and Meloidogyne sp.), soybean cyst (Heterodera glycines), and sting (Belonolaimus longicaudatus) nematodes. These have provided a phylogenetic basis for their designation to a distinct clade within the family Alicyclobacillaceae of the gram-positive endospore-forming bacteria. Two apparent biotypes of P. penetrans demonstrating a host preference for different Meloidogyne spp. showed identical 16S rDNA sequences, suggesting host-recognition evolves within a given species. The sequences of genes encoding sporulation transcription factors, sigE and sigF, from P. penetrans biotype P-20 show different phylogenetic relationships to other endospore-forming bacteria, supporting their application to further discriminate Pasteuria spp. and biotypes. Distribution of an adhesin-associated epitope on polypeptides from different Pasteuria isolates provides an immunochemical approach to differentiate species and biotypes with specific host preferences. Application of bioinformatics to genomic data, as well as further characterization of the biochemical basis for host recognition, will facilitate development of Pasteuria spp. as benign alternatives to chemical nematicides.

Two synaptonemal complexes (SCs) were present in the pachytene nuclei of Ascaris megalocephala. The SC was tripartite and comprised of two lateral elements (25 nm) with a striated central element (25 nm) and a central region of 65 nm. Spherical recombination nodules were observed to be associated only with the central element, although they are non-existent in the related A. lumbricoides var. suum (Goldstein, 1977). The SCs were attached to the nuclear envelope at only one end, while the other end was free in the nucleoplasm. This lack of bouquet formation of the chromosomes is consistent with all other nematodes studied. Morphologically distinct sex chromosomes were not observed, which differs from the presence of five Y-chromosomes present in A. lumbricoides var. suum (Goldstein and Moens, 1976). Centrioles (0.2 microm wide) reproduced by budding off the parental centriole. The centrioles consisted of nine singlet microtubules connected by an electron-dense proteinaceous ring. This structure is consistent with centrioles described in other nematodes, yet distinctly different from the centriole structure observed in most organisms in which it consists of nine triplet microtubules without any connecting ring. Multiple synaptonemal complexes, or polycomplexes, are found in A. megalocephala and A. lumbriocoides var. suum. They appear as stacked SC and are present inside the nucleus during zygotene and in the cytoplasm at pachytene.

Seasonal and spatial variation in soil nematode communities was investigated in a field study conducted on a loessial plain in the northern Negev Desert, Israel. Soil samples from 0- to 50-cm depths were collected seasonally during 2001 under the canopy of Atriplex halimus and Hammada scoparia, and between shrubs (control). Total population abundance ranged from 8 to 887 individuals per 100 g soil, represented by 32 genera from 16 families. Significant temporal and vegetation effects were elucidated using most ecological indices applicable to express nematode community composition. Of those indices computed, only the Shannon index, the modified maturity index, and genus dominance distinguished differences in vertical distribution.