Top-down impact of bacterivorous nematodes on the bacterial community structure: a microcosm study

The influence of bacterivorous nematodes (Diplolaimelloides meyli, Diplolaimelloides oschei, Diplolaimella dievengatensis, Panagrolaimus paetzoldi) on the development of a bacterial community growing on decaying cordgrass detritus was studied in laboratory microcosm experiments. Cordgrass leaves were incubated on a sediment surface with a natural bacterial mixture containing bacteria from sediment, cordgrass detritus and habitat water. The four nematode species were applied separately to the microcosms; controls remained without nematodes. Samples were taken seven times over a 65-day period. The bacterial community structure was analysed by means of DGGE of the 16S rRNA genes. Multi Dimensional Scaling showed grouping of the samples per treatment. Analysis of Similarities indicated that the differences between treatments were significantly larger than differences within treatments. Our results suggest that nematodes can have a significant structuring top-down influence on the 'pool' of bacteria growing on the detritus, even at low densities. Dissimilarities were similar between all treatments. Differences in bacterial community composition within the treatments with monhysterids (D. meyli, D. oschei and D. dievengatensis) can be explained by species-specific food preferences. Panagrolaimus paetzoldi on the other hand feeds unselectively, and thus affects the bacterial community differently. A top-down effect of the nematodes on the diversity of the bacterial community was only evident under high grazing pressure, i.e. in the presence of P. paetzoldi.     

Effects of Nematophagous Fungi on Numbers and Death Rates of Bacterivorous Nematodes in Arable Soil

In a series of microcosm experiments with an arable, sandy loam soil amended with sugarbeet leaf, the short-term (8 weeks) dynamics of numbers of nematodes were measured in untreated soil and in γ-irradiated soil inoculated with either a field population of soil microorganisms and nematodes or a mixed population of laboratory-propagated bacterivorous nematode species. Sugarbeet leaf stimulated an increase in bacterivorous Rhabditidae, Cephalobidae, and a lab-cultivated Panagrolaimus sp. Differences were observed between the growth rates of the nematode population in untreated and γ-irradiated soils, which were caused by two nematophagous fungi, Arthrobotrys oligospora and Dactylaria sp. These fungi lowered the increase in nematode numbers due to the organic enrichment in the untreated soil. We estimated the annually produced bacterivous nematodes to consume 50 kg carbon and 10 kg nitrogen per ha, per year, in the upper, plowed 25 cm of arable soil.     

Sunn Hemp Cover Cropping and Organic Fertilizer Effects on the Nematode Community Under Temperate Growing Conditions

Field experiments were conducted in Maryland to investigate the influence of sunn hemp cover cropping in conjunction with organic and synthetic fertilizers on the nematode community in a zucchini cropping system. Two field treatments, zucchini planted into a sunn hemp living and surface mulch (SH) and zucchini planted into bare-ground (BG) were established during three field seasons from 2009 to 2011. In 2009, although SH slightly increased nematode richness compared with BG by the first harvest (P < 0.10), it reduced nematode diversity and enrichment indices (P < 0.01 and P < 0.10, respectively) and increased the channel index (P < 0.01) compared to BG at the final harvest. This suggests a negative impact of SH on nematode community structure. The experiment was modified in 2010 and 2011 where the SH and BG main plots were further split into two subplots to investigate the added influence of an organic vs. synthetic fertilizer. In 2010, when used as a living and surface mulch in a no-till system, SH increased bacterivorous, fungivorous, and total nematodes (P < 0.05) by the final zucchini harvest, but fertilizer type did not influence nematode community structure. In 2011, when incorporated into the soil before zucchini planting, SH increased the abundance of bacterivorous and fungivorous nematodes early in the cropping season. SH increased species richness also at the end of the season (P < 0.05). Fertilizer application did not appear to influence nematodes early in the season. However, in late season, organic fertilizers increased enrichment and structure indices and decreased channel index by the end of the zucchini cropping cycle.     

食细菌线虫对土壤微生物量和微生物群落结构的影响

线虫与微生物的相互作用研究往往是在悉生培养体系 (gnotobiotic microcosm)中进行 ,为了研究在自然或开放土壤条件下土壤线虫与微生物的相互作用 ,作者在开放盆栽体系中接种土壤食细菌线虫 (原小杆线虫 ,Protorhabditis sp) ,研究在小麦不同生育期、在有和无根系作用下食细菌线虫对土壤微生物量和微生物群落结构的影响。结果表明 :接种线虫分别使 SMBC、SMBN、SMBP提高了 2 6 .4 %、32 .9%、2 1.8% ,这种促进作用除个别无根系土和非根际土处理外 ,均达到显著性差异。根际土中的 SMBC、SMBN、SMBP>非根际土 >无根系土。从方差解释比例 v来看 ,SMBN受线虫的影响最大 (v=2 4 % )、其次是 SMBC(v=16 % )、然后是 SMBP(v=12 % ) ,线虫对 SMBC的促进作用在根际土中最突出。接种线虫对土壤细菌、真菌和放线菌的数量有明显的影响。在苗期的无根系土和根际土中 ,接种线虫显著降低了细菌的数量、特别在根际土中尤为突出 ,但在其它处理中却增加了细菌的数量。接种线虫对真菌和放线菌数量的促进作用比对细菌更为明显 ,接种线虫后真菌和放线菌数量的总平均值分别比未接种提高了 4 8.5 %和 6 8.2 % ,而细菌数量的总平均值没有变化。细菌数量与微生物量 C相关散点图表明二者相关性在根际土、非根际土和无根系土中均未     

Differential effects of food availability on population growth and fitness of three species of estuarine, bacterial-feeding nematodes

The significance of bottom-up controls on biological communities has been a long-standing topic of interest in ecology. However, before environmental effects on communities can be properly assessed, a thorough knowledge of the individual species' responses is required. We studied effects of food availability on population development and on different life-history traits in three species of bacterial-feeding nematodes, Diplolaimelloides oschei, Diplolaimelloides meyli (both Monhysteridae) and Pellioditis marina (Rhabditidae), which co-occur on macrophyte detritus in the Westerschelde Estuary (SW Netherlands). The bacteria Escherichia coli was offered in five food-availability treatments corresponding to initial cell densities from 3 × 10¹⁰ cells ml^− 1 to 3 × 10⁷ cells ml^− 1. The three bacterial-feeding nematode species studied here showed differential responses to food availability, which agreed with the general idea that Rhabditidae have extreme colonization abilities under very high food availability, while Monhysteridae tend to have a somewhat slower population development and comparatively lower food requirements. Several life-history traits, including juvenile mortality and development time, did not exhibit a clear food-availability dependence, but bioenergetics-related parameters did. Results on the F1 generation may, however, be affected by strong maternal effects on life-history traits of their progeny. Patterns of food-availability dependence of population increase and size at maturity were similar in P. marina. Both Diplolaimelloides species, however, exhibited a large body size at maturity but a very low population increase at the highest food availability, suggesting a trade-off between biomass and reproduction. Comparison with published data on other nematode species reveals that nematode responses to food availability as well as to other environmental factors are highly species-specific.     

Parasitism of Nematodes by the Fungus Hirsutella rhossiliensis as Affected by Certain Organic Amendments

Experiments were conducted to determine whether the addition of organic matter to soil increased numbers of bacterivorous nematodes and parasitic activity of the nematophagous fungus Hirsutella rhossiliensis. In a peach orchard on loamy sand, parasitism of the plant-parasitic nematode Criconemella xenoplax by H. rhossiliensis was slightly suppressed and numbers of C. xenoplax were not affected by addition of 73 metric tons of composted chicken manure/ha. In the laboratory, numbers of bacterivorous nematodes (especially Acrobeloides spp.) and fungivorous nematodes increased but parasitism of nematodes by H. rhossiliensis usually decreased with addition of wheat straw or composted cow manure to a loamy sand naturally infested with H. rhossiliensis. These results do not support the hypothesis that organic amendments will enhance parasitism of nematodes by H. rhossiliensis.     

Variability of water temperature may influence food-chain length in temperate streams

Nutrient enrichment may alter population dynamics of species in different ways depending on their life strategies. The aim of this study was to test the effect of different nutrient concentrations on the population development of two bacterivorous freshwater nematodes, Bursilla monhystera and Plectus aquatilis. Microcosms with autoclaved natural sand from a pristine stream (Fuirosos, NE of Spain) were enriched with different levels of phosphate, nitrate and ammonia as inorganic nutrients and glucose as a biodegradable dissolved organic carbon source. Although leaching of carbon and nutrients from the detritus fraction in the sediment initially may have overruled differences between treatments, later samplings revealed bottom-up control, with Bursilla monhystera abundances positively correlated to bacterial abundances at high nutrient concentrations. Nevertheless, there were several indications that nematodes in turn affected microbial abundance, most likely through excretion of ammonia and through grazing. In contrast to B. monhystera, Plectus aquatilis at high nutrient concentrations showed a unimodal abundance curve, while not increasing in abundance at low nutrient concentrations. Glucose enrichment did not have any stimulatory effect on either microbial or nematode abundances, probably as a result of unfavourable C:N:P stoichiometry. P enrichment, by contrast, stimulated microbial and Bursilla abundances. Our results indicate that episodic nutrient enrichment may affect populations of bacterial-feeding nematodes in the short term. Their longer-term dynamics may, however, be more dependent on leaching of carbon and nutrients from the pools of sediment-bound detritus.     

Pathogenicity and Interaction of Three Nematode Species on Six Bermudagrasses

The ring nematode (Criconemoides ornatus), stunt nematode (Tylenchorhynchus martini), and sting nematode (Belonolaimus Iongicaudatus) reproduced readily on six bermudagrasses (Common, ''U-3'', ''Tufcote'', ''Continental'', ''Tiffine'', and ''Tifdwarf''). Populations of a single nematode species influenced the population development of a second and third parasitic nematode species on a particular host plant. Activity of most nematodes adversely affected reproduction of other nematode species in mixed cultures. Generally, the number of fibrous roots produced by plants decreased as the number of nematode species in the treatments increased. Tifdwarf bermudagrass appeared to be more tolerant to C. ornatus and T. martini than other grasses tested.     

Attraction of four entomopathogenic nematodes to four white grub species

To better understand the differences in the efficacy of entomopathogenic nematode species against white grub species, we are studying the various steps of the infection process of entomopathogenic nematodes into different white grub species using nematode species/strains with particular promise as white grub control agents. In this study we compared the attraction of the entomopathogenic nematodes Steinernema scarabaei (AMK001 strain), Steinernema glaseri (NC1 strain), Heterorhabditis zealandica (X1 strain), and Heterorhabditis bacteriophora (GPS11 strain) to third-instars of the scarabs Popillia japonica, Anomala orientalis, Cyclocephala borealis, and Rhizotrogus majalis, and late-instar greater wax moth, Galleria mellonella, larvae. Individual larvae were confined at the bottom of 5.5 cm vertical sand columns, nematodes added to the sand surface after 24 h, and nematodes extracted after another 24 h. Nematode attraction to hosts was strongly affected by nematode species but the effect of insect species varied with nematode species. S. glaseri had a high innate dispersal rate (i.e., in absence of insects) and was strongly attracted to insects without significant differences among insect species. S. scarabaei had a very low innate dispersal rate so that even a strong relative response to insects resulted in low absolute dispersal rates toward insects. S. scarabaei tended to be most attracted to G. mellonella and least attracted to C. borealis. H. zealandica had a high innate dispersal rate but only responded weakly to insects without significant differences among species. H. bacteriophora had limited innate dispersal and only weakly responded to insects with G. mellonella tending to be the most attractive and C. borealis the least attractive insect. It has to be noted that we cannot exclude that the use of different rearing hosts (A. orientalis and P. japonica larvae for S. scarabaei, G. mellonella larvae for the other nematodes) might have had an impact on the nematodes dispersal and relative attraction behavior. This study indicates that host attractiveness and nematode dispersal rates may contribute but do not play a major role in the variability in white grub susceptibility and/or nematode virulence.     

Plant-parasitic Nematode Acetylcholinesterase Inhibition by Carbamate and Organophosphate Nematicides

The sensitivity of acetylcholinesterases (ACHE) isolated from the plant-parasitic nematodes Meloidogyne arenaria, M. incognita, and Heterodera glycines and the free-living nematode Caenorhabditis elegans to carbamate and organophosphate nematicides was examined. The AChE from plant-parasitic nematode species were more sensitive to carbamate inhibitors than was AChE from C. elegans, but response to the organophosphates was approximately equivalent. The sulfur-containing phosphate nematicides were poor inhibitors of nematode acetylcholinesterase, but treatment with an oxidizing agent greatly improved inhibition. Behavioral bioassays with living nematodes revealed a poor relationship between enzyme inhibition and expression of symptoms in live nematodes.     

Antagonists of Plant-parasitic Nematodes in Florida Citrus

In a survey of antagonists of nematodes in 27 citrus groves, each with a history of Tylenchulus semipenetrans infestation, and 17 noncitrus habitats in Florida, approximately 24 species of microbial antagonists capable of attacking vermiform stages of Radopholus citrophilus were recovered. Eleven of these microbes and a species of Pasteuria also were observed attacking vermiform stages of T. semipenetrans. Verticillium chlamydosporium, Paecilomyces lilacinus, P. marquandii, Streptomyces sp., Arthrobotrys oligospora, and Dactylella ellipsospora were found infecting T. semipenetrans egg masses. Two species of nematophagous amoebae, five species of predatory nematodes, and 29 species of nematophagous arthropods also were detected. Nematode-trapping fungi and nematophagous arthropods were common inhabitants of citrus groves with a history of citrus nematode infestation; however, obligate parasites of nematodes were rare.     

The effect of bacterivorous nematodes on detritus incorporation by macrofaunal detritivores: A study using stable isotope and fatty acid analyses

Several studies have investigated the effect of nematodes on microbial degradation of macrophyte detritus, but little is known about the potential effect of bacterivorous nematodes on productivity of macrofaunal detritivores. We investigated the influence of the bacterivorous nematode Rhabditis (Pellioditis) mediterranea on the incorporation rate of isotopically-labelled macroalgal detritus by the amphipod Paramoera chevreuxi in a laboratory microcosm. The fatty acid composition of amphipods, nematodes, and macroalgal detritus was characterized to help determine the pathway of detritus incorporation by amphipods. The potential importance of R. mediterranea as a source of highly unsaturated fatty acids (HUFAs) to higher trophic levels was also investigated.We found no clear evidence for an effect of nematodes on the incorporation rate of fresh macroalgal detritus by amphipods, although there was some indication that the type of detritus (i.e. the green Chaetomorpha sp. vs the red Polysiphonia sp.) is important in determining the nature and extent of the relationship between nematodes and macrofaunal detritivores. Fatty acid data indicated that nematodes did not contribute significantly to the diet of amphipods when detritus was present, and there was no evidence that nematodes affected the pathway of detritus incorporation by amphipods. Amphipods incorporated Chaetomorpha sp. detritus about 10 times faster than Polysiphonia sp. detritus despite the higher C/N ratio and lower HUFA content of the former. This suggests that other factors, such as the presence of grazer deterrent compounds, are important in determining the palatability of macroalgal detritus. Amphipods fed exclusively on nematodes retained high HUFA levels but suffered high mortality. The burrowing behaviour of nematodes is suggested as the most likely factor limiting their availability to predators.Results suggest a limited interaction between amphipods and bacterivorous nematodes in detrital food webs. Further experiments are needed to test the wider applicability of these findings to different nematode and macrofaunal taxa, and for different types of detritus.     

Effects of host nutrition on virulence and fitness of entomopathogenic nematodes: Lipid- and protein-based supplements in Tenebrio molitor diets

Entomopathogenic nematodes, Heterorhabditis indica and Steinernema riobrave, were tested for virulence and reproductive yield in Tenebrio molitor that were fed wheat bran diets with varying lipid- and protein-based supplements. Lipid supplements were based on 20% canola oil, peanut, pork or salmon, or a low lipid control (5% canola). Protein treatments consisted of basic supplement ingredients plus 0, 10, or 20% egg white; a bran-only control was also included. Some diet supplements had positive effects on nematode quality, whereas others had negative or neutral effects. All supplements with 20% lipids except canola oil caused increased T. molitor susceptibility to H. indica, whereas susceptibility to S. riobrave was not affected. Protein supplements did not affect host susceptibility, and neither lipid nor protein diet supplements affected reproductive capacity of either nematode species. Subsequently, we determined the pest control efficacy of progeny of nematodes that had been reared through T. molitor from different diets against Diaprepes abbreviatus and Otiorhynchus sulcatus. All nematode treatments reduced insect survival relative to the control (water only). Nematodes originating from T. molitor diets with the 0% or 20% protein exhibited lower efficacy versus D. abbreviatus than the intermediate level of protein (10%) or bran-only treatments. Nematodes originating from T. molitor lipid or control diets did not differ in virulence. Our research indicates that nutritional content of an insect host diet can affect host susceptibility to entomopathogenic nematodes and nematode fitness; therefore, host media could conceivably be optimized to increase in vivo nematode production efficiency.     

Susceptibility of the Plum Curculio,Conotrachelus nenuphar,to Entomopathogenic Nematodes

The plum curculio, Conotrachelus nenuphar, is a major pest of pome and stone fruit. Our objective was to determine virulence and reproductive potential of six commercially available nematode species in C. nenuphar larvae and adults. Nematodes tested were Heterorhabditis bacteriophora (Hb strain), H. marelatus (Point Reyes strains), H. megidis (UK211 strain), Steinernema riobrave (355 strain), S. carpocapsae (All strain), and S. feltiae (SN strain). Survival of C. nenuphar larvae treated with S. feltiae and S. riobrave, and survival of adults treated with S. carpocapsae and S. riobrave, was reduced relative to non-treated insects. Other nematode treatments were not different from the control. Conotrachelus nenuphar larvae were more susceptible to S. feltiae infection than were adults, but for other nematode species there was no significant insect-stage effect. Reproduction in C. nenuphar was greatest for H. marelatus, which produced approximately 10,000 nematodes in larvae and 5,500 in adults. Other nematodes produced approximately 1,000 to 3,700 infective juveniles per C. nenuphar with no significant differences among nematode species or insect stages. We conclude that S. carpocapsae or S. riobrave appears to have the most potential for controlling adults, whereas S. feltiae or S. riobrave appears to have the most potential for larval control.     

Diagnostic Probes Targeting the Major Sperm Protein Gene That May Be Useful in the Molecular Identification of Nematodes

Discrimination of closely related nematode species is typically problematic when traditional identification characteristics are prone to intraspecific variation. In this study, a molecular approach that can distinguish Pratylenchus penetrans and P. scribneri is described. The approach uses universal primers in conjunction with polymerase chain reaction (PCR) to amplify equivalent fragments of the major sperm protein (msp) gene from any nematode. This gene fragment typically includes an intron of variable sequence. The presence of this highly variable segment in an otherwise conserved gene sequence allows P. penetrans and P. scribneri to be distinguished by either a species-specific amplification or by dot-blot hybridization. The approach is potentially of general utility in species-specific identification of nematodes.     

Management of Rotylenchulus reniformis in Pineapple,Ananas comosus,by Intercycle Cover Crops

The effects of intercycle cover crops on Rotylenchulus reniformis population densities in pineapple were evaluated in one greenhouse and two field experiments. In the greenhouse, Crotalaria juncea, Brassica napus, and Tagetes erecta were planted for 3 months and then incorporated. These treatments were compared to weedy fallow with or without 1,3-dichloropropene (1,3-D) in three soils (Makawao fallow, Wahiawa fallow, and Wahiawa pineapple) naturally infested with R. reniformis. All cover crop incorporation suppressed R. reniformis numbers in cowpea more than did the weedy treatment in the Makawao (P < 0.05) but not in the Wahiawa soils. Crotalaria juncea treatment increased bacterivorous nematodes and nematode-trapping fungal population densities more than the other treatments in Makawao fallow and Wahiawa pineapple-planted soils. The field trials included the same plants as well as Sinapis alba. Treatments with Crotalaria juncea and 1,3-D maintained lower R. reniformis population densities on pineapple longer than other cover crops or weedy fallow treatments. Crotalaria juncea could have suppressed R. reniformis because it is a poor host and because it enhances nematode-trapping fungi when incorporated into soil. Treatment with 1,3-D reduced microbial activities but produced the greatest pineapple yield.     

Aggregative group behavior in insect parasitic nematode dispersal

Movement behavior of foraging animals is critical to the determination of their spatial ecology and success in exploiting resources. Individuals sometimes gain advantages by foraging in groups to increase their efficiency in garnering these resources. Group movement behavior has been studied in various vertebrates. In this study we explored the propensity for innate group movement behavior among insect parasitic nematodes. Given that entomopathogenic nematodes benefit from group attack and infection, we hypothesised that the populations would tend to move in aggregate in the absence of extrinsic cues. Movement patterns of entomopathogenic nematodes in sand were investigated when nematodes were applied to a specific locus or when the nematodes emerged naturally from infected insect hosts; six nematode species in two genera were tested (Heterorhabditis bacteriophora, Heterorhabditis indica, Steinernema carpocapsae, Steinernema feltiae, Steinernema glaseri and Steinernema riobrave). Nematodes were applied in aqueous suspension via filter paper discs or in infected insect host cadavers (to mimic emergence in nature). We discovered that nematode dispersal resulted in an aggregated pattern rather than a random or uniform distribution; the only exception was S. glaseri when emerging directly from infected hosts. The group movement may have been continuous from the point of origin, or it may have been triggered by a propensity to aggregate after a short period of random movement. To our knowledge, this is the first report of group movement behavior in parasitic nematodes in the absence of external stimuli (e.g., without an insect or other apparent biotic or abiotic cue). These findings have implications for nematode spatial distribution and suggest that group behavior is involved in nematode foraging.     

Evaluation of 31 Potential Biofumigant Brassicaceous Plants as Hosts for Three Meloiodogyne Species

Brassicaceous cover crops can be used for biofumigation after soil incorporation of the mowed crop. This strategy can be used to manage root-knot nematodes (Meloidogyne spp.), but the fact that many of these crops are host to root-knot nematodes can result in an undesired nematode population increase during the cultivation of the cover crop. To avoid this, cover crop cultivars that are poor or nonhosts should be selected. In this study, the host status of 31 plants in the family Brassicaceae for the three root-knot nematode species M. incognita, M. javanica, and M. hapla were evaluated, and compared with a susceptible tomato host in repeated greenhouse pot trials. The results showed that M. incognita and M. javanica responded in a similar fashion to the different cover cultivars. Indian mustard (Brassica juncea) and turnip (B. rapa) were generally good hosts, whereas most oil radish cultivars (Raphanus. sativus ssp. oleiferus) were poor hosts. However, some oil radish cultivars were among the best hosts for M. hapla. The arugula (Eruca sativa) cultivar Nemat was a poor host for all three nematode species tested. This study provides important information for chosing a cover crop with the purpose of managing root-knot nematodes.     

Comparison of complete mitochondrial genomes of pine wilt nematode Bursaphelenchus xylophilus and Bursaphelenchus mucronatus (Nematoda: Aphelenchoidea) and development of a molecular tool for species identification

We determined the complete mitochondrial genome sequences for Bursaphelenchus mucronatus, one species of pinewood nematode. The genome is a circular-DNA molecule of 14,583 bp (195 bp smaller than its congener Bursaphelenchus xylophilus) and contains 12 protein-coding genes (lacking atp8), 22 tRNA genes, and 2 rRNA genes encoded in the same direction, consistent with most other nematodes. Based on sequence comparison of mtDNA genomes, we developed a PCR-based molecular assay to differentiate B. xylophilus (highly pathogenic) and B. mucronatus (relatively less virulent) using species-specific primers. The molecular identification system employs multiplex-PCR and is very effective and reliable for discriminating these Bursaphelenchus species, which are economically important, but difficult to distinguish based on morphology. The comparison of the mitochondrial genomes and molecular identification system of the two species of Bursaphelenchus spp. should provide a rich source of genetic information to support the effective control and management (quarantine) of the pine wilt disease caused by pinewood nematodes.     

Characterization of Sialyl and Galactosyl Residues on the Body Wall of Different Plant Parasitic Nematodes

The plant parasitic nematodes Helicotylenchus multicinctus, Meloidogyne javanica, Tylenchulus semipenetrans, and Xiphinema index, differing in their host specificity and parasitic habits, were analyzed as to their cuticle surface sialyl, galaclosyl, and/or N-acetylgalactosaminyl residues. The procedure involved the selective oxidation of sialic acid and galactose/N-acetylgal-actosamine residues using periodate and galactose oxidase, respectively, to form reactive aldehyde groups. These functional groups were coupled directly with a new hydrazide-containing compound, the fluorescent reagent lissamine rhodamine-β-alanine hydrazide, or they were utilized to introduce DPN-groups to the nematode cuticle. The distribution of the DNP-tagged glycoconjugates was visualized by treating the nematodes with rabbit anti-DNP antibody and staining with fluorescein isothiocyanate (FITC)-labeled goat antirabbit IgG. Sialo residues were observed along the entire outer body wall of the first three aforementioned nematodes, but there were some differences in reaction among the various life stages within the species. In X. index, sialo residues were sited in the tail and head areas, mainly on the lips, oral opening, amphid apertures and stylet. Galactose oxidase treatments revealed galactose on N-acytylgalactosamine residues on T. sentipenetrans and X. index, but there were no indications that their presence was dependent on the developmental stage. Trypsin, pronase, and neuraminidase pretreatment completely abolished the fluorescence in T. semipenetrans but did not alter the sialo residue binding reaction in H. multicinctus or M. javanica, indicating possible differences in the outer body wall saccharide structure and composition between these nematodes. The existence and nature of sugar residues on the cuticle surface of nematodes could contribute to an understanding of the specific recognition by phytophagous nematodes of their host, and perhaps also of the virus transmission mechanism in those nematodes which serve as vectors.