Repeated Sampling to Determine the Precision of Estimating Nematode Population Densities

The first phase of this study involved repeated samplings of five fields using composite samples of 10, 20, 40, and 80 soil cores, to determine the precision of nematode assays. The second phase focused on randomly selecting two and four 2-ha subunits (data on Meloidogyne spp.) of 24 fields ranging from 6 to 40 ha and computing the precision of estimated means for these numbers ofsubunits versus the general field mean (based on all 2-ha subunits). Average numbers of nematodes from most samples containing Meloidogyne spp., Heterodera glycines, Helicotylenchus dihystera, Scutellonema brachyurum, and (or) Hoplolaimus galeatus were within 50% of the overall means. Coefficient of variation (CV) values were generally lower for 40 cores than for 10, 20, and 80 cores per sample. When data for all nematodes and fields were combined, this value was lowest for 40 and 80 cores. The CV values were higher for Meloidogyne spp. than for H. glycines. Means of two samplings increased the probability of obtaining numbers nearer the mean for that field than numbers from a single composite sample. For the second phase, population estimates of Meloidogyne spp. based on four 2-ha subunits generally were closer to field means than were those for two subunits. Sampling precision with these subunits diminished greatly in large fields with variable soils and (or) mixed cropping histories. Either two or four subunits gave population estimates within 3-20% of the field mean in most instances. The mean man hours required for sampling ca. 2-ha parcels of 4-20-ha fields was 0.54 hours.     

Effects of Temperature on Pratylenchus neglectus and on Its Pathogenicity to Barley

In a petri-dish study, development of the nematode Pratylenchus neglectus was observed every 4 days, and stage-specific development times were estimated, using a parameter estimation algorithm for a distributed-delay population model. The lower threshold temperature for development of a population of P. neglectus was 7.75 C. Temperatures above 25 C were unfavorable for this population on barley. Total numbers of P. neglectus in barley roots and associated soil in pots were greatest at 25 C and lower at temperatures above and below that level. There was no change in nematode numbers per gram of root as temperature increased between 24 C and 32 C because root weights decreased at higher temperatures. Restricted root mass may contribute to the lower total nematode population levels at higher temperature. Maximum number of nematodes moved through a 2-cm layer of sand on a Baermann funnel at about 20 C; lowest number of nematodes moved at 10 C and 30 C.     

Susceptibility of early larval stages of Pseudaletia unipuncta and Spodoptera exigua (Lepidoptera: Noctuidae) to the entomogenous nematode Steinernema feltiae (Rhabditida: Steinernematidae)

Early stages (neonate to 7- or 8-day-old larvae) of Spodoptera exigua and Pseudaletia unipuncta were exposed to the entomogenous nematode, Steinernema feltiae, at concentrations of 0, 10, 25, 60, 100, or 200 nematodes per larva. Larvae of both species were susceptible to nematode infections. However, neonate larvae of S. exigua were significantly less susceptible to nematode infection than 3- or 8-day-old larvae at or above 50 nematodes per larva. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 68 to 74% while mortalities of 3- and 8-day-old larvae ranged from 91 to 100%. The results with P. unipuncta showed similar trends as described for S. exigua, albeit at a lower mortality level and usually with no statistical differences. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 34 to 44% while mortalities of 7-day-old larvae ranged from 32 to 91%.     

Plant-parasitic Nematode Communities and Their Associations with Soil Factors in Organically Farmed Fields in Minnesota

A survey was conducted to determine the assemblage and abundance of plant-parasitic nematodes and their associations with soil factors in organically farmed fields in Minnesota. A total of 31 soil samples were collected from southeast (SE), 26 samples from southwest (SW), 28 from west-central (WC), and 23 from northwest (NW) Minnesota. The assemblage and abundance of plant-parasitic nematodes varied among the four regions. The soybean cyst nematode, Heterodera glycines, the most destructive pathogen of soybean, was detected in 45.2, 88.5, 10.7, and 0% of organically farmed fields with relative prominence (RP) values of 10.3, 26.5, 0.6, and 0 in the SE, SW, WC, and NW regions, respectively. Across the four regions, other common genera of plant-parasitic nematodes were Helicotylenchus (42.6, RP value, same below), Pratylenchus (26.9), Tylenchorhynchus and related genera (9.4), Xiphinema (5.6), and Paratylenchus (5.3). Aphelenchoides, Meloidogyne, Hoplolaimus, Mesocriconema, and Trichodorus were also detected at low frequencies and/or low population densities. The similarity index of plant-parasitic nematodes between two regions ranged from 0.44 to 0.71 and the similarity increased with decreasing distance between regions. The densities of most plant-parasitic nematodes did not correlate with measured soil factors (organic matter, pH, texture). However, the densities of Pratylenchus correlated negatively with % sand, and Xiphinema was correlated negatively with soil pH.     

Properties of glutathione peroxidase from the hepatopancreas of freshwater prawn Macrobrachium malcolmsonii

Oxidative stress stimulates production of hydrogen peroxide and organic hydroperoxides which are involved in lipid peroxidation and oxidative damage to protein. Glutathione peroxidase protects against oxidative stress. Quantitative data on glutathione peroxidase of crustaceans are very limited and so this study was performed to determine the assay condition of glutathione peroxidase in crustaceans. Particularly, some properties of hepatic selenium dependent glutathione peroxidase and total glutathione peroxidase of the freshwater prawn Macrobrachium malcolmsonii were examined. The optimal pH for Se-GSH-Px was 7.5 and for total GSH-Px was 8.0. The activation energy (E_a) was found to be 9.14 Kcal/mol for Se-GSH-Px and 10.96 Kcal/mol for total GSH-Px. The energy inhibition (E_i) value for Se-GSH-Px was 14.36 Kcal/mol and 13.71 Kcal/mol for total GSH-Px. The Km values were also determined at various concentrations of substrates (GSH, CHP, H₂O₂) for both enzymes.     

Sensitivity of Meloidogyne incognita and Rotylenchulus reniformis to Abamectin

Avermectins are macrocyclic lactones produced by Streptomyces avermitilis. Abamectin is a blend of B_1a and B_1b avermectins that is being used as a seed treatment to control plant-parasitic nematodes on cotton and some vegetable crops. No LD₅₀ values, data on nematode recovery following brief exposure, or effects of sublethal concentrations on infectivity of the plant-parasitic nematodes Meloidogyne incognita or Rotylenchulus reniformis are available. Using an assay of nematode mobility, LD₅₀ values of 1.56 μg/ml and 32.9 μg/ml were calculated based on 2 hr exposure for M. incognita and R. reniformis, respectively. There was no recovery of either nematode after exposure for 1 hr. Mortality of M. incognita continued to increase following a 1 hr exposure, whereas R. reniformis mortality remained unchanged at 24 hr after the nematodes were removed from the abamectin solution. Sublethal concentrations of 1.56 to 0.39 μg/ml for M. incognita and 32.9 to 8.2 μg/ml for R. reniformis reduced infectivity of each nematode on tomato roots. The toxicity of abamectin to these nematodes was comparable to that of aldicarb.     

Distribution of Pratylenchus scribneri between Root and Soil Habitats

The abundance of Pratylenchus scribneri in soil and root habitats was compared in potato and corn plots during 1986-88. Nematodes were extracted from 100-cm³ soil samples and the roots contained within the samples. The percentage of the population recovered from soil, similar among years and crops, averaged ca. 50% at the beginning and end of the growing season and ca. 20% from early to late season. Proportionately more adults and fourth-stage juveniles than younger stages were located outside roots until harvest. In a related study, nematodes were isolated from the roots, root surfaces, and soil associated with roots of whole corn and potato plants sampled from the field. Nematode population estimates calculated from the whole plant samples were generally lower than those based on soil cores, but showed similar patterns of population growth. Nematode density per gram dry weight was highest in roots, intermediate on root surfaces, and lowest in soil. Estimates of the absolute abundance of nematodes in each of the three habitats were highest in roots or soil, depending on the sampling date, and lowest on root surfaces. This study demonstrates that P. scribneri inhabits soil environments even when host roots are present and illustrates the importance of considering all possible habitats when estimating the size of Pratylenchus spp. populations.     

Hirschmanniella spp. in Rice Fields of Vietnam

Root and soil samples from one-crop and two-crop rice fields were collected in a survey for Hirschmanniella spp. in Vietnam during 1978-80. Hirschmanniella spp. were found in 50-78% of the soil samples and 98-100% of the root samples collected. Population densities of nematodes in root systems were lowest at posttransplanting and highest at heading time. Numbers of nematodes inside roots increased 20-22 times from transplanting to heading in fields with both crop sequences. Population densities of Hirschmanniella spp. in two-crop rice fields were more than twice those in one-crop rice fields.     

Interactions between Nematodes and Earthworms: Enhanced Dispersal of Steinernema carpocapsae

Dispersal of the nematode Steinernema carpocapsae (All strain), applied on the top or the bottom of soil columns, was tested in the presence or absence of two earthworm species, Lumbricus terrestris or Aporrectodea trapezoides. Nematode dispersal was estimated after a 2-week period with a bioassay against the greater wax moth, Galleria mellonella. Vertical dispersal of nematodes was increased in the presence of earthworms. When nematodes were placed on the surface of soil columns, significantly more nematodes dispersed to the lower half of the columns when either earthworm species was present than when earthworms were not present. When nematodes were placed on the bottom of soil columns, significantly more nematodes dispersed to the upper half of the columns when L. terrestris was present than when A. trapezoides was present or in the absence of earthworms. Because nematodes were found on the exterior and in the interior of earthworms, nematode dispersal may be enhanced by direct contact with the earthworms.     

Granulosis virus in the intestinal lumen of Neoaplectana carpocapsae: Retention of infectivity after treatment with formaldehyde or high pH

High pH values (>11.0) cause the dissolution of occlusion bodies of the granulosis virus (GV) of Pseudaletia unipuncta and subsequent inactivation of the virus within 24 hr. The GV is also inactivated within 48 hr by 0.04% formaldehyde. The GV is found in the intestinal lumen of infective third stage nematodes (dauer juveniles) of Neoaplectana carpocapsae when development occurs in GV-infected hosts. The GV in these dauer juveniles retains its infectivity even when the nematodes are placed into an alkaline solution with pH values of 11.1 or 12.1 or in 0.04% formaldehyde up to 336 hr. However, significant loss of infectivity of GV occurs when the nematodes are in formaldehyde but not at high pH values. The dauer juveniles are ensheathed by the second stage cuticle. This cuticle probably protects the GV in the intestinal lumen of the nematode from the high pH and formaldehyde.     

Population Densities of Nematodes Under Seven Tillage Regimes

Numbers of plant-parasitic nematodes on corn growing under seven tillage regimes were monitored. Differences among treatments occurred for Helicotylenchus pseudorobustus, Pratylenchus spp., Xiphinema americanum, dorylaimids, and total numbers of nematodes. Except with members of the Tylenchinae, highest densities occurred in no-till ridge plots and lowest numbers occurred in spring- and fall-plowed pots.     

Identification and Inhibitory Mechanism of Angiotensin I-Converting Enzyme Inhibitory Peptides Derived from Bovine Hemoglobin

Angiotensin I-converting enzyme (ACE, EC.3.4.15.1) inhibitory peptide is an efficacious therapy for hypertension. In this study, four dipeptides, TY, FD, FL and FG, were identified from the desalted fraction of bovine hemoglobin hydrolysate, obtained by in vitro simulated gastrointestinal digestion, via liquid chromatography-tandem mass spectrometry (LC-MS/MS). The IC₅₀ value of TY and FL are 96.43?±?6.17 and 290.66?±?57.92 μM, respectively. The result of molecular docking indicated that TY occupied the ACE subsite S1 and S1′ with a lowest estimated binding energy of ?9.96 Kcal/mol, while FL occupied the subsite S5 with a lowest estimated binding energy of ?9.37 Kcal/mol. The subsite S1′ and S2′ are closer to the ACE active center (Zn²⁺) than S5, and the lowest estimated binding energy of TY is lower than that of FL. This work provided new ACE-inhibitory peptides derived from bovine hemoglobin hydrolysate and explained their inhibitory mechanism.     

Effect of Hirschmanniella caudacrena on the Submersed Aquatic Plants Ceratophyllum demersum and Hydrilla verticillata

In vitro pathogenicity tests demonstrated that Hirschmanniella caudacrena is pathogenic to Ceratophyllum demersum (coontail). Symptoms were chlorotic tissue, deformed stems, and, finally, death of the plant. Inoculum densities of 500 nematodes per 5-cm-long cutting in a test tube containing 50 ml of water resulted in death and decay of some of the cuttings within 8 weeks; 100 nematodes killed the plants in 12 weeks, and 50 and 25 nematodes killed them in 16 weeks. The lowest inoculum level of 10 nematodes did not seriously affect the plants at 16 weeks when the experiment was terminated. A second test conducted outdoors in glass jars containing 3 liters of water and two cuttings weighing a total of 15 g fresh weight showed damage, but results were not statistically significant. Hydrilla verticillata inoculated with H. caudacrena was not affected seriously.     

Phosphorus acquisition from phytate depends on efficient bacterial grazing, irrespective of the mycorrhizal status of Pinus pinaster

Background and aims

Phosphorus from phytate, although constituting the main proportion of organic soil P, is unavailable to plants. Despite the well-known effects of rhizosphere trophic relationships on N mineralization, no work has been done yet on P mineralization. We hypothesized that the interactions between phytate-mineralizing bacteria, mycorrhizal fungi and bacterial grazer nematodes are able to improve plant P use from phytate.

Methods

We tested this hypothesis by growing Pinus pinaster seedlings in agar containing phytate as P source. The plants, whether or not ectomycorrhizal with the basidiomycete Hebeloma cylindrosporum, were grown alone or with a phytase-producing bacteria Bacillus subtilis and two bacterial-feeder nematodes, Rhabditis sp. and Acrobeloides sp. The bacteria and the nematodes were isolated from ectomycorrhizal roots and soil from P. pinaster plantations.

Results

Only the grazing of bacteria by nematodes enhanced plant P accumulation. Although plants increased the density of phytase-producing bacteria, these bacteria alone did not improve plant P nutrition. The seedlings, whether ectomycorrhizal or not, displayed a low capacity to use P from phytate.

Conclusions

In this experiment, the bacteria locked up the phosphorus, which was delivered to plant only by bacterial grazers like nematodes. Our results open an alternative route for better utilization of poorly available organic P by plants.     

Penetration of Crotalaria juncea,Dolichos lablab,and Sesamum indicum Roots by Meloidogyne javanica

Penetration of Crotalaria juncea (PI 207657 and cv. Tropic Sun) Dolichos lablab cv. Highworth, and Sesamum indicum by juveniles (J2) of Meloidogyne javanica was assessed to investigate the mechanism by which these plants may reduce nematode numbers in the field. Growth chamber experiments were conducted at 25 C, with vials containing 90 g sand infested with 450 J2; tomato (UC 204 C) was included as a susceptible host. Fifteen days after inoculation, roots were stained and the nematodes within stained roots were counted. Both C. juncea lines were highly resistant to penetration, as they contained significantly fewer nematodes per cm of root and per root system than the other plants. Although containing more nematodes per cm of root than C. juncea, S. indicum and D. lablab had significantly fewer nematodes per root system and per cm of root than tomato. Roots were significantly longer in the plants with the lowest nematode penetration. Although C. juncea, D. lablab, and S. indicum may have potential utility as cover or rotation crops in soil infested with M. javanica, further quantitative information on the reproduction of M. javanica and other nematodes in these plants is needed.     

Horizontal Transmission of Bursaphelenchus xylophilus between Sexes of Monochamus alternatus

Four experiments were conducted using nematode-infested and nematode-free adults of the cerambycid beetle, Monochamus alternatus, to determine horizontal transmission pathways of Bursaphelenchus xylophilus. When nematode-infested beetles of one sex and nematode-free beetles of the opposite sex were paired in containers for 48 or 72 hours, the number of nematodes carried by nematode-free beetles tended to increase with increased number of nematodes carried by nematode-infested beetles. The nematodes acquired by "nematode-free" beetles could be transmitted to pine. A female beetle that received 13 nematodes from a male transmitted one nematode to a Pinus densiflora bolt via an oviposition wound. When the nematode-infested and nematode-free beetles were observed continuously, it was observed that the number of nematodes carried by nematode-free beetles at the end of the first sexual mounting increased as the number of nematodes carried by nematode-infested beetles just before mounting increased. The number of nematodes transferred to nematode-free beetles was positively related to duration time of mounting. There was no difference in transmission efficacy between male-to-female transmission and female-to-male transmission. The horizontal transmission pathways are discussed relative to the persistence of B. xylophilus in resistant pine forests and the control of pine wilt disease.     

Quantitative Digital Imaging of Banana Growth Suppression by Plant Parasitic Nematodes

A digital camera fitted with a hemispherical lens was used to generate canopy leaf area index (LAI) values for a banana (Musa spp.) field trial with the aim of establishing a method for monitoring stresses on tall crop plants. The trial in Uganda consisted of two cultivars susceptible to nematodes, a plantain, Gonja manjaya and an East African Highland banana, Mbwazirume, plus a nematode resistant dessert banana, Yangambi km5. A comparative approach included adding a mixed population of Radopholus similis, Helicotylenchus multicinctus and Meloidogyne spp. to the soil around half the plants of each cultivar prior to field planting. Measurements of LAI were made fortnightly from 106 days post-planting over two successive cropping cycles. The highest mean LAI during the first cycle for Gonja manjaya was suppressed to 74.8±3.5% by the addition of nematodes, while for Mbwazirume the values were reduced to 71.1±1.9%. During the second cycle these values were 69.2±2.2% and 72.2±2.7%, respectively. Reductions in LAI values were validated as due to the biotic stress by assessing nematode numbers in roots and the necrosis they caused at each of two harvests and the relationship is described. Yield losses, including a component due to toppled plants, were 35.3% and 55.3% for Gonja manjaya and 31.4% and 55.8% for Mbwazirume, at first and second harvests respectively. Yangambi km5 showed no decrease in LAI and yield in the presence of nematodes at both harvests. LAI estimated by hemispherical photography provided a rapid basis for detecting biotic growth checks by nematodes on bananas, and demonstrated the potential of the approach for studies of growth checks to other tall crop plants caused by biotic or abiotic stresses.     

Winter Survival of Pratylenchus scribneri

Population densities of Pratylenchus scribneri in a Plainfield loamy sand soil were sampled from 1 October to 1 May for 4 years. From May to October of each year, the site was planted to Russet Burbank potato and Wis 4763 corn. Percentages of change in population densities of nematodes were computed on the basis of number of nematodes present on 1 October. The decline of P. scribneri between growing seasons was nonlinear, with most mortality occurring in the autumn before the soil froze. Winter survival, defined as the percentage of change in population densities from 1 October to 1 May the following year, ranged from 50 to 136% for nematodes in corn plots and from 15 to 86% for nematodes in potato plots. There was no difference in survival of nematodes of different life stages or among root and soil habitats. Winter survival of nematodes was density-dependent in 3 of 4 years in corn plots and in 1 of 4 years in potato plots. Although predators were present, their abundance was not correlated with the winter survival of nematodes. Cumulative and average snow cover was correlated with the survival of nematodes associated with corn but not with potato. No relationships between other climatic factors and survivorship were detected.     

Effect of Crotalaria juncea Amendment on Nematode Communities in Soil with Different Agricultural Histories

Effect of sunn hemp (Crotalaria juncea) hay amendment on nematode community structure in the soil surrounding roots of yellow squash (Cucurbita pepo) infected with root-knot nematodes was examined in two greenhouse experiments. Soils were from field plots treated long-term (LT) with yard-waste compost or no yard-waste compost in LT experiment, and from a short-term (ST) agricultural site in ST experiment. Soils collected were either amended or not amended with C. juncea hay. Nematode communities were examined 2 months after squash was inoculated with Meloidogyne incognita. Amendment increased (P < 0.05) omnivorous nematodes in both experiments but increased only bacterivorous nematodes in ST experiment (P < 0.05), where the soil had relatively low organic matter (<2%). This effect of C. juncea amendment did not occur in LT experiment, in which bacterivores were already abundant. Fungivorous nematodes were not increased by C. juncea amendment in either experiment, but predatory nematodes were increased when present. Although most nematode faunal indices, including enrichment index, structure index, and channel index, were not affected by C. juncea amendment, structure index values were affected by previous soil organic matter content. Results illustrate the importance of considering soil history (organic matter, nutrient level, free-living nematode number) in anticipating changes following amendment with C. juncea hay.     

Evaluating the predatory potential of carnivorous nematodes against Rotylenchulus reniformis and Meloidogyne incognita

Predatory behavior of a nematode is usually determined through gut content observation or prey delimitation counts. In this experiment, Mononchus and Neoactinolaimus predation of Rotylenchulus reniformis or Meloidogyne incognita was determined using a PCR-based nematode gut content analysis. Soil samples naturally infested with Mononchus were placed in tubes and potential prey nematodes R. reniformis, M. incognita, or a mixture of both were introduced. The gut contents of Mononchus were assayed for the DNA from R. reniformis or M. incognita using PCR specific primers. A higher % of Mononchus tested positive for DNA of R. reniformis than for M. incognita when the prey were added alone. However, when provided with both prey species, Mononchus was tested positive for DNA of M. incognita more frequently than for R. reniformis. Percent Mononchus testing positive for DNA of R. reniformis correlated positively with the abundance of R. reniformis, but this relationship was not observed between Mononchus and M. incognita. Neoactinolaimus was added to aqueous solution containing a mixture of free-living nematodes and R. reniformis. More Neoactinolaimus tested positive for DNA of R. reniformis than other predatory or omnivorous nematodes in the same samples. Based on regression analysis, the presence of fungivorous and other predatory nematodes in the soil could distract Neoactinolaimus from predation on R. reniformis. Our results suggested that Prismatolaimus, Mesodiplogasteroides and Eudorylaimus could also prey on R. reniformis. Although less than 40% of the predatory or omnivorous nematodes tested preyed on R. reniformis, this level of predation could contribute to reducing the population densities of plant-parasitic nematodes in the soil.