Effects of Selected Nematicides on Hatching of Heterodera schachtii

Aldicarb, carbofuran, fensulfothion, and phenamiphos were tested in concentrations of 1-100 μg/ml for their effects on hatching of Heterodera schachtii. Exposure of cysts to 1 μg aldicarb or carbofuran/ml stimulated hatch whereas phenamiphos and, to a lesser degree, fensulfothion inhibited hatch. Addition of aldicarb to sugarbeet root diffusate or 4 mM zinc chloride suppressed activities of these hatching agents. Transfer of cysts previously treated with aldicarb or carbofuran to zinc chloride or water rapidly initiated hatch which finally exceeded the hatch from cysts not treated with the nematicides.     

Genetic and metabolic analysis of the carbofuran catabolic pathway in Novosphingobium sp. KN65.2

The widespread agricultural application of carbofuran and concomitant contamination of surface and ground waters has raised health concerns due to the reported toxic effects of this insecticide and its degradation products. Most bacteria that degrade carbofuran only perform partial degradation involving carbamate hydrolysis without breakdown of the resulting phenolic metabolite. The capacity to mineralize carbofuran beyond the benzofuran ring has been reported for some bacterial strains, especially sphingomonads, and some common metabolites, including carbofuran phenol, were identified. In the current study, the catabolism of carbofuran by Novosphingobium sp. KN65.2 (LMG 28221), a strain isolated from a carbofuran-exposed Vietnamese soil and utilizing the compound as a sole carbon and nitrogen source, was studied. Several KN65.2 plasposon mutants with diminished or abolished capacity to degrade and mineralize carbofuran were generated and characterized. Metabolic profiling of representative mutants revealed new metabolic intermediates, in addition to the initial hydrolysis product carbofuran phenol. The promiscuous carbofuran-hydrolyzing enzyme Mcd, which is present in several bacteria lacking carbofuran ring mineralization capacity, is not encoded by the Novosphingobium sp. KN65.2 genome. An alternative hydrolase gene required for this step was not identified, but the constitutively expressed genes of the unique cfd operon, including the oxygenase genes cfdC and cfdE, could be linked to further degradation of the phenolic metabolite. A third involved oxygenase gene, cfdI, and the transporter gene cftA, encoding a TonB-dependent outer membrane receptor with potential regulatory function, are located outside the cfd cluster. This study has revealed the first dedicated carbofuran catabolic genes and provides insight in the early steps of benzofuran ring degradation.     

Inhibition of Acetylcholinesterases from Aphelenchus avenae by Carbofuran and Fenamiphos

Exposure to carbofuran and fenamiphos for 72 hours reduced the numbers of active Aphelenchus avenae in aqueous suspension by > 75%. When nematicides were removed, many A. avenae exposed to carbofuran resumed normal movement but A. avenae treated with fenamiphos did not recover. Acetylcholinesterase (AChE) activity was suppressed by > 95% in nematodes treated with carbofuran or fenamiphos. However, 48 hours after treated nematodes had been placed in water, AChE activity in carbofuran treated populations was 98% of the levels in control nematodes. Nematodes that had been treated with fenamiphos showed only slight AChE recovery. The antidotes, atropine sulfate and 2-PAM, were largely ineffective in counteracting the toxic effects of the nematicides.     

Effect of Foliar Application of Carbofuran and a Related Compound on Plant-parasitic Nematodes under Greenhouse and Growth Chamber Conditions

Greenhouse and growth chamber studies were made to investigate the downward systemic nematicidal activity of carbofuran and its analog 2,3-dihydro-2,2-dimethyl-7-benzofuranyl [(di-n-butyl)-4-aminosulfenyl] carbamate against Meloidogyne incognita, Tylenchorhynchus claytoni, and Pratylenchus penetrans. Oxamyl was used as standard in tests with T. claytoni. Both carbofuran and its analog reduced all three plant-parasitic species when plant foliage was sprayed with chemical suspension of 1,200, 2,400 or 4,800 ppm. The studies show that fewer chemical applications were required to reduce populations of P. penetrans than to reduce populations of T. claytoni. Oxamyl was somewhat less active than either carbofuran or its analog.     

Constitutive but no Triops-induced differences in bet-hedging strategies for hatching in Daphnia

Since cladocerans from the genus Daphnia are known to have evolved several inducible defenses (morphological and life history shifts) against the notostracan predator Triops, we investigated whether hatching was also altered in response to Triops. We tested whether dormant eggs of Daphnia magna are able to detect Triops cancriformis kairomones in the water as a signal of predation pressure and alter their hatching response accordingly to avoid predation. We predicted that, in the presence of Triops kairomones, hatching fractions might be reduced (postponing hatching to a next growing season) and/or that hatching might peak earlier (increasing chances to reproduce before Triops becomes predatory). We also tested whether this response depended on the origin of the population. Ephippia from three D. magna populations, originating from one permanent lake and two temporary pond systems, were exposed to Triops kairomone and control treatments. We observed significant population differences in hatching patterns, both in terms of the fraction of eggs that hatch as well as the timing of hatching, with evidence for within-season bet-hedging through delayed hatching in the populations inhabiting temporary habitats. However, no indication was found that the populations also adjust their hatching pattern to the presence of Triops kairomones.     

Mineralization of Carbofuran by a Soil Bacterium

A bacterium, tentatively identified as an Arthrobacter sp., was isolated from flooded soil that was incubated at 35°C and repeatedly treated with carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranyl N-methylcarbamate). This bacterium exhibited an exceptional capacity to completely mineralize the ring-labeled ¹⁴C in carbofuran to ¹⁴CO₂ within 72 to 120 h in a mineral salts medium as a sole source of carbon and nitrogen under aerobic conditions. Mineralization was more rapid at 35°C than at 20°C. No degradation of carbofuran occurred even after prolonged incubation under anaerobic conditions. The predicted metabolites of carbofuran, 7-phenol (2,3-dihydro-2,2-dimethyl-7-benzofuranol) and 3-hydroxycarbofuran, were also metabolized rapidly. 7-Phenol, although formed during carbofuran degradation, never accumulated in large amounts, evidently because of its further metabolism through ring cleavage. The bacterium readily hydrolyzed carbaryl (1-naphthyl N-methylcarbamate), but its hydrolysis product, 1-naphthol, resisted further degradation by this bacterium.     

A Comparison of the Hatching of Juveniles from Cysts of Heterodera schachtii and H. trifolii

The effects of root diffusates of selected plants within the families Chenopodiaceae and Cruciferae and the hatching agent zinc chloride were tested for their effects on hatching and emergence of juveniles from cysts of Heterodera schachtii and a race of H. trifolii parasitic on Chenopodaceae and Cruciferae in The Netherlands. Although all diffusates strongly stimulated hatching of juveniles of H. schachtii, their effects on H. trifolii were less evident.     

Nematicidal activity of Haplophyllum tuberculatum and Plectranthus cylindraceus oils against Meloidogyne javanica

The potentials of Haplophyllum tuberculatum and Plectranthus cylindraceus oils to control Meloidogyne javanica were investigated in vitro and in a greenhouse. A mixture of Haplophyllum and Plectranthus oils (1:1) was highly toxic to M. javanica in vitro, as it killed all nematode juveniles and inhibited hatching of eggs at 12.5 μg/ml concentration after 24 h exposure time, as did carbofuran at the same concentration. In the green-house, tomatoes grown in soil treated with a combination (1:1) of the two oils developed fewer root galls than those grown in soil treated with higher doses of either oil. The oil mixture, at 2.5 and 5.0 μg/ml of soil, was not phytotoxic to tomato plants as evident from the appearance and height of plants after 12 weeks exposure time, compared to treatment over the same period at lower effective doses. The nematicidal activity of the combined essential oils was suggested by the presence of C₁₀ dienes, C₁₀ trienes and C₁₀ phenol.     

Stimulation of Autotrophic Ammonium Oxidation in Rice Rhizosphere Soil by the Insecticide Carbofuran

The application of the insecticide carbofuran (technical or formulated) to rice rhizosphere soil suspensions at 10 and 100 ppm (μg/ml) of active ingredient distinctly stimulated the autotrophic oxidation of ammonium. Evidence suggested that Nitrosomonas sp. was enriched in the presence of carbofuran. Formulated carbofuran (Furadan 3G) exhibited a more pronounced stimulation of ammonium oxidation than that exhibited by technical-grade (99.5%) carbofuran, a result which was attributed to the CaCO₃ present in the formulation.     

Chemical Control of Nematodes and Soil-borne Plant-Pathogenic Fungi on Cabbage Tranplants

Six general-purpose fumigants and one fungicide were applied by different methods and evaluated for control of nematode-fungus complexes on cabbage grown for transplant production. All chemicals reduced populations of nematodes and soil-borne fungi but varied greatly in effectiveness. Methyl bromide + chloropicrin (98% methyl bromide + 2% chloropicrin) (MBR-CP gas), DD + methyl isothiocyanate (DD-MENCS), methyl bromide + chloropicrin (67% methyl bromide + 31.75% cbloropicrin) (MBR-CP gel), and chloropicrin were more effective than sodium methyl dithiocarbamate (metham), pentachloronitrobenzene (PCNB), and potassium N-hydroxy-methyl-N-methyldithiocarbamate (Bunema) against Meloidogyne incognita. Populations of Pythium spp. and Fusarium spp. were reduced markedly by all treatments except PCNB. Plant growth, uniformity, and yield were greater when nematodes and fungi were controlled.     

Behavioral Effects of Carbofuran and Phenamiphos on Pratylenchus vulnus. III. Penetration and Development

Penetration of bean roots by Pratylenchus vulnus was inhibited by continuous exposure of the nematode to carbofuran and phenamiphos and by drenches of higher concentrations of these chemicals. The inhibition was explicable by inhibition of motility, dispersion, and attraction. If incubated in aerated distilled water, nematodes treated with carbofuran and phenamiphos recovered and reproduced as well as untreated nematodes. Foliar treatments were ineffective. Apparently, no basipetal transport of carbofuran and phenamiphos occurs in beans. Both nematicides arrested nematode development by interfering with egg production and transitions between life stages.     

Egg hatching of two locusts,Schistocerca gregaria and Locusta migratoria,in response to light and temperature cycles

The present study showed that the eggs of the desert locust, Schistocerca gregaria, and the migratory locust, Locusta migratoria, responded to photoperiod by hatching when placed on sand in the laboratory. S. gregaria mainly hatched during the dark phase and L. migratoria during the light phase. The importance of light as a hatching cue depended on the magnitude of the temperature change during the thermoperiod; photoperiod played a more important role in the control of hatching time in both species when the magnitude of the temperature change was small. In addition, the eggs of the two species that were covered with sand did not respond to photoperiod and hatched during both the light and dark phases, indicating that light did not penetrate through the sand. Because locust eggs are normally laid as egg pods and a foam plug is deposited between the egg mass and the ground surface, we tested a possibility that naturally deposited eggs perceived light through the foam plug. The eggs that were deposited and left undisturbed in the sand hatched during the light and dark phases at similar frequencies. These results suggest that the eggs of both locust species responded to light and controlled their hatching timing accordingly but would not use light as a hatching cue in the field. The evolutionary significance of the ability of eggs to respond to light in these locusts was discussed.     

Effect of Aldicarb,Ethoprop, and Carbofuran on Control of Coffee Root-knot Nematode,Meloidogyne exigua

Egg hatch of Meloidogyne exigua was significantly inhibited in 14 days pretreatment with aldicarb, ethoprop, or carbofnran at concentrations higher than 0.1 μg/ml; these eggs were found to delay hatch in 19 days posttreatment in ethoprop. Aldicarb and carbofuran solutions at concentrations greater than 0.1 μg/ml significantly decreased the motility and the life span of the second-stage juveniles; aldicarb was more toxic than carbofuran to the nematode. In a field test, aldicarb (Temik 10G), ethoprop (Mocap 10G), and carbofuran (Furadan 5G and Furadan Liquid 350F) significantly decreased M. exigua populations.     

Effects of Dosage Sequence on the Efficacy of Nonfumigant Nematicides, Plantain Yields,and Nematode Seasonal Fluctuations as influenced by Rainfall

Four nonfumigant nematicides applied three times during the wet season were used to study dosage sequence and nematicide effectiveness. Control of Helicotylenchus multicinctus (Cobb) Thorne and Meloidogyne javanica (Treub) Chitwood increased plantain (Musa AAB) yields. The nematicide (aldicarb, carbofuran, oxamyl, and miral) performance and yield response varied with dosage sequences. Applications of 2, 3, and 2 g ai/tree in March, July, and October (sequence I), respectively, gave greater control of M. javanica than did applications of 3, 2, and 2 g ai/tree in March, June, and September (sequence II), respectively. However, the high initial dose sequence was effective against H. multicinctus. Persistence of the different nematicides differed over the 14-month experimental period. Miral, aldicarb, and carbofuran were the most effective treatments against either species by the end of the wet and dry seasons. Dry season residual nematode populations were significantly lower in nematicide treated than in control plots. Yield increases over controls were 96.9, 90.1, 78.4, and 70.1% for carbofuran applied by sequence II, aldicarb by II and I, and oxantyl by II, respectively. Nematode populations directly fluctuated with rainfall and dropped to low (H. multicinctus) or to undetectable (M. javanica juveniles) levels during the dry season. Of the two nematodes studied, the more serious pest to plantain was H. multicinctus; it was tolerant to drought and survived the dry season in untreated soils.     

Bioaugmentation of carbofuran residues in soil using Burkholderia cepacia PCL3 adsorbed on agricultural residues

Burkholderia cepacia PCL3 (GenBank accession number of EF990634) is a carbofuran degrader isolated from phytoremediated rhizosphere soil in our laboratory. Free and the immobilized PCL3 on corncob and sugarcane bagasse were investigated for their abilities to degrade carbofuran in Basal Salt Medium (BSM) and soil microcosm. The reusability and survival of immobilized PCL3 in comparison to free cells were also examined. Short half-lives (t_1/2) of carbofuran of 3–4 d in BSM were obtained using the isolate PCL3 in both free and immobilized cell forms. Immobilized cells could survive (10⁶–10⁷ cfu ml^?1) through 30 d of incubation, while the number of free cells decreased continuously after 10 d. Immobilized B. cepacia PCL3 could be reused twice without loss in their abilities to degrade carbofuran in BSM, which suggested an advantage of using immobilized cell over free cell. Free and immobilized cells were augmented into soil and showed an effective capability to remediate carbofuran residues, both of which indicated by 5-folds decrease in carbofuran half-lives in augmented soil. Immobilization of PCL3 on corncob and sugarcane bagasse provided the possibilities of reusing the cells as well as improving the cell survival without decreasing carbofuran degradation activity.     

Adaptive difference in daily timing of hatch in two locust species,Schistocerca gregaria and Locusta migratoria: The effects of thermocycles and phase polyphenism

This study examined the effects of temperature and phase polyphenism on egg hatching time in the desert locust, Schistocerca gregaria, and the migratory locust, Locusta migratoria. The two species exhibited differences and similarities in hatching behavior when exposed to different temperature conditions. In 12-h thermocycles of various temperatures, the S. gregaria eggs hatched during the cryoperiod (low temperature period), whereas L. migratoria eggs hatched during the thermoperiod (high temperature period). The eggs of both species hatched during the species-specific period of the thermoperiod in response to a temperature difference as small as 1 °C. Furthermore, the locusts adjusted hatching time to a new thermal environment that occurred shortly before the expected hatching time. In both species, the hatching of the eggs was synchronized to a specific time of the day, and two hatching peaks separated by approximately 1 day were observed at a constant temperature after the eggs were transferred from thermocycles 3 days before hatching. Eggs laid by gregarious females hatched earlier than those laid by solitarious females in S. gregaria but this difference was not observed in L. migratoria.     

Effect of Foliar Applications of Carbofuran and a Related Compound on Plant-parasitic Nematodes under Microplot and Field Conditions

Studies were conducted to investigate the basipetal translocation of nematicidal activity from foliar treatments of carbofuran and its analog, 2,3-dihydro-2,2-dimethyl-7-benzofuranyl ([dibutylamino]thio)methyl carbamate, on corn in microplot studies and on tobacco and potato in field trials. Two and three foliar applications of either product at 2,400 μg/ml (9-20 Kg ai/Ha) significantly reduced populations of Pratylenchus penetrans in roots and populations of Tylenchorhynchus claytoni, Xiphinema americanum, and Hoplolaimus sp. in soil. In most cases there was no difference in control between two or three chemical applications. Foliar treatments with carbofuran were equivalent to, or better than, soil treatment, although rates of applications were different.     

Dispersion and Distribution of Pratylenchus scribneri and Hoplolaimus galeatus in Soybean Fields

Examination of dispersional characteristics of Pratylenchus scribneri and Hoplolaimus galeatus indicated that there were patches within soybean fields in which both survival and reproduction wexe enhanced in spite of apparent homogeneity of soil type and topography. Treatment with carbofuran reduced the patchiness (or increased the dispersion) for H. galeatus while it had the opposite effect for P. scribneri. P. scribneri was less highly dispersed in conventional tillage plots than in the zero tillage plots. Populations from quadrats contained entirely within the patches could be described by the normal distribution (in the case of P. scribneri) or by the Poisson distribution (in the case of H. galeatus), while populations from quadrats contained entirely outside the patches could be described by the Poisson distribution for both nematodes. None of the distributions tested (Poisson, normal, negative binomial, Neyman''s) gave an adequate fit when populations from both inside and outside the patches were considered together. In all instances, log₁₀ and ln transformations reduced the goodness of fit of the data to all of the distributions tested. Even with logarithmic transformations, parametric statistics were not appropriate for analysis of data in most instances.     

Granular Nematicides for Control of the Yam Nematode,Scutellonema bradys, and Relevant Residues in Raw Tubers

Four granular nentaticides were evaluated for control of the yam nematode, Scutellonema bradys (Steiner &LeHew) Andrassy, on Guinea yam, Dioscorea rotundata Poir, under field conditions prevelant in the tropics. A single application of nematicides (sidedressing) at the rate of 2 kg ai/ha as postplanting treatment at the onset of the rainy season depressed numbers of S. bradys attacking yams during the growing season and significantly increased tuber yields over untreated. The efficacy, based on the regression coefficient values of evaluated nematicides, was in the order of miral, carbofuran, aldicarb, and oxamyl (b = -75.9, -75.5, -72.1, and -65.9, respectively). Yam tuber yields increased by 136.9, 90.6, 87.9, and 85.3% over untreated (P = 0.05) in aldicarb, carbofuran, oxamyl, and miral treated plots, respectively. Residues in raw tubers pretreated with aldicarb, carbofuran, or miral were negligible (front less than 0.02 to 0.3 ppm) and far below the established tolerance levels (l.0 and 1.3 ppm for aldicarb and carbofuran, respectively) of a related crop in the United States. This is the first report on residues of systemic pesticides in yams.