A simple extraction method was developed for obtaining the white tip nematode, Aphelenchoides besseyi Christie, from single seeds of rice, Oryza sativa. The method was as follows: Individual rice seeds were split longitudinally and then transferred into single pipet tips. Tips containing a split seed were then singly placed upright in glass vials with water to extract the nematodes. This method was more efficient than the Baermann funnel technique and allowed nearly 100% recovery of living A. besseyi from single rice seeds within 4 hours.
Aphelenchoides besseyi; assay; extraction; methodology; nematode; nematode load; Oryza sativa; rice; white tip disease
To reduce the risks associated with global transport of wood infested with pinewood nematode Bursaphelenchus xylophilus, microwave irradiation was tested at 14 temperatures in replicated wood samples to determine the temperature that would kill 99.9968% of nematodes in a sample of ≥ 100,000 organisms, meeting a level of efficacy of Probit 9. Treatment of these heavily infested wood samples (mean of > 1,000 nematodes/g of sapwood) produced 100% mortality at 56 °C and above, held for 1 min. Because this “brute force” approach to Probit 9 treats individual nematodes as the observational unit regardless of the number of wood samples it takes to treat this number of organisms, we also used a modeling approach. The best fit was to a Probit function, which estimated lethal temperature at 62.2 (95% confidence interval 59.0-70.0) °C. This discrepancy between the observed and predicted temperature to achieve Probit 9 efficacy may have been the result of an inherently limited sample size when predicting the true mean from the total population. The rate of temperature increase in the small wood samples (rise time) did not affect final nematode mortality at 56 °C. In addition, microwave treatment of industrial size, infested wood blocks killed 100% of > 200,000 nematodes at ≥ 56 °C held for 1 min in replicated wood samples. The 3rd-stage juvenile (J3) of the nematode, that is resistant to cold temperatures and desiccation, was abundant in our wood samples and did not show any resistance to microwave treatment. Regression analysis of internal wood temperatures as a function of surface temperature produced a regression equation that could be used with a relatively high degree of accuracy to predict internal wood temperatures, under the conditions of this study. These results provide strong evidence of the ability of microwave treatment to successfully eradicate B. xylophilus in infested wood at or above 56 °C held for 1 min.
Pinewood nematode; quarantine; microwave; dielectric heating; international trade; embargo; eradication; Probit 9; International Standard of Phytosanitary Measures No. 15
A PCR-based diagnostic assay was developed for early detection and identification of Aphelenchoides fragariae directly in host plant tissues using the species-specific primers AFragFl and AFragRl that amplify a 169-bp fragment in the internal transcribed spacer (ITS1) region of ribosomal DNA. These species-specific primers did not amplify DNA from Aphelenchoides besseyi or Aphelenchoides ritzemabosi. The PCR assay was sensitive, detecting a single nematode in a background of plant tissue extract. The assay accurately detected A. fragariae in more than 100 naturally infected, ornamental plant samples collected in North Carolina nurseries, garden centers and landscapes, including 50 plant species not previously reported as hosts of Aphelenchoides spp. The detection sensitivity of the PCR-based assay was higher for infected yet asymptomatic plants when compared to the traditional, water extraction method for Aphelenchoides spp. detection. The utility of using NaOH extraction for rapid preparation of total DNA from plant samples infected with A. fragariae was demonstrated.
Aphelenchoides fragariae; detection; diagnosis; foliar nematode; ITS1; method; NaOH; ornamental host; PCR; rDNA
Population dynamics of A. ritzemabosi and D. dipsaci were studied in two alfalfa fields in Wyoming. Symptomatic stem-bud tissue and root-zone soil from alfalfa plants exhibiting symptoms of D. dipsaci infection were collected at intervals of 3 to 4 weeks. Both nematodes were extracted from stem tissue with the Baermann funnel method and from soil with the sieving and Baermann funnel method. Soil moisture and soil temperature at 5 cm accounted for 64.8% and 61.0%, respectively, of the variability in numbers of both nematodes in soil at the Big Horn field. Also at the Big Horn field, A. ritzemabosi was found in soil on only three of the 14 collection dates, whereas D. dipsaci was found in soil on 12 dates. Aphelenchoides ritzemabosi was found in stem tissue samples on 9 of the 14 sampling dates whereas D. dipsaci was found on all dates. Populations of both nematodes in stem tissue peaked in October, and soil populations of both peaked in January, when soil moisture was greatest. Numbers of D. dipsaci in stem tissue were related to mean air temperature 3 weeks prior to tissue collection, while none of the climatic factors measured were associated with numbers of A. ritzemabosi. At the Dayton field, soil moisture plus soil temperature at 5 cm accounted for 98.2% and 91.4% of the variability in the soil populations of A. ritzemabosi and D. dipsaci, respectively. Aphelenchoides ritzemabosi was extracted from soil at two of the five collection dates, compared to extraction of D. dipsaci at three dates. Aphelenchoides ritzemabosi was collected from stem tissue at six of the seven sampling dates while D. dipsaci was found at all sampling dates. The only environmental factor that was associated with an increase in the numbers of both nematodes in alfalfa stem tissue was total precipitation 1 week prior to sampling, and this occurred only at the Dayton field. Numbers of A. ritzemabosi in stem tissue appeared to be not affected by any of the environmental factors studied, while numbers of D. dipsaci in stem tissue were associated with cumulative monthly precipitation, snow cover at time of sampling, and the mean weekly temperature 3 weeks prior to sampling. Harvesting alfalfa reduced the numbers of A. ritzemabosi at the Big Horn field and both nematodes at the Dayton field.
alfalfa; alfalfa stem nematode; Aphelenchoides ritzemabosi; chrysanthemum foliar nematode; climate; distribution; Ditylenchus dipsaci; Medicago sativa; nematode; sampling; seasonal fluctuations
Aphelenchoides besseyi, the nematode causal agent of white-tip disease of rice, was recovered from 5.5% of 474 seed samples obtained from rice seed warehouses in Louisiana. Laboratory tests in which A. besseyi-infested rice seed was treated with Phostoxin®, a compound used for control of insects in stored grain, indicate that it also has nematicidal properties. In 18-week-duration greenhouse tests, populations of A. besseyi increased 4-5-fold on the cultivars Saturn and Melrose and 3-fold on Nova '76. Green weights of Nova '76 plants inoculated with A. besseyi and Sclerotium oryzae, the causal agent of rice stem rot, were significantly reduced below those of plants inoculated with either organism alone or with distilled water. Weights of Melrose plants were reduced significantly by treatments with A. besseyi alone and A. besseyi plus S. oryzae, but not by S. oryzae alone. Saturn plant weights were not reduced significantly by either organism alone or by the two in combination.
Oryza sativa; host suitability; nematode-fungus interaction; white-tip disease; chemical control
The sugar-flotation-sieving (SFS) and Baermann-funnel (BF) methods were compared for nematode extraction efficiency. The SFS method recovered nematodes from more trophic groups whereas greater total numbers of individuals were recovered by BF. In a test to validate the efficiency of SFS, virtually 100% of the nematodes added to desert soil prior to extraction were recovered by four consecutive SFS washings of each soil sample. Estimations of nematode biomass in desert soils based on numbers of nematodes extracted by the two methods were similar unless there was large reserve of eggs in the soil. The biomass of nematodes from a Colorado desert soil was 0.9 g/m² as determined by both methods, whereas BF gave 0.17 g/m² for nematodes from a Mojave desert soil as compared to 0.9 g/m² with SFS.
In a survey of potato and rotational crops on Prince Edward Island, Canada, the cyst stage of the clover cyst nematode, Heterodera trifolii, was found in 43 of 63 sites sampled; however, only 12% of the cysts contained eggs. The root lesion nematode, Pratylenchus penetrans, was the dominant plant parasitic nematode and was found in 56 sites. Extraction of cysts from soil was similar using either the Schuiling centrifuge or the Fenwick can method, although the former was more convenient to use. The modified Baermann funnel method was not efficient for detecting the clover cyst nematode in soil.
Baermann funnel; clover cyst nematode; genwick can; Heterodera trifolii; method; nematode; Pratylenchus penetrans; root lesion nematode; Schuiling centrifuge; survey
Experiments to determine the effects of extraction techniques and the influence of shipping on extraction of Heterodera glycines life stages gave variable results. Shipping did not significantly affect numbers of nematodes extracted. More second-stage juveniles (J2) were extracted with Baermann funnels than with an elutriator, probably because incubation of encysted eggs on the Baermann funnel for 1 week allowed hatching to occur. Sieving was more efficient than elutriation for extracting cysts. Adding air agitation to the water pressure during elutriation increased extraction efficiency of cysts but not J2. Sample sizes of 250 cm³ and 500 cm³ did not influence extraction efficiency of cysts; however, sample size did influence extraction of J2.
Baennann funnel; centrifugal flotation; cyst; egg; elutriation; extraction; Heterodera glycines; methods; nematode; second-stage juvenile; shipping; soybean cyst nematode
Guidelines are suggested to implement the introduction of beneficial insect-parasitic nematodes into the United States from abroad. These suggestions result from experiences and research with these and other biological control agents and from the current need for procedures to import nematodes. Subjects considered are need to import, foreign exploration, taxonomy, shipment, quarantine facilities, permits, host range tests, release, and documentation. Nematodes covered under these suggested guidelines include entomopathogenic species of mermithids, sphaerulariids, aphelenchids, steinernematids, and heterorhabditids. Host specificity and safety tests are discussed. Concern over the possible concomitant introduction of plant-parasitic nematodes, insects, or other pests is expressed. Current information on the treatment of insect-parasitic nematodes by the Environmental Protection Agency and USDA's Animal and Plant Health Inspection Service is presented. These suggested guidelines are presented to stimulate the development of workable protocols for safe introduction of beneficial insect-parasitic nematodes into the United States from aboard.
biological control; entomogenous nematode; guideline; insect parasite; introduction; nematode introduction; permit form; procedure
The Pacific islands have a diverse range of food and cash crops with indigenous and introduced nematode problems. The staple food crops have serious nematode pests, such as Meloidogyne spp. on sweet potato, Hirschmanniella miticausa causing corm rot of taro, and Pratylenchus coffeae and Radopholus sp. producing tuber dry rot of yams. Bananas are infested with P. coffeae or R. similis, citrus with Tylenchulus semipenetrans, rice with Aphelenchoides besseyi, and ginger with Meloidogyne spp. and R. similis. Rotylenchulus reniformis, P. zeae, P. brachyurus, and Helicotylenchus spp. are important on all of these and other crops, such as sugarcane, passion fruit, pawpaw, and cassava. Meloidogyne spp. cause serious damage to local and introduced leaf and fruit vegetables and other crops, such as tobacco, sugarcane, pawpaw, black pepper, and pyrethrum. Many other plant-parasitic genera and species, some undescribed, occur in the Pacific, and there are many islands still to be investigated.
Pacific islands; plant-parasitic nematode
Bed bugs are hematophagous insects responsible for a re-emerging and challenging indoor pest in many countries. Bed bugs infestations may have health consequences including nuisance biting, cutaneous and systemic reactions. This resurgence can probably be attributed to factors such as increased international travel and development of resistance against insecticides. Resistance against pyrethroids has been reported several times from the USA and rarely in Europe. In France, very few data on bed bugs are available. The present study aimed to assess the infestation by bed bugs of a complex of two high-rise apartment buildings in the suburb of Paris and to evaluate their susceptibility to pyrethroid insecticides. We inspected for bed bugs 192 out of 198 apartments units (97%) and interviewed their residents. 76 (39.6%) apartments were infested. Among the 97 residents living in infested apartments, 53 (54.6%) reported bed bug bites. A total of 564 bed bugs were collected in the infested units. Bioassays showed that 54 out of 143 bed bugs were resistant to pyrethroids (37.8%; 95% confidence interval: 29.9-45.7%). DNA sequencing showed that all bed bugs tested (n = 124) had homozygous L925I kdr-like gene mutation. The level of pyrethroid resistance found indicates that this phenomenon was already established in the site and prompts the need to reevaluate the wide use of pyrethroids to control bed bugs.
bed bugs; Cimex lectularius; pyrethroids; insecticide; resistance; punaises de lit; Cimex lectularius; pyréthrinoïdes; insecticide; résistance
We have developed a simple PCR assay protocol for detection of the root-knot nematode (RKN) species Meloidogyne arenaria, M. incognita, and M. javanica extracted from soil. Nematodes are extracted from soil using Baermann funnels and centrifugal flotation. The nematode-containing fraction is then digested with proteinase K, and a PCR assay is carried out with primers specific for this group of RKN and with universal primers spanning the ITS of rRNA genes. The presence of RKN J2 can be detected among large numbers of other plant-parasitic and free-living nematodes. The procedure was tested with several soil types and crops from different locations and was found to be sensitive and accurate. Analysis of unknowns and spiked soil samples indicated that detection sensitivity was the same as or higher than by microscopic examination.
Detection; diagnosis; Meloidogyne arenaria; Meloidogyne incognita; Meloidogyne javanica; PCR; root-knot nematode; soil
The red palm weevil Rhynchophorus ferrugineus Olivier (Curculionidae/Rhynchophoridae/Dryophthoridae) is a lethal pest of young coconut palms, Cocos nucifera L. (Arecales: Arecaceae), with a highly aggregated population distribution pattern. R. ferrugineus is managed in several coconut growing countries using area-wide pheromone based programmes that need a substantial commitment of funds over a period of time. Often, decisions to implement area-wide management of R. ferrugineus are based on pheromone trap captures in surveillance traps and or infestation reports. Implementing area-wide management of this pest on the basis of such data can be inaccurate, as it may either under or over estimate the pest intensity in the field. This study presents sampling plans for rapid and accurate classification of R. ferrugineus infestation in coconut plantations of India by inspecting palms to detect infestation in a sequence until a decision to either implement or not to initiate area-wide management of R. ferrugineus can be made. The sampling plans are based on a common aggregation index of 3.45, assumed action threshold values of either 1.0 (plan A) or 0.5 (plan B) per cent infested palms and a risk factor of making the wrong decision set at 0.05. Using plans A and B, if the cummulative number of infested palms in a young 1 hectare coconut plantation is zero out of 150 palms for both plans, then area-wide management is not required, while on the other hand, if the cummulative number of infested palms for the same area is 6 (plan A), or 5 (plan B), then area-wide management of R. ferrugineus is essential. The proposed sampling plans are efficient tools in decision making, particularly at very low and high levels of infestation and can also be used to assess the performance of R. ferrugineus IPM programmes that are in progress. These plans not only save time and money as only a small area needs to be sampled to arrive at a correct decision, but are also efficient in rating the infestation level accurately.
sequential sampling; Cocos nucifera
The sedentary semi-endoparasitic nematode Rotylenchulus reniformis, the reniform nematode, is a serious pest of cotton and soybean in the United States. In recent years, interest in the molecular biology of the interaction between R. reniformis and its plant hosts has increased; however, the unusual life cycle of R. reniformis presents a unique set of challenges to researchers who wish to study the developmental expression of a particular nematode gene or evaluate life stage–specific effects of a specific treatment such as RNA-interference or a potential nematicide. In this report, we describe a simple method to collect R. reniformis juvenile and vermiform adult life stages under in vitro conditions and a second method to collect viable parasitic sedentary females from host plant roots. Rotylenchulus reniformis eggs were hatched over a Baermann funnel and the resultant second-stage juveniles incubated in petri plates containing sterile water at 30°C. Nematode development was monitored through the appearance of fourth-stage juveniles and specific time-points at which each developmental stage predominated were determined. Viable parasitic sedentary females were collected from infected roots using a second method that combined blending, sieving, and sucrose flotation. Rotylenchulus reniformis life stages collected with these methods can be used for nucleic acid or protein extraction or other experimental purposes that rely on life stage–specific data.
host-parasitic relationship; life stages; reniform nematode; Rotylenchulus reniformis; technique
Failure to detect a disease agent or vector where it actually occurs constitutes a serious drawback in epidemiology. In the pervasive situation where no sampling technique is perfect, the explicit analytical treatment of detection failure becomes a key step in the estimation of epidemiological parameters. We illustrate this approach with a study of Attalea palm tree infestation by Rhodnius spp. (Triatominae), the most important vectors of Chagas disease (CD) in northern South America.
The probability of detecting triatomines in infested palms is estimated by repeatedly sampling each palm. This knowledge is used to derive an unbiased estimate of the biologically relevant probability of palm infestation. We combine maximum-likelihood analysis and information-theoretic model selection to test the relationships between environmental covariates and infestation of 298 Amazonian palm trees over three spatial scales: region within Amazonia, landscape, and individual palm. Palm infestation estimates are high (40–60%) across regions, and well above the observed infestation rate (24%). Detection probability is higher (∼0.55 on average) in the richest-soil region than elsewhere (∼0.08). Infestation estimates are similar in forest and rural areas, but lower in urban landscapes. Finally, individual palm covariates (accumulated organic matter and stem height) explain most of infestation rate variation.
Individual palm attributes appear as key drivers of infestation, suggesting that CD surveillance must incorporate local-scale knowledge and that peridomestic palm tree management might help lower transmission risk. Vector populations are probably denser in rich-soil sub-regions, where CD prevalence tends to be higher; this suggests a target for research on broad-scale risk mapping. Landscape-scale effects indicate that palm triatomine populations can endure deforestation in rural areas, but become rarer in heavily disturbed urban settings. Our methodological approach has wide application in infectious disease research; by improving eco-epidemiological parameter estimation, it can also significantly strengthen vector surveillance-control strategies.
Blood-sucking bugs of the genus Rhodnius are major vectors of Chagas disease. Control and surveillance of Chagas disease transmission critically depend on ascertaining whether households and nearby ecotopes (such as palm trees) are infested by these vectors. However, no bug detection technique works perfectly. Because more sensitive methods are more costly, vector searches face a trade-off between technical prowess and sample size. We compromise by using relatively inexpensive sampling techniques that can be applied multiple times to a large number of palms. With these replicated results, we estimate the probability of failing to detect bugs in a palm that is actually infested. We incorporate this information into our analyses to derive an unbiased estimate of palm infestation, and find it to be about 50% – twice the observed proportion of infested palms. We are then able to model the effects of regional, landscape, and local environmental variables on palm infestation. Individual palm attributes contribute overwhelmingly more than landscape or regional covariates to explaining infestation, suggesting that palm tree management can help mitigate risk locally. Our results illustrate how explicitly accounting for vector, pathogen, or host detection failures can substantially improve epidemiological parameter estimation when perfect detection techniques are unavailable.
Among ocular vector-borne pathogens, Onchocerca volvulus, the agent of the so-called “river blindness”, affects about 37 million people globally. Other Onchocerca spp. have been sporadically reported as zoonotic agents. Cases of canine onchocerciasis caused by Onchocerca lupi are on the rise in the United States and Europe. Its zoonotic role has been suspected but only recently ascertained in a single case from Turkey. The present study provides further evidence on the occurrence of O. lupi infesting human eyes in two patients from Turkey (case 1) and Tunisia (case 2). The importance of obtaining a correct sample collection and preparation of nematodes infesting human eyes is highlighted.
In both cases the parasites were identified with morpho-anatomical characters at the gross examination, histological analysis and anatomical description and also molecularly in case 1.
The nematode from the first case was obviously O. lupi based on their morphology at the gross examination, histological analysis and anatomical description. In the second case, although the diagnostic cuticular characters were not completely developed, other features were congruent with the identification of O. lupi. Furthermore, the morphological identification was also molecularly confirmed in the Turkish case.
The results of this study suggest that O. lupi infestation is not an occasional finding but it should be considered in the differential diagnosis of other zoonotic helminths causing eye infestation in humans (e.g., D. immitis and Dirofilaria repens). Both cases came from areas where no cases of canine onchocerciasis were previously reported in the literature, suggesting that an in depth appraisal of the infestation in canine populations is necessary. Physicians and ophthalmologists are advised on how to preserve nematode samples recovered surgically, to allow a definitive, correct etiological diagnosis.
Onchocerca lupi; Zoonosis; Ocular infestation; Dog; Turkey, Tunisia
Four procedures were compared in their efficacy to extract juveniles of Anguina agrostis from commercial grass seed. The procedures included those currently used by the state regulatory laboratories of Oregon and California, as well as new tests developed to determine juvenile viability for the phytosanitary certification of fumigated grass seed. Eleven seed lots of Agrostis tenuis (bentgrass) and Dactylis glomerata (orchardgrass) naturally infested with varying levels of juveniles of Anguina were individually analyzed. Only one procedure, a new live recovery test, yielded nematodes in all 11 samples and is recommended as the best method for use by regulatory agencies. In comparison, although the other three extraction procedures resulted in greater numbers of Anguina agrostis juveniles per gram of seed, they failed to yield any nematodes in as many as four seed lots with low infection levels.
Anguina agrostis; assay; bentgrass; extraction procedures; nematode; orchardgrass; phytosanitary certification; regulatory nematology; seed-borne nematodes; technique
Chagas disease prevention critically depends on keeping houses free of triatomine vectors. Insecticide spraying is very effective, but re-infestation of treated dwellings is commonplace. Early detection-elimination of re-infestation foci is key to long-term control; however, all available vector-detection methods have low sensitivity. Chemically-baited traps are widely used in vector and pest control-surveillance systems; here, we test this approach for Triatoma spp. detection under field conditions in the Gran Chaco.
Using a repeated-sampling approach and logistic models that explicitly take detection failures into account, we simultaneously estimate vector occurrence and detection probabilities. We then model detection probabilities (conditioned on vector occurrence) as a function of trapping system to measure the effect of chemical baits. We find a positive effect of baits after three (odds ratio [OR] 5.10; 95% confidence interval [CI95] 2.59–10.04) and six months (OR 2.20, CI95 1.04–4.65). Detection probabilities are estimated at p≈0.40–0.50 for baited and at just p≈0.15 for control traps. Bait effect is very strong on T. infestans (three-month assessment: OR 12.30, CI95 4.44–34.10; p≈0.64), whereas T. sordida is captured with similar frequency in baited and unbaited traps.
Chemically-baited traps hold promise for T. infestans surveillance; the sensitivity of the system at detecting small re-infestation foci rises from 12.5% to 63.6% when traps are baited with semiochemicals. Accounting for imperfect detection, infestation is estimated at 26% (CI95 16–40) after three and 20% (CI95 11–34) after six months. In the same assessments, traps detected infestation in 14% and 8.5% of dwellings, whereas timed manual searches (the standard approach) did so in just 1.4% of dwellings only in the first survey. Since infestation rates are the main indicator used for decision-making in control programs, the approach we present may help improve T. infestans surveillance and control program management.
Triatoma infestans is the main vector of Chagas disease in southern South America. Dwelling-infesting populations are controlled through insecticide-spraying campaigns; however, dwellings are often re-infested when insecticide effects wane, and this leads to the re-establishment of disease transmission. Detecting and eliminating re-infestation foci is therefore crucial to prevent new cases. Unfortunately, available vector detection methods all have low sensitivity. Here, we show that simple sticky traps baited with widely available chemicals are significantly more sensitive than either unbaited traps or active manual searches by trained staff — the standard method used in control programs. Increased trap sensitivity (about 500% higher), together with an analytical approach that takes detection failures into account, allows us to estimate dwelling infestation rates at about 20–26%; in contrast, just 0–1.4% of dwellings were identified as infested by manual searches. This large difference highlights the importance of enhancing surveillance systems, and reveals how crude infestation indices may mislead decision-makers. We conclude that chemically baited sticky traps can help improve T. infestans surveillance systems and thus strengthen vector control program management.
Rotylenchulus reniformis is rapidly becoming the most economically important pest associated with cotton in the southeastern United States. Incentive programs have been implemented to support sampling of production fields to determine the presence and abundance of R. reniformis. These sampling programs have dramatically increased the number of soils samples submitted to nematology laboratories during autumn. The large numbers of samples overwhelm most labs and require placement in cold storage until extraction. Therefore, the objective of this study was to examine the length of time soils infested with R. reniformis can be stored before nematode extraction without compromising the accuracy of estimates of population densities. A sandy loam and a silty loam were the two cotton production soils used in this study. Rotylenchulus reniformis numbers decreased 61%during the first 180 days of storage in both soils. Rotylenchulus reniformis numbers from the initial sampling through 180 days decreased as a linear function. The decline of R. reniformis numbers during storage was estimated as 0.28% of the population lost daily from the maximum population through 180 days. The diminution of nematode numbers from 180 through 1,080 days in storage continued, but at a slower rate. Numbers of R. reniformis declined to less than 89%, 93%, and 99% of the initial population within 360, 720, and 1,080 days, respectively, of storage. The reduction of R. reniformis numbers over 180 days can be adjusted, allowing a more accurate estimation of R. reniformis levels in soil samples stored at 4 °C.
Rotylenchulus reniformis; soil storage; population density
Storage of nematodes in soil at -15 C for 1 to 16 weeks greatly increased nematode recovery by a sugar-flotation-sieving procedure. One week of exposure to -15 C killed all nematodes except Pratylenchus zeae and Tylenchorhynchus claytoni which were recoverable in decreasing numbers up to 10 weeks by the Baermann funnel method. Optimum storage temperature for survival of most nematode species was 13 C. The numbers of Meloidogyne incognita, T. claytoni, Belonolaimus Iongicaudatus, and P. zeae recoverable by either extraction method remained constant or increased when stored at 13-24 C for 16 weeks. This was also true for Helicotylenchtts dihystera and Xiphinema americanum extracted by the Baermann funnel technique, whereas the numbers retrieved by the sugar-flotation-sieving method decreased slightly. All species except T. claytoni decreased appreciably in soil stored at 36 C.
Baermann funnels were modified to eliminate or reverse the small temperature gradient (1-2 C/cm) across the soil layer that normally results from water evaporation. Effects of modifications on extraction efficiency were examined at various ambient temperatures and after overnight adaptation of three nematode species at 20 and 30 C. Extraction of Meloidogyne incognita from sandy loam, Tylenchulus semipenetrans from sandy clay loam, and Rotylenchulus reniformis from silt was greatly accelerated simply by covering funnels to prevent evaporation. In most cases, covering increased the nematodes extracted by 10-100 times after 5.5-48 hours. Faster and more efficient extraction of R. reniformis occurred over a wide range of ambient temperature (18-29 C). Effects of ambient temperature and temperature gradient direction on Baermann funnel extraction of R. reniformis were partly inconsistent with the behavior of R. reniformis in agar. Nematodes in agar moved toward cold at some ambient temperatures and toward heat at other temperatures. They always appeared to move toward cold on Baermann funnels. Differences were not attributable to blockage of gas exchange by covers. In agar and in funnels, the patterns of response to ambient temperature were shifted in the direction of the storage temperature.
Baermann funnel; behavior; Meloidogyne incognita; nematode extraction; Rotylenchulus reniformis; thermal adaptation; thermotaxis; Tylenchulus semipenetrans
Spatial heterogeneity in nematode population densities presents an obstacle to the precise determination of infestation levels. Three field plots were intensively sampled for soybean cyst nematode (Heterodera glycines Ich.) cysts before and after spring cultivation to quantify the spatial attributes of the population. Population density strata were detected running parallel to plant rows. Highest population densities before cultivation were found in the plant row and the middle furrow, Population density in the plant row averaged 26% higher and 4% lower than the whole-plot mean before and after cultivation, respectively. Cysts containing fewer than 25 eggs were not stratified, indicating that most were produced before the previous season. Sample population counts were fit to the negative binomial distribution model before cultivation, but distributions differed among plots. The Neyman type A and negative binomial distributions both fit the data after cultivation disturbed the soil. Population clusters 1-3 m long were detected in plant beds before cultivation. Heterogeneity in population density increased with plant row length after cultivation. Optimum plot length for minimal spatial heterogeneity in four-row mechanically tended field plots was estimated at 6 m after trimming plot ends.
dispersion; Heterodera glycines; soybean cyst nematode; population ecology; stratification
Aedes aegypti and Ae. albopictus are the vectors of dengue, the most important arboviral disease of humans. To date, Aedes ecology studies have assumed that the vectors are truly absent from sites where they are not detected; since no perfect detection method exists, this assumption is questionable. Imperfect detection may bias estimates of key vector surveillance/control parameters, including site-occupancy (infestation) rates and control intervention effects. We used a modeling approach that explicitly accounts for imperfect detection and a 38-month, 55-site detection/non-detection dataset to quantify the effects of municipality/state control interventions on Aedes site-occupancy dynamics, considering meteorological and dwelling-level covariates. Ae. aegypti site-occupancy estimates (mean 0.91; range 0.79–0.97) were much higher than reported by routine surveillance based on ‘rapid larval surveys’ (0.03; 0.02–0.11) and moderately higher than directly ascertained with oviposition traps (0.68; 0.50–0.91). Regular control campaigns based on breeding-site elimination had no measurable effects on the probabilities of dwelling infestation by dengue vectors. Site-occupancy fluctuated seasonally, mainly due to the negative effects of high maximum (Ae. aegypti) and minimum (Ae. albopictus) summer temperatures (June-September). Rainfall and dwelling-level covariates were poor predictors of occupancy. The marked contrast between our estimates of adult vector presence and the results from ‘rapid larval surveys’ suggests, together with the lack of effect of local control campaigns on infestation, that many Aedes breeding sites were overlooked by vector control agents in our study setting. Better sampling strategies are urgently needed, particularly for the reliable assessment of infestation rates in the context of control program management. The approach we present here, combining oviposition traps and site-occupancy models, could greatly contribute to that crucial aim.
The long-horned beetle, Dectes texanus LeConte (Coleoptera: Cerambycidae), is a stem-boring pest of soybeans, Glycine max (L.) Merrill (Fabales: Fabaceae). Soybean stems and stubble were collected from 131 counties in Arkansas, Mississippi, Missouri, and Tennessee and dissected to determine D. texanus infestation rates. All states sampled had D. texanus present in soybeans. Data from Tennessee and Arkansas showed sample infestations of D. texanus averaging nearly 40%. Samples from Missouri revealed higher infestation in the twelve southeastern counties compared to the rest of the state. Data from Mississippi suggested that D. texanus is not as problematic there as in Arkansas, Missouri, and Tennessee. Infestation rates from individual fields varied greatly (0–100%) within states. In Tennessee, second crop soybeans (i.e. soybeans planted following winter wheat) had lower infestations than full season soybeans. A map of pest distribution is presented that documents the extent of the problem, provides a baseline from which changes can be measured, contributes data for emergency registration of pesticides for specific geographic regions, and provides useful information for extension personnel, crop scouts, and growers.
Glycine max; distribution
The development of Bursaphelenchus xylophilus in pine wood infested with and free of Monochamus carolinensis was investigated. Formation of third-stage dispersal juveniles occurred in the presence and absence of pine sawyer beetles. The proportion of third-stage dispersal juveniles in the total nematode population was negatively correlated with moisture content of the wood. Formation of nematode dauer juveniles was dependent on the presence of the pine sawyer beetle. Dauer juveniles were present in 3 of 315 wood samples taken from non-beetle-infested Scots pine bolts and 81 of 311 samples taken from beetle-infested bolts. Nematode densities were greater in wood samples taken adjacent to insect larvae, pupae, and teneral adults compared with samples taken from areas void of insect activity. Nematodes recovered from beetle larvae, pupae, and teneral adults were mostly fourth-stage dauer juveniles, although some third-stage dispersal juveniles were also recovered. Dauer juvenile density was highest on teneral adult beetles.
Bursaphelenchus xylophilus; dauerjuvenile; Monochamus carolinensis; nematode; pine sawyer; pinewood nematode