Presence-absence sequential sampling plan for northern fowl mite, Ornithonyssus sylviarum (Acari: Macronyssidae), on caged-layer hens

Caged-layer hens were scored as infested or uninfested by visual examination of the vent region, and the number of northern fowl mite, Ornithonyssus sylviarum (Canestrini & Fanzago), per hen was estimated. The proportion infested and average number of mites per hen were shown to have a highly significant, positive relationship (r = 0.936). Sampling among houses within a flock, and rows and sections within houses were analyzed to determine the reliability of sampling a representative portion of a flock. Low- and moderate-tolerance treatment thresholds, based on percentage of hens infested with mites, were developed from sampling 1 wk before and 1 wk after acaricide treatments determined necessary by the producer. These thresholds were used to compare a fixed (single) sampling plan, a curtailed procedure of the fixed sampling plan, and a sequential sampling plan based on a sequential probability ratio test, by sampling 174 hens (the maximum number needed for the single sampling plan). The sequential sampling plan required fewer hen examinations on average to reach a treatment decision than did the other plans, depending on the infestation tolerance limits. Using a low tolerance approach in which infestations below 15% are considered noneconomic (safe threshold) and infestations above 25% are considered economically important (action threshold), as few as 5 hens required examination to reach a treatment decision. Sequential sampling plan graphs are presented for 2 tolerance threshold scenarios (a 15% safe-threshold paired with a 25% action threshold and a 35% safe-threshold paired with a 45% action threshold). These sequential sampling plans using presence absence assessments should greatly facilitate monitoring and treatment decisions for this important pest.     

Economic injury level and sequential sampling plan for Bemisia tabaci in outdoor tomato

Abstract:  This work aimed to determine the economic injury levels and to establish sequential sampling plans for nymphs and adults of the whitefly Bemisia tabaci Genn. (Sternorrhyncha: Aleyrodidae) in tomato fields. Densities of nymphs and adults, as well as crop yield were evaluated in 13 commercial tomato fields to determine the economic injury levels. The whitefly nymphs were sampled by direct counting in a leaf from the lower part of the canopy and the adults were sampled by beating an apical leaf against a white plastic tray. The sequential sampling plan was based on data collected in eight commercial tomato fields. The validation of the sequential sampling plan was carried out based on the curves of operational characteristics and average sample numbers. The decisions reached with the conventional and the sequential sampling plans in 21 commercial fields were compared for the intended validation of the sequential plan. The economic injury levels were four nymphs per leaf and one adult per tray. The decisions taken based on the sequential sampling plan were similar to those obtained through the conventional sampling plan. Most of the decisions taken with the sequential sampling plan were obtained through the minimum number of seven samples per field for nymphs and 11 samples per field for adults, with reductions of 84.44% and 54.17% in the number of samples required to reach a decision with the sequential sampling plan compared with the conventional sampling plan.     

Sample size for collecting seeds in germplasm conservation: the case of the Lima bean (Phaseolus lunatus L.)

 The design of optimum sampling strategies integrating criteria of efficiency relevant to multilocus models and many target populations has been investigated with respect to the number of plants and the number of seeds per plant to be sampled for a Lima bean (Phaseolus lunatus L.) gene pool. This study, using five populations and six polymorphic enzyme loci, shows that the number of plants rather than the number of seeds collected per plant primarily determines the success of seed sampling, suggesting that plant number plays an essential part in maintaining the allelic multiplicity of predominantly selfing species like Lima bean. According to the results, it appears that among Lima bean populations an efficient sampling procedure is achieved by collecting 1–4 seeds from 200 to 300 plants. These sample sizes will retain 8–10 alleles, regardless of their frequencies. When we consider polymorphism at the 5% level, it is expected that sampling 10–80 plants will collect combinations of 4–8 alleles. Based on data from genetic and demographic studies, we suggest an efficient sampling scheme for Lima bean germplasm at both population and geographical levels. Received: 10 March 1998 / Accepted: 1 April 1998     

Sequential hypothesis testing with spatially correlated presence-absence data

A pest management decision to initiate a control treatment depends upon an accurate estimate of mean pest density. Presence-absence sampling plans significantly reduce sampling efforts to make treatment decisions by using the proportion of infested leaves to estimate mean pest density in lieu of counting individual pests. The use of sequential hypothesis testing procedures can significantly reduce the number of samples required to make a treatment decision. Here we construct a mean-proportion relationship for Oligonychus perseae Tuttle, Baker, and Abatiello, a mite pest of avocados, from empirical data, and develop a sequential presence-absence sampling plan using Bartlett's sequential test procedure. Bartlett's test can accommodate pest population models that contain nuisance parameters that are not of primary interest. However, it requires that population measurements be independent, which may not be realistic because of spatial correlation of pest densities across trees within an orchard. We propose to mitigate the effect of spatial correlation in a sequential sampling procedure by using a tree-selection rule (i.e., maximin) that sequentially selects each newly sampled tree to be maximally spaced from all other previously sampled trees. Our proposed presence-absence sampling methodology applies Bartlett's test to a hypothesis test developed using an empirical mean-proportion relationship coupled with a spatial, statistical model of pest populations, with spatial correlation mitigated via the aforementioned tree-selection rule. We demonstrate the effectiveness of our proposed methodology over a range of parameter estimates appropriate for densities of O. perseae that would be observed in avocado orchards in California.     

Transect sampling to enhance efficiency of corn rootworm (Coleoptera: Chrysomelidae) monitoring in corn

Crop monitoring for adult corn rootworms, Diabrotica virgifera virgifera LeConte and Diabrotica barberi Smith and Lawrence, remains the best means to assess fields at risk from this pest if replanted to corn, Zea mays (L.). Improvements in sampling methodology, including the development of a sequential sampling plan, have reduced the minimum sampling time required to make a management decision to 20 min or less per field per visit. However, many growers and crop consultants still find this time commitment a constraint to repeated scouting. A common currently used sampling method involves systematically covering most of the field following a "W" pattern. The feasibility of replacing the current sampling pattern with a simpler and less time-consuming transect (straight line) pattern was assessed. When sampling methods were compared, computer simulations demonstrated that treatment decisions based on transect sampling would have an acceptably low error rate averaging 10% over a range of realistic corn rootworm densities (0-2 adults per plant). This error rate represented a decrease in accuracy of <1% compared with systematic sampling. Field trials using transect, systematic, and random sampling in each field were used to compare the categorization of adult corn rootworm densities into "above" or "below" threshold with a sequential sampling plan. Efficiency measured in time to reach a decision, number of corn plants evaluated, and time divided by plants observed were compared between sampling methods. The three methods did not differ significantly in the number of plants evaluated or in the categorization of corn rootworm populations. Transect sampling resulted in a significantly shorter time divided by plants observed (38 s), than either systematic (78 s), or random sampling methods (166 s). Based on these results transect sampling reduces sampling time 51% compared with systematic sampling and thus could be used to reduce total sampling times substantially.     

Cost-effective binomial sequential sampling of western bean cutworm, Striacosta albicosta (Lepidoptera: Noctuidae), egg masses in corn

Striacosta albicosta (Smith) (Lepidoptera: Noctuidae), is a native pest of dry beans (Phaseolus vulgaris L.) and corn (Zea mays L.). As a result of larval feeding damage on corn ears, S. albicosta has a narrow treatment window; thus, early detection of the pest in the field is essential, and egg mass sampling has become a popular monitoring tool. Three action thresholds for field and sweet corn currently are used by crop consultants, including 4% of plants infested with egg masses on sweet corn in the silking-tasseling stage, 8% of plants infested with egg masses on field corn with approximately 95% tasseled, and 20% of plants infested with egg masses on field corn during mid-milk-stage corn. The current monitoring recommendation is to sample 20 plants at each of five locations per field (100 plants total). In an effort to develop a more cost-effective sampling plan for S. albicosta egg masses, several alternative binomial sampling plans were developed using Wald's sequential probability ratio test, and validated using Resampling for Validation of Sampling Plans (RVSP) software. The benefit-cost ratio also was calculated and used to determine the final selection of sampling plans. Based on final sampling plans selected for each action threshold, the average sample number required to reach a treat or no-treat decision ranged from 38 to 41 plants per field. This represents a significant savings in sampling cost over the current recommendation of 100 plants.     

Spontaneous plant regeneration in transformed roots and calli from Tylophora indica: changes in morphological phenotype and tylophorine accumulation associated with transformation by Agrobacterium rhizogenes

We examined the effects of genetic transformation by Agrobacterium rhizogenes on the production of tylophorine, a phenanthroindolizidine alkaloid, in the Indian medicinal plant, Tylophora indica. Transformed roots induced by the bacterium grew in axenic culture and produced shoots or embryogenic calli in the absence of hormone treatments. However, hormonal treatment was required to regenerate shoots in root explants of wild type control plants. Transformed plants showed morphological features typically seen in transgenic plants produced by A. rhizogenes, which include, short internodes, small and wrinkled leaves, more branches and numerous plagiotropic roots. Plants regenerated from transformed roots showed increased biomass accumulation (350–510% in the roots and 200–320% in the whole plants) and augmented tylophorine content (20–60%) in the shoots, resulting in a 160–280% increase in tylophorine production in different clones grown in vitro.     

Sequential sampling plans for western flower thrips (Thysanoptera: Thripidae) on greenhouse cucumbers

The development of cost-effective and reliable sampling programs for the management of western flower thrips, Frankliniella occidentalis (Pergande), on greenhouse cucumbers is important for getting growers to adopt economic injury levels and economic thresholds. The objectives of this study were to develop two sequential sampling plans. A fixed-precision sequential sampling plan was designed for estimating F. occidentalis adult density at a fixed-precision level on cucumber flowers. Also, a sequential sampling plan for classifying thrips population levels as below or above economic thresholds was developed to assist in decision making for the timing of pesticide applications. Both sequential sampling plans were validated using a resampling simulation technique on nine independent data sets ranging in density from 1.25 to 12.95 adults per flower. With the fixed-precision sampling plan, average means obtained in 100 repeated simulation runs were within the 95% CI of the estimated mean for all data sets. Appropriate levels of precision for the different population densities were recommended based on the simulation results. With sequential sampling for classifying the population levels of thrips in terms of an economic threshold, it has the advantage of requiring smaller sample sizes to determine the population status when the population densities differ greatly from the critical density (i.e., economic threshold). However, this plan needs a great number of samples when population density is close to the critical density. In this case, use of a combination of both sampling plans is recommended.     

Enumerative and binomial sequential sampling plans for soybean aphid (Homoptera: Aphididae) in soybean

Since the discovery of the soybean aphid, Aphis glycines Matsumura, in midwestern U.S. soybean, Glycine max L., in 2000, the aphid has become a significant economic pest. Basic information about estimating population density within fields is unknown. Therefore, we developed two sampling plans to efficiently characterize A. glycines densities. Enumerative and binomial sequential plans were developed using 89 data sets collected from 10 commercial fields sampled during 2001-2003. Re-sampling software was used to validate the enumerative plan on whole plant counts, based on Taylor's power law parameters (a = 9.157 and b = 1.543). For research applications, the enumerative plan was modified to provide an actual precision level of 0.10 (SE/mean), which resulted in an average sample number of 310 individual plants. For integrated pest management (IPM) purposes, we developed an enumerative plan with an actual precision of 0.25, which resulted in an average sample number of 38 individual plants. For IPM applications, the binomial plan will likely be more practical. Binomial plans were developed using two tally thresholds at five action thresholds. Final analysis of the operating characteristic curve for each plan indicated that the tally threshold of > or = 40 aphids per plant, and an action threshold of 0.837 (84% of the plants infested) provided the most correct treat (4%) and no-treat (95%) decisions, with very low incorrect treat (0.5%) and no-treat (0.5%) decisions. A tally threshold of > or = 40 aphids per plant and action thresholds of 84% of plants infested is equivalent to a mean density of 250 aphids per plant, a recently recommended economic threshold. Using this threshold, the minimum required sample number for the binomial plan was 11 plants.     

Diagnosing sulfur deficiency in field-grown oilseed rape (Brassica napus L.) and wheat ( Triticum aestivum L.)

The effects of increasing sulfur applications on field-grown oilseed rape (Brassica napus L.) and wheat (Triticum aestivum L.) were investigated in 1998 and 1999, and the critical values and efficiency of several diagnostic indicators for S deficiency were determined. Critical values for leaf concentrations of total S, sulfate and glutathione changed over time and were not suitable for diagnosing S deficiency early in the growth season. The N:S ratio was more reliable but involved two analytical measurements. A practical and reliable indicator for S deficiency was the malate:sulfate peak area ratio as measured by ion chromatography, which required only a single analysis and was independent of the time of sampling or calibration of the samples. A malate:sulfate ratio ≤ 1 indicated S sufficiency at the time of sampling, whereas a ratio > 1 suggested S deficiency at the time of sampling. The malate:sulfate ratio was reliable at growth stage 3.6–3.7 (flower stalks extending to first flowers yellow) for oilseed rape and at growth stage 22–25 (main stem and 2–5 tillers) for wheat, which was sufficiently early in the growth season to apply remedial sulfur fertilizer, if necessary. This revised version was published online in June 2006 with corrections to the Cover Date.     

Soil sampling for <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> spores in Queensland sugarcane fields

Three sugarcane fields in Bundaberg and four fields in each of the Burdekin, Tully and Innisfail (Queensland, Australia) were sampled for spores of Metarhizium anisopliae (Metchnikoff) (Deuteromycotina: Hyphomycetes). This entomopathogenic fungus is the active ingredient in the biocide “BioCane^TM”, which was developed for the management of the greyback canegrub Dermolepida albohirtum (Waterhosue) (Coleoptera: Scarabaeidae) and other scarabs in cane fields. Fields sampled were of different crop ages and all had a history of BioCane^TM treatment in Plant Cane in past years. Soil samples were taken in each field from four depths (0–10, 10–20, 20–30 and 30–40 cm below soil level) with the use of an auger. Spore levels were highest at the depths of 10–20 and 20–30 cm. Spore levels differed between locations with Innisfail and Tully recording the highest spore counts. Spores were also found in the inter-row space in plots sampled in Tully. Sampling statistics were determined for M. anisopliae spores at the four soil depths with 0.1 and 0.25 precision levels. Three sampling methods were compared (use of marker beads; use of 100 mm auger and 150 mm auger). Samples that relied on marker beads resulted in higher spore counts, however, an auger can still be used since BioCane^TM does not normally contain coloured markers. Results obtained demonstrate the ability of the pathogen to translocate in soil profile and across rows, most likely due to grub movements and other soil fauna. Sampling for M. anisopliae spores provides good monitoring of their levels in soil. The implications of this on grub management decisions are discussed.     

Calibration of sampling techniques and determination of sample size for the estimation of egg and larval populations of Helicoverpa spp. (Lepidoptera: Noctuidae) on irrigated soybean

Abstract  Informed spray decisions require accurate assessments of the target pest's density. Currently, no advice is provided to farmers on the best method for sampling soybean for insect pests, although spray thresholds for Helicoverpa larvae are provided. This article describes the results of a trial designed to calibrate relative sampling techniques for Helicoverpa larvae; visual inspection of plants in situ in the field, beat cloth, sweep net and D-vac sampling were compared with an absolute measure of population density. The absolute measure was derived from the bagging and removal of whole plants in the field, followed by subsequent examination in the laboratory. Analysis of the distribution of Helicoverpa larvae collected by the different samples was then used to calculate the number of samples required to determine whether the economic threshold had been reached to different levels of certainty and accuracy. Significant relationships were detected between all the relative sampling techniques and the absolute, suggesting that all could be used to estimate field populations. However, due to the high variance and therefore increased sample sizes required, or the length of time taken to collect samples, only beat-cloth sampling appeared to offer a realistic method for farmers in the field. The results also suggest that the current best practice of sampling six locations per crop provides an adequate assessment of the field populations at the currently accepted threshold level of 6 larvae m⁻². However, if the economic spray was reduced, the number of samples required to determine an accurate population estimate would increase dramatically.     

High-frequency callus induction and plant regeneration in Tripsacum dactyloides (L.)

Summary A protocol for high-frequency callus, somatic embryogenesis, and plant regeneration for Tripsacum is described. Plants were regenerated from complete shoot meristems (3–4 mm) via organogenesis and embryogenesis. In organogenesis, the shoot meristems were cultured directly on a high cytokinin medium comprising 5–10 mgl⁻¹ (22.2–44.4 μM) 6-benzyladenine (BA). The number of multiple shoots varied from six to eight from each meristem. The time required for production of plants from organogenesis was rapid (4–6 wk). In contrast, callus was induced on an auxin medium and continuously cultured on an auxin medium for production of somatic embryos. Prolific callus with numerous somatic embryos developed within 3–4 wk when cultured on an auxin medium containing 5 mgl⁻¹ (22.6μM), 2,4-dichlorophenoxyacetic acid (2,4-D). The number of shoots induced varied from two to five per callus. Regardless of the cultivars used, the frequency of callus induction and plant regeneration was between 48% and 94%. The seed germination procedures also were modified and resulted in a maximum of 60–80% seed germination. Finally, the rate of T-DNA transfer to complete shoot meristems of Tripsacum was high on the auxin medium and was independent of whether super-virulent strains of Agrobacterium were used or not.     

Assessing diversity of arbuscular mycorrhizal fungi in a local community: role of sampling effort and spatial heterogeneity

Diversity of arbuscular mycorrhizal fungi (AMF) was assessed in two 9.2 × 9.2-m plots planted with landscape trees and shrubs at an experimental site in Phoenix, AZ, USA. Twenty-five soil samples were collected in a regular grid pattern from each plot, and AMF species were identified using trap cultures. A total of 12 species were detected, with 7 species detected in one plot and 11 in the other. We found that sampling effort had a major impact on assessing species richness and composition in this local community. Fifteen samples would be necessary to detect 70–80% of species present in each plot. A limited number of additional undetected species are likely to be present in both plots, based on the sampling effort curves and jackknife estimates. Only two species, Glomus eburneum and Glomus microaggregatum, were detected in over 50% of the samples from both plots, and rank–frequency plots revealed a lognormal species distribution. Despite the patchiness of plants in the plots, the number of species detected per point exhibited spatial structuring only at the smallest sampling scale in a single plot, and only a single species in each plot was not randomly distributed. These results indicate that sampling effort and strategy can affect perceptions of AMF community structure.     

A Feasibility Study to Monitor the Macroinvertebrate Diversity of the River Nile Using Three Sampling Methods

The River Nile is one of the world’s major rivers. Its’ catchment in Egypt has a population of 75,000,000. River flow is highly regulated and there are known discharges of pollutants. 1035 km of the river downstream of the Aswan high dam was studied to test the hypothesis that representative qualitative samples can be used to estimate macroinvertebrate biodiversity. Benthic macroinvertebrates are difficult to sample in large rivers and a reliable sampling strategy is required to evaluate their ecological status. Three methods for sampling have been investigated. Ekman Grab, macrophyte sweep netting and Artificial Substrate Samplers (ASS) were used to sample 15 sites from Aswan to Cairo between September 2001 and June 2002. Organisms were identified to species level where possible. Taxon accretion curves indicated that the all taxa present at a site should be collected using either 15 grab samples, 10 macrophyte samples or 5 ASS. The best time to sample was May–June. The biodiversity of macroinvertebrates in the Nile was recorded as 50 taxa with values of 7–31 at individual bank-side sites. Mid-stream biodiversity was much lower (0–19). Lowest biodiversity occurred at sites with known pollution inputs whilst highest occurred at sites with high levels of sedimentation. A regular programme for biomonitoring is recommended which will allow current status to be confirmed and future changes detected.     

Efficient micropropagation of Robinia ambigua var. idahoensis (Idaho Locust) and detection of genomic variation by ISSR markers

Robinia ambigua var. idahoensis, presumably originated from interspecific hybridization of R. pseudoacacia L. and R. hispida L., is a multipurpose tree. Several reports have showed that in vitro micropropagation is a feasible method to produce large quantities of ‘clonal’ plants from R. pseudoacacia, however, no information is available on micropropagation of R. ambigua or the other assumed parental species, R. hispida. Here, we report on a tissue culture system for efficient micropropagation of R. ambigua plants by enhanced branching of axillary buds taken from a single branch of a donor tree. The culture system consists of sequential use of three media, namely, the bud-induction medium (MS medium supplemented with 0.8–1.4 mg l⁻¹ 6-BA, 0.05–0.08 mg l⁻¹ NAA and 0.07–0.1 mg l⁻¹ GA), elongation medium (MS medium added with 0.35–0.5 mg l⁻¹ 6-BA, 0.05–0.08 mg l⁻¹ NAA and 0.07–0.1 mg l⁻¹ GA) and root-induction medium (1/4 MS medium fortified with 1.7–2.5 mg l⁻¹ IAA and 0.1–0.5 mg l⁻¹ IBA). In addition, we investigated the genetic stability (relative to the donor plant) of a sample of 41 morphologically normal plants randomly taken from ca. 13,000 micropropagated plants, by using the inter-simple sequence repeat (ISSR) marker with 32 selected primers. We found that of the 226 reproducible bands scored, 24 were polymorphic (10.62%), thus pointing to the occurrence, though at a relatively low level compared with an earlier study on R. pseudoacacia, of genomic variation in these micropropagated plants. Further sequencing on seven loci underlying the variations showed that two had significant homology to known or predicted plant genes.     

Population densities of herbivorous lepidopterans in diverse cruciferous cropping habitats: Effects of mixed cropping and using a living mulch

Two field experiments were conducted in Hawaii to determine the responses of lepidopteran pests and their associated natural enemies to broccoli (Brassica oleracea), grown in monoculture or interplanted with chili pepper (Capsicum annuum), or yellow sweetclover (YSC) (Melilotus officinalis). The population responses of Artogeia rapae and Trichoplusia ni to habitat types were not consistent during the field trials. Additionally, their reactions to habitat diversification differed according to non-host plant species. For example, Trichoplusia ni densities were significantly lower in the broccoli–YSC habitat contrasted with the broccoli–pepper habitat on most sampling dates. Similarly, the proportion of plants damaged by Hellula undalis was significantly lower in the broccoli–YSC habitat contrasted with broccoli–pepper habitat. Findings from the study, suggest that biological control agents (e.g., spiders, parasitoids, and␣entomopathogens) are the most significant factors influencing the abundance of A.␣rapae in habitat type and that these agents inflict greater mortality on A. rapae in diculture than monoculture habitats.     

Variation in soil seed bank species composition of a dry coniferous forest: spatial scale and sampling considerations

In order to assess the effects of disturbance on soil seed bank spatial structure, variation in seed bank species composition was analyzed at two spatial scales in mature (undisturbed) and recently disturbed stands of Interior Douglas-fir forest in south-central British Columbia. Coarse-scale analysis among stands 10–100 km apart using Multi-Response Permutation Procedures showed that individual mature stands were mainly characterized by unique seed bank species compositions. Disturbance had an homogenizing effect on seed bank species composition at the coarse scale––there was less variability among stands following disturbance by both low and high severity fires, and by non-salvage logging. In contrast, finer scale analysis among sampling units c. 10 m apart showed that heterogeneity in seed bank species composition was greater on severely burned and logged sites, commensurate with greater levels of soil disturbance, than on lightly burned and undisturbed sites. Despite the high intensity of seed bank sampling (sixty 25 cm² soil samples from each of 16 sites), species-area curves leveled off only when infrequent species were removed from the dataset. The number of seed bank samples required to account for the common species ranged from 84 on severely burned sites, to 196 on undisturbed sites; more than the 240 samples collected per disturbance class in this study would have been required to account for the remaining infrequent species. Overall, this study highlights the importance of sampling intensively within multiple stands to capture the variation in species composition inherent to these dry coniferous forest soil seed banks.     

Efficiency gain of marker-assisted backcrossing by sequentially increasing marker densities over generations

Expenses for marker assays are the major costs in marker-assisted backcrossing programs for the transfer of target genes from a donor into the genetic background of a recipient genotype. Our objectives were to (1) investigate the effect of employing sequentially increasing marker densities over backcross generations on the recurrent parent genome (RPG) recovery and the number of marker data points (MDP) required, and (2) determine optimum designs for attaining RPG thresholds of 93–98% with a minimum number of MDP. We simulated the introgression of one dominant target gene for genome models of sugar beet (Beta vulgaris L.) and maize (Zea mays L.) with varying marker distances of 5–80 cM and population sizes of 30–250 plants across BC₁ to BC₃ generations. Employing less dense maps in early backcross generations resulted in savings of over 50% in the number of required MDP compared with using a constant set of markers and was accompanied only by small reductions in the attained RPG values. The optimum designs were characterized by increasing marker densities and increasing population sizes in advanced generations for both genome models. We conclude that increasing simultaneously the marker density and the population size from early to advanced backcross generations results in gene introgression with a minimum number of required MDP.