Simulating effects of environmental factors on biological control of Tetranychus urticae by Typhlodromus pyri in apple orchards

Successful biological control of mites is possible under various conditions, and identifying what are the requirements for robust control poses a challenge because interacting factors are involved. Process-based modeling can help to explore these interactions and identify under which conditions biological control is likely, and when not. Here, we present a process-based model for population interactions between the phytophagous mite, Tetranychus urticae, and its predator, Typhlodromus pyri, on apple trees. Temperature and leaf nitrogen concentration influence T. urticae rates of development and reproduction, while temperature and rate of ingestion of prey and pollen influence T. pyri rates of survival and reproduction. Predator and prey population dynamics are linked through a stage structured functional response model that accounts for spatial heterogeneity in population density throughout the trees. T. urticae biomass-days (BMD’s), which account for sizes of larvae, nymphs and adults, indicate level of mite-induced leaf damage. When BMD’s exceed 290 per leaf, there are economic losses. When BMD’s exceed 350 per leaf, T. urticae population growth is curbed and eventually the population decreases. Simulations were run to determine which conditions would lead to current year economic loss and increased risk of loss in the following year, i.e. where more T. urticae than T. pyri are present at the end of September. Risk was high with one or more of the following initial conditions: a high prey: predator ratio (10:1 or more); a low to intermediate (0.04–0.2 T. urticae per leaf) initial density; T. urticae with a higher initial proportion of adult females than T. pyri; and a delayed first detection of mites, whether in late July, or sometimes in late June, but not in early June. Warm summer weather, higher leaf nitrogen and T. urticae immigration into trees were also risk factors. Causes for these patterns based on biological characteristics of T. urticae and T. pyri are discussed, as are counter measures which can be taken to reduce risk.     

Laboratory evaluation of resistance to pesticides in the phytoseiid predator Typhlodromus pyri from English apple orchards

Typhlodromus pyri, reared on plate cultures and fed on pollen of Vicia faba, were bioassayed using a taped-slide technique. Mite stocks from isolated unsprayed orchards were used to establish base-line susceptibility to azinphos-methyl, parathion, carbaryl and permethrin. Stocks from English orchards with a spray history of organophosphates and carbaryl showed resistance of 4 to 6x to azinphos-methyl, c 50 x to parathion, >20x to carbaryl, and no resistance to permethrin. An orchard population derived from T. pyri imported from New Zealand in 1977 responded similarly. In other tests, mites exposed to spray residues on glass and on apple leaves were killed by lower concentrations than in taped-slide tests (glass < apple leaf < taped-slide technique); but for all three assay techniques the resistance factors to azinophos-methyl and carbaryl, comparing two stocks, were similar. All stocks from sprayed orchards were resistant to both azinphos-methyl and carbaryl, suggesting cross-resistance; and resistance to both these pesticides appeared to be stable in the field when selection pressure was relaxed. The results are discussed in relation to earlier work on T. pyri and two other resistant orchard-inhabiting phytoseiid species.     

Inter- and intraspecific predation on immatures by adult females in Euseius finlandicus, Typhlodromus pyri and Kampimo dromus aberrans (Acari: Phytoseiidae)

In a series of experiments, the interspecific predation and cannibalism on immatures by the adult females of Euseius finlandicus, Kampimodromus aberrans and Typhlodromus pyri were examined under laboratory conditions. The three species showed differing tendencies to prey on each other's motile immature stages. Euseius finlandicus females consumed more larvae and protonymphs than the females of T. pyri and K. aberrans. In cages without free water E. finlandicus ate a range of 6.51 larvae or 5.31 protonymphs of T. pyri and 5.27 larvae or 5.95 protonymphs of K. aberrans per female per day. Kampimodromus aberrans and T. pyri females exhibited a greater tendency to prey on heterospecifics than on conspecifics. When feeding on phytoseiid immatures and without free water, T. pyri females survived longer and laid more eggs than the females of E. finlandicus and K. aberrans. Adult females of E. finlandicus and T. pyri having free water and preying on heterospecific protonymphs were able to maintain egg laying during the whole experimental period of 12 days. The females of all three species had difficulties in piercing phytoseiid eggs, and the number of sucked eggs per female per day was low. Kampimodromus aberrans females ate 0.48 eggs of T. pyri daily, which was the highest recorded number. The great tendency to interspecific predation on motile immature phytoseiids by the females of E. finlandicus is discussed with regard to the dominance of this species on deciduous trees and bushes in Austria.     

The analysis and interpretation of DNA distributions measured by flow cytometry

A principal use of flow cytometers is for the measurement of fluorescence distributions of cells stained with DNA specific dyes. A large amount of effort has been and is being expended currently in the analysis of these distributions for the fractions of cells in the G1, S, and G2 + M phases. Several methods of analysis have been proposed and are being used; new methods continue to be introduced. Many, if not most, of these methods differ only in the mathematical function used to represent the phases of the cell cycle and represent attempts to fit exactly distributions with known phase fractions or unusual shapes. In this paper we show that these refinements probably are not necessary because of cell staining and sampling variability. This hypothesis was tested by measuring fluorescence distributions for Chinese hamster ovary and KHT mouse sarcoma cells stained with Hoechst-33258, chromomycin A3, propidium iodide, and acriflavine. Our results show that: a) single measurements can result in phase fraction estimates that are in error by as much as 40% for G2 + M phase and 15-20% for G1 and S phases; b) different dyes can yield phase fraction estimates that differ by as much as 40% due to differences in DNA specificity; c) the shapes of fluorescence distributions and their interpretation are very dependent on the dye being used and on its binding mechanism.     

几种常见桔园螨类Wolbachia基因检测与序列分析

应用共生菌沃尔巴克氏体Wolbachia的wsp、ftsZ和16SrDNA基因特异引物,通过PCR扩增法对采自江西省南昌、九江和赣州等3地桔园内的东方钝绥螨、江原钝绥螨、尼氏真绥螨、巴氏新小绥螨、桔全爪螨和酢浆草如叶螨等6种常见螨类进行了检测。结果表明,在采自南昌桔园内的酢浆草如叶螨体内检测到Wolbachia感染,其wsp、ftsZ和16SrDNA基因长度为593、758和896bp,Genbank登录号依次为GQ162482、GQ162483和GQ162484,而其他5种螨体内均未检测出Wolbachia的感染;同时通过对酢浆草如叶螨体内感染的Wolbachia的wsp基因序列进行系统发育分析,得出其与B大组的Epo组的Wolbachia株十分相近,可能是相近或相同的株。     

Estimating gene flow between refuges and crops: a case study of the biological control of Eriosoma lanigerum by Aphelinus mali in apple orchards

Parasitoid disturbance populations in agroecosystems can be maintained through the provision of habitat refuges with host resources. However, specialized herbivores that feed on different host plants have been shown to form host-specialized races. Parasitoids may subsequently specialize on these herbivore host races and therefore prefer parasitizing insects from the refuge, avoiding foraging on the crop. Evidence is therefore required that parasitoids are able to move between the refuge and the crop and that the refuge is a source of parasitoids, without being an important source of herbivore pests. A North-South transect trough the Chilean Central Valley was sampled, including apple orchards and surrounding Pyracantha coccinea (M. Roem) (Rosales: Rosacea) hedges that were host of Eriosoma lanigerum (Hemiptera: Aphididae), a globally important aphid pest of cultivated apples. At each orchard, aphid colonies were collected and taken back to the laboratory to sample the emerging hymenopteran parasitoid Aphelinus mali (Hymenoptera: Aphelinidae). Aphid and parasitoid individuals were genotyped using species-specific microsatellite loci and genetic variability was assessed. By studying genetic variation, natural geographic barriers of the aphid pest became evident and some evidence for incipient host-plant specialization was found. However, this had no effect on the population-genetic features of its most important parasitoid. In conclusion, the lack of genetic differentiation among the parasitoids suggests the existence of a single large and panmictic population, which could parasite aphids on apple orchards and on P. coccinea hedges. The latter could thus comprise a suitable and putative refuge for parasitoids, which could be used to increase the effectiveness of biological control. Moreover, the strong geographical differentiation of the aphid suggests local reinfestations occur mainly from other apple orchards with only low reinfestation from P. cocinnea hedges. Finally, we propose that the putative refuge could act as a source of parasitoids without being a major source of aphids.     

Comparison of soil tests and leaf analysis as methods of diagnosing Zn deficiency in British Columbia apple orchards

Soil Zn extracted by 0.25M MgCl₂ or DTPA as an index of Zn availability was compared to Zn or P:Zn concentration in leaves for 40 commercial apple orchards in southern British Columbia. Sampled trees included a wide range of ages and cultivar/rootstock combinations. Leaves were sampled from the midportion of current season's shoots at four times; May–June 1984, July 1984, May 1985 and July 1985. At the same time the severity of the four Zn deficiency symptoms, rosetting, blind bud, little leaf and chlorosis was assessed. Little relationship was found between soil and leaf Zn measures, except in July 1984, when both terminal leaf Zn and P:Zn concentrations varied directly with 0.25M MgCl₂ extractable soil Zn. Soil Zn extracted by 0.25M MgCl₂ was more closely related to severity of deficiency symptoms on the trees at all four samples dates than was DTPA-soil Zn or leaf Zn concentration. Blindbud was the most useful indicator of deficiency in samples collected in May whereas chlorosis was the most useful one in July.     

Exposure risks to pesticide applicators posed by the use of electric backpack sprayers and stretcher-mounted sprayers in orchards

Abstract

The introduction of the pesticide registration system in pesticide risk assessments has promoted the scientific and safe use of pesticides, and the assessment of occupational exposure risk. In the present study, we performed an experiment in a citrus orchard subject to normal orchard management practices. By measuring the exposure of applicators’ (farmers and technicians) body parts to 45% malathion emulsifiable concentrate during its application using electric backpack sprayers (0.25?KPa) and stretcher-mounted sprayers (1.5?KPa), the unit exposure (UE) was determined. The risks of exposure for pesticide applicators who adopted five different protective measures (A: no personal protective equipment (PPE), i.e., no clothes, no gloves, no caps, and no socks; B: short-sleeved top and shorts; C: short-sleeved top, shorts, and a single pair of gloves; D: short-sleeved top, shorts, a single pair of gloves, and a cap; E: long-sleeved top, long pants, a single pair of gloves, and a cap) were also assessed. The results were as follows: 1) The total levels of exposure for pesticide applicators using electric backpack sprayers and stretcher-mounted sprayers were 3613.63?µg and 5654.28?µg, respectively. When electric backpack sprayers were used, the body parts that had the highest exposure were the head (13.8%), hands (19.9%) and back (14.0%), and when stretcher-mounted sprayers were used, the hands (32.5%) and lower legs (21.1%) had the highest level of exposure; 2) In the absence of PPE, the UE values for farmers who used electric backpack sprayers and farmers who used stretcher-mounted sprayers were significantly different. However, when PPE was used, the difference in UE values between the farmers using the two different types of sprayers was not significant; 3) When protective measure A was adopted, the risk quotient (RQ) values of the farmers and technicians who used electric backpack sprayers for the application of malathion were 1.44 and 0.54, respectively; the corresponding RQ values when protective measure B was adopted were 0.97 and 0.28, respectively. When stretcher-mounted sprayers were used for the application of chlorpyrifos, the RQ value of the farmers who adopted protective measure E was 0.43 while other types of PPE use resulted in RQ values greater than 1. In contrast, the RQ value for technicians was 1.62 when protective measure A was used and 1.02 when protective measure B was adopted, whereas other types of PPE use resulted in RQ values less than 1. Therefore, besides increasing the awareness of personal protection among pesticide applicators, improvement in the management of pesticide use and the enhancement of standard operations are of practical significance for controlling occupational exposure to pesticides.     

Potential of the luminol reaction in the sensitive detection of pesticide residues by flow injection analysis.

This study presents the first analytical application of the luminol chemiluminescence (CL) reaction for the sensitive detection of carbamate residues. Some experiments have been carried out to check the influence of the presence of traces of a N-methylcarbamate (carbaryl) on the CL emission produced from the oxidation of luminol using different oxidants, showing a significant enhancing effect on the CL emission when the oxidation of luminol is produced by potassium permanganate in alkaline medium, this enhancement being proportional to the carbaryl concentration. This fact has permitted the establishment of a sensitive chemiluminescence flow-injection (CL-FIA) method for the direct determination of carbaryl. The optimization of instrumental and chemical variables influencing the CL response has been carried out by applying experimental designs. Under the optimal conditions, the CL intensity was linear for a carbaryl concentration over the range 5-100 ng/mL with a detection limit of 4.9 ng/mL. This luminol-KMnO4-based FIA-CL system in basic medium shows an easy, fast and cheap alternative detection mode for the analysis of carbaryl residues in environmental water samples.     

Gene flow and mating system in five <Emphasis Type=Italic>Cryptomeria japonica</Emphasis> D. Don seed orchards as revealed by analysis of microsatellite markers

We investigated gene flow in five Cryptomeria japonica D. Don seed orchards of two different types (common and miniature) at widely spaced locations using microsatellite markers. The quality of a seed crop is determined by many factors, including pollen contamination from outside sources, self-fertilization, and the proportion of contributions from constituent clones. Contamination rates were found to vary among ramets both within seed orchards (10.0–76.7% in the most variable seed orchard) and among seed orchards (35.0–65.8% on average). Among ramets, there were significant negative correlations between pollen contamination rate and their distance from the orchard edge; among seed orchards, there were significant positive correlations between the pollen contamination rate and the C. japonica forest area nearby. Some proportion of the pollen (10.7% of total contamination) also migrated from parts of the orchards that had not been treated with gibberellin to induce flowering. Self-fertilization rates varied among seed orchards (1.4–4.4% on average), and there were significant positive correlations between self-fertilization rate and the number of ramets per clone in both types of seed orchard. Contributions as pollen donors differed significantly among clones in all seed orchards. The distance between planted ramets, flowering phenology, and relative pollen fecundity may also have contributed to observed differences in paternal contribution. The influence of these factors on genetic potential did not differ greatly between the two types of orchards. This work was supported by grants from the Pioneer Special Studies Program of the Japanese Ministry of Agriculture, Fishery, and Forestry; the Program for Promotion of Basic Research Activities for Innovative Biosciences; and research fellowships from the Japan Society for the Promotion of Science for Young Scientists.     

Selection and genetic analysis of permethrin resistance in Amblyseius fallacis (Garman) (Acari: Phytoseiidae) from Ontario apple orchards

Gravid female phytoseiid mites, Amblyseius fallacis (Garman), were selected for resistance to permethrin using a table-top Petri dish (PD) technique. After 55 selections, the LC₅₀ of the R population increased 964-fold to 12 241 p.p.m. (PD) and 3.6-fold to 167 p.p.m. by a slide-dip (SD) method. A genetic analysis was conducted to examine the mode of inheritance by a series of single-pair reciprocal interstrain crosses and backcrosses of female F₁ progeny. Concentration-mortality regressions (PD) for parental populations showed a 69.4-fold difference between R and S strains. Regressions for F₁ females resulting from both interstrain crosses were intermediate between parental strains and not significantly different, showing no obvious maternal effect. Resistance in the R strain was not completely dominant or recessive and the estimated dominance, D, for the F₁ females from combined data of both reciprocal crosses was-0.18, for the female R x male S cross it was 0.24 and for the female S x male R cross it was-0.01. We concluded that more than one gene was responsible for the resistance in the selected R strain. No incompatibility was observed in any reciprocal interstrain crosses or backcrosses between the S strain and R strain.     

Surface antigen changes occurring in short-term cultures of activated human T lymphocytes: analysis by flow cytometry

Surface antigens of activated and cultured human T cells were studied using peripheral blood lymphocytes activated with conditioned medium from phytohemagglutinin-activated leukocytes and maintained in liquid culture for 2 weeks with conditioned medium containing Interleukin 2. The ensuing cell population was tested for kinetic changes in cell size and for the expression of surface antigens by immunofluorescence staining with a panel of monoclonal antibodies and analysis by flow cytometry. Upon activation, the cell population progressively increased in size to large blasts, with the rapid appearance on all of the large dividing cells of the antigen recognized by OKT9, the transferrin receptor. Cells within the population continued to express the common peripheral T-cell antigens bound by OKT3 and UCHT1, and also the antigen bound by 3A1, but never the antigen bound by OKT6, a thymic cell marker. From the time of activation an increasing proportion of the T cells, up to 80%, expressed the antigen detected with OKIa and FMC4, which recognise nonpolymorphic Ia determinants. This sequence of events was followed by a general decrease in size of the cell population, a process accompanied by further phenotypic changes. The percentage of cells expressing Ia antigens decreased, but most striking was the rapid change in the OKT4:OKT8 ratio of cells within the population, from 60:40 to 40:60. Thereafter the proportions of OKT4⁺ to OKT8⁺ cells within the cultures remained relatively stable and it is suggested that these data provide evidence for a possible change in phenotype of cultured human T lymphoblasts, from OKT4 to OKT8.     

Signal transduction in monocytes and granulocytes measured by multiparameter flow cytometry.

The novel calcium indicator fura red and the oxidative burst indicator dihydrorhodamine (both excited at 488 nm) were used in combination with multiparameter flow cytometry to allow simultaneous kinetic measurements of calcium fluxes and oxidative bursts in monocytes and granulocytes. Using this method it was possible to obtain direct evidence for the following cell type- and stimulus-specific differences in signal transduction pathways: 1) n-formyl-methionyl-leucyl-phenylalanine (FMLP)/cytochalasin B-induced oxidative burst is several-fold higher in granulocytes than in monocytes although the calcium fluxes have similar amplitudes in the two cell types; 2) stimulus-induced calcium fluxes in granulocytes are mainly due to release from intracellular stores, whereas monocytes mobilize calcium mainly by influx from the medium; 3) the FMLP/cytochalasin B-induced calcium flux in monocytes is less sensitive to the G-protein inhibitor pertussis toxin than the flux in granulocytes; 4) in contrast to FMLP/cytochalasin B, the protein kinase C activator phorbol myristate acetate (PMA) induces an oxidative burst that is not preceded by a cytoplasmic calcium flux; 5) the PMA-induced oxidative burst can be triggered in monocytes and granulocytes that are depleted of intracellular calcium ions, whereas that induced by FMLP/cytochalasin B can not; 6) the G-protein inhibitor pertussis toxin blocks an early event in the signal transduction pathway of FMLP/cytochalasin B, as shown by inhibition of both calcium fluxes and oxidative burst; and 7) 100 nM of the protein kinase inhibitor staurosporine blocks the FMLP/cytochalasin B-induced respiratory burst by interfering with a step downstream to cytoplasmic calcium fluxes, whereas only 10-20 nM is necessary to block PMA-induced oxidative burst.     

Laboratory studies on the feeding habits,reproduction and development of three phytoseiid species,Typhlodromus pomi,Phytoseius macropilis andAmblyseius finlandicus (Acari: Phytoseiidae), occurring on abandoned apple trees in Ontario,Canada

Laboratory studies on predation, reproduction and development were conducted on three phytoseiid species,Typhlodromus pomi (Parrott),Phytoseius macropilis (Banks) andAmblyseius finlandicus (Oudemans). These species were dominant phytoseiid mites on the abandoned apple trees with which ecological experiments were carried out in southern Ontario, Canada. Throught the studies, a rust mite species (Diptacus gigantorhynchus sp. complex), a single consistent prey in the orchard, was used as food. Results showed that the three predators developed well with the rust mites as food, and thatA. finlandicus developed most rapidly and had the highest predation rate. However, this species showed a poor tolerance to low prey densities. These observations suggest that this species is a predacious species and the other two species were more or less general feeders. All species showed the same pre-mating behaviour (Amblyseius-Typhlodromus type), but they exhibited a variety of mating and oviposition behaviours.     

Comparison of S-phase fractions measured by flow cytometry and autoradiography in human transplant tumors

The 3H-thymidine labeling index (TLI) and the percentage of cells in the S-phase have been determined by autoradiography and by flow cytometry, (FCM), respectively, in six malignant tumors of human origin transplanted on athymic nude mice. The Dean and Jett model and the graphical model were used to determine the percent of S-phase cells by FCM. Cell cycle analysis was performed using 1) no correction for background; 2) an algebraic function for background correction; and 3) an exponential function for background subtraction. Each of these three data sets was evaluated using both the Dean and Jett model and a graphical model for the evaluation of DNA histograms. The S-phase fractions (SPF) were compared to the corresponding labeling index results. SPF without background correction were 1.54 times higher than the TLI. SPF, after correction using the algebraic model, were 1.29-fold higher than the TLI, whereas SPF obtained after background subtraction according to the exponential model were only 1.05-fold higher than the TLI. Student's t-test revealed significant differences between the mean TLI values (16.25 +/- 9.06) and the mean SPF obtained by FCM without background correction (mean 25.0 +/- 9.36, P less than 0.01), but not between the mean TLI values and the mean SPF percentages after algebraic (mean 21.0 +/- 10.29) and exponential background correction (mean 17.11 +/- 11.59), P greater than 0.05 each. There was no difference between the results obtained using the Dean and Jett model and those obtained using the graphic evaluation.(ABSTRACT TRUNCATED AT 250 WORDS)