Dispersal of Amblyseius swirskii (Acari: Phytoseiidae) on High-Tunnel Bell Peppers in Presence or Absence of Polyphagotarsonemus latus (Acari: Tarsonemidae).

Amblyseius swirskiiAthias-Henriot (Acari: Phytoseiidae) is a predatory mite used to control thrips (Thysanoptera), whiteflies (Bemisia tabaci Genn., Hemiptera: Aleyrodidae), and broad mites (BMs) (Polyphagotarsonemus latus Banks, Acari: Tarsonemidae). Dispersal of A. swirskii, using the ornamental pepper "Explosive Ember" as a banker plant was evaluated for control of BMs in high-tunnel peppers. Open-canopy plants (5 weeks old) versus closed-canopy plants (10-weeks old) were used to evaluate the effect of plant connectedness in A. swirskii dispersal, in the presence (two females per plant) and absence of BMs. Plots consisted of a single central banker plant and four bell peppers extending linearly north and south. Sets of all treatments were destructively sampled 1, 4, and 7 days after releasing A. swirskii Within 24 h, A. swirskii dispersed four plants away from the banker plants (1 m), regardless of the state of the canopy. Canopy connectedness did increase the presence of A. swirskii on the crop plants. Predatory mite numbers on closed-canopy treatments doubled within the 7-day sampling period, whereas no significant increase was observed on open-canopy treatments. The presence of BMs had no significant effect on the movement of A. swirskii The results suggest further experiments with A. swirskii and banker plants for control of BMs is warranted.

Acute Toxicity and Sublethal Effects of Fenpyroximate to Amblyseius swirskii (Acari: Phytoseiidae).

Knowledge about the effects of pesticides on biological control agents is required in order to successfully implement integrated pest management programs. The predatory mite Amblyseius swirskii Athias-Henriot has been used to control thrips, whiteflies, and broad mites in vegetable production; however, effects of fenpyroximate, an acaricide and insecticide used in vegetable crops, on A. swirskii have not been evaluated. The effect of four residual concentrations of fenpyroximate on A. swirskii females was measured under laboratory conditions including its effect on their fecundity and larval survival. Fresh residues of fenpyroximate were significantly toxic to adult females and larvae. Mortality increased and fecundity decreased as the concentration (0.026-0.208 ml/50 ml of water) and time after treatment (24-120 h) increased. Fifty percent of the larvae survived on the two lower concentrations (0.026 and 0.052 ml/50 ml of water) after 120 h.

A molecular phylogenetic analysis of the vampire moths and their fruit-piercing relatives (Lepidoptera: Erebidae: Calpinae).

Within butterflies and moths, adult hematophagy is limited to species within the vampire moth genus Calyptra. These moths are placed within the subfamily Calpinae, whose other members are known to exhibit a broad range of feeding behaviors including those that can be considered 'piercers' of fruits or other hosts and 'tear feeders'. Here, we reconstruct a phylogenetic hypothesis of Calpinae using molecular data to test whether hematophagy in Calyptra arose from plant or animal-related behaviors. We use a Bayesian method of ancestral state reconstruction to determine the most likely feeding behaviors for the subtribes and genera within this lineage.

First Report of Tar Spot on Orange Geiger, Cordia sebestena, Caused by Diatractium cordianum in Florida.

In July 2007, tar spot symptoms were observed on the leaves of orange Geiger, Cordia sebestena L. (Boraginaceae), in the landscape and a commercial nursery in Homestead, FL. The disease appears to be spreading and is locally severe. Symptoms were circular, slightly hypertrophied spots approximately 5 to 8 cm in diameter, which were slightly chlorotic on the abaxial surface and had numerous circular blackened stroma, 0.2 to 0.4 mm in diameter, on the adaxial surface. As leaves aged and yellowed, the areas around the spots remained pale green. Embedded in the stroma were numerous perithecia, 173 to 312 µm in diameter, circular to irregular in shape, with lateral necks as much as 200 µm long and 73 to 104 µm in diameter. Asci, 77 to 92 × 11 to 13 µm, contained elongate, two-celled ascospores, 50 to 61 × 3 to 5 µm that had a conspicuous constriction at the dividing cell wall. These dimensions and the pathogen's appearance matched closely with those published for Diatractium cordianum (Ellis & Kelsey) Syd (1). Young, symptomless leaves of C. sebestena were sprayed to runoff with a suspension of ascospores approximately 104 ml-1 that were harvested from affected leaves. Inoculated leaves were placed on water-saturated paper towels in petri plates and maintained in a growth chamber at 25°C with fluorescent light at 10 h day-1. Symptoms similar to those observed on affected trees in the landscape began to develop after 21 days and perithecia were evident after 28 days. An ITS 1, ITS 2, and 5.8s rDNA sequence was deposited in GenBank (Accession No. EU541488). A herbarium specimen was deposited at the U.S. National Fungus Collections (BPI No. 878441). This is a new host record for D. cordianum and is the first time the pathogen has been reported in the United States. Previous records were from Venezuela and several Caribbean islands, including Cuba and Jamaica. Symptoms of this disease have not been observed on Texas wild olive, Cordia boissieri, in close proximity to affected C. sebestena. P. F. Cannon (1) indicated that the disease had no economic impact. However, the conspicuous nature of symptoms on C. sebestena and the importance of this tree in the South Florida ornamental trade (2) suggest that this disease may become significant on the latter host. References: (1) P. F. Cannon. Mycol. Res. 92:327, 1989. (2) E. F. Gilman and D. G. Watson. Fact Sheet ST-182. Univ. Fla, Fla Coop Ext. Serv., 1993.

Morphological and molecular characterization of a new microsporidian species from the predatory mite Metaseiulus occidentalis (Nesbitt) (Acari, Phytoseiidae).

A new microsporidian species is described from the predatory mite Metaseiulus (formerly Typhlodromus or Galendromus) occidentalis (Nesbitt) (Acari, Phytoseiidae). The ultrastructure of this new species is presented together with the first molecular characterization for a microsporidium of mites. All stages of this new microsporidium are haplokaryotic and develop in direct contact with the host-cell cytoplasm. Sporogony is disporoblastic and spores are formed in eggs, immature stages, and adults of M. occidentalis. There are two morphological classes of spores, one with a short polar filament (3-5 coils) that measured 2.53 x 1.68 microm and one with a longer polar filament (8-9 coils) that measured 3.14 x 1.77 microm. Horizontal transmission of this new species occurs by cannibalism of eggs and other stages and perhaps involves the spores with the long polar filament. Spores with the short polar filament may play a role in autoinfection and vertical (transovarial) transmission that is highly efficient in transferring the microsporidium from adults to progeny. Analysis of the small subunit ribosomal DNA indicated that this species from M. occidentalis is most closely related to the Nosema/Vairimorpha clade of microsporidia. A conflict between the morphological and molecular data is discussed. The species is compared to previously described microsporidia of arachnids resulting in creation of Oligosporidium occidentalis n. sp. in the family Unikaryonidae.

Long PCR improves Wolbachia DNA amplification: wsp sequences found in 76% of sixty-three arthropod species.

Bacteria belonging to the genus Wolbachia are associated with a variety of reproductive anomalies in arthropods. Allele-specific polymerase chain reaction (= Standard PCR) routinely has been used to amplify Wolbachia DNA from arthropods. While testing the two-spotted spider mite Tetranychus urticae and other arthropods known to be infected with Wolbachia, Standard PCR frequently produced false negatives, perhaps because the DNA from the arthropod host interfered with amplification by Taq DNA polymerase. Long PCR, which uses two enzymes (Taq and Pwo), consistently amplified Wolbachia DNA and a sensitivity analysis indicated that Long PCR was approximately six orders of magnitude more sensitive than Standard PCR in amplifying plasmid DNA spiked into insect genomic DNA. A survey indicated that 76% of sixty-two arthropod species and two subspecies in thirteen orders tested positive for the Wolbachia wsp sequence by Long PCR, which is considerably higher than the rate of 16.9% obtained previously for the ftsZ sequence using Standard PCR (Werren, J.H., Windsor, D. and Gao, L. (1995a) Proc R Soc Lond B 262: 197-204). A subsample of Long PCR products from fourteen arthropod species and two subspecies were sequenced, both directly and after cloning. Two A- and eleven B-Wolbachia strains were detected and their wsp sequences displayed a maximum of 23.7% sequence divergence at this locus. Two new groups (named Fus and Ten) were identified in addition to nineteen reported earlier (Zhou, W., Rousset, F. and O'Neill, S.L. (1998) Proc R Soc Lond B 265: 1-7; van Meer, M.M.M., Witteveldt, J. and Stouthamer, R. (1999) Insect Mol Biol 8: 399-408), because they displayed more than 2.5% sequence divergence from other Wolbachia wsp sequences. PCR products from seventeen of twenty-nine (59%) arthropod species analysed could not be sequenced directly due to apparent infection by multiple Wolbachia strains. The wsp sequences cloned from two such species (Plutella xylostella and Trichoplusia ni) indicated both A- and B-Wolbachia were present in a single individual. Hence, superinfection also may be more widespread than the 1.2% incidence previously estimated.

Field evaluation of integrated pest management-compatible pesticides for the citrus leafminer Phyllocnistis citrella (Lepidoptera: Gracillariidae) and its parasitoid Ageniaspis citricola (Hymenoptera: Encyrtidae).

Potentially selective and integrated pest management (IPM)-compatible pesticides for the citrus leafminer Phyllocnistis citrella Stainton and its parasitoid Ageniaspis citricola Logvinovskaya were compared under nursery field conditions at Gainesville, FL. In 1996, replicated blocks of young grapefruit trees were treated with 2% petroleum oil and 1 x the lowest recommended field rate (LRFR) of diflubenzuron + oil (0.4%). Untreated and treated controls (avermectin at 1 x the LRFR + oil) were included. In 1997, blocks were treated with oil (3%), 1 x the LRFR of azadirachtin + oil, 1 x the LRFR of diflubenzuron + oil and 0.1 x the LRFR of avermectin + oil. Untreated and treated controls were again included. Oil at 3%, azadirachtin at 1 x the LRFR + 0.4% of oil, and diflubenzuron at 1 x the LRFR + 0.4% of oil were shown to be IPM-compatible pesticides. In 1997, these blocks had fewer mines per leaf and P. citrella pupae parasitized by A. citricola per total leaves sampled compared with the untreated control but more than the treated control (alpha = 0.05). Avermectin at 0.1 x the LRFR + 0.4% of oil was not considered an IPM-compatible pesticide because, while it reduced the number of P. citrella mines per leaf, it reduced the number of A. citricola to levels as low as the treated control. Actual P. citrella infestation levels had no detectable effect on tree growth and vigor. Pesticide applications were not justified when P. citrella infestations were < 1 mine per leaf and the biological control agent A. citricola was present.

Transgenic arthropods for pest management programs: risks and realities.

The ability to genetically engineer arthropods using recombinant DNA methods opens new opportunities for improving pest management programs but also creates new responsibilities, including evaluation of the potential risks of releasing transgenic arthropods into the environment. It is now becoming easier to transform diverse species of arthropods by a variety of recombinant DNA methods. Useful genes and genetic regulatory elements are being identified for pest arthropods, but less effort is being expended to identify genes that could improve the efficacy of beneficial arthropods. A transgenic strain of the natural enemy Metaseiulus (= Typhlodromus or Galendromus) occidentalis (Acari: Phytoseiidae) was developed using a method termed maternal microinjection. This transgenic strain was released into an experimental site on the campus of the University of Florida in 1996 after extensive reviews by the University of Florida Biosafety Committee, Florida Department of Agriculture and Consumer Services, United States Department of Agriculture Animal and Plant Health Inspection Service, and the United States Fish and Wildlife Service. The short term releases established a precedent for releasing a transgenic arthropod but, at present, no guidelines are available that would allow transgenic arthropods to be released permanently into the environment. Several scientific, environmental, and policy issues must be resolved before transgenic pests or beneficial arthropods can be deployed in practical pest management programs.

Myths, models and mitigation of resistance to pesticides.

Resistance to pesticides in arthropod pests is a significant economic, ecological and public health problem. Although extensive research has been conducted on diverse aspects of pesticide resistance and we have learned a great deal during the past 50 years, to some degree the discussion about 'resistance management' has been based on 'myths'. One myth involves the belief that we can manage resistance. I will maintain that we can only attempt to mitigate resistance because resistance is a natural evolutionary response to environmental stresses. As such, resistance will remain an ongoing dilemma in pest management and we can only delay the onset of resistance to pesticides. 'Resistance management' models and tactics have been much discussed but have been tested and deployed in practical pest management programmes with only limited success. Yet the myth persists that better models will provide a 'solution' to the problem. The reality is that success in using mitigation models is limited because these models are applied to inappropriate situations in which the critical genetic, ecological, biological or logistic assumptions cannot be met. It is difficult to predict in advance which model is appropriate to a particular situation; if the model assumptions cannot be met, applying the model sometimes can increase the rate of resistance development rather than slow it down. Are there any solutions? I believe we already have one. Unfortunately, it is not a simple or easy one to deploy. It involves employing effective agronomic practices to develop and maintain a healthy crop, monitoring pest densities, evaluating economic injury levels so that pesticides are applied only when necessary, deploying and conserving biological control agents, using host-plant resistance, cultural controls of the pest, biorational pest controls, and genetic control methods. As a part of a truly multi-tactic strategy, it is crucial to evaluate the effect of pesticides on natural enemies in order to preserve them in the cropping system. Sometimes, pesticide-resistant natural enemies are effective components of this resistance mitigation programme. Another name for this resistance mitigation model is integrated pest management (IPM). This complex model was outlined in some detail nearly 40 years ago by V. M. Stern and colleagues. To deploy the IPM resistance mitigation model, we must admit that pest management and resistance mitigation programmes are not sustainable if based on a single-tactic strategy. Delaying resistance, whether to traditional pesticides or to transgenic plants containing toxin genes from Bacillus thuringiensis, will require that we develop multi-tactic pest management programmes that incorporate all appropriate pest management approaches. Because pesticides are limited resources, and their loss can result in significant social and economic costs, they should be reserved for situations where they are truly needed--as tools to subdue an unexpected pest population outbreak. Effective multi-tactic IPM programmes delay resistance (= mitigation) because the number and rates of pesticide applications will be reduced.

Complete sequence of a mariner transposable element from the predatory mite Metaseiulus occidentalis isolated by an inverse PCR approach.

Degenerate primers designed and synthesized based on two conserved regions of the mariner transposase open reading frame were used to amplify a 454 bp DNA fragment from M. occidentalis. Two inverse primers were then synthesized and used to amplify flanking genomic DNA fragments from M. occidentalis by a ligation-mediated inverse PCR. The complete mariner element (Moc1) was 1284 bp long, including the imperfect 28 bp inverted terminal repeat sequences, and shared 59% similarity to an active 1286 bp long D. mauritiana mariner element (Mos1). Insertions, deletions and substitutions were observed in the Moc1 sequence at several positions. No intact open reading frame was detected and the Moc1 element is considered inactive. Stringent Southern blot hybridizations revealed at least twelve copies of mariner sequences similar to Moc1 in the colonies tested.

Transmission of injected DNA sequences to multiple eggs of Metaseiulus occidentalis and Amblyseius finlandicus (Acari: Phytoseiidae) following maternal microinjection.

The persistence of DNA injected into two species of adult female phytoseiids and its transmission to serial eggs deposited by them was assessed by the polymerase chain reaction (PCR). The effect of DNA concentration on persistence and transmission was examined in Metaseiulus occidentalis. M. occidentalis females were microinjected with plasmid DNA at three different concentrations (250, 500, 750 ng microL-1) and allowed to deposit one to five eggs before the females and their last eggs were analyzed. Plasmid DNA was found in 82% of the females assayed and in 70% of all the eggs analyzed (including the fifth eggs produced after microinjection). Transmission of DNA to multiple eggs was also examined in Amblyseius finlandicus. Females of this species are less traumatized by microinjection allowing analysis of transmission over a more extended number of eggs. Females were microinjected and allowed to deposit eggs until their death. DNA from every fifth egg was analyzed by the PCR. PCR products were amplified from 51% of the eggs and from all egg classes except the 30th egg. The persistence and presence of plasmid DNA in both eggs and females suggests that (1) maternal microinjection is a more efficient method for DNA delivery than traditional egg microinjection, (2) it may be possible to isolate transformants from fewer maternally-microinjected females than originally expected, and (3) maternal microinjection could be useful as a DNA delivery system in other phytoseiids.

Transient expression of a Drosophila melanogaster hsp70 promoter/lacZ construct injected into larvae of two species of predatory mites (Acari: Phytoseiidae).

The Drosophila melanogaster heat shock 70 promoter (hsp70) was used to regulate expression of the Escherichia coli beta-galactosidase gene (lacZ) in transiently-transformed predatory mite larvae. A construct containing the hsp70 promoter upstream of the D. melanogaster alcohol dehydrogenase (adh) translational start site and Escherichia coli lacZ gene fusion (adh/lacZ) was injected into larvae of Metaseiulus occidentalis and Amblyseius finlandicus. LacZ expression was compared to expression of a similar construct lacking any upstream regulatory sequence. Expression from the hsp70 promoter was strong and heat shock-dependent in both species. The Drosophila hsp70 promoter therefore appears useful for regulating expression of exogenous DNA in both phytoseiid species and may be broadly applicable in the Phytoseiidae. Furthermore, the lacZ gene is a useful gene for analysis of expression in both species. Larval microinjection provides a method of assessing transient expression and of examining native regulatory sequences in these two phytoseiids and will likely be useful in other phytoseiid mites with only minor modifications.

Stable genetic transformation of a beneficial arthropod, Metaseiulus occidentalis (Acari: Phytoseiidae), by a microinjection technique.

A microinjection technique has resulted in stable transformation of the western predatory mite Metaseiulus occidentalis. Early preblastoderm eggs within gravid females were microinjected. The needle was inserted through the cuticle of gravid females into the egg, or the tissue immediately surrounding the egg. This maternal injection method resulted in relatively high levels of survival and transformation. Transformation was achieved without the aid of any transposase-producing helper plasmid. The predatory mite was transformed with a plasmid containing the Escherichia coli beta-galactosidase gene (lacZ) regulated by the Drosophila hsp70 heat-shock promoter. Putatively transformed lines were isolated based on beta-galactosidase activity in first-generation larvae. Transformation was confirmed in the sixth generation by polymerase chain reaction amplification of a region spanning the Drosophila/E. coli sequences. Amplification of a nested region, also spanning the interspecific boundary, provided further evidence for stable transformation. Maternal microinjection may be adaptable to other beneficial arthropods, particularly other phytoseiid mites. Genetic transformation of M. occidentalis may improve its efficiency as a biological control agent as well as provide a method for investigating details of its physiology and ecology.

Heterochromatinization, chromatin elimination and haploidization in the parahaploid mite Metaseiulus occidentalis (Nesbitt) (Acarina: Phytoseiidae).

Embryogenic mitoses, mitoses in females and spermatogenesis are described in the predatory mite Metaseiulus occidentalis (Nesbitt). At 22 degrees C, egg development lasts approximately 4 days. Six chromosomes are seen in mitotic metaphases and anaphases of 0--24 h eggs. Toward the end of this period some embryo squashes have patches of cells containing nuclei which are partially heteropycnotic. These patches of cells apparently increase in size with the age of the embryo. In approximately 1/2 of all 24--48 h-old eggs they encompass all or most cells of the embryo. In these embryos metaphases involved 6 chromosomes, anaphases 3. Either prior to, or following metaphase, a pairing of chromosomes appeared to take place to form 3 units which resembled meiotic diplotene chromosomes where there is opening out between homologues. At metaphase, two sets of 3 chromosomes were slightly differentially stained. One, designated the H set, was darker and slightly more contracted than the other, the E set. At anaphase, 3H and 3E chromosomes segregated in a reductional division retaining the differential contraction until telophase. No cytokinesis appeared. The H set appeared to remain contracted while the E set decontracted to assume the appearance of an interphase nucleus. Both of these entities, side-by-side, created the partially heteropycnotic nucleus mentioned above. The H set then appeared to be excluded from the cell. Mitotic meta- and anaphases involving 6 chromosomes were noted in female deutonymphs. Spermatogenesis appeared to encompass an equational division of 3 chromosomes, which the formation of a binucleate spermatid. Two tail structures appeared juxtaposed at the edge of each spermatid and thereafter a separation into two individual sperms occurred.--While mitosis was not studied in known males, we believe that the embryos exhibiting heterochromatinization and elimination of chromosomes in most or all cells were in fact demonstrating parahaploidization.

Rapid response to selection for a nondiapausing gypsy moth.

By genetic selection a gypsy moth strain was obtained within eight generations that could be reared continuously in the insectary without its normal "obligatory" diapause. This strain should be useful for making physiological and genetical comparisons with normal diapausing gypsy moths. It could also be used in a genetic control program. However, caution should be exercised, for its release could create a new environmental hazard.

The GP - doctor or middle-man? A study of his paper work.

The convention of using medical certificates to assess benefit claims has grown over the years to a complex situation. The certificate serves as written testimony of the patient's state of health (be it illness, fitness or a need which is based on those grounds). While the practitioner is presumed to be serving only his patient, this study suggests that he is required to act as arbitrator in many situations that are unnecessarily defined as medical.