Vector competence of Rhipicephalus sanguineus sensu stricto for Anaplasma platys.

Anaplasma platys is a Gram-negative, obligate intracellular bacteria that causes canine infectious cyclic thrombocytopenia in dogs. The brown dog tick Rhipicephalus sanguineus sensu lato is presumed to be the vector of A. platys based on the overlap in distribution of R. sanguineus and A. platys infections, detection of A. platys DNA in both flat and engorged field-collected R. sanguineus, and the fact that dogs are primary hosts of both brown dog ticks and A. platys. However, the only study evaluating the vector competence of R. sanguineus for A. platys under controlled laboratory conditions reported an apparent inability of ticks to acquire or maintain the pathogen. In 2016, engorged female Rhipicephalus sanguineus sensu stricto ticks were collected off dogs to start a laboratory tick colony. After one generation of colony maintenance on tick-naïve and pathogen-free New Zealand White rabbits, a rabbit used to feed F1 adults seroconverted to Anaplasma phagocytophilum antigen. PCR and subsequent DNA sequencing identified the presence of A. platys in both the adult ticks fed on this rabbit and their resulting F2 progenies. Retrospective testing of all previous (P and F1) life stages of this colony demonstrated that the infection originated from one field-collected A. platys-infected female whose progeny was propagated in the laboratory and produced the PCR-positive F1 adults. Over the following (F2-F4) generations, the prevalence of A. platys in this colony reached 90-100 % indicating highly efficient transovarial and horizontal transmission, as well as transstadial maintenance, of this pathogen by R. sanguineus s.s.

To Acclimate or Not to Acclimate? Simultaneous Positive and Negative Effects of Acclimation on Susceptibility of Tribolium confusum (Coleoptera: Tenebrionidae) and Oryzaephilus surinamensis (Coleoptera: Silvanidae) to Low Temperatures.

Laboratory tests on acclimated and nonacclimated life stages of Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) (adults, pupae, larvae, and eggs) and Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae) (adults, larvae, and eggs) were conducted at 0, -5, -10, and -15°C to evaluate effects of acclimation on susceptibility to cold treatment. Acclimation of all tested life stages for 7 d at 15°C affected susceptibility of both species to the cold temperatures. After 1 d exposures for ≥2 h, acclimated adults had a noticeable increase in cold tolerance compared with nonacclimated adults for both tested species. Nonacclimated pupae of T. confusum were equally susceptible to cold compared with acclimated pupae at short exposures to low temperatures. Exposure of nonacclimated life stages of T. confusum, at -10°C for 1 d gave 0% survival. Similarly, almost all (99.6%) nonacclimated individuals of O. surinamensis died at -10°C. At 0°C, nonacclimated larvae were more cold tolerant than acclimated larvae, but this trend was reversed when larvae were exposed to -5°C. Mixed results were obtained for larvae of O. surinamensis because in some of the combinations tested, nonacclimated larvae were more tolerant, even at temperatures that were lower than 0°C. In contrast to O. surinamensis, eggs of T. confusum that had not been exposed to cold were not affected by acclimation, while exposure to cold showed increased cold hardiness in acclimated eggs. Results show that individual stored-product insect species may have mixed susceptibility to cold temperatures, which must be taken into account when using cold treatment as a management strategy.

Susceptibility of Different Life Stages of Tribolium confusum (Coleoptera: Tenebrionidae) and Oryzaephilus surinamensis (Coleoptera: Silvanidae) to Cold Treatment.

Laboratory tests were carried out to examine the efficacy of different exposure intervals (2 h, 4 h, 8 h, 1 d, 2 d, 3 d, and 7 d) on different life stages (adults, pupae, larvae, and eggs) of Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae), the confused flour beetle, and Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae), the saw-toothed grain beetle (adults, larvae, and eggs) to 0, -5, -10, and -15°C. Larvae and pupae of T. confusum were more cold-tolerant than eggs or adults. Exposure to temperatures of -10°C for 1 d will kill nearly 100% of all life stages of T. confusum. O. surinamensis was more cold-tolerant than T. confusum. Adults of O. surinamensis were not killed when exposed for 1 d at -5°C, but egg hatch was drastically reduced after 2 h of exposure at the same temperature. Eggs and adults of O. surinamensis were more cold-tolerant than larvae. Our study indicates that target insect species and life stage, temperature, and exposure interval should all be considered when cold treatment is selected as a control strategy against T. confusum and O. surinamensis. Facility managers can use these data in planning cold treatments.

A nuptially transmitted ichthyosporean symbiont of Tenebrio molitor (Coleoptera: Tenebrionidae).

The yellow mealworm, Tenebrio molitor, harbors a symbiont that has spores with a thick, laminated wall and infects the fat body and ventral nerve chord of adult and larval beetles. In adult males, there is heavy infection of the epithelial cells of the testes and between testes lobes with occasional penetration of the lobes. Spores are enveloped in the spermatophores when they are formed at the time of mating and transferred to the female's bursa copulatrix. Infection has not been found in the ovaries. The sequence of the nuclear small subunit rDNA indicates that the symbiont is a member of the Ichthyosporea, a class of protists near the animal-fungi divergence.

Phenoloxidase in larvae of Plodia interpunctella (Lepidoptera: Pyralidae): molecular cloning of the proenzyme cDNA and enzyme activity in larvae paralyzed and parasitized by Habrobracon hebetor (Hymenoptera: Braconidae).

Phenoloxidase (PO) is a major component of the insect immune system. The enzyme is involved in encapsulation and melanization processes as well as wound healing and cuticle sclerotization. PO is present as an inactive proenzyme, prophenoloxidase (PPO), which is activated via a protease cascade. In this study, we have cloned a full-length PPO1 cDNA and a partial PPO2 cDNA from the Indianmeal moth, Plodia interpunctella (Hubner) (Lepidoptera: Pyralidae) and documented changes in PO activity in larvae paralyzed and parasitized by the ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae). The cDNA for PPO1 is 2,748 bp and encodes a protein of 681 amino acids with a calculated molecular weight of 78,328 and pI of 6.41 containing a conserved proteolytic cleavage site found in other PPOs. P. interpunctella PPO1 ranges from 71-78% identical to other known lepidopteran PPO-1 sequences. Percent identity decreases as comparisons are made to PPO-1 of more divergent species in the orders Diptera (Aa-48; As-49; and Sb-60%) and Coleoptera (Tm-58; Hd-50%). Paralyzation of host larvae of P. interpunctella by the idiobiont H. hebetor results in an increase in phenoloxidase activity in host hemolymph, a process that may protect the host from microbial infection during self-provisioning by this wasp. Subsequent parasitization by H. hebetor larvae causes a decrease in hemolymph PO activity, which suggests that the larval parasitoid may be secreting an immunosuppressant into the host larva during feeding.