A degree-day model for forecasting adult phenology of Popillia japonica (Coleoptera: Scarabaeidae) in a temperate climate.

Japanese beetle, Popillia japonica (Newman), was first detected in the United States in New Jersey in 1916. The beetle gradually spread to the Midwest U.S. region, and was first confirmed in Minnesota in the late 1960's. Popillia japonica has subsequently become a major invasive insect pest in turfgrass and several agricultural crops. As P. japonica continues to spread throughout the U.S., and other countries, it is important to develop efficient ways to monitor adult populations, and where possible, forecast the phenology of adult population dynamics. During 2019-2021, field trials were conducted to develop a degree-day model that can be used to forecast P. japonica adult phenology under Minnesota, and Midwest summer climatic conditions in. We used commercially available traps and lures to monitor adult flight phenology, specifically beetle trap-catch, along with weather data at four locations in Minnesota, to relate ambient field temperatures to population phenology. The concordance correlation coefficient (CCC), an index of both precision and accuracy, was used to develop a final degree-day model. Model development included evaluation of simple and sine-wave degree-day calculation methods, start dates between 1 Jan. and 1 April, and a range of lower (0-15 °C) and upper (20-37 °C) thresholds. The optimum model was found to be a simple degree-day calculation, using a biofix date of 1 Jan, and lower and upper thresholds of 15 and 21.7 °C, respectively, for predicting 10% beetle trap-catch. The model will aid in future integrated pest management (IPM) and regulatory strategies by providing a tool for prediction of P. japonica adult flight phenology.

Degree-Day Prediction Models for the Flight Phenology of Western Bean Cutworm (Lepidoptera: Noctuidae) Assessed with the Concordance Correlation Coefficient.

Western bean cutworm, Striacosta albicosta (Smith) (Lepidoptera: Noctuidae), is a native, univoltine pest of corn and dry beans in North America. The current degree-day model for predicting a specified percentage of yearly moth flight involves heat unit accumulation above 10°C after 1 May. However, because the moth's observed range has expanded into the northern and eastern United States, there is concern that suitable temperatures before May could allow for significant S. albicosta development. Daily blacklight moth catch and temperature data from four Nebraska locations were used to construct degree-day models using simple or sine-wave methods, starting dates between 1 January and 1 May, and lower (-5 to 15°C) and upper (20 to 43.3°C) developmental thresholds. Predicted dates of flight from these models were compared with observed flight dates using independent datasets to assess model performance. Model performance was assessed with the concordance correlation coefficient to concurrently evaluate precision and accuracy. The best model for predicting timing of S. albicosta flight used simple degree-day calculations beginning on 1 March, a 3.3°C (38°F) lower threshold, and a 23.9°C (75°F) upper threshold. The revised cumulative flight model indicated field scouting to estimate moth egg density at the time of 25% flight should begin when 1,432 degree-days (2,577 degree-days °F) have accumulated. These results underscore the importance of assessing multiple parameters in phenological models and utilizing appropriate assessment methods, which in this case may allow for improved timing of field scouting for S. albicosta.

Thermocouple design for measuring temperatures of small insects.

Contact thermocouples often are used to measure surface body temperature changes of insects during cold exposure. However, small temperature changes of minute insects can be difficult to detect, particularly during the measurement of supercooling points. We developed two thermocouple designs, which use 0.51 mm diameter or 0.127 mm diameter copper-constantan wires, to improve our ability to resolve insect exotherms. We tested the designs with adults from three parasitoid species: Tetrastichus planipennisi, Spathius agrili, and S. floridanus. These species are <3 mm long and <0.1 mg. Mean exotherms were greater for fine-gauge thermocouples than thick-gauge thermocouples for the smallest species tested, T. planipennisi. This difference was not apparent for larger species S. agrili and S. floridanus. Thermocouple design did not affect the mean supercooling point for any of the species. The cradle thermocouple design developed with the fine gauge wire was reusable and allowed for easy insect recovery after cold exposure.

Synthesis and structure determination of 6-methylbenzo[a]pyrene-deoxyribonucleoside adducts and their identification and quantitation in vitro and in mouse skin.

Activation of the moderate carcinogen 6-methylbenzo[a]pyrene (6-CH(3)BP) by one-electron oxidation to form DNA adducts was studied. Iodine oxidation of 6-CH(3)BP in the presence of dGuo produces BP-6-CH(2)-N(2)dGuo, BP-6-CH(2)-N7Gua and a mixture of 6-CH(3)BP-(1&3)-N7Gua, whereas in the presence of Ade the adducts BP-6-CH(2)-N1Ade, BP-6-CH(2)-N3Ade, BP-6-CH(2)-N7Ade and 6-CH(3)BP-(1&3)-N1Ade are obtained. Furthermore, for the first time an aromatic hydrocarbon radical cation afforded an adduct with dThd, the stable adduct BP-6-CH(2)-N3dThd. Formation of these adducts indicates that the 6-CH(3)BP radical cation has charge localized at the 6, 1 and 3 position. When 6-CH(3)BP was activated by horseradish peroxidase in the presence of DNA, two depurinating adducts were identified, BP-6-CH(2)-N7Gua (48%) and 6-CH(3)BP-(1&3)-N7Gua (23%), with 29% unidentified stable adducts. In the binding of 6-CH(3)BP catalyzed by rat liver microsomes, the same two depurinating adducts, BP-6-CH(2)-N7Gua (22%) and 6-CH(3)BP-(1&3)-N7Gua (10%), were identified, with 68% unidentified stable adducts. In 6-CH(3)BP-treated mouse skin, the two depurinating adducts, BP-6-CH(2)-N7Gua and 6-CH(3)BP-(1&3)-N7Gua, were identified. Although quantitation of these two adducts was not possible due to coelution of metabolites on HPLC, they appeared to be the major adducts found in mouse skin. These results show that 6-CH(3)BP forms depurinating adducts only with the guanine base of DNA, both in vitro and in mouse skin. The weaker reactivity of 6-CH(3)BP radical cation vs. BP radical cation could account for the weaker tumor-initiating activity of 6-CH(3)BP in comparison to that of BP.

Synthesis of adducts formed by iodine oxidation of aromatic hydrocarbons in the presence of deoxyribonucleosides and nucleobases.

Polycyclic aromatic hydrocarbons (PAH) undergo two main pathways of metabolic activation related to the initiation of tumors: one-electron oxidation to give radical cations and monooxygenation to yield bay-region diol epoxides. Synthesis of standard adducts is essential for identifying biologically formed adducts. Until recently, radical cation adducts were synthesized by oxidation of the PAH in an electrochemical apparatus, not readily available in many organic chemistry laboratories. We have developed a convenient and efficient method for synthesizing PAH-nucleoside adducts by using I2 as the oxidant. Adducts of benzo[a]pyrene (BP), dibenzo[a, l]pyrene (DB[a,l]P), and 7,12-dimethylbenz[a]anthracene were synthesized with deoxyguanosine (dG), deoxyadenosine, guanine (Gua), or adenine in either Me2SO or dimethylformamide (DMF) with or without AgClO4. When, for example, the potent carcinogen BP was dissolved in DMF in the presence of 3 equiv of I2, 5 equiv of dG, and 1 equiv of AgClO4, 45% of the BP was converted to BP-6-N7Gua. When BP was placed under the same reaction conditions in the absence of AgClO4, the extent of formation of BP-6-N7Gua decreased to 30%. When the potent carcinogen DB[a,l]P was dissolved in DMF in the presence of 3 equiv of I2, 5 equiv of dG, and 1 equiv of AgClO4, 43% of the DB[a,l]P was converted to DB[a,l]P-10-N7Gua. In the more polar solvent Me2SO under the same reaction conditions, however, the yield of DB[a,l]P-10-N7Gua was only 20%. Synthesis of adducts with the oxidant I2 is more convenient and, in some cases, more efficient than synthesis by electrochemical oxidation. This method simplifies the synthesis of PAH-nucleoside and nucleobase adducts that are essential for studying biologically formed PAH-DNA adducts.