On the natural history of Pseudochlamys megalostomoides Lacordaire, 1848 (Coleoptera: Chrysomelidae: Cryptocephalinae)

The objective of this study was to determine the number of instars of Monochamus alternatus Hope (Coleoptera: Cerambycidae) larvae by comparing their head capsule widths (HCW) published in previous studies, as well as additional laboratory experiments. Larvae of M. alternatus showed repeated molting in the laboratory. Most larvae ceased their development at the 10th instar stage. Frequency distributions of HCW for the first, second, and third instar larvae were clearly separated while those of the fourth through 11th instar larvae largely overlapped between successive instars in our results. The HCW values for the first, second, and third instar larvae directly measured for each instar in our study indicated that they were more precise than those of previous reports based on field-collected HCW which might have missed HCW of the first instar larvae or wrongly determined HCW for some instars. Unlike the reports of four instars of previous studies, M. alternatus larvae passed five instars in the field, which was confirmed by the discovery of five pairs of mandibles in the feeding gallery and pupal chamber. Also, the comparative study for the frequency distributions of HCW revealed that most M. alternatus larvae passed five instars. Consequently, the average sizes of HCW for their first, second, and third instar larvae are newly suggested to be 0.896 ± 0.069, 1.291 ± 0.131, and 1.707 ± 0.165 mm (mean ± SD) .

Bursaphelenchusxylophilus, a pathogenic pine wood nematode (PWN), is responsible for pine wilt disease (PWD), which has caused significant economic and ecological damage worldwide, particularly in East Asia. Multiple biological factors, such as the beetle vector Monochamus, symbiotic bacteria and associated fungi, are involved in the disease infection cycle. This study isolated and identified the fungal communities of Monochamusalternatus larval galleries and pupal chambers from different instars through field investigation, morphological observation and multi-locus DNA sequence analyses in Zhejiang Province, China. A total of 255 and 454 fungal strains were isolated from M.alternatus galleries and pupal chambers infected with PWN, from the 2nd-3rd and 4th-5th instar larvae, respectively. A total of 18 species of fungi were identified, 14 species were isolated from the 2nd-3rd instar larval galleries and six species from the galleries and pupal chambers of the 4th-5th instar larvae. Amongst them were six species belonging to four genera of ophiostomatalean fungi, including two novel species, Graphilbumxianjuensis sp. nov. and Ophiostomataizhouense sp. nov. and four known species, Ceratocystiopsisweihaiensis, Ophiostomaips, Sporothrixzhejiangensis and S.macroconidia. The findings revealed that the fungal diversity and abundance of the 2nd-3rd instar larvae differed markedly from those of the 4th-5th instar larvae. This difference could be the result of fungal succession. This study provides a thorough understanding of the fungi associated with PWD and lays the groundwork for future research.

Except for one unconfirmed case, chironomid larvae have been reported to pass through four larval instars between egg and pupal stages. We have observed a fifth larval instar to be a standard life-cycle feature of the podonomine Trichotanypus alaskensis Brundin 1966 in tundra ponds on the Arctic Coastal Plain near Barrow, Alaska. T. alaskensis has a one-year life cycle in these arctic ponds. Adults emerge in June ~2-3 weeks after pond thaw, then mate and oviposit; most newly-hatched larvae reach instar IV by October when pond sediments freeze. Overwintering larvae complete instar IV within a few days of thaw, then molt again to a fifth larval instar. Imaginal discs, normally seen only during instar IV in Chironomidae, develop across both instars IV & V prior to pupation and adult emergence. While monitoring larval development post-thaw in 2014, we noticed freshly-molted T. alaskensis larval exuviae a week or more prior to any pupation by that species. In 2015-16 we reared overwintering instar IV larvae from single pond sources, individually with daily monitoring, through molts to instar V, pupa, and adult. Some overwintering instar II and III larvae were reared as well, but were few in number. During 2016 we also reared T. alaskensis progeny (from eggs) through instar II, thus documenting head capsule size ranges for all five instars in a single pond’s population. Without individual rearings, the fifth larval instar was not readily apparent for two reasons: 1) The molt itself occurs immediately after thaw and is so synchronous it is difficult to discern in daily field samples. 2) The head capsule size increment between instars IV-V is much lower than the ratio predicted by the Brooks-Dyar Rule. Up through instar IV, the Brooks-Dyar ratio for T. alaskensis ranged 1.30-1.61, but during the IV-V molt head capsule dimensions (sexes pooled) increased by a ratio of 1.09 – comparable to the magnitude of sexual dimorphism in head capsule size within each of the final two larval instars. Individual rearings coupled with 2014-2016 field surveys in nine other ponds suggest that five larval instars is an obligatory trait of this species at this location. As this is the first confirmed case of five larval instars in a chironomid, the phylogenetic uniqueness of this trait needs further investigation.

Synaptotagmin is a synaptic vesicle protein required for efficient neurotransmitter release, yet its exact role in the synaptic vesicle cycle is unclear. Drosophila presents an ideal organism for studies aimed at determining the in vivo functions of proteins. However, synaptotagmin studies have been limited by the early (embryonic or first instar) lethality previously reported for Drosophila synaptotagmin I null (syt(null)) mutants. Here we report a new culturing technique that enhances survival of severely uncoordinated mutants thereby permitting Drosophila syt(null) mutants to survive through early adulthood. We examined synapses in syt(null) third instar larvae by electrophysiology and found that they exhibit severely decreased and asynchronous evoked neurotransmitter release, as well as an increased rate of spontaneous neurotransmitter release, as previously seen in first instar syt(null) larvae. The ability to examine severe synaptotagmin mutants as third instar larvae, a stage where electrophysiological and morphological analyses are more easily accomplished, will facilitate structure/function studies.

The behaviour and development of Elasmosoma sp., near pergandei Ashm. (Neoneurinae: Braconidae: Hymenoptera), a parasite of the ant, Formica obscuriventris clivia Creighton, was studied in a coastal sand dune habitat in central Oregon over a 2-year period. Female wasps oviposited into the abdomen of major worker ants after alighting briefly on the host. Embryonic development of the wasp occurred inside the serosal membrane or trophamnion of the egg in the abdomen of the ant host. First instar wasp larvae possessed a large head capsule with falcate mandibles, 11 ventral body lobes, an elongate tail and a dorsally located anus. Second instar larvae were unspecialized and lacked the large head capsule, elongate tail and paired ventral body lobes. Third instar larvae were hymenopteriform with a distinct head capsule. Cocoon formation and adult emergence is described. Observations on the percentage parasitism and altered behaviour of parasitized ants are presented. The association between neoneurine wasps and ants is a well-balanced system that has been in existence for at least 40 million years.

In 1979 five elm bolts, each 8 inches long by 4 inches diam and in separate 5-gal screened lard cans, were each infested with 25 male and 25 female adults. The cans were in a continuously lighted room maintained at 85°F. After 3 wk these parent adults were removed from the logs and cans with a vacuum cleaner, thus leaving the logs infested only with progeny as eggs and larvae under the bark. One log was set aside as an untreated check and each of the other logs was sprayed to run-off with a particular insecticide using a laboratory paint gun sprayer. The insecticides were used at the equivalent rate of 16 lb AI/100 gal spray except Pydrin was used at 3.2 lb AI/100, which is 16X the recommended agricultural rate. Sprays were buffered to pH 6.0. After drying the sprayed, infested logs were again confined in the lard cans at ca 75°F. From 3 to 11 wk after treatment progeny adults emerged from the logs and were counted. After adult emergence ceased the bark on each log was examined under a 10-power microscope and the bark routed to the depth of the bark beetle pupal chamber with a drill bit in a Moto-Tool. The dead adults in the bark were counted. Their locations varied from the pupal chamber to partial protrusion from the bark, with the more toxic insecticides stopping movement nearer the pupal chambers. This experiment was repeated again in 1980 and in 1981. With each year serving as a replicate the 3 years data were subjected to a 2-way ANOVA.

Considering the cryptic life cycle of Hylesinus taranio (Dantoine), the prediction of adult emergence is the first step to establish the optimum timing for its management. The fieldwork was carried out over four years (2016 - 2019) in Lécera (Zaragoza, Spain). H. taranio was monitored from early April in larval phase until adult emergence in June when all pupal chambers were empty. The H. taranio development and the olive grove phenology were the variables considered to test whether there is a relationship between them. The strength of this relationship and the uncertainty coefficient were assessed. The most significant relationships (p < 0.001) were examined, being the association pupae/corolla coloration change and pupal chamber empty/freshly formed fruits those which show the highest contribution percentage to the relationship. The results substantiate that the olive grove phenology predicts the H. taranio development, becoming an easy strategy for the farmer to predict the adults on release, which could to decide the optimum timing of H. taranio management by only watching for olive grove phenophases. Accordingly, some management measures are suggested for reducing the density of H. taranio in the olive groves.

Ontogeny of sexual size dimorphism in the hornless rose chafer Pachnoda marginata (Coleoptera: Scarabaeidae: Cetoniinae)

First, second and third instar larvae of the blowfly, Chysomya chloropyga collected from laboratory stock were exposed to a mixture of ground rice and fish supplemented with 1, 2, 3, 4, 6 and 10 % concentrations of neem seed (NS) and neem seed kernel(NSK) powders respectively.Twenty pairs of laboratory-reared adult males and females were also maintained separately on diets treated with 1, 2, 4, 6 and 10% NS and NSK for survival, longevity, fecundity and progressive weights of adult male and female at 5-day interval. The NS-as well as NSK- treated diets prolonged the development of second instar larvae but not significantly different (p > 0.05) from the duration of first and third instar larvae as well the pupal period in different concentrations of treated diets. Percent adult emergence decreased significantly with increase in concentration NS treated-diets from 1-6 % but there was no emergence of adults in 10 % treated diets. There was no significant difference in the emergence of adults from first, second and third instar larvae treated with NSK. Percent survival of male and female populations in NS-treated diets were 45 and 60% at day 40 of exposure but female in 10%-treated diet was 30%. Control male and female were at 80 and 75%. There was no significant difference (p>0.05) in the survival of male and female exposed to diffrent concentrations of NSK-treated diets throughout the 40-day exposure with percentage survival of male and female at 66 and 79% respectively. Females survived better than males at all concentrations of NS- and NSK-treated diets.Maximum longevity of male and female was 45 and 47 days respectively in 10% NS-treated diet and significantly different (p<0.05) from longevity of male and female at 62 and 70 days respectively in 10% NSK-treated diet. Mean fecundity and total number of eggs laid decreased with increase in concentration of powders of NS and NSK. Females exposed to control diets deposited significantly more eggs than eggs laid by females exposed to different concentrations of NS- and NSK- treated diets (p < 0.05). Mean weights of males and females in different concentrations of NS- and NSK-treated-diets generally increased from days 0 to 10 and fluctuated thereafter but there was no significant difference in the weights of males and females exposed to different concentrations throughout the 40 days of exposure. In conclusion, NS and NSK powders incorporated in diets prolonged the develpoment of larval instars and subsequent emergence of adults and their weights but the NS was more effective in reducing the survival, longevity and the fecundity of C. chloropyga.

Impact of Triclopyr Amine onGalerucella calmariensisL. (Coleoptera: Chrysomelidae) and a Step toward Integrated Management of Purple LoosestrifeLythrum salicariaL.

In 1712 the poet Elkanah Settle (1648–1724) published a funeral poem, Threnodia Apollinaris , dedicated to the memory of Dr Martin Lister.[1][1] Settle had a good deal of material to draw upon, because Lister had been Vice-President of the Royal Society, a physician to Queen Anne, and the first

Studies in the lab have demonstrated that evaluating the effect of soil moisture and other variables is essential for understanding the importance of environmental factors influencing the Heliothinae pupal stage, but simulated field studies are conducted infrequently. We compared the pupation of Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) under saturated and unsaturated conditions across 3 distinct soil types (coarse sand, high organic muck, and fine-textured clay) and observed adult emergence, as well as pupal depth and weight. The interaction between soil type and moisture had a significant effect on adult emergence. Fewer adults emerged from dry fine-textured clay soil than from dry coarse sand and high organic muck. However, there was no effect of soil type and no interaction between soil and moisture on pupal depth. Soil moisture was the primary driver of pupal depth, suggesting prepupae use moisture to mediate their position within the pupal chamber. In addition, pupal weight was unrelated to soil type, moisture, or their interaction. Our study demonstrates that soil moisture can be a greater driver of H. zea pupation than soil type, but additional observations are necessary to understand the mechanism by which moisture impacts pupation.

Treated The second larval instar by residual method indicated that, chlorpyrifos-methyl and methomyl formulations expect Goldben (90.48 % mortality) caused 100 % mortality, while the lowest mortality percent occurred in emamectin benzoate and lufenuron formulations.Except Broact formulation0, there are significant differences between all the tested formulations (ranged from 0.00 to 64.29 % pupation and to 53.57% adult emergence) and untreated control in pupation (85.71 %) and adult emergence (78.57%) On the contrary, there are no significant differences between two formulations of each active ingredient except two formulations of emamectin benzoate in pupation percentage only.Treated The second larval instar by feeding method indicated that,chlorpyrifos-methyl formulations is the highly toxic (100 % mortality), when methomyl, emamectin benzoate and lufenuron formulations were low toxic (ranged from 3.70 to 27.27 % mortality) and lufenurononly was high toxic by ingested than leaf residual exposure.. Also, there are highly significant differences between chlorpyrifos-methyl formulations (0.00 % for pupation and adult emergence) with other treatments (ranged from 60.71 to 78.57% for pupation and from 57.14 to 71.43 % for adult emergence).Treated egg by dipping method showed a significant reduction in hatchability rates for formulations of methomyl and chlorpyrifosmethyl ranged from 65.05% (Goldben) to 76.69% (Reldan), however, there are no significant reductions in hatchability rates for formulations of emamectin benzoate and lufenuron, ranged from 86.08% (Broact) to 95.88% (Match) compared to the control (98.33%), but there are not significant differences between two formulations of each active ingredient.

Sahlbergella singularis is one of the insects that causes the most damage to cocoa trees in Togo. The management of this pest is mainly based on chemical control, with its negative consequences for human health and the environment. In order to find an alternative to the use of synthetic pesticides, the insecticidal and repellent activities of neem oil were tested in vitro on fifth instar larvae and adults of S. singularis. Larvae were collected from a cocoa plot. The fifth instar larvae were separated from the other stages. Some of the larvae were reared in the laboratory until adult emergence. The concentrations of neem oil used were 0.5, 1, 1.5, 2 and 2.5 µl.ml-1. Insecticidal tests were carried out by spraying the solutions directly onto the insects. The repellency test was carried out using the preferential zone method on filter paper. The results showed an increasing mortality rate with increasing concentrations of neem oil for both fifth instar larvae and adults. The calculated LC50’s for larvae and adults were 0.89 and 0.98 µl.ml-1 respectively. Neem oil had a weak repellent effect on both fifth instar larvae and adults, with average repellency rates of 23.60% and 28.8% respectively. These results show that neem oil can be an alternative to the use of synthetic pesticides in the control of S. singularis.

Coleoptera of the family Melolonthidae plays an important role in decomposition and nutrient recycling in the environment, however the information of this group is scarce in Brazil; therefore studies were developed with the objective of understanding the biological aspects of saproxylophagous. The studies were developed in the experimental farm of the State University of Mato Grosso do Sul, in Aquidauana, from March 2011 to April 2012. In a forest fragment of the cerrado, when encountering fallen tree trunks, they were cut into small pieces for extraction and acquisition of immature and adult Melolonthidae. Larvae were taken to the laboratory where they were reared until emergence of adults, obtaining biological and biometric data. A total of 145 larvae and 11 adults were collected. Third instar larvae of Strategus surinamensis Burmeister, 1847, collected in this phase lasted 230 days and 31.3 days in the pupal stage. Third instar larvae of Phileurus valgus (Linnaeus, 1758), in this phase lasted 49 days and 13 days in the pupal stage. Third instar larvae Pelidnota fulva Blanchard, 1850, after being collected remained for 61 days in this phase and 25.3 days in the pupal stage. AdultsPelidnota aff. vazdemeloi were collected inside tree trunks. ToMacraspis festiva Burmeister, 1844, the third instar larvae collected lasted for 39 days at this stage, and pupae lasted 17 days. Third instar larvae of Rutela lineola(Linnaeus, 1767), in this phase lasted 23 days, and pupae lasted 16.3 days. ToHoplopyga liturata (Olivier, 1789), third instar larvae collected lasted 21 days and pupae lasted 18 days. To Hoplopyga boliviensis (Moser, 1918), third instar larvae lasted 33 days and pupae 16 days. Second and third larval instarMarmarina maculosa (Olivier, 1789) collected lasted respectively, 105 and 171.3 days and pupae lasted 26 days. The species collected in greatest quantity were M. festiva with 75 larvae,R. lineola with 20 larvae and H. boliviensis with 18 larvae. Larvae were collected during several months, where in July 32 larvae were sampled and in August 39 larvae, the largest quantities encountered.