Mirosław Nakonieczny

Digestive enzymes activity in larvae of Cameraria ohridella (Lepidoptera: Gracillariidae)

This article presents the activity of carbohydratases and proteases in the midgut of Cameraria ohridella larvae--an oligophagous pest whose preferred feeding is horse chestnuts leaves. Optimal media pH of the assayed enzymes were similar to those of other Lepidopterans. Relatively high amylase activity, as well as maltase and sucrase activities, indicates that starch and sucrose are the main digested saccharides. Trehalase activity was similar to that described in other Lepidopterans. Activities of glycosidases were significantly lower than those of disaccharidases what suggests that neither cellulose nor glycosides are important for C. ohridella. Trypsin is the main endoprotease of this pest. Like in other leaf-eaters carboxypeptidase activity was higher than that of aminopeptidase. The activity of the majority of examined enzymes increased in the following successive pest generations, which could be explained by the decreased nutritional value of older leaves. Probably this phenomenon in hydrolases activity in Cameraria is a nonspecific mechanism present at this stage of co-evolution of the horse chestnut and its pest.

Parnassius apollo last-instar larvae development prediction by analysis of weather condition as a tool in the species' conservation.

Parnassius apollo (Lepidoptera: Papilionidae) has already disappeared or is under threat of extinction in many of its former habitats. It has been documented that weather conditions--anomalies in particular--contributed to this process. In this study, we combined developmental data obtained previously for the last-instar Apollo larvae (collected in 1996, 1997, and 2003) with corresponding meteorological data to assess the effects of ambient temperature and rainfall episodes on the duration and the completion of the instar. For comparing the temperature effect, we applied the degree-day concept. We found significant positive correlation between the number of rainy days during the instar development (x) and its duration time (y): y=8.293+0.936x (±2.813) (r=0.662, P<10(-7)). Logarithmic transformation of the growth curves of the last-instar Apollo larvae revealed that there was no difference in growth among females; however, there was slower growth of males in 2003 in comparison to 1996. Growth (y) of female Apollo larvae as a function of instar duration (x) can by described by one common equation, irrespectively of the year: y=317.6+502.3 lnx (±263.3) (r=0.82, P<10(-4)).

Digestive enzymes activity in subsequent generations of Cameraria ohridella larvae harvested from horse chestnut trees after treatment with imidacloprid

In the present study we describe the effect of chloronicotinoid pesticide (imidacloprid) on the digestive enzymes activity of the Cameraria ohridella larvae after lasting 1 year sublethal exposure to imidacloprid pesticide. Caterpillars - L4 stage (fourth instar, hyperphagic tissue-feeding phase) - were collected from chemically protected white horse chestnut trees 1 year after imidacloprid treatment, and compared with caterpillars collected from non-treated trees in a previous study. Enzymes activity of α-amylase, disaccharidases, glycosidases and proteases was assayed. The presence of pesticide in ingested food changed the digestive enzymes profile of caterpillars. The analysis of correlations between different digestive enzymes showed many significant correlations (P<0.05) among glycolytic activities like β-glucosidase and α-galactosidase activities. Statistically significant correlations for proteolytic activity were found between trypsin and chymotrypsin activity and aminopeptidase activity that occurred only in the 1st generation. PCA distinguished five primary components with eigenvalues higher than 1, from which the first two explain almost 59% of analyzed results. Surprisingly, in the pesticide treated groups significantly higher activities of sucrase and lactase in relation to control were found. In general, glycosidase (α-glucosidase, β-glucosidase and β-galactosidase) activities showed a similar pattern of activity in different generations. These results contrast with those obtained with control larvae, where significant differences in activities of α-glucosidase, β-glucosidase and β-galactosidase may result from the different quantity and quality food intake by subsequent generations of larvae. No inter-generation differences in total proteolytic activity were observed in treated larvae. The absolute value of total proteolytic activity was higher than that in the control group. The pesticide present in the vascular system of the horse chestnut tree significantly affected some of the digestive enzymes activities and - in consequence - also interrelationships between enzymes, what may affect the food digestion.

Occurrence of sarmentosin and other hydroxynitrile glucosides in Parnassius (papilionidae) butterflies and their food plants.

Sequestration of plant secondary metabolites is a widespread phenomenon among aposematic insects. Sarmentosin is an unsaturated γ-hydroxynitrile glucoside known from plants and some Lepidoptera. It is structurally and biosynthetically closely related to cyanogenic glucosides, which are commonly sequestered from food plants and/or de novo synthesized by lepidopteran species. Sarmentosin was found previously in Parnassius (Papilionidae) butterflies, but it was not known how the occurrence was related to food plants or whether Parnassius species could biosynthesize the compound. Here, we report on the occurrence of sarmentosin and related compounds in four different Parnassius species belonging to two different clades, as well as their known and suspected food plants. There were dramatic differences between the two clades, with P. apollo and P. smintheus from the Apollo group containing high amounts of sarmentosin, and P. clodius and P. mnemosyne from the Mnemosyne group containing low or no detectable amounts. This was reflected in the larval food plants; P. apollo and P. smintheus larvae feed on Sedum species (Crassulaceae), which all contained considerable amounts of sarmentosin, while the known food plants of the two other species, Dicentra and Corydalis (Fumariaceae), had no detectable levels of sarmentosin. All insects and plants containing sarmentosin also contained other biosynthetically related hydroxynitrile glucosides in patterns previously reported for plants, but not for insects. Not all findings could be explained by sequestration alone and we therefore hypothesize that Parnassius species are able to de novo synthesize sarmentosin.

Elemental Distribution in Reproductive and Neural Organs of the Epilachna nylanderi (Coleoptera: Coccinellidae), a Phytophage of Nickel Hyperaccumulator Berkheya coddii (Asterales: Asteraceae) by micro-PIXE

ABSTRACT. The phenomenon of metal hyperaccumulation by plants is often explained by a pathogen or herbivore defense hypothesis. However, some insects feeding on metal hyperaccumulating plants are adapted to the high level of metals in plant tissues. Former studies on species that feed on the leaves of Berkheya coddii Roessler 1958 (Asteraceae), a nickel-hyperaccumulating plant, demonstrated several protective mechanisms involved in internal distribution, immobilization, and elimination of Ni from the midgut and Malpighian tubules. These species are mainly coleopterans, including the lady beetle, Epilachna nylanderi (Mulsant 1850) (Coleoptera: Coccinellidae), collected from the ultramafic ecosystem near Barberton in South Africa. By performing particle-induced X-ray emission microanalysis elemental microanalysis (PIXE), this study examined whether Ni may be harmful to internal body systems that decide on insect reactivity (central nervous system [CNS]), their reproduction, and the relationships between Ni and other micronutrients. Data on elemental distribution of nine selected elements in target organs of E. nylanderi were compared with the existing data for other insect species adapted to the excess of metals. Micro-PIXE maps of seven regions of the CNS showed Ni mainly in the neural connectives, while cerebral ganglia were better protected. Concentrations of other bivalent metals were lower than those of Ni. Testis, compared with other reproductive organs, showed low amounts of Ni. Zn was effectively regulated at physiological dietary levels. In insects exposed to excess dietary Zn, it was also accumulated in the reproductive organs. Comparison of E. nylanderii with other insects that ingest hyperaccumulating plants, especially chrysomelid Chrysolina clathrata (Clark) (Coleoptera: Chrysomelidae), showed lower protection of the CNS and reproductive organs.

Population dynamics of Western corn rootworm beetles on different varieties of maize identified using pheromone and floral baited traps

P. K. BERES ́1*, S. DRZEWIECKI, M. NAKONIECZNY, M. TARNAWSKA, J. GUZIK AND P. MIGULA 1 Institute of Plant Protection – National Research Institute, Regional Experimental Station in Rzeszów, Langiewicza 28, PL 35-101 Rzeszów, Poland 2 Institute of Plant Protection – National Research Institute, Sosńicowice Branch, Gliwicka 29, PL 44-153 Sosńicowice, Poland Department of Animal Physiology and Ecotoxicology, University of Silesia, Bankowa 9, PL 40-007 Katowice, Poland

The Glycolytic Enzymes Activity in the Midgut of Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) adult and their Seasonal Changes

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) is an important pest of maize. The diet of the D. virgifera imago is rich in starch and other polysaccharides present in cereals such as maize. Therefore, knowledge about enzymes involved in digestion of such specific food of this pest seems to be important. The paper shows, for the first time, the activities of main glycolytic enzymes in the midgut of D. virgifera imago: endoglycosidases (α-amylase, cellulase, chitinase, licheninase, laminarinase); exoglycosidases (α- and β-glucosidases, α- and β-galactosidases) and disaccharidases (maltase, isomaltase, sucrase, trehalase, lactase, and cellobiase). Activities of α-amylase, α-glucosidase, and maltase were the highest among assayed endoglycosidases, exoglycosidases, and disaccharidases, respectively. This indicates that in the midgut of D. virgifera imago α-amylase, α-glucosidase and maltase are important enzymes in starch hydrolysis and products of its digestion. These results lead to conclusion that inhibition of most active glycolytic enzymes of D. virgifera imago may be another promising method for chemical control of this pest of maize.