Further studies on acclimatization to low temperatures in the grain weevils Sitophilus oryzae (L.) and Sitophilus granarius (L.)

The effect of thermal acclimatization on the resting metabolism of adult grain weevils was examined by closely restraining the weevils during respirometry. Sitophilus oryzae (L.) acclimatized to 15°C consumed less oxygen over 11–23°C than 27°C-acclimatized weevils but like amounts over 25–35°C. Similar results were observed with S. granarius (L.). Respiration R–T curves of unrestrained S. oryzae and S. granarius showed no evidence of rotation at temperatures close to their chill-coma thresholds but warm- and cold-acclimatized weevils consumed similar amounts of oxygen when both categories of weevils were in chillcoma. Acclimatizing S. oryzae from 27 to 13.5°C through four, weekly 4.5°C step-transfers lowered the median chill-coma thresholds of the weevils from 8.9 to 6.2°C, increased their life span at 9°C, an unfavourable temperature for this species, from 2 to 8 weeks and lowered their oxygen uptake at 9°C by 19%.     

Purification,partial characterization,and postembryonic levels of amylases from Sitophilus oryzae and Sitophilus granarius

Amylases from adults of Sitophilus oryzae (L.) and S. granarius (L.) were purified by using a sequential procedure of ammonium sulfate precipitation, glycogen-complex formation, and ion exchange chromatography. Amylase of S. oryaze was purified 47.4-fold to a specific activity of 478 units/mg protein. One amylase unit equals 1 mg maltose hydrate produced/min at 30°C. Amylase of S. granarius was purified 85.4-fold to a specific activity of 453 units/mg protein. Amylase of S. oryzae had a Km of 0.173% for soluble starch and consisted of two anionic isozyrnes with isoelectric points of pH 3.70 and pH 3.76. Amylase of S. granarius had a Km of 0.078% for starch and was a single protein with an isoelectric point of pH 3.76. Purified amylases of both species had molecular weights of 56,000 estimated by sodium dodecyl sulfatepolyacrylamide gel electrophoresis, were activated by chloride, and had double energies of activation calculated from Arrhenius plots. Based on fresh weights of adults feeding on whole wheat through 10 weeks of age, S. oryzae contained three-fold and eight-fold more amylase than S. granarius and S. zeamais Motschulsky, respectively. High amylase levels in S. oryzae may provide this species with an adaptive advantage when feeding on cereals containing naturally occurring amylase inhibitors.     

Vitamin requirements of larvae of Sitophilus oryzae

Improvements were made in the meridic larval diet for Sitophilus oryzae by replacing the minerals and vitamins supplied by dietary brewer's yeast and wheat germ with mineral and vitamin mixtures. The effects of different concentrations of individual vitamins were studied with the improved diet containing 20% casein. In later tests the dietary casein was replaced with a mixture of 15 amino acids. The results indicated that these larvae, which contain an associated bacteria-like micro-organism, required thiamine, nicotinic acid, pyridoxine, folic acid, and biotin in the diet. No requirement for riboflavin or pantothenic acid could be demonstrated on either the 20% casein diet or the amino acid diet possibly due to contamination of the cornstarch with these two vitamins. In addition, the larvae did not require choline or inositol for the growth of one larval generation.The asymbiotic larvae of S. granarius failed to develop on the improved casein diet indirectly implicating the symbiotes in a nutritional rôle; however, the symbiotes present in S. oryzae apparently do not provide B vitamins. Larvae of S. oryzae failed to develop when the concentration of casein was reduced to 10% while growth was maintained with a 10% concentration of the amino acid mixture. Casein is not an optimal source of amino acids for this species.     

A comparison of the capacity for increase at a low temperature of foreign and Australian populations of Sitophilus oryzae (L.) and S. granarius (L.)

The capacity for increase (r_c) of three populations of S. oryzae and three populations of S. granarius from Great Britain and Canada was determined over 15 and 24 weeks respectively at 15°C in wheat of 14% moisture content. The fertility of these populations at 27°C, their body weight and chill-coma threshold was also measured. With both S. oryzae and S. granarius, values of r_c for the foreign populations fell within the range previously observed when several Australian populations were reared under the same conditions. The cohort generation times (T_c) of the foreign S. oryzae populations were significantly shorter than those of the Australian populations because of the shorter immature development periods (D) of the former. Two of the foreign S. granarius populations had lower values for D than all but one of their Australian counterparts but their values of T_c were within the range previously observed for Australian weevils. When the Australian and foreign populations were considered together, the value of r_c was more strongly correlated with fertility at 27°, an optimal temperature, than with body weight in both S. oryzae and S. granarius. With S. oryzae, r_c was correlated with the chill-coma threshold of cold-acclimated weevils only. There were no correlations between r_c and chill-coma thresholds in S. granarius. It was concluded that the observed differences in r_c were related to differences in the vigour of the populations rather than to physiological differences in cold-tolerance. The likelihood of grain weevils becoming cold-tolerant as a result of grain aeration is discussed and the practical significance of differences in r_c considered.     

Utilization of fatty acids by Pieris brassicae reared on artificial and natural diets

The fatty acid composition of Pieris brassicae was measured from larvae reared on four different diets. Pieris can alter the composition of fatty acids in the diet through selective incorporation and synthesis. Oleate is preferentially accumulated on artificial diets (15·9 per cent in diet, 43·8 per cent in neutral lipid (NL) of fifth instar larvae), but not equally on natural diets (18·1 per cent in Brassica napus, 25·6 per cent in the NL of fifth instar larvae). Incorporation of linolenate appears to depend on the concentration of both linolenate and linoleate in the diet. With dietary levels of 35·7% linolenate and 32·2% linoleate, fifth instar larvae contain 12·2 and 16·0 per cent, respectively, of these acids. With 45·8% linolenate and 12·5% linoleate in the diet, fifth instar larvae contain 44·1 and 11·6 per cent of these acids, respectively, in the NL. Palmitoleate is actively synthetized on the artificial diets; with trace amounts of dietary palmitoleate, fifth instar larvae have 9·3 per cent of this acid in the NL. Pieris regulates the uptake of linoleate from the diet at the intestinal wall as was shown by linoleic acid-1-¹⁴C, and is unable to convert dietary linoleate to any of the 18-carbon analogues. The female apparently accumulates linolenate into egg phospholipids on the artificial diet, but in general the fatty acid composition of the eggs resembles that of the fat body.     

Water exchange and effect of water vapour activity on metabolic rate in the dust mite, Dermatophagoides.

Oxygen consumption by Dermatophagoides farinae was found to vary with temperature and water vapour activity. The relationship between temperature and O₂ consumed/hr per mite was first order. Q₁₀'s were equal to exp (k 10°C) and were found to be 3·04 and 2·49 for 1 to 6 and 6 to 22 hr monitoring periods respectively. A significant difference between O₂ consumed/hr per mite for 1 to 6 and 6 to 22 hr monitoring periods was found. Inactivity of mites explained 29·6 and 31·8 per cent of this reduction. Dehydrating conditions and reduced permeability of the water and gas exchange surface explained a further reduction to 58·7 and 60·8 per cent.     

Susceptibility of Sitophilus granarius and S. Oryzae (Coleoptera: Curculionidae) to high temperature after exposure to supra-optimal temperature

S. granarius and S. oryzae were exposed to 32° and 35° for 2, 7 and 14 days and 1, 2 and 7 days, respectively, and mortality tested at 40°. Under these conditions maximum acclimation of S. granarius was achieved after 14 days at 32° and 2 days at 35°, while S. oryzae maximum acclimation occurred after 2 days at 32° and 1 day at 35°. Longer exposure at 32° for S. oryzae and for both species at 35°, appeared to reverse the process. Rearing S. granarius for three and five generations at 30°, raised its tolerance to the test temperature. Rearing S. oryzae for three generations at 30° did not have any significant effect on that species.

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Zusammenfassung Imagines von S. granarius und S. oryzae wurden physiologisch bei 32° und 35° für 2, 7 und 14 Tage, oder für 1, 2 und 7 Tage akklimatisiert und ihre Sterblichkeit bei 40° geprüft. Unter diesen Bedingungen wurde die maximale Akklimatisierung bei S. granarius nach 14 Tagen bei 32° und nach 2 Tagen bei 35° erreicht, während bei S. oryzae die maximale Akklimatisierung nach 2 Tagen bei 32° und 1 Tag bei 35° erreicht wurde. Längere Akklimatisierungs-Perioden bei 32° für S. oryzae und für beide Arten bei 35° schienen den Prozess umzukehren. Die Züchtung von S. granarius für drei und fünf Generationen, und von S. oryzae für drei Generationen bei 30° vergrösserte die Toleranz von S. granarius gegen die experimentelle Temperatur, hatte aber keinen bedeutsamen Effekt bei S. oryzae.Die Daten beweisen die Fähigkeit von S. granarius und S. oryzae, sich an höhere Temperaturen zu akklimatisieren, wobei der Grad der vergrösserten Toleranz bei den zwei Arten verschieden war. Es zeigte sich auch, dass in diesen zwei Arten der physiologische Akklimatisierungsprozess bei stufenweisem Wechsel zu einer höheren Temperatur effektiver war als der Prozess der Akklimatisierung bei Dauerzucht in einer erhöhten Temperatur. Die gegenwärtigen Daten beziehen sich nur auf die veränderte Empfindlichkeit hinsichtlich der Mortalität bei den akklimatisierten Insekten in hoher Temperatur.

     

Voracity of the main stored-product insect pests on wheat grain

The results of an experimental study of voracity of adults and larvae of seven main stored-product insect pests are described. One adult of Sitophilus oryzae, S. granarius, Rhizopertha dominica, Tribolium castaneum, T. confusum, and Oryzaephilus surinamensis consumed 0.38, 0.48, 0.89, 0.15, 0.16, and 0.11 mg of wheat grain per day, respectively. During their development, larvae of these insects consumed 7.60, 13.77, 7.92, 3.32, 3.60, and 2.32 mg of wheat grain, respectively. One larva of Sitotroga cerealella consumed 17.46 mg of wheat grain. The coefficients of harmfulness of these species are calculated, and a method for estimating the “total infestation density” of grain is proposed.     

Auswirkungen von Diatomeenerde auf den Wasserhaushalt des Kornkäfers: Sitophilus granarius (L.) (Col., Curculionidae) und möglicher Einsatz innerhalb des Vorratsschutzes

Effects of diatomoceous earth on water content of Sitophilus granarius (L.) (Col., Curculionidae) and its possible use in stored product protection Increased problems caused by application of chemical pesticides within pest protection has led to the demand for non‐polluting, biologically specific active countermeasures. This study aimed to test the possible use of diatomaceous earth against Sitophilus granarius. It will also examine the physiological effects of amorphous diatomaceous earth against this insect. After 2 weeks, 100% mortality was observed of the adults treated with 2 g Fossil Shield^® per kilogram wheat, at 11–12% corn moisture content and relative humidity of about 62 ± 2%. The rise in grain moisture and relative humidity decreased the effectiveness of the Fossil Shield^®. It was observed, that the diatomaceous earth reduced the number of adult granary weevils and increased the mortality rate at development stages, but they did not completely prevent the hatching of the next generation. After contact with diatomaceous earth, the individuals of S. granarius had lost weight and reduced their water content, respectively. The adults significantly lost weight and body water after 1 day exposure to Fossil Shield^®. This showed that silica dusts break the water barrier of the cuticle. However, it increases the transpiration rate of water across the cuticle and several insects died through dehydration. At higher relative humidity diatomaceous earth was less effective toward S. granarius, because there was a lower permeability for water across the cuticle. This delays or rather prevents the drying action of diatomaceous earths. Furthermore, the addition of food affected the effectiveness of dusts against S. granarius. A lot of stored product beetles are able to produce metabolic water in the feed substrates. This ability protects them and works contrary to the loss of water.     

Self-selection of an optimum diet from a mixture of wheat fractions by the larvae of Tribolium confusum

Tribolium confusum was reared from the beginning of the first instar to the pupa on wheat bran, endosperm or germ or on a 1 : 1 : 1 mixture of these three fractions. The food intake of the larvae reared on the mixture included more or less similar proportions of the three wheat fractions, averaging 1·5 per cent bran, 17·1 per cent endosperm and 81·4 per cent germ. Past work on the nutrition of stored-product insects has involved the assumption that the insects cannot feed selectively from a finely powdered diet. Our findings show that this assumption may not be warranted. The mixed diet supported growth better than any one of the pure fractions or any wheat diet previously tested by us. It was superior to germ, the best of the pure fractions, because it was more digestible and because the digested portion was more efficiently utilized for growth. Endosperm, the poorest diet of the fractions, was highly digestible but was not efficiently utilized. Bran, better than endosperm but inferior to germ, was relatively indigestible although the digested portion was efficiently utilized. Germ was both digestible and efficiently utilized.     

The feeding habits of Sargochromis Codringtonii in Lake Kariba, Zimbabwe

The food of adult Sargochromis codringtonii in Lake Kariba was dominated by prosobranch snails with pulmonate snails forming an insignificant dietary component. S. codringtonii does not show particular preference for any snail species, feeding on those species that are readily available. Morphometrical analysis of the feeding structures of S. codringtonii showed that the fish is well adapted to handling a molluscivorous diet. The pharyngeal jaws with their flat crowned teeth are well adapted to crushing molluscs. The daily food consumption rate of S. codringtonii was estimated over different seasons. Estimates of food consumption indicate that the fish consumed 13.96% of their dry body weight per day in summer falling to 4.46% in winter. The low consumption of pulmonate snails suggests that S. codringtonii can not be a successful biological control agent against the vectors of schistosomiasis.     

Dietary cholesterol requirements of housefly, Musca domestica, larvae.

Quantitative analyses have been made of the dietary cholesterol requirement for the growth of the larvae of Musca domestica. The larvae will not grow on diets to which no cholesterol is added, a few pupae and adults are obtained when the concentration of cholesterol is 0·05 μmol/g of diet, but the concentration has to be raised to 0·36 μmol/g of diet before the maximum numbers of pupae and adults are obtained. Further addition of cholesterol above 0·36 μmol/g diet did not have any significant effect on the weight and growth of the larvae. However, the ratios of the cholesterol to phospholipid fractions recovered from the larvae increased rapidly when the concentration of cholesterol in the diet was raised from 0·05 to 0·56 μmol/g of diet. Above this concentration only a slight increase in the ratios was observed. Larvae reared on diets containing 0·05 μmol cholesterol/g of diet contain only 25 per cent of the cholesterol content of the larvae reared on the diets containing more than 0·28 μmol of cholesterol/g of diet, the cholesterol content being expressed relative to the weight of the larvae,The absence of cholesterol synthesis has been demonstrated in the larvae by feeding [4-¹⁴C] cholesterol. The specific activity of the cholesterol recovered from the larvae is the same as that of cholesterol added to the diet. Irrespective of the cholesterol concentration of the larval diet, approximately 97 per cent of the radioactivity recovered from the larvae behaved as free cholesterol, less than 1 per cent as cholesterol esters and the rest as unidentified ‘polar sterols’. The results are compared with those from similar studies on other insects.     

Characterization of chitinase from Shewanella inventionis HE3 with bio-insecticidal effect against granary weevil,Sitophilus granarius Linnaeus (Coleoptera: Curculionidae)

A 40 kDa chitinase from Shewanella inventionis HE3 was purified (ChiA-Si40) and characterized. Using fermentor with an optimized medium for 48 h at 37 °C, enzyme activity was enhanced by 10-times compared to those using shaking-flask-culture. Purified chitinase is a homogenous monomer with molecular mass of 40 kDa. Its N-terminal residues revealed significant identity with glycoside hydrolase family 18 (GH18) chitinases from Gammaproteobacteria. Using colloidal chitin as a substrate, its finest activity was accomplished at pH 4 and a temperature of 70 °C. Its catalytic efficiency (k_cat/K_m) was superior to that of some bacterial GH18 chitinases and commercial enzyme, Chitodextrinase®. For scale-up and with regards to the improvement of ChiA-Si40 with PEG 6000 storage stability (6 months), the atomizing process was more pronounced than that of lyophilizing. Bio-assay of ChiA-Si40 against grain weevil Sitophilus granarius, indicates that it had an efficient insecticidal effect. About 10–100 % mortality rates were obtained 1-h after insect came in contact with ChiA-Si40. Histological study clearly demonstrated that luxury larval mid-gut, peritrophic-membrane, and epithelial-cells have been affected considerably after ChiA-Si40 treatment. These properties make ChiA-Si40 a potential bio-insecticidal agent for the biological control of S. granarius that is popular among insect pests of stored grains in Algeria.     

Rôle of labelled dietary fatty acids and acetate in phospholipids during the metamorphosis of Pieris brassicae

The incorporation of labelled dietary palmitic, linoleic, and linolenic acids into neutral (NL) and phospholipids (PL) during the metamorphosis of Pieris brassicae was studied, and the ability of the fat body to incorporate acetate into PL determined. Thirty-three per cent of total lipid in early fifth instar larvae (minus haemolymph) is PL, while the corresponding value in female 4-day pupae is 13·0 per cent and in the fat body of 4-day pupae 6·3 per cent. Incorporation of label into PL was studied more closely and in all cases the label was recovered from phosphatidylcholine (PTC) and phosphatidylethanolamine (PTE). The label from palmitate was also found in sphingomyelin and possibly phosphatidylserine. Specific activity of PL in the case of palmitic and linolenic acids was greatest in late fifth instar larvae. In early fifth instar larvae on palmitic acid-1-¹⁴C 39·0 per cent of label was in PTC, 52·8 per cent in PTE, and 2·0 per cent in sphingomyelin. In late fifth instar 45·0 per cent was in PTC, 45·5 per cent in PTE, and 6·5 per cent in sphingomyelin, while in 4-day female pupae 45·2 per cent was in PTC, 41·3 per cent in PTE, and 13·5 per cent in sphingomyelin. The label from linolenic acid only varied a little from early fifth instar to 4-day pupae, 51·8 per cent being in PTC and 48·2 per cent in PTE in early fifth instar larvae. The label from linoleic acid is incorporated in fat body PL almost exclusively in PTC and PTE, 55·8 and 43·2 per cent respectively in 4-day female pupae. Injected acetate is distributed after 1 hr between PTC (58·6 per cent), PTE (24·4 per cent), and sphingomyelin (17·0 per cent). It was concluded that the polyunsaturated acids are proportionately more common in PTE than in other PL types, and that the fatty acids of sphingomyelin are mainly those that the insect is capable of synthesizing from acetate. Palmitic acid is desaturated by Pieris to palmitoleic acid and the latter possibly utilized in PTE to compensate for a deficiency of linolenic acid in the artificial diet. No saturation of linoleic or linolenic acid was found. The rates of PL and NL synthesis during development and the rôle of the investigated fatty acids in the biosynthesis of PL are discussed.     

Effect of particle size on the anthelmintic efficacy of mebendazole against Nippostrongylus brasiliensis in the rat.

The effect of reduction in particle size on the anthelmintic efficacy of mebendazole (methyl 5 (6)- benzoyle 1–2 benzimidazole carbamate) was studied in rats undergoing a primary infection with N. brasiliensis. A single oral treatment with fine ground mebendazole (particle size spectrum—54·95 per cent of particles less than 10·62 μ dia.; 86·06 per cent less than 21·27 μ) removed more than 98 per cent of adult worms from the intestine at a dose rate of 12·5 mg/kg body wt. On the other hand the best result achieved with coarse ground mebendazole (18·47 per cent of particles less than 10·62 μ dia; 42·26 per cent less than 21·27 μ dia) was 58 per cent of adult worms removed at a dose rate of 100 mg/kg body wt. It was also shown that fine ground mebendazole adversely affected migrating third stage larvae of N. brasiliensis.     

Chemical composition and rate of synthesis of the larval salivary secretion of the fly Rhynchosciara americana

The salivary secretion of Rhynchosciara americana was chemically analysed. The secretion shows a yellow colour, with a pH of 7·5 and protein as its major component (94·5 per cent of the secretion dry weight). Carbohydrates are minor components of the secretion which amount to 3·4 per cent of the secretion dry weight, of which 2·3 per cent are neutral carbohydrates and 1·1 per cent are galactosamine. The major amino acids present in the secretion proteins are aspartic acid, glycine, serine, and glutamic acid. The salivary secretion proteins can be separated into eleven protein fractions by urea-acrylamide gel electrophoresis from which nine fractions are PAS positive. The salivary pigment moves together with the protein fraction No. 8, which is quantitatively the most important one, and has spectral characteristics identical to a haemolymph pigment. The higher rate of gland protein labelling by ¹⁴C-phenylalanine determined in vivo and in vitro occurs around the middle of the spinning stage at the same time as the appearance of the large chromosomal puffs. The rôle of the salivary secretion in cocoon production is discussed.     

The chemical basis of honeybee, Apis mellifera, caste formation. Partial purification of queen bee determinator from royal jelly

On royal jelly, 1- to 2-day-old honeybee worker larvae have been reared in vitro to adults in a yield of 67±18 per cent. Up to 100 per cent and, on an average, 60 per cent of them were queens and intercaster. The preparation of a basic food from royal jelly by extensive alcohol extraction is described. With this control food, a survival rate of 47±18 per cent was achieved; 15 per cent of the adults were determined, 4·3 per cent of them were queens. Rearing of 1- to 2-day-old worker larvae on a basic food, to which unknown fractions may be added, was used as a biological test for the partial purification of queen bee determinator from royal jelly. By chromatography of the ethanol extract, previously treated with charcoal, on the cation exchanger Dowex 50 WX4 and rechromatography on silica gel, a 10⁵-fold purification of determinator was achieved. Chemical properties of the highly hydrophilic, low molecular active fraction are described.     

Effect of parasitism by Cardiochiles nigriceps on food consumption and utilization by Heliothis virescens

Parasitism of Heliothis virescens larvae by the endoparasitoid Cardiochiles nigriceps resulted in a reduction in the amount of food consumed by parasitized larvae. This effect was attributed in part to inoculation of material from the accessory reproductive glands of the female at the time of oviposition. Injection of solutions of either the calyx fluid or the poison gland (0·04 gl/larva) into non-parasitized larvae resulted in a reduction in the amount of food consumed by these larvae. A 1 : 1 mixture of these glands (total of 0·04 gl/larva) appeared to be more active than either of the two glands alone. Both of these glands were essential for total activity since larvae parasitized by females lacking the poison gland (poi gl^? female) continued to eat and consumed more food than did those parasitized by a normal female.Parasitism resulted in a slower rate of crop-emptying. This effect was, however, shown to be a result of the quantity of food consumed. Inhibition of gut movement was therefore not considered the cause for the reduction in the amount of food consumed by parasitized larvae.The effect of parasitism on the ability of H. virescens larvae to utilize ingested food was partially reduced by parasitism. Larvae parasitized by a normal female were less efficient than non-parasitized larvae in digesting food. Those larvae parasitized by a poi gl^? female did not convert as much of their food to body substance as did non-parasitized larvae. Injection of solutions of accessory glands into non-parasitized larvae did not cause these effects.     

Lipids and fatty acids of the southwestern corn borer,Diatraea grandiosella

The lipids of the adults and of several immature stages of the southwestern corn borer, Diatraea grandiosella, were studied after they were fed natural corn stalks or artificial diets. Linoleic acid (18:2) was the major fatty acid of the neutral lipids in both the natural and the artificial diets, but aleic acid (18:1) was the principal neutral lipid in all insect stages. Also, linoleic acid and oleic acid were the principal acids in the insect phospholipids of all stages. The content of linoleic acid in the natural diet was also high, but that in the artificial diet appeared to be much too low for insect requirements. Phosphatidyl choline (PC) and phosphatidyl ethanolamine (PE) were the major phospholipids in all growth stages. Thus, in larvae diapausing in the field, the unsaturated fatty acid content of PC was 59·3 per cent, primarily 16:1 and 18:1, and PE was 87·4 per cent, primarily 18:1, 18:2, and 18:3, and the fatty acids in the number 1- and 2-positions of PC were 53·6 and 97·2 per cent unsaturated, respectively. The haemolymph of diapausing southwestern corn borer larvae contained primarily glycerides but also had some PC and PE. Fat body from diapausing larvae contained primarily 16:0, 16:1, and 18:1 in a ratio of 1 : 1 : 2. Thus lipids of the southwestern corn borer do not reflect dietary lipids as closely as do other insects studied.