Chemical and immunological properties of lipophorins from seven insect orders

The major insect hemolymph lipoprotein, lipophorin, was isolated from adults of eight insect species representing seven insect orders. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to compare their respective apoprotein components. In all species examined lipophorin was composed of at least two apoproteins, apolipophorin I (M_r ～ 250,000) and apolipophorin II (M_r ～ 78,000), and two species had a third apoprotein, apolipophorin III (M_r ～ 17,000). The density of each isolated lipophorin was determined from the refractive index of KBr following density gradient centrifugation. Immunoblotting with anti-larval Manduca sexta apolipophorin I and II of the apoproteins separated by SDS-PACE indicated cross reactivity between anti-M sexta apoLp-ll and apoLp-ll in all species tested. Anti-M sexta apoLp-l exhibited no cross reactivity for any species tested. Fluorescent lectin staining of the apoproteins separated on SDS-PAGE gels revealed the presence of covalently bound carbohydrate residues.     

Effect of water content and temperature on seed longevity of seven Brassicaceae species after 5 years of storage

Maximising seed longevity is crucial for genetic resource preservation and longevity of orthodox seeds is determined by environmental conditions (water content and temperature). The effect of water content (down to 0.01 g·H₂O·g^?1) on seed viability was studied at different temperatures for a 5‐year storage period in taxonomically related species. Seeds of seven Brassicaceae species (Brassica repanda, Eruca vesicaria, Malcolmia littorea, Moricandia arvensis, Rorippa nasturtium‐aquaticum, Sinapis alba, Sisymbrium runcinatum) were stored at 48 environments comprising a combination of eight water contents, from 0.21 to 0.01 g·H₂O·g^?1 DW and six temperatures (45, 35, 20, 5, ?25, ?170 °C). Survival curves were modelled and P50 calculated for those conditions where germination was reduced over the 5‐year assay period. Critical water content for storage of seeds of six species at 45 °C ranged from 0.02 to 0.03 g·H₂O·g^?1. The effect of extreme desiccation at 45 °C showed variability among species: three species showed damaging effects of drying below the critical water content, while for three species it was neither detrimental nor beneficial to seed longevity. Lipid content could be related to longevity, depending on the storage conditions. A variable seed longevity response to water content among taxonomically related species was found. The relative position of some of the species as long‐ or short‐lived at 45 °C varied depending on the humidity at which storage behaviour was evaluated. Therefore, predictions of survival under desiccated conditions based on results obtained at high humidity might be problematic for some species.     

Light use efficiency of dry matter gain in five macro-lichens: relative impact of microclimate conditions and species-specific traits

Relations between irradiance (I) and lichen growth were investigated for five macro‐lichens growing at two sites in Sweden. The lichens represented different mycobiont–photobiont associations, two morphologies (foliose, fruticose) and two life forms (epiphytic, terricolous). The lichens were transplanted at two geographically distant sites in Sweden (1000 km apart) from Sept 1995 to Sept 1996 in their typical microhabitats, where microclimate and growth were followed. Between April/May and Sept 96, the terricolous species had a dry matter gain of 0·2 to 0·4 g (g DW)^–1 and the epiphytes 0·01 to 0·02 g (g DW)^–1. When related to area, growth amounted to 30 to 70 g m^?2 for the terricolous species and to 1 to 4 g m^?2 for the epiphytes. There was a strong correlation between growth and intercepted irradiance when the lichens were wet (I_wet), with 0·2 to 1·1 g lichen dry matter being produced per MJ solar energy. Across the 10 sets of transplants, light use efficiencies of dry matter yield (e) ranged between 0·5 and 2%, using an energy equivalent of 17·5 kJ g^?1 of lichen dry matter. The higher productivity of the terricolous species was due to longer periods with thallus water contents sufficient for metabolic activity and because of the higher mean photon flux densities of their microhabitat. A four‐fold difference in photosynthetic capacity among the species was also important. It is concluded that lichen dry matter gain was primarily related to net carbon gain during metabolically active periods, which was determined by light duration, photon flux density and photosynthetic capacity.     

Heritability of bulb shape in some north European onion varieties

Heritabilities for shape index (the ratio of bulb height to diameter), based on parent—offspring regressions, were calculated for north European onion cultivars and inbred lines derived from them. Heritability estimates of 0·46 and 0·47 respectively were obtained for the two groups, S₀ parent bulbs giving S₁ progenies, and S₁ parent bulbs giving S₂ progenies. Within each offspring progeny, the regression of bulb shape index on log_e bulb weight was significant. The regressions were used to estimate the mean shape indices of the progenies at the same mean bulb weight as their parents, and a series of ‘corrected’ progeny shape means thus obtained. Recalculation of the heritabilities using these ‘corrected’ progeny means gave increased estimates (0·78 and 0·84). By using this regression approach, the breeder can achieve high heritabilities when selecting for a specific mean shape index.     

ANALYSIS OF INTESTINAL CELL PROLIFERATION AFTER GUANETHIDINE-INDUCED SYMPATHECTOMY

Rats were treated with guanethidine-sulphate every 48 hr from birth until 15 days and then maintained until young adulthood. Sympathectomy was verified by dissection and light microscopic preparation of the superior cervical and coeliac ganglia which showed at least a 78% reduction in the number of perikarya. The effect of the chemical sympathectomy was a decrease in the amplitude of the circadian mitotic rhythm from 44·7 to 27·1%, 67·0 to 25·3% and 54·9 to 24·7%, in the duodenum, jejunum, and ileum, respectively. The shape of the mitotic index curve was altered and the mean mitotic index was significantly decreased (P < 0·01) in all three segments of the small intestine. The mean mitotic index of control intestinal epithelium was 3·2 ± 0·1%, 3·6 ± 0·1% and 4·0 ± 0·1% in the duodenum, jejunum, and ileum, respectively, and 2·3 ± 0·1%, 2·4 ± 0·1%, and 2·5 ± 0·1% in guanethidine-treated rats. Stathmokinetic estimates of cycle time were obtained by use of the metaphase arrest agent, colchicine. The longest cell generation cycle time (T_c) and lowest mitotic index occurred between 12.00 and 16.00 hours and the shortest T_c and highest mitotic index occurred between 00.00 and 04.00 hours, in all three segments of the small intestine. Guanethidine-treatment lengthens T_c throughout the small intestinal epithelium and reduces the range of variation in T_c over a 24-hr period. It is suggested that norepinephrine depletion induced by guanethidine may be the cause of the inhibition in circadian periodicity and that norepinephrine and the sympathetic nervous system may be essential for the maintenance of circadian rhythms in mitotic activity.     

The effects of gibberellic acid and nutrient sprays on growth and yield of brussels sprout

Young plants of brussels sprout, cv. Cambridge Special, growing in pots in a glasshouse, were sprayed on ten occasions with gibberellic acid (GA) at 0, 25, 100 and 400 p.p.m. and ammonium nitrate (NH₄NO₃) at 0, 0·0125, 0·025 and 0·05 M concentrations in all combinations. In 24 days both GA and NH₄NO₃ increased leaf area, leaf number, dry weights of leaf, stem and root, and fresh weights of leaf and stem. GA increased stem height and decreased fresh weight/unit leaf area (leaf thickness), whereas NH₄NO₃ did not affect stem height and increased leaf thickness. Of the GA treatments, 100 p.p.m. gave the largest plants as judged by fresh weight of the whole plant and leaf area, and of the N treatments 0·05 M NH₄NO₃ increased growth most. The best treatment combination was 0·05 M NH₄NO₃ with 100 p.p.m. GA, which gave the greatest fresh and dry weights of the whole plant, leaf area and leaf dry weight as well as increasing leaf thickness. Significant interactions were found between GA and N for dry weight of leaf, fresh and dry weight of root, and leaf thickness but not for leaf area or stem growth (fresh weight, dry weight, height). The combination of levels of GA and N that can be expected to increase leaf area, leaf dry weight and leaf thickness simultaneously, may lie within narrow limits for a particular crop. In a second experiment plants were sprayed with all combinations of GA (0 and 100 p.p.m.) seven times under glass in pots and ten times in the field, and three levels of KH₂PO₄ on twelve occasions in the field. Sprouts (axillary buds) were harvested in October and February. At the first harvest GA did not affect fresh weight or number, but increased both the total number of sprouts picked (with GA = 80·7; without GA = 69·8 per plant) and the total fresh weight of saleable sprouts (with GA = 2·59; without GA = 2·33 lb/plant). KH₂PO₄ also increased the weight and number of sprouts at the final harvest and the number of small sprouts at the first. There were interactions between GA and KH₂PO₄ (P= < 0·001) for both tota weight and number of saleable sprouts.     

Potential effects of climate warming on the survivorship of adult Monochamus galloprovincialis

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Occurrence of Gobiidae larvae in a tropical Brazilian estuary,with particular emphasis on the use of size classes to categorize species guilds

The structure and seasonal dynamics of larvae of the Gobiidae family in the Mucuri Estuary (Bahia, Brazil) were studied for nine consecutive years. Sampling was conducted at three stations in the lower estuary channel, between 2002 and 2010, in relation to season, day and night and tidal variations. A total of 5802 Gobiidae larvae, representing 15 taxa (12 species and three morphotypes), were collected in the Mucuri Estuary during this time. The highest mean ± s.d. density of fish larvae, 54·7 ± 79·8 larvae 100 m^?3, was recorded during the flood tide and night sampling. Ctenogobius boleosoma was the most abundant species (68%), being dominant in the rainy and dry seasons and had a long reproductive period. This species was classed as a marine estuarine‐opportunist because it was observed at high frequencies and active larvae entering the estuary between 6·1 and 12·0 mm standard length (L_S). Gobionellus oceanicus, second in abundance (12%), occurred only in later larval stages but did not use the estuary as a preferred location for spawning, being classed a marine estuarine dependent. Microgobius carri (11%) was represented in all L_S classes and was resident in the estuary for spawning, remaining there throughout their life cycle. The other species were considered rare due to their low densities and could not be classified in any guild.     

Offspring diet supersedes the transgenerational effects of parental diet in a specialist herbivore Neolema abbreviata under manipulated foliar nitrogen variability

Diet quality influences organismal fitness within and across generations.For herbivorous insects,the transgenerational effecets of diet remain relatively underexplored.Usinga3×3×2 factorial experiment,we evaluated how N enrichment in parental diets of Neolemd abbreviata(Larcordaire)(C oleoptera:Chrysomelidae),a biological control agent for Tradescantia fluminensis Vell.(Commelinaceae),may influence life history and performance of Fi and F2 offspring under reciprocal experiments.We found limited transgenerational effects of foliar nitrogen variability among life-history traits in both larvae and adults.Larval weight gain and mortality were responsive to parental diet contrary to feeding damage,pupal weight and duration taken to pupate.There were significant parental diet x test interactions in larval feeding damage,weight gain,pupal weight and time to pupation.Generally,offspring from parents under high N plants performed better even under low N test plants.Adult traits including oviposition selection,feeding weight and longevity did not respond to the efects of parental diet nor its interaction with test diet as was the case in the larval stage.However,the main efects of test diet were more important in determining adult performance in both generations suggesting limited sensitivity to parental diet in the adult stage.Our results show conflicting responses to parental diet between larvae and adults ofthe same generation among an insec species with both actively feeding larual and adult life stagee These tranegeneratinonal efferte,or lack thereof,may have implications on the field performance of N.abbrevita under heterogencous nutritional landscapes.     

THE ISOLATION AND LIPID COMPOSITION OF MYELIN-FREE AXONS FROM RAT CNS1

—Myelin-free axons were isolated from rat CNS using a modification of the method of De Vries et al. (1972). On a dry weight basis, the axons contained 15·2% lipid composed of 19·4% cholesterol, 56·9% phospholipid and 23·7% galactolipid with a weight ratio of cerebroside to sulfatide of 3·6-1. The phospholipid was composed of 11·0% ethanolamine phosphatides (44·4% in the plasmalogen form), 21·0% choline phosphatides (9·3% in the plasmalogen form), 4·5% sphingomyelin, 4·5% phosphatidyl serine, 4·3% phosphatidyl inositol, 3·0% diphosphatidyl glycerol and 8·5% unidentified phospholipid. The rat axons contained 0·18 μg ganglioside NeuNAc/mg dry wt. In addition to the 4 major brain gangliosides, the rat axons contained gangliosides G_D2 and G_D3. The axonal galactolipid could not be accounted for by myelin contamination as revealed by electron microscopy, absence of the characteristic ratio of myelin specific proteins in the axonal protein profile as shown by polyacrylamide gel electrophoresis, and the axonal level of the myelin marker enzyme 2′,3′-cyclic nucleotide-3′-phosphohydrolase. The relationship between lipids of axons isolated from rat and bovine CNS, and rat whole brain and CNS myelin is discussed.     

A continental-scale study of seed lifespan in experimental storage examining seed,plant, and environmental traits associated with longevity

Management of seed banks conserving the biodiversity of phylogenetically diverse species requires insight into seed longevity. This study determined the seed longevity of 172 species sourced from across the mega-diverse flora of the Australia continent. Seeds were aged via a controlled ageing experiment through storage at 45 °C and 60 % RH, or 60 °C and 60 % RH, and regularly tested for germination. Relative seed longevity between species was determined by comparing the time to 50 % viability loss (p ₅₀), calculated via probit analysis of seed survival curves. Seed, plant, and environmental traits were examined for associations with longevity. The p ₅₀ values varied between species from 3.0 to 588.6 days. Serotinous species, and woody trees and shrubs, had significantly longer-lived seeds than geosporous species, and species of herbaceous habit. Seeds that possess physical dormancy, and seeds with large embryos with little endosperm, were also long-lived. There was a weak, but significant, positive correlation between seed mass and longevity. Seeds sourced from regions of higher mean annual temperature and rainfall were significantly longer-lived than seeds from cooler and drier regions, although both environmental factors were weakly associated with longevity. Compared with species from other regions of the world, prolonged longevity is a feature of many Australian species. Nevertheless, seed life-spans vary substantially between species and close consideration of seed traits along with biotic and abiotic components of the plants and their environment can assist to differentiate between potentially long- and short-lived seeds.     

Interactions of Adult Stoneflies (Plecoptera) with Riparian Zones I. Effects of Air Temperature and Humidity on Longevity

The effects of constant air temperature and relative humidity on the longevity of three species of gripopterygid stonefly adults from New Zealand were investigated in laboratory experiments, and the results were compared to field measurements of air temperature and humidity obtained during summer. Greatest longevity for Zelandobius furcillatus, Zelandoperla decorata and Acroperla trivacuata was recorded in cool humid conditions (10°C, 100% humidity) for adults fed water and a 5% sucrose solution. Absence of feeding reduced survival by 37–73% at 17°C and 100% humidity. Survival decreased significantly with increasing constant air temperature (10, 17 and 25°C) and decreasing mean relative humidity (100, 81 and 15%). Males survived significantly longer than females in all temperature treatments for Z. furcillatus, but longevity was not influenced by gender in other species or in the humidity experiments. Interpolated LT 50 values over 96 h for female stoneflies in the temperature treatments averaged of 22–23°C. Field measurements at near-ground level and 1.5 m above the streambank during summer indicated that these air temperatures were exceeded for 25% of the time in a pasture catchment compared to &lt;0.1% of the time in a native forest catchment. These findings implicate air temperature as a factor potentially influencing the longevity of adult stoneflies, and suggest that maintenance of appropriate microclimate conditions should be a consideration in riparian management.     

Composition and Productivity of the Benthic Macroinvertebrate Community of a Subtropical Reservoir

Physical-chemical conditions, phytoplankton productivity, community structure and productivity of the macroinvertebrate benthic community were determined during 1976–77 in a subtropical reservoir. Physical-chemical results revealed high nitrate and phosphate concentrations with highest values in the riverine segment. Large phytoplankton populations were present during most of the year. Phytoplankton productivity was high, producing an annual mean of 87 mg C · m⁻³ · h⁻¹ (12 hours light day). High turbidity in the riverine segment limited phytoplankton productivity during winter and spring. Macrobenthos was dominated by chironomids (Chironomus, Procladius, Coelotanypus and Tanypus) and oligochaetes (Limnodrilus). The annual mean benthic population was estimated at 1,626 · m⁻² with a mean dry weight of 0.66 g · m⁻². Mean benthic species diversity was 1.80. A lacustrine-riverine community gradient was revealed. Benthic productivity was 6.8 g · m⁻² · yr⁻¹ (dry weight) with a P: B ratio of 10. A low correlation was observed between benthic and phytoplankton productivity, and between phytoplankton standing crop and benthic macroinvertebrate numbers throughout the reservoir. Algal food supplies had little impact on the benthic community which was composed predominately of species which fed mostly on organic detritus. Stressful conditions caused by low dissolved oxygen concentrations probably inhibited development of the benthic community throughout the reservoir during summer months, while high sedimentation rates limited development in the head waters.     

Effect of chlorfluazuron and flufenoxuron on development and reproductive performance of the black cutworm,Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae)

The black cutworm, Agrotis ipsilon, is an insect pest with great agricultural importance worldwide. In Egypt, it has acquired resistance against most of the conventional pesticides, particularly organophosphate insecticides, applied against it. Therefore, the present study aimed to investigate the effect of two benzoylureas (chitin synthesis inhibitors) namely chlorfluazuron and flufenoxuron on some developmental and reproductive parameters of Agrotis ipsilon that survived treatment of newly molted fourth instars with the LC₅₀ of these compounds (1.00 and 4.68 mg/L, respectively) using leaf–dip method. These two compounds significantly enhanced the mean larval and pupal durations, whereas they significantly declined the mean percentage pupation, adult emergence, pupal weight, adult longevity, fecundity, and fertility compared to untreated insects. The mean pre-oviposition period was not significantly affected, whereas the mean oviposition period was significantly decreased. Both chlorfluazuron and flufenoxuron significantly decreased the mean total protein, carbohydrate, and lipid content in the ovaries compared to untreated insects, except for insignificant decrease in the total protein in flufenoxuron-treated ones. Additionally, the total ovariolar length was significantly atrophied. In conclusion, chlorfluazuron and flufenoxuron interfere with vitellogenesis of A. ipsilon. Moreover, they could suppress the population size of this insect species by affecting its development and reproductive potential.     

Estimating the survival rate and mean longevity for adults in a field population of the green rice leafhopper,Nephotettix cincticeps Uhler,by the application ofHokyo andKiritani’s method

Application of the female dissection method proposed byHokyo andKiritani (1967) was attempted in both 1968 and 1969 to estimate the daily survival rate and the mean longevity for the adult population of the green rice leafhopper, Nephotettix cincticeps, in a paddy field. The estimated mean longevity for females was far shorter than the physiological longevity of this species, ranging from 4 to 7 days with some variation between different generations. This could explain the remarkable discontinuity among successive generations which proved to form an important feature of the pattern of seasonal population changes of this insect. Between the two years, the estimated survival rate (and hence the mean longevity also) was negatively correlated to the estimated population size of adults that emerged in either of the two successive generations. This apparent density dependence suggests the possibility that the adult survival, including the effect of dispersal, plays some critical role in bringing about the remarkable population stability from year to year shown by the field population of N. cincticeps.     

Cultivation of Lemna gibba under desert conditions. I: Twelve months of continuous cultivation in open ponds

Duckweed, Lemna gibba, was grown in 12 m² shallow ponds in the Negev desert, during 12 months of continuous cultivation, beginning April 1984. Average monthly growth rates varied with the season of the year. The lowest daily yield, 2·6±0·4 g dry weight m⁻² day⁻¹, was obtained during January. Highest daily yields, 7·9±2·6 g dry weight m⁻² day⁻¹ and 7·0±1·2 g dry weight m⁻² day⁻¹, were obtained during September and May. A 35% decline of the yield was seen during midsummer (July), 4·8±1·2 g dry weight m⁻² day⁻¹. The average rate for the year was 5·15±1·7 g dry weight m⁻² day⁻¹. The protein content of the plants ranged from 30 to 38% per unit dry weight.Growth performance is discussed in relation to the prevailing climatic conditions.     

Upper thermal tolerance of closely related Danio species

The main finding of this study was that measuring maximum heart rate during incremental warming was an effective tool to estimate upper thermal limits in three small cyprinid Danio species, which differed significantly. Arrhenius breakpoint temperature for maximum heart rate, purportedly an index of optimum temperature, was 21·2 ± 0·4, 20·1 ± 0·4 and 18·9 ± 0·8° C (mean ± s.e .) for zebrafish Danio rerio, pearl danio Danio albolineatus and glowlight danio Danio choprae, respectively. The temperature where cardiac arrhythmias were first induced during warming (T_arr) was 36·6 ± 0·7, 36·9 ± 0·8 and 33·2 ± 0·8° C (mean ± s.e .) and critical thermal maximum (T_Cm) was 39·9 ± 0·1, 38·9 ± 0·1 and 37·2 ± 0·1° C (mean ± s.e .) for D. rerio, D. albolineatus and D. choprae, respectively. The finding that T_arr was consistently 3–4° C lower than T_Cm suggests that collapse of the cardiac life support system may be a critical trigger for upper temperature tolerance. The upper thermal limits established here, which correlate well with a broad natural environmental temperature range for D. rerio and a narrow one for D. choprae, suggest that upper thermal tolerance may be a genetic trait even among closely related species acclimated to common temperatures.     

Pre‐zygotic parental environment modulates seed longevity

The potential for the pre‐zygotic plant growth environment to play a role in determining seed longevity was investigated for a species that inhabits arid to semi‐arid Australia. Seed longevity is particularly important for wild populations in fluctuating environments because the longer a seed‐lot is able to survive in the soil seed bank the more likely it is to buffer the population from unpredictable environments. Thus Wahlenbergia tumidifructa plants received wet or dry soil moisture within a warm or cool glasshouse until flowering. Seeds subsequently produced by flowers that opened on the day that plants were moved to a common environment were collected at maturity and longevity assessed by controlled ageing at 60% relative humidity and 45°C. Mean seed longevity was similar for seeds produced by plants that grew in warm‐wet, warm‐dry and cool‐dry conditions (P₅₀ of about 20 days), but extended for plants in cool‐wet conditions (P₅₀ = 41.7 days). Cool temperatures resulted in seeds with a wider distribution of lifespans (σ = 20 days) than warm conditions (σ = 12 days); the large σ caused the extended P₅₀ for cool‐wet plants, but not cool‐dry as a result of a concomitant reduction in initial seed germination (K_i). After moving to the common environment, all plants generated new vegetative material, which went on to produce seeds with similar longevity (P₅₀ approx. 20 days) irrespective of original environment. Visible phenotypic responses of the parent to environmental conditions correlated with longevity and quality parameters of the progeny seeds, suggesting that a parental effect modified seed longevity. Our study provides novel empirical data showing that environmental conditions expected under climate change scenarios may potentially cause seed longevity to decline for a species that inhabits arid to semi‐arid Australia. These negative impacts on population buffering may weaken the storage effect mechanism of species coexistence in fluctuating environments.     

A laboratory energy budget for the ormer Haliotis tuberculata L.

Each component of the energy budget (ingestion, egestion,somatic growth, reproductive investment, respiration, excretion, and mucus production) was measured for Haliotis tuberculata L. held at 15°C in a 12-h light: 12-h dark regime. Ulva lactuca L. was used as the food source throughout, and the budget was assessed over the whole size range of the animal. Ingestion rates ranged from 1.94 to 997.2 cal·animal⁻¹·day⁻¹ in 0.01 to 50 g dry wt (3.71 × 10⁻³ to 17.3 g dry flesh wt) animals, respectively. The major component of the energy budget was somatic growth (37.5% of I) in a 0.01-g dry wt animal while it was respiration (31.1% of I) in a 50-g dry wt animal. Mucus production formed a large part of the budget (from 23.3% of I in a 0.01-g dry wt animal to 29.l% of I in a 50-g dry wt animal). Scope for growth, I − (E + R + U + M), was calculated as ranging from 24.5% of ingestion in a 50-g dry wt animal to 36.8% in a 0.01-g dry wt animal.Each component was measured independently and allometric relationships with animal dry weight calculated. Exponents ranged from 0.60 (somatic growth) to 1.06 (reproductive investment).The calorific value of food was 3419 cal·g dry wt⁻¹ and for faeces was 2817 cal·g dry wt⁻¹. Absorption as a percentage of ingestion in terms of dry weight ranged from 78% for 95-mm length animals (50 g dry wt) to 81% in 6-mm animals (0.01-g dry wt⁻¹). Gross and net growth efficiencies (K₁ and K₂) were calculated on an energy basis and both were logarithmically related to animal dry weight.     

Patterns of nitrogen utilization between the ambrosia beetle Xyleborus dispar and its symbiotic fungus

Some parameters of nitrogen utilization between the ambrosia beetle Xyleborus dispar in mutualistic association with its symbiotic fungus Ambrosiella hartigii, were examined. Qualitative and quantitative analyses of the major nitrogenous excretory products were made on the various life stages of X. dispar. The main nitrogenous product found in excreta and hindguts of beetles, larvae, and pupae, was uric acid (range 7·6–14·8 μg uric acid/beetle). No ninhydrin-positive compounds were located in excreta of the beetles. The concentration of ammonia-nitrogen in the various life stages averaged between 0·70 and 1·13 μg NH₃-N/beetle.Total nitrogen determinations were made on sapwood samples of Malus sylvestris (0·34 ± 0·005% N by dry weight), attacked wood, ‘pre-brood’ (0·31 ± 0·005% N by dry weight), and attacked wood ‘post-brood’ (0·17 ± 0·02% N). Similar determinations of the artificial medium (l-asparagine medium) indicated that a nitrogen requirement of about 0·08–0·1% N by dry weight was necessary before oviposition could occur.Fixation of atmospheric nitrogen by individual X. dispar beetles in vitro was not indicated using the acetylene ethylene reductase method. Proteolytic enzyme activity was not found on examination of diapause beetles, their excreta, larval and pupal excreta, and the ambrosial and mycelial forms of A. hartigii.Comparative concentrations of soluble proteins and free amino acids suggested that fungus in the mycangia was built up from free amino acids of the insects. At the period of emergence, flight, and attack of new hosts, the females were found to have a concentration of soluble proteins more than double that found in the beetles during the remainder of the year. The free amino acids were the lowest values recorded during this period (March–October).