Quantitative requirements of the saw-toothed grain beetle,Oryzaephilus surinamensis,for dietary l-isoleucine and l-valine

Larvae of the saw-toothed grain beetle, Oryzaephilus surinamensis, were reared individually on meridic diets from hatching to adult emergence at 32±0·25°C and 75±5% r.h. On diets devoid of l-isoleucine or of l-valine, these larvae developed little, if at all, and died before reaching the larval-pupal ecdysial stage. Graded concentrations of l-isoleucine up to 6·0 mg/g or of l-valine up to 5·0 mg/g increased the rate of development and survival of this insect. Concentrations of either amino acid beyond these levels was neither beneficial nor harmful to O. surinamensis, in terms of rate of development or survival.     

DISTRIBUTION OF INTRACELLULAR NITROGEN IN MARINE MICROALGAE: BASIS FOR THE CALCULATION OF SPECIFIC NITROGEN-TO-PROTEIN CONVERSION FACTORS

The utilization of nitrogen-to-protein conversion factors (N-Prot factors) is a widely accepted and practical way to determine total protein content. The accuracy of protein determination depends on the establishment of specific N-Prot factors, since the conventional factor of 6.25 may be unsuitable for all species. This study was designed to determine the concentrations of the main nitrogenous compounds and to establish N-Prot factors specific for the following marine microalgae: Chlorella minutissima, Dunaliella tertiolecta, Hillea sp., Isochrysis galbana, Nannochloropsis oculata, Phaeodactylum tricornutum, Prorocentrum minimum, Skeletonema costatum, Synechococcus subsalsus, and Tetraselmis gracilis. Cultures were maintained under a 12-h photoperiod (300 μmol photons·m^?2·s^?1) at temperatures of 20.0°± 1.0° C (dark) to 23.0°± 2.0° C (light) in Walne’s culture medium without additional external carbon sources. The distribution of intracellular nitrogen was studied by determining total nitrogen (TN, by CHN [carbon, hydrogen, and nitrogen] analysis), protein N (PN, by analysis of total amino acids), and nonprotein N (NPN, determined by analysis of DNA, RNA, chlorophylls (chl) a,b, and c, and intracellular inorganic nitrogen—NO₃^?, NO₂^?, and NH₃+ NH₄⁺) in logarithmic and stationary growth phases of cultures. Variations occurred in both accumulation and distribution of PN and NPN among the species, as well as in each species during the different growth phases. Inorganic nitrogen compounds were observed to be the most important NPN source (from 6.4 ± 0.1% to 41.8 ± 4.2% of total N) in all species (except D. tertiolecta), followed by nucleic acids (from 0.8 ± 0.1% to 26.1 ± 2.4% of TN) and chlorophylls (from 0.2 ± 0.0% to 3.1 ± 0.3% of TN). Total amino acid residues ranged from 63.1 ± 4.6% up to 88.1 ± 11.2% of TN, which is in agreement with the presence of high NPN concentrations. N-Prot factors are proposed for each growth phase in the studied species, based on the ratio of amino acid residues to TN, establishing specific N-prot factors ranging from 3.60 ± 0.27 to 4.99 ± 0.64. The mean N-Prot factor for all species/growth phases was 4.58 ± 0.11. The present study shows that the use of the traditional factor 6.25 is not suitable for these marine microalgae, and possibly for other species, because it overestimates their actual protein content.     

Analysis of Tiphia parasitoids preovipositional behaviors and of their scarab host defensive responses

Tiphia vernalis Rohwer and Tiphia popilliavora Rohwer were introduced as biocontrol agents against Japanese beetles (Popillia japonica Newman) and oriental beetles (Anomala orientalis Waterhouse). Studies have shown that under field conditions, T. vernalis parasitize Japanese beetles more often than oriental beetles. This study was done to understand how tiphiid wasps handle the two different host species and the influence of host defensive behaviors on the oviposition process of tiphiid wasps. The preovipositional behaviors performed by Tiphia wasps included: stinging, examining, moving soil, kneading, host-feeding, and host scraping. The frequency, sequence, and total time spent on each behavior before oviposition were scored and compared between two host species. The sequence and frequency of preovipositional behaviors performed by both Tiphia wasps did not show a difference between the two host species. However, female T. vernalis spent significantly longer time trying to sting oriental beetles than Japanese beetles in order to paralyze them. The time T. popilliavora spent on prestinging behaviors did not show a difference between Japanese and oriental beetles. The defensive behaviors performed by Japanese and oriental beetle grubs included: vigorous movements, rubbing their abdomen or head against the wasp’s abdomen, and biting at the attacking wasp. The frequency and total time spent on each defensive behavior was scored and compared between two host species. Overwintered, third instar oriental beetle grubs spent significantly longer time on defensive behaviors when they were attacked by T. vernalis which likely cause wasps to spend longer time trying to sting oriental beetle grubs. The active host resistance gained through behavioral defenses could make oriental beetle grubs less susceptible to T. vernalis attack than Japanese beetle grubs, especially under field conditions. Younger grubs attacked by T. popilliavora did not exhibit these differences.     

Ambrosia beetle response to ethanol concentration and host tree species

Ethanol emitted by stressed trees is an olfactory cue used by ambrosia beetles (Coleoptera; Curculionidae; Scolytinae) to locate susceptible hosts to colonize. In addition, ethanol enhances the growth of ambrosia beetle fungal symbionts, improving colonization. Whether host selection and colonization are affected also by the amount of ethanol produced by stressed trees and by tree species is unclear. To investigate these mechanisms, we induced attacks by ambrosia beetles in bolts of eight tree species by coring and filling them with either 5% or 90% ethanol solutions in water. For each ethanol concentration, bolts of the eight different tree species were replicated six times in a randomized complete block design. Entry holes were used as a proxy for host selection whereas gallery development stage was used as a proxy for colonization. Ethanol concentration differentially affected host selection of the three ambrosia beetles that were active during this study. Anisandrus dispar Fabricius preferentially attacked bolts with 90% ethanol concentration, Xylosandrus crassiusculus (Motschulsky) preferentially attacked bolts with 5% ethanol concentration, and Xyleborinus saxesenii (Ratzeburg) attacked bolts irrespective of ethanol concentration. Colonization of X. crassiusculus reflected the same pattern observed for entry holes. The effect of host tree species on host selection was most prominent for X. saxesenii, while X. crassiusculus established a higher number of developed galleries in Ostrya carpinifolia Scopoli bolts than on five of the other tested tree species. Our results suggest that ethanol concentration and host tree species may influence ecological niche partitioning among ambrosia beetle species.     

Aspects of the nutrition of adult female Glossina morsitans during pregnancy

The adult female Glossina morsitans fed on goat takes, in terms of dry weight, approximately 37·6 mg of blood during a pregnancy period, and at least 84% of this dry blood comprises proteins. In this diet leucine is the most abundant, isoleucine and methionine contents are markedly small, while values for other amino acids differ and are intermediate. Shortly after pupariation the third instar larva produced by goat-fed females contains 67·3% water, 15·3% residual dry weight, 11·0% fat, and 6·4% puparium. The amount of each of the amino acids present in the larva and its puparium comprises a small proportion of that taken by the female fly during a pregnancy period. The fat content of the fully-fed larva is also very small compared with the total nutrient intake by the pregnant female. These results are discussed in terms of the nutrition of intra-uterine larva in relation to feeding by its female parent.     

Quantitative dietary requirements of the saw-toothed grain beetle,Oryzaephilus surinamensis,for l-leucine

Larvae of the saw-toothed grain beetle, Oryzaephilus surinamensis, were reared individually on meridic diets from hatching to adult emergence at 32±0·25°C and 75% r.h. These larvae developed little, if at all, and failed to carry out larval-pupal ecdysis on a diet devoid of leucine. Survival increased as the concentration of dietary l-leucine increased from 0·0 to 3·0 mg/g and rate of development, as l-leucine increased to 8·0 mg/g. With concentrations of dietary l-leucine greater than 8·0 mg/g, the rate of development of Oryzaephilus surinamensis decreased.     

Raised nutrient levels change heathland into grassland

In 1969 N and P fertilizer experiments were carried out on the Westerheide (The Netherlands) to investigate the growth of Calluna vulgaris. In 1981 the floristic composition of the experimental plots was analyzed. Repeated nitrogen treatment of 28 kg · ha^-1 · yr^-1 resulted in dramatic replacement of Calluna vulgaris by Festuca ovina as a dominant. Phosphorus treatments did not result in such a change. The amounts of nitrogen applied in this experiment are similar to those that will be available during the first years following the dying off of Calluna vulgaris as result of a heather beetle infestation. It is hypothesized that a heather beetle infestation alone may result in a similar replacement of Calluna vulgaris by grass species.     

Adult feeding affects fecundity of the predator,Calosoma sycophanta (Col.: Carabidae)

Calosoma sycophanta L. adults were fed either gypsy moth (Lymantria dispar L.) larvae or split grapes for set periods of time while their reproduction was monitored. Few female beetles reproduced unless fed gypsy moth larvae during the first week after they ended hibernation. Even females initially fed grapes that were later fed larvae had reduced reproduction. The implications these results have for relationships between beetle and gypsy moth populations are discussed.     

Ileal digestibility of nutrients and amino acids in low quality soybean meal sources treated with β-mannanase for growing pigs

Apparent ileal digestibility (AID) of energy, dry matter (DM), nitrogen (N) and amino acids and standardized ileal digestibility (SID) of amino acids in low quality soybean meals with different CP concentration (SBM 44% CP and SBM 48% CP) with or without 400 U β-mannanase/kg supplementation were evaluated in 20 cannulated barrows ((Landrace×Yorkshire)×Duroc) with an average BW of 25.08±3.42 kg. A N-free diet was used to determine basal endogenous losses of amino acids. The supplementation of β-mannanase improved (P<0.05) AID of DM compared with non-supplemented diet. However, enzyme supplementation did not improve (P>0.05) AID of N and energy. The type of SBM (SBM 44% CP v. SBM 48% CP) had no effect on AID of DM, N and energy. β-mannanase improved (P<0.05) AID of sum of essential amino acids, arginine, histidine, lysine, valine and glycine. The SID of lysine was higher (P<0.05) in enzyme supplemented than in non-supplemented diets. Larger AID and SID of threonine and proline (P<0.05) were observed in SBM 48% CP than in SBM 44% CP. In conclusion, the supplementation of enzyme improved AID of arginine, histidine, lysine, valine and glycine, but it did not cause marked difference in SID of these amino acids except for lysine. The low nutrient digestibility of the SBM sources used in the present experiment might have favoured the positive effect of β-mannanase supplementation.     

The effect of food composition on ingestion,development, and survival of a harpacticoid copepod, Tisbe cucumariae Humes

Feeding experiments were carried out on the benthic harpacticoid copepod Tisbe cucumariae Humes, using seven different diets of various dried and ground macroalgae and marsh grass, algal Aufwuchs, diatoms, polychaete meat, and cereal. In short-term experiments (1 h), ¹⁴C-labelled foods were used to measure ingestion rate of non-ovigerous adult females (individual dry wt = 5.57 ± 2.49 μg). No significant difference was found among the rates at which all foods, except polychaete meat, were ingested (12.7 to 17.3 × 10^?2μg dry wt· ind^?1· h^?1). Polychaete meat was consumed faster (23.9 × 10^?2μg dry wt·ind ^?1· h^?1). The nutritional value of the foods was estimated in long-term experiments (22 days) by measuring development time and survival of T. cucumariae. Both these variables were significantly correlated with the nitrogen, protein content, and C:N ratio of the foods. No relation was found, however, with the amount of carbon, calories and available calories in the diets. Thus, nitrogen (protein) content of the food was the factor limiting secondary production of the copepods.     

Which wavelength does the cigarette beetle, Lasioderma serricorne (Coleoptera: Anobiidae), prefer? Electrophysiological and behavioral studies using light-emitting diodes (LEDs)

The cigarette beetle, Lasioderma serricorne (Fabricius), is an important pest insect that consumes a variety of dry foods. It is known that UV light traps attract this species. However, less attention has been paid to its preferred wavelength. First, we investigated the spectral sensitivity of the compound eye. Next, we compared the attraction efficiency of LEDs of different colors (wavelengths). Our results showed that ultraviolet (UV, 375 nm) and blue (470 nm) LEDs attracted the most cigarette beetles of both sexes, irrespective of mating or oviposition status, although the UV LED consistently tended to attract the most beetles. Although the primary sensitivity peak of the compound eye was 520 nm, the green LED (520 nm) scarcely attracted beetles. Although the reason for the difference between the peaks in spectral sensitivity and attraction of beetles awaits further studies, whether UV and/or blue LEDs is more effective as a practical light trap for controlling L. serricorne beetle is discussed in this study.     

Increased synthesis of α‐tocopherol,paramylon and tyrosine by Euglena gracilis under conditions of high biomass production

Aims: To analyse the production of different metabolites by dark‐grown Euglena gracilis under conditions found to render high cell growth. Methods and Results: The combination of glutamate (5 g l^?1), malate (2 g l^?1) and ethanol (10 ml l^?1) (GM + EtOH); glutamate (7·15 g l^?1) and ethanol (10 ml l^?1); or malate (8·16 g l^?1), glucose (10·6 g l^?1) and NH₄Cl (1·8 g l^?1) as carbon and nitrogen sources, promoted an increase of 5·6, 3·7 and 2·6‐fold, respectively, in biomass concentration in comparison with glutamate and malate (GM). In turn, the production of α‐tocopherol after 120 h identified by LC‐MS was 3·7 ± 0·2, 2·4 ± 0·1 and 2 ± 0·1 mg [g dry weight (DW)]^?1, respectively, while in the control medium (GM) it was 0·72 ± 0·1 mg (g DW)^?1. For paramylon synthesis, the addition of EtOH or glucose induced a higher production. Amino acids were assayed by RP‐HPLC; Tyr a tocopherol precursor and Ala an amino acid with antioxidant activity were the amino acids synthesized at higher concentration. Conclusions: Dark‐grown E. gracilis Z is a suitable source for the generation of the biotechnologically relevant metabolites tyrosine, α‐tocopherol and paramylon. Significance and Impact of the Study: By combining different carbon and nitrogen sources and inducing a tolerable stress to the cell by adding ethanol, it was possible to increase the production of biomass, paramylon, α‐tocopherol and some amino acids. The concentrations of α‐tocopherol achieved in this study are higher than others reported previously for Euglena, plant and algal systems. This work helps to understand the effect of different carbon sources on the synthesis of bio‐molecules by E. gracilis and can be used as a basis for future works to improve the production of different metabolites of biotechnological importance by this organism.     

Quantitative microanalysis of cell walls of Saprolegnia diclina humphrey and Tremella mesenterica fries

Cell walls of the fungi Saprolegnia diclina Humphrey and Tremella mesenterica Fries were analyzed quantitatively. Particular attention was paid to the hydrolysis and analysis of neutral sugars, amino sugars and amino acids. These components, together with total lipids, total uronic acids and the ashed residue, accounted for more than 90% by weight of the original dry cell wall preparation. There were substantial losses of amino acids during hydrolysis; however, analytical recovery approached 100% when total protein was calculated from the total nitrogen analysis. The analytical procedures were reproducible (±3% for amino acids and amino sugars, and ±5–10% for other components) when applied to individual cell wall preparations. However, even under carefully standardized conditions, different cell wall preparations from the same species showed variable composition.Glucose was the predominant neutral sugar in the cell wall polymers of both species. The amino acid compositions were remarkable in that neither species contained detectable levels of cyst(e)ine. Hydroxyproline was detected in both species. The report from Tremella mesenterica is the first for this imino acid from the cell wall of a Basidiomycete.     

Nitrogenous excretory materials from the American cockroach

Excreta obtained from both sexes of the American cockroach, Periplaneta americana, maintained on three different diets was examined for the presence of numerous possible nitrogenous materials in an effort to establish what substances are excreted by this insect. It was found that none of the classical terrestrial insect excretory products, including uric acid, could be detected by the methods employed. Rather, ammonia, amino groups, three tryptophan metabolites, unidentified water soluble materials, and water insoluble components constitute most of the total nitrogen excreted. Ammonia seems to be the largest single component, and it was quantitated in fresh and dried excreta. Upon lyophilization of the excreta more than half of the ammonia is lost.     

Overwintering ecology of <Emphasis Type="Italic">Philonthus rotundicollis</Emphasis> (Coleoptera,Staphylinidae) in the nest of the carpenter ant <Emphasis Type="Italic">Camponotus herculeanus</Emphasis> on the coast of the Sea of Okhotsk

A widespread rove beetle species, Philonthus rotundicollis, whose distribution range stretches across different climatic zones, including the coldest regions of the Asian northeast, was discovered as an inquiline within the nests of the carpenter ant Camponotus herculeanus on the coast of the Sea of Okhotsk in winter. It remained unclear if the beetles had significant cold-hardiness and whether they overwintered deep in the soil or were confined to particularly warm habitats. To clarify these aspects, the following metrics of cold-hardiness were measured: supercooling point (SCP), freezing point (FP), supercooling capacity (SCP-FP), and temperature minima at the beetles’ overwintering sites. In Ph. rotundicollis, mean SCP was -11.1 ± 0.7°C (ranging from - 7.9 to -18.8°C, n + 15), which was insufficient for successful overwintering even on the coast, since temperature minima in leaf litter during a snow-deficient winter fell to -14°C at the depth of 5 cm and -12°C at 20 cm. The beetles could not burrow deep into stiff soil and made use of crevices in dry peat-like soil layers as well as tunnels of soil- and rootdwelling animals, including carpenter ants. The presence of this rove beetle species in the ant nest was probably due to feeding on ant larvae because, at near-zero temperatures, the activity threshold of the beetles was lower than that of the ants that guarded the larvae.     

Encounters of old foes on a new battle ground for an invasive tree, Albizia julibrissin Durazz (Fabaceae)

Studies of invasive species often demonstrate that exotic or invasive plants have reduced levels of predation when first introduced into novel geographic areas. In some systems, native predators may subsequently be introduced and the plant-predator association is re-established. In this study, we analyzed a recent reassociation between the introduced tree, Albizia julibrissin, and its native seed predator, Bruchidius terrenus, which has also been introduced, in the vicinity of Athens, GA, USA. We documented a rapid increase in the levels of bruchid beetle predation on seeds of A. julibrissin from 2001 to 2002 and substantial predation in the following 5 years. From a six-year survey, we found that relative susceptibility to beetle predation among individual trees remained consistent over time. Furthermore, we found that phenology had significant effects on beetle predation on fruits and seeds, with fruits developed from early flowers having higher levels of beetle attacks than ones developed from later flowers in the season. Finally, seeds and fruits attacked by beetles were more likely to be attacked by a Fusarium fungus, probably due to coinfestation of the two predators. Host-predator reassociation that causes substantial loss of fertility or shows a relationship with reproductive phenology may have population level demographic effects or evolutionary consequences and deserve further research attention.     

Interactions in the uptake of amino acids, ammonium and nitrate ions in the Arctic salt-marsh grass, Puccinellia phryganodes

The uptake of amino acids and inorganic nitrogen by roots of Puccinellia phryganodes was examined to assess the potential contribution of soluble organic nitrogen to plant nitrogen uptake in Arctic coastal marshes, where free amino acids constitute a substantial fraction of the soil‐soluble N pool. Short‐term excised root uptake experiments were performed using tillers grown hydroponically under controlled conditions in the field. The percentage reductions in ammonium uptake at moderate salinity (150 mm NaCl) compared with uptake at low salinity (50 mm NaCl) were double those of glycine, but glycine uptake was more adversely affected than ammonium uptake by low temperatures. Glycine uptake was higher at pH 5·7 than at pH 7·0 or 8·2. The glycine uptake was up‐regulated in response to glycine, whereas ammonium uptake was up‐regulated in response to ammonium starvation. Nitrate uptake was strongly down‐regulated when tillers were grown on either ammonium or glycine. In contrast to N‐starved roots, which absorbed ammonium ions more rapidly than glycine, the roots grown on glycine, ammonium and nitrate and not N‐starved prior to uptake absorbed glycine as rapidly as ammonium and nitrate ions combined. Overall, the results indicate that amino acids are probably an important source of nitrogen for P. phryganodes in Arctic coastal marshes.     

High Nitrogen and Phosphorous Acquisition by Belowground Parts of Caulerpa prolifera (Chlorophyta) Contribute to the Species' Rapid Spread in Ria Formosa Lagoon,Southern Portugal

Despite worldwide proliferation of the genus Caulerpa and subsequent effects on benthic communities, little is known about the nutritional physiology of the Caulerpales. Here, we investigated the uptake rates of ammonium, nitrate, amino acids, and phosphate through the fronds and rhizoids + stolon, the internal translocation of nitrogen, and developed a nitrogen budget for the rapidly spreading Caulerpa prolifera in Ria Formosa lagoon, southern Portugal. Caulerpa prolifera acquired nutrients by both aboveground and belowground parts at similar rates, except nitrate, for which fronds showed 2-fold higher uptake rates. Ammonium was the preferential nitrogen source (81% of the total nitrogen acquisition), and amino acids, which accounted for a significant fraction of total N acquisition (19%), were taken up at faster rates than nitrate. Basipetal translocation of ¹⁵N incorporated as ammonium was nearly 3-fold higher than acropetal translocation, whereas ¹⁵N translocation as nitrate and amino acids was smaller but equal in either direction. The estimated total nitrogen acquisition by C. prolifera was 689 μmol · m⁻² · h⁻¹, whereas the total nitrogen requirement for growth was 672 μmol · m⁻² · h⁻¹. The uptake of ammonium and amino acids by belowground parts accounted for the larger fraction of the total nitrogen acquisition of C. prolifera and is sufficient to satisfy the species nitrogen requirements for growth. This may be one reason explaining the fast spreading of the seaweed in the bare sediments of Ria Formosa where it does not have any macrophyte competitors and the concentration of nutrients is high.     

Effects of nitrogen supplementation of the culture medium on the growth, total lipid content and fatty acid profiles of three microalgae (Tetraselmis subcordiformis, Nannochloropsis oculata and Pavlova viridis)

To improve the properties of microalgae as sources for biodiesel production, three microalgae (Tetraselmis subcordiformis SHOU-S05, Nannochloropsis oculata SHOU-S14 and Pavlova viridis SHOU-S16) were cultured in media supplemented with different amounts of nitrogen (0, 0.22, 0.44, 0.88 and 1.76 mmol N·L^?1). The growth, total lipid contents, lipid classes and fatty acid profiles of the three microalgae were assayed after 10 days of cultivation. The results indicated that the specific growth rates of T. subcordiformis, N. oculata and P. viridis were lowest (0.014, 0.033 and 0.018, respectively) in the treatments without nitrogen supplementation and increased significantly with increasing nitrogen supplementation. The microalgae treated with 0.22 mmol N·L^?1 had the highest total lipid contents, which were 29.77, 35.85 and 32.10 % in T. subcordiformis, N. oculata and P. viridis, respectively. The total lipid contents as well as the proportions of neutral lipid in the three microalgae decreased significantly with increasing nitrogen supplementation between 0.22 and 1.76 mmol N·L^?1. The fatty acid profiles of the three microalgae were significantly different. The obvious characteristic of the fatty acid profile of T. subcordiformis was a high amount (17.68–22.22 %) of 18:3n3. However, EPA (C20:5n3) and C16 fatty acids were significantly high in N. oculata and P. viridis, respectively. N. oculata and P. viridis accumulated more 16-carbon fatty acids and fewer polyunsaturated fatty acids in nitrogen-limited media. It is therefore suggested that a limited nitrogen treatment is effective for improving the properties of P. viridis and N. oculata as sources for biodiesel.