Macronutrient regulation in the Rasberry crazy ant (<Emphasis Type="Italic">Nylanderia</Emphasis> sp. nr. <Emphasis Type="Italic">pubens</Emphasis>)

Animals grow and optimize performance when they collect foods in amounts and ratios that best meet their species-specific nutritional requirements. For eusocial organisms like ants, where only a small fraction of the colony members collect food, increasing evidence demonstrates that strong macronutrient regulation occurs at the colony level. Here, we explored regulation of protein and carbohydrate (p:c) intake in the Rasberry crazy ant, Nylanderia sp. nr. puben. We did this using dry artificial foods (14–42% total macronutrient content) having a range of fixed p:c ratios in a series of choice and no-choice laboratory experiments and used worker mortality to gauge colony-level costs associated with active nutrient regulation. Choice experiments revealed that colonies preferred carbohydrate-rich foods and self-selected a diet having a p:c ratio ~1:2. No-choice experiments demonstrated that food p:c ratio only moderately affected worker food collection behavior, likely because colonies regulated the intake of only the non-limiting nutrients. Absolute worker mortality was generally high, but lowest in colonies feeding on the food having a p:c ratio of 1:2 (the p:c ratio ants self-selected in the choice experiment), although mortality was not significantly affected by food p:c ratio. The self-selected p:c ratio in our study is consistent with that observed in other recent ant nutrient regulation studies. Taken together, the results from these combined studies reveal emerging commonalities among ants in macronutrient regulation strategies, and similarities in foraging behaviors and costs associated with macronutrient regulation. Finally, from a methodological perspective, the high mortality observed in our study, when compared with other recent studies, suggests that ant nutrient regulation studies should be conducted using foods having high moisture and total macronutrient content.     

The Nutritional Balancing Act of a Large Herbivore: An Experiment with Captive Moose (Alces alces L)

The nutrient balancing hypothesis proposes that, when sufficient food is available, the primary goal of animal diet selection is to obtain a nutritionally balanced diet. This hypothesis can be tested using the Geometric Framework for nutrition (GF). The GF enables researchers to study patterns of nutrient intake (e.g. macronutrients; protein, carbohydrates, fat), interactions between the different nutrients, and how an animal resolves the potential conflict between over-eating one or more nutrients and under-eating others during periods of dietary imbalance. Using the moose (Alces alces L.), a model species in the development of herbivore foraging theory, we conducted a feeding experiment guided by the GF, combining continuous observations of six captive moose with analysis of the macronutritional composition of foods. We identified the moose’s self-selected macronutrient target by allowing them to compose a diet by mixing two nutritionally complementary pellet types plus limited access to Salix browse. Such periods of free choice were intermixed with periods when they were restricted to one of the two pellet types plus Salix browse. Our observations of food intake by moose given free choice lend support to the nutrient balancing hypothesis, as the moose combined the foods in specific proportions that provided a particular ratio and amount of macronutrients. When restricted to either of two diets comprising a single pellet type, the moose i) maintained a relatively stable intake of non-protein energy while allowing protein intakes to vary with food composition, and ii) increased their intake of the food item that most closely resembled the self-selected macronutrient intake from the free choice periods, namely Salix browse. We place our results in the context of the nutritional strategy of the moose, ruminant physiology and the categorization of food quality.     

Nutrient self-selection by the omnivorous cockroach Supella longipalpa

Larvae of the brown-banded cockroach, Supella longipalpa, grown from the beginning of the third instar to the adult moult, self-selected a 15.5:85.5 casein:glucose ratio when given a choice between two diet cubes that were nutritionally complete except that one lacked protein (casein) and the other lacked carbohydrate (glucose). The self-selectors were superior in all utilization parameters to control larvae placed in similar arenas and fed on two nutritionally complete diet cubes, each containing equal amounts of casein and glucose. However, when forced to feed throughout their entire larval life on a diet containing a 20:80 casein:glucose ratio (approximating the self-selected ratio) incorporated into a single cube, larvae grew very poorly compared to others offered a 50:50 cube or an opportunity to self-select. Measurements of casein and glucose consumption throughout the entire first- and last-larval stadia showed that in each case carbohydrate intake was high at first, but decreased through the stadium, finally equalling the level of protein intake, which remained low and constant throughout the stadium. Thus, a 20:80 diet may be nutritionally unsuitable because it does not match closely enough the varying needs of the insect throughout the stadium.     

Diet Balancing in the Cockroach Rhyparobia madera: Does Serotonin Regulate this Behavior?

Animals, including insects, have the ability to self-select an optimal diet from a choice of two or more incomplete diets that lack an essential nutrient. This paper demonstrates that nymphs of the cockroach Rhyparobia madera also have this ability. The nymphs chose a protein:carbohydrate (P:C) ratio of approximately 25:75 when faced with a choice between one cube of protein (casein) and another of carbohydrate (sucrose). This self-selected ratio was shown to promote growth as well or better than other diets tested. When given a wide range of P:C choices, the R. madera nymphs consistently selected a P:C ratio of approximately 25:75, suggesting that they have the ability to diet-balance. Finally, injections of various serotonergic drugs into self-selecting nymphs influenced their choice of diets. Serotonin promoted a decrease in carbohydrate feeding, while injection of the antagonist -methyltryptophan caused the nymphs to overfeed on carbohydrate. The results suggest that serotonin may help alter the carbohydrate feeding response in cockroaches.     

Dietary ratio of protein to carbohydrate induces plastic responses in the gastrointestinal tract of mice

Some vertebrates change the size of their digestive system in response to quantity and fibre content of ingested food, but the effects of dietary nutrients on gut structure remain poorly understood. Here we investigate how the protein to carbohydrate ratio of diets affects the mass of the gastrointestinal tract in mice. We fed 6-week-old male mice one of five isocaloric diets differing only in protein to carbohydrate ratio (the “no-choice” treatments), while a further four treatment groups received nutritionally complementary food pairings from which they could self-select a diet (the “choice” treatments). After 32 days, we measured the resulting dry mass of stomachs, intestines, caeca and colons. In the no-choice treatments, the stomachs were heavier in the mice fed diets containing more protein and less carbohydrate, indicating that larger stomachs may be needed for efficient digestion of the protein-rich food. In contrast, intestines, caeca and colons were heavier when diets contained more carbohydrates and less protein. This response may function to increase the digestive rate of carbohydrates when the dietary content of this macronutrient increases, but it may also indicate a compensatory response to increase amino acid uptake from a protein-deficient food. Mice in the choice treatments self-selected a diet with a protein to carbohydrate ratio of 0.46, and had gut dimensions similar to the expectation derived from no-choice treatments for this diet composition. Our results provide an example of plasticity in the differential allocation of resources to organ function, which is triggered by variation in resource quality.     

Dietary selection and nutritional regulation in a common mixed‐feeding insect herbivore

The geometric framework provides a way for understanding the multi‐dimensional nutritional relationships between consumers and their food. We use this approach to further our understanding of the feeding and nutritional ecology of a ubiquitous mixed‐feeding insect herbivore that consumes a variety of host plants spanning a wide range of nutritional composition. Our overall objective was to examine feeding decisions, resulting performance, and post‐ingestive consequences in a common mixed‐feeding insect herbivore, Melanoplus bivittatus (Say) (Orthoptera: Acrididae), when presented with paired diets differing in protein:carbohydrate (p:c) ratio. Intake p:c of M. bivittatus differed among all but two treatments and in many cases was farther than expected from the previously identified p:c intake target for this species. Despite this variability in intake of protein and carbohydrate, we found few effects of the diet treatments on performance or post‐ingestive processing. However, our results suggest that when feeding on high‐quality diets, nutrients consumed in excess may be stored rather than excreted.     

Sex-specific differences in nutrient regulation in a capital breeding caterpillar, Spodoptera litura (Fabricius)

Nutrient requirements by male and female insects are likely to differ, but relatively little is known regarding how sexes differ in their regulation of macronutrient acquisition. The present study reports the results from a laboratory experiment in which behavioural and physiological components of nutrient regulation were compared between male and female caterpillars of Spodoptera litura (Fabricius). When provided with choices between two nutritionally complementary foods (one is a protein-biased diet and the other a carbohydrate-biased diet), both males and females adjusted their food selection to defend an intake target. However, the composition of diet preferred by the two differed, with females selecting significantly more protein than males with no difference in carbohydrate intake between the two. When confined to single diets with varying mixtures of protein and carbohydrate [P:C ratios, expressed as the percentage of diet by dry mass: protein 42%:carbohydrate 0% (p42:c0), p35:c7, p28:c14, p21:c21, p14:c28, p7:c35], females consumed more macronutrients than did males across on all P:C diets except the extremely carbohydrate-biased diet (p7:c35). Under both choice and no-choice feeding condition, such sex differences in nutrient intake were not expressed until late in the feeding stage of the final stadium. Sexes also differed in post-ingestive utilization of ingested nutrients. Females utilized ingested protein for body growth with greater efficiency compared to males, presumably reflecting provisioning their adult needs for protein to develop eggs, whereas males were more efficient at depositing lipids from carbohydrate intake than females.     

Plant carbohydrate content limits performance and lipid accumulation of an outbreaking herbivore

Locusts are major intermittent threats to food security and the ecological factors determining where and when these occur remain poorly understood. For many herbivores, obtaining adequate protein from plants is a key challenge. We tested how the dietary protein : non-structural carbohydrate ratio (p : c) affects the developmental and physiological performance of 4th-5th instar nymphs of the South American locust, Schistocerca cancellata, which has recently resurged in Argentina, Bolivia and Paraguay. Field marching locusts preferred to feed on high carbohydrate foods. Field-collected juveniles transferred to the laboratory selected artificial diets or local plants with low p : c. On single artificial diets, survival rate increased as foods became more carbohydrate-biased. On single local plants, growth only occurred on the plant with the lowest p : c. Most local plants had p : c ratios substantially higher than optimal, demonstrating that field marching locusts must search for adequate carbohydrate or their survival and growth will be carbohydrate-limited. Total body lipids increased as dietary p : c decreased on both artificial and plant diets, and the low lipid contents of field-collected nymphs suggest that obtaining adequate carbohydrate may pose a strong limitation on migration for S. cancellata. Anthropogenic influences such as conversions of forests to pastures, may increase carbohydrate availability and promote outbreaks and migration of some locusts.     

Dietary self‐selection and rules of compromise by fifth‐instar Vanessa cardui

We examined dietary self‐selection and rules of compromise for protein (P) and digestible carbohydrate (C) intake by fifth‐instar Vanessa cardui L. (Lepidoptera: Nymphalidae: Nymphalini). We presented six fat‐free diet pairs to larvae in a choice trial to determine the ‘intake target’. In addition, we fed larvae seven fat‐free single diets differing in dietary nutrient ratio in no‐choice trials to determine the rules of compromise they exhibit when constrained to a singular, sub‐optimal dietary source. In choice trials, caterpillars regulated nutrient intake to a ratio of 1 protein to 1.09 carbohydrate (1P:1.09C), exhibiting tighter regulation of protein than of carbohydrate. Furthermore, larvae from different diet pair treatments did not differ in pupal mass or stadium duration. In no‐choice experiments, larvae reduced consumption on increasingly protein‐biased diets and increased consumption on increasingly carbohydrate‐biased diets, relative to a 1P:1C ratio diet. Differences in carbohydrate consumption were much greater between no‐choice treatments than differences in protein consumption. Dietary nutrient ratio affected pupal mass when accounting for initial larval mass. Pupal mass decreased as nutrient ratio was shifted off of 1P:1C, but to a greater extent when the ratio was skewed toward carbohydrate. Stadium duration increased as nutrient ratio diverged from 1P:1C, being more pronounced when shifted toward carbohydrate than toward protein. Regulation to near 1P:1C is consistent with results found for other Lepidoptera, and the rule of compromise exhibited by V. cardui is consistent with that expected for a generalist herbivore.     

Dietary self-selection by Heliothis zea larvae: roles of metabolic feedback and chemosensory stimuli

The effects on dietary self-selection of phagostimulation and metabolic feedback from nutrients were seperated by using two unusual carbohydrates alone or in combination: mannitol, which is nutritive but not phagostimulatory for H. zea larvae; and sorbose, which is weakly phagostimulatory but not nutritive. Last instar H. zea larvae recognized and responded positively to mannitol by metabolic feedback when it was included in a diet with casein. When offered a choice of two diets, one containing casein but lacking a utilizable carbohydrate and another lacking casein but including either mannitol or sorbose, they did not self-select for either mannitol or sorbose. However, in a similar experiment, in which the carbohydrate diet contained a mixture of mannitol and sorbose, there was a positive response, demonstrating that both metabolic feedback and phagostimulation are required to elicit self-selection.

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Résumé Les chenilles d'H. zea peuvent choisir un mélange nutritif optimal parmi deux régimes séparés, répondant à des besoins alimentaires différents. Cette aptitude à l'autosélection pose la question des voies de sélection d'un régime optimal par l'insecte. L'autosélection est probablement réalisée par un feedback métabolique des aliments; la stimulation sensorielle par les aliments pourrait jouer elle aussi un rôle. Notre objectif est de séparer ces deux facteurs et d'en déterminer l'importance relative.Des L 5 venant de muer ont pu s'alimenter sur 2 régimes nutritionnellement complets, mais dont l'un contenait des carbohydrates (sucrose) mais non des protéines, tandis que l'autre contenait des protéines (caséine) mais non des carbohydrates. L'auto-sélection caséine/sucrose a été 81/19; elle correspond à la portion idéale pour le dernier stade des chenilles H. zea établie lors d'expériences antérieures (Waldbauer et al., 1984a). Pour comparer l'importance relative du feedback métabolique et de la stimulation sensorielle, 2 carbohydrates ont été substitués au sucrose, qui est à la fois nutritif et phagostimulant: le mannitol, nutritif mais ne stimulant pas de prise de nourriture, et le sorbose, faiblement stimulant mais non nutritif. Quand elles ont le choix, les chenilles ne sélectionnent ni mannitol, ni sorbose, mais consomment presque exclusivement le régime alternatif à la caséine. Quand sorbose et mannitol sont mélangés et proposés ensemble, la réponse est positive, montrant que le feedback métabolique et la stimulation sensorielle sont impliqués tous deux dans l'autosélection alimentaire.Des expériences en deux étapes d'autosélection ont montré que le feedback métabolique est de première importance. Dans la première, les chenilles avaient le choix entre caséine seule et caséine et mannitol. Les chenilles ont consommé significativement plus d'aliment avec mannitol que sans carbohydrate, bien que les goûts des deux régimes aient du être identiques, puisque le mannitol n'a pas de goût pour H. zea. La caséine a été vraisemblablement assez phagostimulante pour maintenir suffisamment les chenilles sur l'aliment avec mannitol et ainsi permettre au feedback métabolique du mannitol d'influer sur leur maintien sur cet aliment sans en rechercher un autre.Dans d'autres expériences, les chenilles ont eu le choix entre un régime alimentaire classique avec de la caséinne et d'autres n'ayant plus leur caséine mais avec des quantités décroissantes de sucrose. Elles n'ont pas réagi en consommant plus d'aliments avec du sucrose dilué, mais en consommant plus de caséine. Cependant, si du mannitol, pourtant non phagostimulant, est ajouté à des aliments avec du sucrose dilué pour remplacer le sucrose manquant, les chenilles ne compensent plus en consommant plus d'aliments avec de la caséine, mais de la même façon qu'elles le feraient avec un aliment contenant la concentration adéquate de sucrose, montrant qu'elles perçoivent ainsi le feedback métabolique du mannitol, qui est pourtant sans goût.

     

Characterization of the complete mitochondrial genome of Bombyx mori strain H9 (Lepidoptera: Bombycidae)

The complete mitochondrial genome (mitogenome) of Bombyx mori strain H9 (Lepidoptera: Bombycidae) is 15,670 base pairs (bp) in length, encoding 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and a control region. The nucleotide composition of the genome is highly A + T biased, accounting for 81.31%, with a slightly positive AT skewness (0.059). The arrangement of 13 PCGs is similar to that of other sequenced lepidopterans. All the PCGs are initiated by ATN codons, except for the cytochrome c oxidase subunit 1 (cox1) gene, which is proposed by the TTAG sequence as observed in other lepidopterans. Unlike the other PCGs, the cox1 and cytochrome c oxidase subunit 2 (cox2) genes have incomplete stop codons consisting of just a T. All tRNAs have typical structures of insect mitochondrial tRNAs, which is different from other sequenced lepidopterans. The structure of A + T-rich region is similar to that of other sequenced lepidopterans, including non-repetitive sequences, the ATAGA binding domain, a 18 bp poly-T stretch and a poly-A element upstream of transfer RNA M (trnM) gene. Phylogenetic analysis shows that the domesticated silkmoth B. mori originated from the Chinese Bombyx mandarina.     

A host-plant specialist, Helicoverpa assulta, is more tolerant to capsaicin from Capsicum annuum than other noctuid species

Plant secondary compounds not only play an important role in plant defense, but have been a driving force for host adaptation by herbivores. Capsaicin (8-methyl-N-vanillyl-6-nonenamide), an alkaloid found in the fruit of Capsicum spp. (Solanaceae), is responsible for the pungency of hot pepper fruits and is unique to the genus. The oriental tobacco budworm, Helicoverpa assulta (Lepidoptera: Noctuidae), is a specialist herbivore feeding on solanaceous plants including Capsicum annuum, and is one of a very few insect herbivores worldwide capable of feeding on hot pepper fruits. To determine whether this is due in part to an increased physiological tolerance of capsaicin, we compared H. assulta with another specialist on Solanaceae, Heliothis subflexa, and four generalist species, Spodoptera frugiperda, Heliothis virescens, Helicoverpa armigera, and Helicoverpa zea, all belonging to the family Noctuidae. When larvae were fed capsaicin-spiked artificial diet for the entire larval period, larval mortality increased in H. subflexa and H. zea but decreased in H. assulta. Larval growth decreased on the capsaicin-spiked diet in four of the species, was unaffected in H. armigera and increased in H. assulta. Food consumption and utilization experiments showed that capsaicin decreased relative consumption rate (RCR), relative growth rate (RGR) and approximate digestibility (AD) in H. zea, and increased AD and the efficiency of conversion of ingested food (ECI) in H. armigera; whereas it did not significantly change any of these nutritional indices in H. assulta. The acute toxicity of capsaicin measured by injection into early fifth instar larvae was less in H. assulta than in H. armigera and H. zea. Injection of high concentrations produced abdominal paralysis and self-cannibalism. Injection of sub-lethal doses of capsaicin resulted in reduced pupal weights in H. armigera and H. zea, but not in H. assulta. The results indicate that H. assulta is more tolerant to capsaicin than the other insects tested, suggesting that this has facilitated expansion of its host range within Solanaceae to Capsicum after introduction of the latter to the Old World about 500 years ago. The increased larval survival and growth due to chronic dietary exposure to capsaicin suggests further adaptation of H. assulta to that compound, the mechanisms of which remain to be investigated.     

The effect of dietary protein on the growth and digestive physiology of larval Heliothis zea and Spodoptera exigua

We compared the ability of larval H. zea (Boddie) and S. exigua (Hubner) to digest and utilize dietary protein by: (a) determining the ability of different concentrations of dietary protein to support larval growth, and (b) determining the effect of different concentrations of dietary protein on the digestive physiology of the organisms, as measured by in vivo digestion of protein and proteolytic activity. Using an artificial diet containing casein as the primary source of protein, we found that H. zea was able to grow at very low levels of casein (≤0.6%), while optimal growth occurred at 1.2% casein. For S. exigua, dietary casein levels of >0.6% were required for growth, and optimal growth occurred at ≥1.2% casein. However, optimal growth in both species was not correlated with the degree of in vivo digestion of protein. The level of in vivo digestion of protein and tryptic activity in S. exigua was proportional to the concentration of dietary protein (under both acute and chronic exposure), and not the amount of food in the gut, suggesting that enzyme synthesis and/or secretion is controlled by a secretagogue mechanism. H. zea only demonstrated a secretagogue mechanism of control of tryptic activity while under acute exposure to different concentrations of casein; under chronic exposure, tryptic activity was uniform regardless of the concentration of dietary casein. When comparing the two species of noctuid, H. zea, which is the larger of the two species, produced less tryptic activity on a unit weight basis, and also digested less of the available dietary protein than S. exigua. Hence, these closely related organisms are processing dietary protein at different efficiencies.     

Serial passage of Heliothis zea singly embedded nuclear polyhedrosis virus in a homologous cell line

This paper describes the replication and serial passage of Heliothis zea nuclear polyhedrosis virus (NPV) in a H. zea cell line. It was demonstrated that long-term serial passages of the H. zea NPV in homologous host cell culture decreased both the total number of polyhedral inclusion bodies (PIBs) produced and the infectivity of the supernatant as measured by TCID₅₀. The growth curve indicated that infectious material was released from cells 24 hr postinfection (p.i.) and approached a maximal titer 3 days p.i. The kinetics of H. zea NPV decay at 4°, 27°, and 37°C were determined. Infectivity was not detected after 3 weeks at 37°C, but approximately 10^3.5 TCID₅₀/ml activity was still present after 3 and 8 weeks storage at 27° and 4°C, respectively. Electron microscopy confirmed the presence of single embedded virions in the inoculated cells.     

Production and Characterization of Bacillus thuringiensis Cry1Ac-Resistant Cotton Bollworm Helicoverpa zea (Boddie)

Laboratory-selected Bacillus thuringiensis-resistant colonies are important tools for elucidating B. thuringiensis resistance mechanisms. However, cotton bollworm, Helicoverpa zea, a target pest of transgenic corn and cotton expressing B. thuringiensis Cry1Ac (Bt corn and cotton), has proven difficult to select for stable resistance. Two populations of H. zea (AR and MR), resistant to the B. thuringiensis protein found in all commercial Bt cotton varieties (Cry1Ac), were established by selection with Cry1Ac activated toxin (AR) or MVP II (MR). Cry1Ac toxin reflects the form ingested by H. zea when feeding on Bt cotton, whereas MVP II is a Cry1Ac formulation used for resistance selection and monitoring. The resistance ratio (RR) for AR exceeded 100-fold after 11 generations and has been maintained at this level for nine generations. This is the first report of stable Cry1Ac resistance in H. zea. MR crashed after 11 generations, reaching only an RR of 12. AR was only partially cross-resistant to MVP II, suggesting that MVP II does not have the same Cry1Ac selection pressure as Cry1Ac toxin against H. zea and that proteases may be involved with resistance. AR was highly cross-resistant to Cry1Ab toxin but only slightly cross-resistant to Cry1Ab expressing corn leaf powder. AR was not cross-resistant to Cry2Aa2, Cry2Ab2-expressing corn leaf powder, Vip3A, and cypermethrin. Toxin-binding assays showed no significant differences, indicating that resistance was not linked to a reduction in binding. These results aid in understanding why this pest has not evolved B. thuringiensis resistance, and highlight the need to choose carefully the form of B. thuringiensis protein used in experiments.     

A geometric analysis of nutrient regulation in the generalist caterpillar Spodoptera littoralis (Boisduval)

Nutritional regulatory responses of the generalist caterpillar Spodoptera littoralis (Boisduval) were explored, in choice and no-choice experiments, using the Geometric Framework. In the choice experiment, newly moulted final instar larvae were provided with one of three protein-biased foods (PB-food: p35:c7, p28:c5.6 or p21:c4.2) and one of three equal protein-carbohydrate ratio foods (ER-food: p21:c21, p16.8:c16.8 or p12.6:c12.6). On five of the nine treatments, caterpillars independently regulated protein and carbohydrate intake to a mixture of 57 and 43%, respectively. However, when the concentration of the ER-food decreased and that of the paired PB-food increased, caterpillars progressively abandoned regulation and ate more of the PB-food. Despite these regulatory differences, performance (survivorship, growth and development) was similar across all nine treatments. In the no-choice experiment, caterpillars were given one of five foods (p35:c7, p28:c14, p21:c21, p14:c28 and p7:c35). Results indicated that caterpillars moved to a point in protein-carbohydrate space that was consistent with the Equal Distance Rule of compromise, a pattern previously predicted for generalist feeders. The insects on the two extreme foods, p35:c7 and p7:c35, showed reduced pupal mass and longer development, respectively. There was also strong evidence for post-ingestive regulation of nutrient utilisation, notably for protein.     

The biochemical composition of phytoplankton in an upwelling region off south Africa

Variations in the concentrations of chlorophyll a, ATP, protein, and carbohydrates in phytoplankton have been investigated in a nearshore upwelling region off the Cape Peninsula. During active upwelling temperatures <10 °C, high nutrient concentrations and low concentrations of the biochemical constituents were measured. When upwelling lessened and conditions stabilized temperatures increased and blooms of phytoplankton appeared. High concentrations of chlorophyll a and ATP and a high protein/carbohydrate ratio were then recorded. At very low nutrient levels chlorophyll a and ATP concentrations were still high but an increase in the acid-soluble carbohydrate content and a corresponding decrease in the protein/carbohydrate ratio was observe. It was concluded that the ratio of protein to carbohydrate was a suitable indicator of the physiological state of a phytoplankton community in the local upwelling region.     

Responses of representative midgut detoxifying enzymes from Heliothis zea and Spodoptera frugiperda to trichothecenes

The relative levels of representative midgut detoxifying enzymes were measured in last instar larvae of Heliothis zea and Spodoptera frugiperda after the insects had been fed for 48 h on diets containing representative trichothecenes, which are protein synthesis inhibitors. 4-Nitroanisole O-demethylation was induced (as indicated by increased enzyme activity) by 1.6-fold in H. zea and by 6.1-fold in S. frugiperda when fed 250 ppm of deoxynivalenol, a dose that severely retarded growth of the insects. Induction of this activity was also noted with lower levels of deoxynivalenol in both species, and with diacetoxyscirpenol and T-2 toxin in H. zea. 1-Chloro-2,4-dinitrobenzene glutathione transferase conjugation was induced by c. 1.3-fold with deoxynivalenol at 250 ppm and 25 ppm, and with T-2 toxin at 25 ppm in H. zea; but was slightly inhibited (c. 10–20%) in S. frugiperda in some cases.1-Naphthyl acetate hydrolysis was generally unaffected by all trichothecenes tested at all doses in H. zea, but was inhibited by c. 30% in S. frugiperda by lower doses of all three trichothecenes. However, a new 1-naphthyl acetate esterase band was noted after exposure to 250 ppm of deoxynivalenol, and to some extent lower doses of deoxynivalenol and the other trichothecenes in both insect species. The hydrolysis of a radiolabeled model trichothecene, 4-monoacetoxyscripenol, was induced by 3.1-fold in H. zea and 2.7-fold in S. frugiperda with 250 ppm of deoxynivalenol (the only compound and dose tested). This activity could be inhibited by nearly 100% with 10⁻⁴ M paraoxon, suggesting a serine-hydroxyl esterase was involed. Although this activity could not be recovered from gels, indirect competitive inhibition assays with 1-naphthyl acetate suggested the induced band could be responsible for some of the hydrolysis of the 4-monoacetoxyscirpenol.