Multifarious activities of gut epithelium in an appendicularian (Oikopleura dioica: Tunicata)

The functions of the various gut regions of Oikopleura dioica (oesophagus, left and right gastric lobes, vertical intestine, mid-intestine and rectum) were investigated by means of histochemical, histoenzymatic and immunohistochemical techniques at light and electron microscopes. Ciliary food progression is evidenced by the presence of ATPases on cilia, along the entire gut, with the exception of the cardiac valve, a passive device controlling food direction. Absorptive processes (alkaline phosphatase), active transport (ATPases) and nitrogen excretion (D-amino acid oxidase) occur along the entire gut, in both ciliated microvillar and globular cells. The latter, typical of the left gastric lobe and rectum, are also involved in endocytotic processes (exogenous peroxidase as tracer) and intracellular digestion (!-amylase, aminopeptidase M, acid phosphatase, 5'-nucleotidase, non-specific esterase). The giant cells of the gastric band participate in extracellular digestion; they contain secretory granules positive to various hydrolytic enzymes, the activity of which is also recognisable in faecal pellets inside the intestinal lumen. Lipid storage occurs mainly in the right gastric lobe and vertical intestine, whereas protein storage takes place in the rectal granular cells. Epithelial transport and possible osmoregulation occur along the entire gut, especially at the level of diffuse baso-lateral interdigitations, which increase the plasmalemma surface enormously, are often associated with mitochondria and possess numerous ATPase pumps. Data extend previous histological observations and hypotheses on the physiological role of the various gut regions. The remarkable and specific location of enzymatic activities and nutrient storage are in agreement with the high capacity of O. dioica to process a great quantity of food very rapidly and efficiently.     

House formation and feeding behaviour of<Emphasis Type="Italic"> Fritillaria borealis</Emphasis> (Appendicularia: Tunicata)

Numerous marine animals use elaborate filtration mechanisms to feed on particles suspended in the sea. Video-microscopy reveals that the common appendicularian Fritillaria borealis do so in a unique way. They live inside acellular, mucous houses, where their tail undulations act like a peristaltic pump in a close-fitting tail chamber to force water through a complex set of filters. Suspended particles, between ~0.2 and 30 µm in diameter, are retained in this process and propagated towards the mouth of the organism to be ingested. Coarser particles are back-washed out of the house during brief intervals of tail arrest, when the elastic recoil of the house walls reduces its volume to one-eighth of its inflated state. At the same time, the bifurcated tip of the animal's tail slides into a sleeve-like extension of the house. This sleeve acts like a flipper to propel the animal and collapsed house several millimetres to a few centimetres through the sea as soon as muscular activity of the tail is resumed. After a second tail arrest, a special muscular twist brings the tail out of the sleeve, whereupon undulatory pumping movements are resumed to reinflate and process particle-laden water through the house. The complete behavioural cycle lasts ~30 s. The repetitive inflation and deflation cycles of the house are probably of importance for unclogging of the coarse "inlet" filters downstream to the tail pump, as well as for agglutination of the smallest food particles trapped in the house to ease their recapture in the endostylar pharyngeal filter during the ingestion process. The unique tail sleeve and flipper at the same time enable the animal to move away from the previously filtered water and rejected coarse particles, before the house is reinflated for resumed food filtration.     

Measurement of in situ clearance rates of Oikopleura vanhoeffeni (Appendicularia: Tunicata) from tail beat frequency, time spent feeding and individual body size

In situ observations (1994 to 1996) of the behavior of Oikopleura vanhoeffeni Lohmann in combination with a previously published model can be used to give a good approximation of clearance rates. The model is based on tail beat frequency, time spent feeding and subsequent measurements of individual size (trunk length). These estimated clearance rates are in close agreement with clearance rates determined in the laboratory under static and flow-through conditions. A comparison of various techniques (such as measurement of gut pigment, and various particle removal and particle uptake methods) with the behavior-model approach, revealed a convergence of all rates within a threefold range. Most of this discrepancy can be explained by the number of non-feeding individuals and the deterioration of the feeding filters in the field. The main reasons for the similarity of the various clearance rate estimates are the low variability of the behavior of appendicularians in response to environmental variables, such as temperature and particle concentration, and their non-selective retention of particles. The suggested mechanistic approach has great value for estimating the flux of material and energy through populations of appendicularians for which only size and abundance data exist, or for appendicularian species that cannot be assessed empirically. Received: 26 August 1997 / Accepted: 28 September 1998     

Elemental and biochemical composition of salps (Tunicata: Thaliacea)

Carbon and nitrogen content have been measured in the solitary and aggregate generations of 11 species of salps. Regression equations for each species and generation permit estimation of carbon or nitrogen content as a function of length of live individuals. Different species of the same length may have nearly tenfold differences in carbon content. Fractionation and biochemical analysis of some samples revealed that the organic content of salps is approximately 80% protein. Ash-free dry weights average 27% of dry weights; mean carbon content is 29% of ash-free dry weight. Excess ash-free dry weight not accountable as organic material is thought to be water of hydration.     

Field behavior and adaptive strategies of appendicularians (Chordata: Tunicata)

The behavior of 7 species of appendicularians from the family Oikopleuridae was observed using SCUBA in the Gulf of California and the Florida Current. The frequency and orientation of feeding and the pattern of swimming while within the house varied considerably among species. Appendicularians expanded new houses in 1 1/2 to 5 min. House-expansion behavior was complex and variable among species. Appendicularians rarely abandoned the house in response to predation. Predators included the sergeant major (Abudefduf sp.), medusae, chaetognaths and ctenophores. The abandonment of the house was based on a cost-benefit behavior strategy which maximized time spent inside the house while minimizing predation and energy investment in house secretion. Strategies of predator avoidance are discussed.     

Field observations on the feeding behavior of salps (Tunicata: Thaliacea)

Observations of living salps by SCUBA diving in the Florida Current and Gulf of California (Mexico) indicate that they filter feed with a continuously renewed mucus net. Ninety-one percent of 1420 salps (6 species) examined were feeding in this manner. Particles collected range from about 1 mm to less than 1 μm. Compact fecal pellets appear to contain much undigested material and may carry significant amounts of carbon out of surface waters.     

Spermatogenesis inPyrosoma atlanticum (Tunicata: Thaliacea: Pyrosomatida): Implications for tunicate phylogeny

Colonies ofPyrosoma atlanticum were collected by submersible in October 1988 in the Caribbean Sea, and testes were studied by electron microscopy. Spermatogonia, spermatocytes and early spermatids have two centrioles. The proximal centriole subsequently disappears, its remains apparently persisting in the spermatozoon as dense material adjacent to the distal centriole, which gives rise to the axoneme. At the tip of early spermatids are several 50 nm proacrosomal vesicles, which disappear leaving no trace in early elongating spermatids. The spermatozoon lacks an acrosome and has a head 35µm long. The head is differentiated into a bulbous posterior portion 5µm long × 1µm wide, a thinner anterior portion 25µm long tapering from a width of 0.7µm to a width of 0.4µm, and a very thin anterior extension 5µm long × 0.5µm wide. At the start of elongation, the anterior extension begins to form just lateral to the proacrosomal vesicles as a spiral projection comprising part of the nucleus, covered by a thin sheath of cytoplasm. This sheath of cytoplasm undergoes a complex differentiation. Ultimately, the nucleus in the anterior extension is overlain by two membrane-bound sheaths of cytoplasm connected by a spiral flange of cytoplasm. Between these two sheaths is a spiral space, open to the exterior through a subterminal pore near the sperm tip. In early spermatids the mitochondria fuse into a single mitochondrion, which remains lateral to the nucleus. The cristae become modified late in spermatogenesis. Throughout elongation of the spermatid there are patches of dense material between the nucleus and mitochondrion. A manchette of microtubules transiently encircles the thin anterior portion of the nucleus during the last phase of elongation. A manchette is not present during most of elongation. In the spermatozoon the mitochondrion, which has reticulate cristae, spirals a few times about the nucleus and extends from the junction between the bulbous portion and the thinner anterior portion of the nucleus to the junction between the thinner anterior portion and the nuclear extension. Spermatogenesis inP. atlanticum, compared to that in other tunicates, most closely resembles that in colonial ascidians, and supports the majority view that pyrosomes arose from aplousobranch ascidians that lost their attachment to the substratum. Pyrosome sperm are more highly derived than doliolid sperm, which have an acrosome that is probably capable of exocytosis. When salp and pyrosome sperm are compared, both are highly derived, but neither shares any apomorphies with the other that it does not share with at least one other tunicate order. Thus, sperm morphology does not support the majority view that pyrosomes gave rise to doliolids and neither confirms nor denies the idea that pyrosomes are intermediate between aplousobranch ascidians and salps. Therefore, it is likely that the class Thaliacea is polyphyletic, with doliolids arising very early from the ascidian lineage and with salps and pyrosomes arising somewhat later.     

The presence of riboflavin and two pterins in ascidians (Tunicata) and their excretion into sea water

Individuals belonging to 6 species of ascidians were kept in experimental vessels filled with filtered sea water, for a defined period. Pterins and flavins were identified in the bodies of each species and in the ambient sea water by the method of Momzikoff (1974). The possible causes of the recorded variations in composition are discussed; these differences seem related to the ordinal position of the species. Numerous unidentified substances are excreted together with pterins (2-amino-4-hydroxypteridine and isoxanthopterin) and riboflavin into the surrounding medium. Ascidians may be the source of a significant quantity of these substances in the sea.     

Comparison of gut morphology and gut microflora of seven species of mud shrimp (Crustacea: Decapoda: Thalassinidea)

The gut morphology is described for seven species of Thalassinidea; Callianassasubterranea (Montagu), Jaxea nocturna Nardo, Axius stirhynchus Leach, Calocaris macandreae Bell, Upogebia pusilla (Petagna), U. deltaura (Leach) and U. stellata (Montagu). Differences were observed in the structure of the dorsal and lateral teeth of the gastric mill. Callianassa subterranea and A. stirhynchus had simple robust dorsal teeth and proportionally larger lateral teeth with flexible spines. The three upogebiids had complex dorsal teeth that were highly ridged and proportionally smaller lateral teeth with inflexible spines. The dorsal and lateral teeth of J. nocturna were intermediate in form. The dorsal tooth of Calocaris macandreae, although relatively simple in design, possessed accessory projections, or “wings”, on either side of the main structure. The lateral teeth were proportionally quite large, but lacked ridges and spines. The epithelia surfaces of the digestive gland, midgut and posterior diverticulum were similar for all seven species. Differences were observed in the morphology of the hindgut. The hindgut lining was completely smooth for the three upogebiid species. The other thalassinidean species had four distinct rows of cuticular projections situated on top of the ridges formed by the longitudinal convolutions of the hindgut. The arrangement of these projections varied between species. A gut flora was observed for six of the seven species, the exception being C. macandreae. The differences observed in the structure of the gut are discussed in relation to feeding and thalassinidean phylogeny. Received: 3 July 1998 / Accepted: 8 October 1998     

Fine structure of spermatids and sperm of Dolioletta gegenbauri and Doliolum nationalis (Tunicata: Thaliacea): implications for tunicate phylogeny

In Dolioletta gegenbauri and Doliolum nationalis, collected in 1987 in the bay of Villefranche-sur-mer (French Mediterranean Sea), spermiogenesis is essentially the same. Early spermatids have a round head, a flagellum arising from a single centriole with short microtubules at 45° to its base, several mitochondria, and an acrosome 50 nm thick and 250 nm long with its long axis parallel to the plasma membrane. The acrosomal contents are dense, with a central denser plate. The nuclear envelope next to the acrosome is thickened and concave. In elongating nuclei, strands of chromatin become oriented parallel to the length of the nucleus and then twist helically. Although the mitochondria surround the nucleus, they remain relatively short and do not fuse into a single mitochondrion as in sperm of other tunicates. In very late doliolid spermatids, the acrosome undergoes exocytosis, and exposes fibrous material that stays associated with the tip of the sperm; no acrosomal tubule forms. Exocytosis at this stage may be triggered by fixation. If so, exocytosis probably occurs naturally at some time before fusion of sperm and egg. Sperm have elongate heads (1 m×10 m), with the anterior two-thirds of the nucleus surrounded by mitochondria. Spermiogenesis in doliolids, compared to that in other tunicates, is most like that in solitary members of the class Ascidiacea, except that in the latter the sperm mitochondria fuse and the acrosome appears incapable of exocytosis. In contrast, previous work has shown that salps (class Thaliacea) and colonial didemnid ascidians have an acrosomeless sperm with a spiral mitochondrion, while the class Appendicularia has a sperm with a midpiece, a compact head and an acrosome capable of exocytosis and acrosomal tubule formation. By outgroup comparison with echinoderms and acraniates, appendicularian sperm are plesiomorphic within the Tunicata. Thus, gamete morphology indicates that (1) solitary ascidians and doliolids had a common ancestor, (2) the popular idea that doliolids gave rise to appendicularians is incorrect, and (3) the Thaliacea are polyphyletic, doliolids having arisen very early from the ascidian lineage and salps having arisen later.     

Estimation in situ d'une population d'Oikopleura longicauda (Appendicularia) à l'aide de deux filets de maille différente

A population of Oikopleura longicauda was sampled daily for 10 days, a period longer than their life cycle, using two nets of different mesh size (53 and 200 m). Analysis of variance revealed that the number of specimens collected by the two nets was not significantly different for individuals larger than 300 m in trunk length. A grand mean regression plotted for log of length versus log of number of specimens indicated that the number of small-sized O. longicauda (<300 m), which are not quantitatively sampled by nets, can be estimated from the collections by use of a coefficient of correction.     

Symbiontic bacteria in the gut of the blood-sucking Antarctic fish parasite Gnathia calva (Crustacea: Isopoda)

Structure and ultrastructure of the digestive tract of the ectoparasitic stages of the Antarctic isopod Gnathia calva, collected in the Weddell Sea and around the Antarctic Peninsula in the 1984–1985 season, are briefly described. There are only two digestive glands, with an ultrastructure similar to that of other isopods. The gut is divided into six regions: oesophagus, stomach, dilatable reservior for sucked-in blood, sphincter, rectal vesicle with symbiontic bacteria, and rectum. The highly dilatable anterior hindgut (reservoir) takes part in the resorption of nutrients and stores lipids and glycogen. The rectal vesicle has an epithelium with all the features of an organ with high metabolism and the capacity for the transportation of small molecules. The surface is increased by irregular microvilli. A very thin intima and a basal labyrinth are present. The presence of symbiontic bacteria is discussed in correlation with the haematophagous nutrition of G. calva.     

Quantification of copepod gut content by differential length amplification quantitative PCR (dla-qPCR)

Quantification of feeding rates and selectivity of zooplankton is vital for understanding the mechanisms structuring marine ecosystems. However, methodological limitations have made many of these studies difficult. Recently, molecular based methods have demonstrated that DNA from prey species can be used to identify zooplankton gut contents, and further, quantitative gut content estimates by quantitative PCR (qPCR) assays targeted to the 18S rRNA gene have been used to estimate feeding rates in appendicularians and copepods. However, while standard single primer based qPCR assays were quantitative for the filter feeding appendicularian Oikopleura dioica, feeding rates were consistently underestimated in the copepod Calanus finmarchicus. In this study, we test the hypothesis that prey DNA is rapidly digested after ingestion by copepods and describe a qPCR-based assay, differential length amplification qPCR (dla-qPCR), to account for DNA digestion. The assay utilizes multiple primer sets that amplify different sized fragments of the prey 18S rRNA gene and, based on the differential amplification of these fragments, the degree of digestion is estimated and corrected for. Application of this approach to C. finmarchicus fed Rhodomonas marina significantly improved quantitative feeding estimates compared to standard qPCR. The development of dla-qPCR represents a significant advancement towards a quantitative method for assessing in situ copepod feeding rates without involving cultivation-based manipulation.     

Influence of temperature changes on oxygen uptake and ammonia and phosphate excretion,in relation to body size and weight,in Oikopleura dioica (Appendicularia)

The relationship between the rates of oxygen consumption, ammonia and phosphate excretion of a pelagic tunicate, the larvacean Oikopleura dioica Fol, 1872 were assessed as a function of size, dry weight and ash-free dry weight at 15°, 20° and 24°C. O. dioica has higher respiration and excretion rates than copepods of similar weight, but the weight exponent of the allometric power function: Y=aX ^b is similar to that of other poikilotherms. Temperatures above 20°C have a depressing effect on respiration and ammonia excretion. 90% of the variance in metabolic rates is explainable by body mass and temperatures Q₁₀ values for oxygen consumption, ammonia and phosphate excretion, respectively, are 2.45, 1.86 and 1.75 between 15° and 20°C, and 3.75, 2.90 and 3.60 between 20° and 24°C. Metabolic quotients (O:N, O:P, N:P) indicate a protein-oriented diet. The results of this study suggest weak metabolic regulation in O. dioica, an energetic strategy which allows an immediate response to favourable changes in feeding conditions.     

Foraging, prey capture, and gut contents of the mesopelagic narcomedusa Solmissus spp. (Cnidaria: Hydrozoa)

Narcomedusae are the most common group of medusae in the mesopelagic depths of Monterey Bay, California. Numerous capture events of various prey taxa were recorded in situ and analyzed using the Monterey Bay Aquarium Research Institute's remotely operated vehicle "Ventana". In situ video analysis of the stomach contents of 82 Solmissus incisa and S. marshalli revealed 88 identifiable prey items. Most (88%) were gelatinous animals. Of these gelatinous prey, 60.3% were ctenophores, 20.5% were cnidarians, 12.8% were salps, 3.8% were chaetognaths, and 2.6% were polychaetes. Euphausiids accounted for 11.4% of the diet, but they were probably captured adventitiously, as the gut contents of ctenophore prey. The tentacle-first foraging behavior of the narcomedusae is an effective way to capture large, comparatively fast-moving prey, because the fluid disturbance caused by swimming is decoupled from the area of prey capture. This behavior contrasts with the prevailing models of feeding behavior in medusae. Stealth predation may be the dominant mode of capturing prey by medusae in the mesopelagic depths of the oceans.     

Bacterial diversity of the sediments transiting through the gut of Holothuria scabra (Holothuroidea; Echinodermata)

This work analyzes bacterial diversity of sediments transiting through the gut of Holothuria scabra which is an important bioturbator in tropical shallow waters. This edible holothurian species has a social and economic importance for coastal populations in many developing countries. Bacterial biodiversity was analyzed by sequencing the 16S rRNA of bacterial cultures and clones. DAPI and FISH methods were used to determine and compare the number of bacteria found in the various gut compartments. A total of 116 phylotypes belonging to the γ-Proteobacteria (60.5 %), α-Proteobacteria (24.5 %), Bacteroidetes (6 %), Actinobacteria (2.75 %), Fusobacteria (1.75 %), Firmicutes (1.75 %), Cyanobacteria (1.75 %) and δ-Proteobacteria (1 %) were identified. The number of bacteria is significantly greater (1.5×) in the foregut than in the ambient sediments. The number of bacteria significantly decreases in the midgut and remains stable until defecation. Some γ-Proteobacteria, especially Vibrio, are less affected by digestion than other bacterial taxa. The season has an impact on the bacterial diversity found in the sediments transiting through the gut: in the dry season, γ-Proteobacteria are the most abundant taxon, while α-Proteobacteria dominate in the rainy season. Vibrio is the most frequent genus with some well-known opportunistic pathogens like V. harveyi, V. alginolyticus and V. proteolyticus. Findings show that sediment-associated microbial communities are significantly modified by H. scabra during their transit through the gut which supports the view that holothurians play a substantial role in the structuring of bacterial communities at the sediment–seawater interface.     

Microbial activity, oxygen status and fermentation in the gut of the irregular sea urchin Echinocardium cordatum (Spatangoida: Echinodermata)

Microbial activity in the gut of the detrivore irregular sea urchin Echinocardium cordatum (Pennant) was studied by measurements of oxygen profiles, fermentation end products, C/N ratios, pH and concentrations of sulphide and sulphate in the gut segments, including the nodules of the intestinal caecum. The highest oxygen flux and consumption, and highest concentrations of short-chained fatty acids, and the lowest pH values occurred in the anterior stomach segments, including the anterior caecum. The C/N ratios indicate synthesis of microbial biomass in the caeca. The concentration of sulphate was high in the anterior stomach segments, whereas sulphide was only detectable in the nodules of the intestinal caecum. The anterior and intestinal caeca were the major sites for microbial activity, and oxidation of acetate and propionate produced in the two caeca corresponded to at least 9% of the total respiration of the sea urchin. Microbial fermentation, especially in the anterior caecum, seems to be important for the metabolism of E. cordatum, allowing the sea urchin to utilise refractile carbohydrates. The functions of the intestinal caecum are probably both residual fermentation and oxidation of accumulated sulphide. Received: 25 February 1998 / Accepted: 30 June 1998     

肠道微生物群和水生毒理学: 环境健康的新概念

高通量测序技术极大地方便了深度考察不同种群中宿主相关微生物群的组成和功能。研究表明微生物在动物包括人类的健康和疾病发病中扮演着必要的角色。微生物已成为环境毒理学中新兴的重要研究主题。这是因为微生物在免疫系统中起着重要的交互的作用,同时在化学解毒中也有作用。污染物扰动肠道微生物,引起胃肠组织的病理生理变化,导致一系列系统效应,致使营养摄入变差和肠道发炎。本文检索了关于环境污染物对水生物种微生物影响的文献,重点关注了肠道微生物。我们强调了脊柱动物宿主中的一些已知的肠道上皮细胞的主要蛋白,这些均是化合物破坏的靶标,这些蛋白可以与微生物直接对话。我们提出了一个有害结局路径(adverse outcome pathway)的总体框架,将肠道生态失调作为有害效应终点事件的主要贡献因子。我们展示了两个案例研究,分别是(1)纳米材料;(2)碳氢化合物,我们参考了Deepwater Horizon港口的石油泄漏事件, 生态失调在案例中展示了微生物的考察如何改善有害结局的研究。最后,我们提出了一些策略以建立化合物诱导的肠道生态失调与有害结局的关联。我们通过实验建立了特定微生物与肠道生态失调的关联。对毒物与微生物关系的深入研究将成为改善动物及人类健康的重大突破。 精选自Ondrej Adamovsky, Amanda N. Buerger, Alexis M. Wormington, Naomi Ector, Robert J. Griffitt, Joseph H. Bisesi Jr., Christopher J. Martyniuk. The gut microbiome and aquatic toxicology: An emerging concept for environmental health. Environmental Toxicology and Chemistry,2018,37:2758-2775.
详情请见 https://doi.org/10.1002/etc.4249
     

Living within constraints: irreversible chemical build-up and seasonal temperature-mediated die-off in a highly acidic (H2SO4) annual seaweed (Desmarestia viridis)

Seaweeds exhibit a number of adaptations to cope with strong selective pressures imposed by shallow marine environments. The exceptional ability of the annual, brown seaweed Desmarestia viridis, to produce and store high concentrations of sulfuric acid (H₂SO₄) in intracellular vacuoles, makes it a particularly compelling model for studies of causes and consequences of acid production in seaweeds. We used two laboratory experiments, as well as measurements of intracellular pH of sporophytes and sea salinity and temperature over an entire growth season at one site in Newfoundland and Labrador (Canada), to test hypotheses about effects of salinity and temperature on acid production and die-off in D. viridis. We show that the acid is continuously and irreversibly accumulated (resulting in an intracellular pH as low as 0.53) as the seaweed grows from recruit to adult (March–June) and that this build-up inevitably culminates into dramatic mass releases of acid and die-offs. Progressive, synchronous death among individuals is under the predominant control of sea temperature (death systematically occurred around 12 °C in both laboratory and field), which suggests the evolution of a life-history strategy in which death occurs at a time when individuals reach a size that may correspond to reproductive maturity. The seaweed exhibits a low tolerance to changes in salinity (death was inevitable below 29 psu), which likely imposes severe limitations on its distribution range throughout its existence as a sporophyte. Our findings suggest that major phenological events and survival in D. viridis are intimately linked to synergistic effects of oceanographic controls, which either impair the ability of the seaweed to retain the acid in its tissues (seasonal sea warming), or contribute to dissipating the acid that is released to the environment (wave and current action). They also provide novel insights into the ecophysiological and evolutionary constraints within which marine organisms adapt.     

Effects of carbaryl on the aminotransferases in the mid gut of blister beetle,Mylabris pustulata (Thunb)

The activity levels of aminotransferases such as alanine aminotransferases (AlAT), aspartate aminotransferases (AAT), isoleucine aminotranferases (IlAT), leucine aminotransferases (LAT), valine aminotrasferases (VAT) were measured in the mid gut of the blister beetle; Mylabris pustulata (Thunb) exposed to sublethal doses of carbaryl. The insecticide markedly inhibited the activity levels of all aminotransferases exposed for either short or prolonged treatments. The beetles regained normal enzymic activity after withdrawal from the treatment. It is possible that chronic exposures of beetles to carbaryl may induce impairment in protein synthesis in the mid gut of the beetle.