Fine structure of the triturating devices in the food pump of Notonecta (Heteroptera: Notonectidae)

The cibarial food pump of heteropteran insects conveys fluid food from the piercing stylets to the pharynx. In aquatic Heteroptera the pump also grinds and filters particulate matter in the food stream. The pump's sclerotized triturating devices differ from one family to the next and are often quite elaborate: because of their small size they are best studied by means of the scanning electron microscope. In Notonecta the main triturating devices occur on the transverse plates of the epipharyngeal roof of the pump. They consist of a complex anterior zone with raised nodes and bifurcating longitudinal ridges, and a simpler posterior zone with small nodules. Additional triturating surfaces occur on the hypopharyngeal floor of the pump. The oblique folds of the epipharynx, which lie anterior to the transverse plates, play only an accessory role. The fine structure of the grinding surfaces on the transverse plates of Gelastocoris (Gelastocoridae), Ambrysus (Naucoridae), and Aphelocheirus (Aphelocheiridae) is here briefly described and compared with that of Notonecta.     

单增李斯特菌耐药外排泵研究进展

单增李斯特菌是一种重要的人兽共患食源性胞内致病菌,广泛存在于自然环境中且易污染动物性食品,人及动物感染后可引起严重的李斯特菌病,死亡率高达30%。单增李斯特菌通常对多种药物敏感,然而,因不合理使用抗菌药或消毒剂形成的选择压力导致李斯特菌多重耐药情况的报道日渐增多。外排泵蛋白是细菌中一类重要的蛋白,可参与机体多种生物学过程,包括影响细菌对抗生素敏感性、促进有毒化合物泵出、影响细菌毒力等。本文综述了近年来关于单增李斯特菌耐药外排泵的功能及调控机制的研究进展,为深入理解李斯特菌耐药等环境适应机制及有效控制该病原污染传播和筛选抗感染药物新靶点提供理论基础。     

Plastron respiration in bugs and beetles

In the naucorid bug, Alphelocheirus, the plastron hairs are twice as thick and nearly twice as dense (ca. 4 × 10⁶/mm²) as they had been thought to be by previous workers. From experiments and calculations it seems clear that when the plastron is subjected to excess pressures it is wetted long before there is any question of the collapse of the hair pile itself. The plastron of Aphelocheirus is thus like the plastrons of other insects in that it is wetted long before the structures supporting the air film collapse.A plastron has been independently evolved in at least five subfamilies of the Naucoridae. A plastron is here recorded for the first time in bugs of the family Helotrephidae.It has been claimed that the plastron-bearing elmid beetles are unable to fly. Many, if not most, of these beetles fly after they emerge from their pupal cells. However, once they have begun to live under water they cease to be able to fly: the flight muscles degenerate, and this degeneration seems to be irreversible.The structure of the plastron scales of several kinds of weevils is described. The resistance of the plastron of the rice water weevil, Lissorhoptrus, to wetting at excess pressures is examined. An explanation is advanced to account for the fact that weevils and other plastron-bearing beetles that live in still waters can often swim whereas those, like elmids and dryopids, that live in running waters cannot swim.     

Prey Selection by the Water Bug Aphelocheirus aestivalis Fabr. (Heteroptera: Aphelocheiridae)

Feeding experiments were conducted in situ in flow through boxes to determine the vulnerability of different prey types (the mayfly larvae Baetis, Ephemerella and Ecdyonurus spp., the caddisfly larva Hydropsyche spp. and the crustacean Gammarus spp.) to the predaceous water bug Aphelocheirus aestivalis and to estimate the predation impact of Aphelocheirus on prey populations. Experiments with equal densities of mixed prey and experiments where each prey was tested individually revealed that Baetis and Ephemerella were most vulnerable to Aphelocheirus predation: Hydropsyche, Ecdyonurus and Gammarus were little/not preyed upon. The present results suggest that vulnerability of prey depends mainly upon prey mobility and microhabitat overlap between predator and prey and that feeding behaviour of Aphelocheirus resembles more that of megalopterans than of stoneflies. Experiments with different prey densities (120–720 prey m⁻²) showed that the predation rate of Aphelocheirus increased with increasing prey density. Experiments with different substrates documented that mortality rates of prey decreased with increasing substrate complexity. When substrate conditions were complex mortality rates in the control and in the experimental boxes were the same which suggest little importance of Aphelocheirus predation on mayflies in the study site.     

In-silico interaction studies suggest RND efflux pump mediates polymyxin resistance in Acinetobacter baumannii

Bacterial efflux pumps have emerged as antibiotic resistance determinants and confers multi-drug resistance to a broad range of antimicrobials as well as non-antibiotic substances. A study about translocation of antibiotic molecules through the efflux transporter, will contribute in determining substrate specificity. In the present study, we have explored RND family efflux pump extensively found in Acinetobacter baumannii i.e. AdeABC. Besides, another well studied RND efflux pump, AcrAB-TolC together with a non-RND efflux pump, NorM was investigated for comparative analysis. We employed a series of computational techniques ranging from molecular docking to binding free energy estimation and molecular dynamics simulations to determine the binding affinity for different classes of drugs, namely aminoglycosides, polymyxins, β-lactams, tetracyclines, glycylcyclines, quinolones and metronidazole with AdeB, AcrB, and NorM efflux proteins. Our results revealed that class polymyxins has the highest binding affinity with the RND efflux pumps i.e. AcrAB-TolC and AdeABC as well as non-RND efflux pump, NorM. The experimental validation study demonstrated bigger zone of inhibition in presence of efflux pump inhibitor than polymyxin alone thus unveiling its specificity toward efflux pump. The reported experimental data comprising of minimum inhibitory concentration of antibiotics toward these efflux pumps also support our finding based on in silico approach. To recapitulate the outcome, polymyxins shows maximum specificity toward RND as well as non-RND efflux pump and may unlatch the way to rationally develop new potential antibacterial agents as well as efflux pump inhibitors in order to combat resistance.     

A methodological constraint influencing measurement of food intake rates in sucking predators

Summary In a number of studies which provide food extraction curves for sucking predators, data were obtained by separating the predator from the prey (i.e. by artificially interrupting feeding) at predetermined intervals within the total feeding time. The amount of food the predator had extracted at these time intervals was then determined by measuring either the mass gain in the predator or the mass loss in the prey. An implicit assumption of this method is that at the time feeding is interrupted, the food extracted by the predator is contained within its own digestive system and no part of the food has been released back into the prey. I found this was not the case with the crab spider Diaea sp. indet. feeding on the fruit fly Drosophila immigrans. The food Diaea extracts from prey is retained in its own digestive system only at times when the spider changes feeding sites on the prey and when it discards the prey when finished feeding. At other times it cycles the extracted food between itself and the prey (i.e. a sucking phase is alternated with a relaxing phase during which the extracted food is released back into the prey). Unless feeding is interrupted as close as possible to the end of the sucking phase, the mass change measured in the spider will be an underestimate of the actual amount of food extracted at this time. My results suggest that understanding how a sucking predator's feeding mechanism affects the transfer of food is necessary, not only in identifying constraints affecting feeding efficiency, but also in identifying how these constraints relate to the design of empirical tests. The precision of these tests will, in part, reflect the degree to which these mechanisms are incorporated into the test design.     

Comparative mechanics of filter feeding in Anas platyrhynchos,Anas clypeata and Aythya fuligula (Aves,Anseriformes)

Summary The straining of seeds ranging in size from 0.01 to 4.6 mm was investigated in the filtering mechanisms of three anseriform species, Anas platyrhynchos, Anas clypeata, and Aythya fuligala. The morphology of the filtering mechanisms was studied by dissection and from microsections. The kinematic work envelope of the beaks was measured by stereotactic manipulation. The actual beak kinematics were analyzed by calculating the changes of the mouth volume from computer simulations and comparing the results with the amounts of water expelled. The performances of the filter and the water pump were measured with special collection boxes. The influence of the lamellae was estimated by removing them in whole or in part and measuring the performances again. The results show that if the size of the strained seeds increases, the gapes and amplitudes also increase (cf. bill rotations). Beak rotation is elevated for small kernels, even more elevated for slightly larger ones, but depressed for large kernels. However, neither frequencies nor phase shifts change between different moving elements. Simulation of these data by stereotactic manipulation of fresh material and by measuring gape sizes and mesh widths shows that the beak kinematics during straining keep mesh widths and gapes carefully adjusted to the seed size. The pumping and the filtering mechanisms influence each other mutually in setting boundary conditions on mouth kinematics. Beyond a certain kernel size, the necessity for mesh-width control sets a boundary on the increase of the water flow. Conversely, phase shifts, frequency changes and change of lingual amplitudes were restricted by the pumping mechanism. Drastic shortening of either dorsal mandibular or maxillary lamellae in Anas platyrhynchos and Aythya fuligula does not effect either the filter performances, the pump performances, or the beak kinematics, but these modifications do change the performances in Anas clypeata. The usually accepted assumption concerning food sieving was thus verified for Anas clypeata and falsified for the other two anseriform species. Therefore, a vortex model in addition to the sieving model, was developed, based on the assumption that local vortices cause inertia of particles so that small kernels are filtered by centrifugation and translation out of the main water flow. Combined effects of sieving and centrifugation occur when larger kernels are filtered. The kinematics of the filtering mechanism are highly flexible; therefore, a wide tolerance for seed sizes exists. However, the morphology of the filtering mechanism differs in the three species. Testing whether the anseriform filter mechanism is so plastic that the mechanism in each species has developed in such a way that the staple food is gathered most economically has shown that filtering of the staple food does indeed occur at the most efficient levels of maxillary rotation.     

Discovery of inhibitors of Pseudomonas aeruginosa virulence through the search for natural-like compounds with a dual role as inducers and substrates of efflux pumps

Multidrug efflux pumps are ancient elements encoded in every genome, from bacteria to humans. In bacteria, in addition to antibiotics, efflux pumps extrude a wide range of substrates, including quorum sensing signals, bacterial metabolites, or plant-produced compounds. This indicates that their original functions may differ from their recently acquired role in the extrusion of antibiotics during human infection. Concerning plant-produced compounds, some of them are substrates and inducers of the same efflux pump, suggesting a coordinated plant/bacteria coevolution. Herein we analyse the ability of 1243 compounds from a Natural Product-Like library to induce the expression of P. aeruginosa mexCD-oprJ or mexAB-oprM efflux pumps' encoding genes. We further characterized natural-like compounds that do not trigger antibiotic resistance in P. aeruginosa and that act as virulence inhibitors, choosing those that were not only inducers but substrates of the same efflux pump. Four compounds impair swarming motility, exotoxin secretion through the Type 3 Secretion System (T3SS) and the ability to kill Caenorhabditis elegans, which might be explained by the downregulation of genes encoding flagellum and T3SS. Our results emphasize the possibility of discovering new anti-virulence drugs by screening natural or natural-like libraries for compounds that behave as both, inducers and substrates of efflux pumps.     

Use of resazurin to detect mefloquine as an efflux-pump inhibitor in Pseudomonas aeruginosa and Escherichia coli

Multi-drug-resistant bacteria can cause serious infections that are extremely difficult to treat. Bacterial efflux pumps are known to contribute to multi-drug resistance and, thus, constitute a promising target for novel antibacterial agents. Resazurin is widely used to monitor bacterial growth because resazurin is reduced to the fluorescent resorufin by live cells. We have shown by flow cytometric analysis and by accumulation studies with wild type and efflux deficient strains that resazurin is a substrate of efflux pumps in Escherichia coli and Pseudomonas aeruginosa. Our investigations showed that the conversion rate of resazurin to resorufin is affected by efflux pumps. This finding was used to design an assay useful to detect efflux pump activity and to find potential efflux-pump inhibitors in a microtiter plate format. Mefloquine was detected as efflux-pump inhibitor when a panel of selected chemical compounds was tested for assay validation purposes.     

Regulation of plant elongation growth by surface and xylem proton pumps

The roles of plasmalemma electrogenic proton pumps in elongation growth of plant stems are discussed on the basis of growth-electrophysiological studies on hypocotyl segments ofVigna unguiculata. Plant stems usually have two spatially separated electrogenic proton pumps: the surface proton pump which is located on the surface membrane of the symplast and the xylem proton pump, on the cell membrane of the symplast/xylem apoplast boundary. The surface proton pump excretes protons into the surface cell wall layer and causes the loosening of the cell wall. The xylem proton pump excretes protons into the xylem apoplast and drives the uptake of solute and water into the symplastvia secondary and/or tertiary active mechanisms: the proton cotransport system and the apoplast canal system. Both the surface and the xylem proton pumps are active during elongation growth because both the yielding of cell wall loosening and the uptake of water are necessary for continued elongation growth.     

Immunolocalization of Na,K-ATPase in blowfly photoreceptor cells

The Na,K-ATPase (sodium pump) plays a central role in the physiology of arthropod photoreceptors as it re-establishes gradients for Na⁺ and K⁺ after light stimulation. We have mapped the distribution of the Na,K-ATPase in the photoreceptors of the blowfly (Calliphora erythrocephala) by immunofluorescent and immunogold cytochemistry, and demonstrate that the distribution pattern is more complex than previously presumed. High levels of sodium pumps have been detected consistently in all photoreceptors R1-8 on the nonreceptive surface, but no sodium pumps are found on the microvillar rhabdomere. Within the nonreceptive surface of the cells R1-6, however, the sodium pumps are confined to sites juxtaposed to neighboring photoreceptor or glial cells; no sodium pumps have been detected on the parts of the nonreceptive surface exposed to the intra-ommatidial space. In R7 and R8, the sodium pumps are found over the entire nonreceptive surface. The cytoskeletal protein spectrin colocalizes with the sodium pumps suggesting that linkage of the pump molecules to the spectrin-based submembrane cytoskeleton contributes to the maintenance of the complex pattern of pump distribution.     

Current noise generated by electrogenic ion pumps

Active ion transport by ATP-or light-driven pumps involves a sequence of elementary steps such as binding and release of ions, as well as conformational transitions of the pump protein. At the microscopic level the individual reaction steps occur at random intervals, and therefore the current generated by electrogenic pumps fluctuates around a mean value. In this paper, a theoretical treatment of the electrical noise associated with active ion transport is given. The analysis, which is based on the calculation of the correlation function, yields the spectral intensity S ₁ of current noise as a function of frequency, f. The shape of S _I(f) contains information on the rate constants as well as on the magnitude of the charge displacements occuring during single reaction steps. The contribution of electrogenic pumps to the total voltage noise of the cell may be estimated from S _I(f) and from the frequency-dependent impedance of the cell membrane.     

The proton pumps of the plasmalemma and the tonoplast of higher plants

Studies on intact cells, membrane vesicles, and reconstituted proteoliposomes have demonstrated in higher plants the existence of an ATP-driven electrogenic proton pump operating at the plasmalemma. There is also evidence of a second ATP-driven H⁺ pump localized at the tonoplast. The characteristics of both these ATP-driven pumps closely correspond to those of the plasmalemma and tonoplast proton pumps ofNeurospora and yeasts.     

Fine structure and sensory apparatus of the mouthparts of the pear psyllid,Cacopsylla chinensis (Yang et Li) (Hemiptera: Psyllidae)

The pear psyllid, Cacopsylla chinensis (Yang et Li) (Hemiptera: Psyllidae), is one of the most significant economic pests of pear in China, causing direct damage through feeding by the highly specialized piercing–sucking mouthparts. The ultrastructural morphology and sensory apparatus of the mouthparts of the adult were examined using scanning and transmission electron microscopy. The piercing–sucking mouthparts of C. chinensis are composed of a three-segmented labium with a deep groove in the anterior side, a stylet fascicle consisting of two mandibular and two maxillary stylets, and a pyramid-shaped labrum. Proximal to the labium, the stylet fascicle forms a large loop within a membranous crumena. Mandibles, with more than ten teeth on the external convex region, can be seen on the distal extremity. Smooth maxillary stylets are interlocked to form a larger food canal and a smaller salivary canal. One dendritic canal housing 2 dendrites is also found in each mandible. Two types of sensilla trichodea, four types of sensilla basiconica, single as well as groups of sensilla campaniformia, and oval flattened sensilla occur in different locations on the labium, whereas a kind of sensilla basiconica is at the junction of the labrum and anteclypeus. Sensilla trichodea and sensilla campaniformia, always present with denticles, are present on the middle labial segment. Three types of sensilla basiconica, two types of sensilla trichodea and two oval flattened sensilla are located on the distal labial segment. The mouthpart morphology and abundance of sensilla located on the labium in C. chinensis are illustrated, along with a brief discussion of their taxonomic and putative functional significance.     

Feeding behaviour and morphology of filtering combs of Daphnia galeata

The population of Daphnia galeata Sars from the fish pond Velký Pálenec (Blatná, Czechoslovakia) living in high food conditions (7 mgC l^–1) was characterized by a small size of the filtering comb on the thoracic limb 3, measured as seta length, length of the base of the comb and number of setae (population 1). One month cultivation of this population in low food conditions (1.5 mgC l^–1) in the laboratory resulted in twofold increase in size of the filtering comb (population 2). Filtering and ingestion rates of both populations were measured at eight concentrations of food (approximately 0.025–3.2 mgC l^–1) using ¹⁴C labeled Scenedesmus acutus. The results show that size of the filtering combs influences considerably feeding behavior of Daphnia. The comparison of animals with the same body length suggests that the population with a large comb feeds at concentration of food below 0.4 mgC l^–1 more intensively and reaches the maximum of the filtering rate at a lower concentration than the population with a small comb. The situation is opposite at concentration above 0.4 mgC l^–1. The higher values of theoretical flow in population with a small projection of filtering area suggest that this population has to compensate disadvantage of a small comb with the higher appendages beat frequency.     

The triradiate sucking pharynx in animal phylogeny

Independent sucking/swallowing pharynges, as found in members of at least ten animal phyla, are pharynges that are opened by muscles that are not attached to the outer body wall. Their musculature is derived from either mesoderm or ectoderm, the latter taking the form of a myoepithelium. I review results of previous work on the morphology of independent sucking/swallowing pharynges among invertebrates and provide new information on the ultrastructure of the pharynx of the cyclostome bryozoan Crisia eburnea. The various morphologies of this type of pharynx have been used in some phylogenetic considerations, but only the myoepithelial sucking pharynx with a triradiate lumen was considered significant in analyses of relationships between phyla. However, I argue that this shape is the only one that makes an efficient suction pump, and this, together with different orientations of the myoepithelial pharyngeal pump and its phylogenetic distribution, indicates that the triradiate myoepithelial pharynx has evolved convergently in a number of lineages.