Structure of the olfactory receptor organs,their GABAergic neural pathways,and modulation of mating behavior,in the giant freshwater prawn,Macrobrachium rosenbergii

In the giant male prawn, Macrobrachium rosenbergii, the olfactory system is thought to be the main pathway for modulating sexual behavior through pheromone perception. In this report, we first used gross anatomical, histological, and SEM methods to describe the structures of the olfactory receptors (sensilla setae), their neural pathways, and possible role in modulating mating behavior. On the surfaces of antennule and antenna filaments there are four types of sensory receptors, viz single spike‐like setae, single flagellum‐like setae, multiple flagella‐like setae, and aesthetascs (ASs). The ASs, which had previously been proposed to be odor receptor setae, are found only on the short filament of lateral antennule (slAn). Each AS on the slAn connects with olfactory receptor neurons (ORNs), whose axons form an outer central antennule nerve (ocAnNv), which then connects with the olfactory neutrophil (ON) of the brain. Thus, the slAn is the major olfactory organ that conveys sensory inputs from each AS to the ON within the deutocerebrum. GABA immunoreactivity was present in ASs, neurons of ORNs, inner central antennular, lateral tegumentary nerve, ocAnNv and the ON, inferring that GABA is the likely neurotransmitter in modulating olfaction. Disruption of the slAn by ablation or covering with Vaseline, resulted in significant reduction of mating behavior, indicating that this organ is crucial for sex pheromone perception. Identification of the active pheromones and further bioassays are now being performed. Microsc. Res. Tech. 76:572–587, 2013. © 2013 Wiley Periodicals, Inc.     

Oogenesis of Diadegma semiclausum (Hymenoptera: Ichneumonidae) and its associated polydnavirus

We present a detailed ultrastructural analysis of ovary and calyx cell differentiation of Diadegma semiclausum. Numerous gametangia in various developmental stages were examined with electron microscopy to characterize the ultrastructure features of oogenesis, the most important of which is the development of nurse cells. In the germarium, the undifferentiated germ cells diversify, and one of the central cells of the cluster differentiates into an oocyte while the remaining become nurse cells. Germ cells continue developing in the vitellarium until mature and then enter into the calyx region. The calyx epithelium contains typical ichneumonid polydnaviruses with the following characteristics: (1) the virus particles assemble in the nuclei of calyx cells where they acquire an initial (inner) membrane, then migrate through the cytoplasm and budd out into the lumen of the ovary, at which time they acquire a second envelope (outer membrane); (2) the particle has a genome comprising several DNA segments. However, this new polydnavirus (Diadegma semiclausum polydnavirus) in the genus ichnovirus was not attached to the surface of the egg chorion.     

Ultrastructure of multicellular dwarf males with external gametangium in Oedogonium macrandrium (Oedogoniales,Chlorophyta)

This is the first comprehensive ultrastructural study on dwarf males with external gametangia in the ge in the genus Oedogonium, from androspore germination to the liberation of mature male gametes. The ultrastructure of the process in O. macrandrium Wittrok is similar to that of Bulbochaete hiloensis (Nordstedt) Tiffany, butwith two remarkable differences. In O. macrandrium : 1) instead of a true transverse wall, only condensed mucilage appears between the gametes of each antheridial cell, and 2) the cell wall between the basal cell and the basal most antheridial cell has simple plasmodesmata similar to those present in the transverse walls of vegetative cells, which are absent in B. hiloensis.     

Ultrastructure of sensory equipments on the heads of Kallitaxila granulata (Stål) (Hemiptera: Fulgoromorpha: Tropiduchidae)

The polyphagous planthopper, Kallitaxila granulata (Stål) (Hemiptera: Fulgoromorpha: Tropiduchidae), has been recently introduced into southeastern China, the Philippine islands, and Hawaii, where it has done significant damage to agricultural and forest ecosystems. The external morphology of the heads of adult male and female K. granulata was examined using scanning electron microscopy. Seven types of sensilla were reported: trichoid sensilla and campaniform sensilla on the antennal pedicel, antennal scape and maxilla; plate organs on the antennal pedicel; coeloconic sensilla in Bourgoin's organ on the expanded flagellar base; ampullaceal sensilla on the antennal pedicel; wavy‐pit sensilla on the antennal pedicel and antennal scape; and coin‐shaped sensilla on each lateral side of the labium. Evans' organ was described as placoid sensilla sunk into shallow cuticular cavities below the antennae. The external morphology, distribution, and abundance of sensilla located on antennae, maxillae, and labium in K. granulata were illustrated, with a brief discussion of their taxonomic, phylogenetic, and putative functional significance. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.     

Ultrastructure and distribution of antennal sensilla of the chilli thrips Scirtothrips dorsalis hood (Thysanoptera: Thripidae)

The chilli thrips, Scirtothrips dorsalis Hood, is a serious pest of numerous important vegetable and ornamental crops. Various signals, especially phytochemical cues, determine the behavior of the phytophagous thrips at host selection. The sensory abilities of S. dorsalis are poorly understood although the antennae of adult are known to possess important sensory structures in orther insects. In this study, the morphology, distribution, and ultrastructure of the antennal sensilla of the S. dorsalis were examined by using scanning and transmission electron microscopy. Microscopy observations revealed that adult male and female S. dorsalis possess filiform antennae. Each antenna comprises a scape, a pedicel, and a flagellum composed of six segments without clear sexual dimorphism in the number and distribution of antennal sensilla. The scape and pedicel exhibit Böhm's bristles, sensilla chaetica, and sensilla campaniform. The external structures of these organs reveal their mechanosensory function. In the flagellum, the most represented sensilla are the multiporous sensilla basiconica, which can be divided into three types of single‐walled olfactory sensilla; three types of sensilla chaetica with mechanosensory and gustatory functions; sensilla coeloconica, which possess hollow cuticular spoke channels and represent double‐walled olfactory sensilla; sensilla capitula and sensilla cavity with thermo‐hygrosensory functions; and aporous sensilla trichodea with smooth cuticula and mechanosensory function. The putative function of described sensilla is discussed in ralation to host plant selection behavior of S. dorsalis.     

Ultrastructure of oocytes of the Urostreptus atrobrunneus (Diplopoda,Spirostreptida, Spirostreptidae): A potential urban centers plague

The knowledge of the process of egg formation is indispensable for understanding the mechanisms involved in the reproduction of different species. In this context, the objective of this work was to describe the ultrastructure of the oocytes of Urostreptus atrobrunneus (Spirostreptida), a potential plague of urban centers in different locations of São Paulo State. The lack of knowledge about the morphology, physiology, and the reproductive behavior of the species have hindered an effective control of it. The oocytes of U. atrobrunneus presented three development stages: young oocyte or type I; intermediary oocyte or type II; and mature oocyte or type III. During the oocyte development, the cytoplasm become filled with several globules of protein, drops of lipids, and sphaerocrystals, and it was not observed in many organelles in the oocytes with exception of mitochondria, abundant, principally in young oocytes. The vitelline membrane is also deposited in a discontinuous form and the chorion does not present differentiation of layers. The follicular epithelium alters its shape according to the development phase of the oocyte. Part of the vitellus is from exogenous origin and part is endogenous. Before this, only two studies about the ultrastructural analysis of the female germ cells of diplopods were published. Microsc. Res. Tech. © 2012 Wiley Periodicals, Inc.     

Ultrastructure of the male reproductive system of Cotesia vestalis (Hymenoptera: Braconidae) with preliminary characterization of the secretions

The morphology and ultrastructure of testes and accessory glands along with the characterization of their secretions were investigated for a braconid species Cotesia vestalis (Hymenoptera: Braconidae) using light and electron microscopes. The male internal reproductive system of this species is distinguished by a pair of testes, one vas deferens, and a pair of male accessory glands. The testes are separate, and the accessory glands are oval and not fused. It was observed that the secretory cells of testes have characteristic smooth and rough endoplasmic reticulum, and that the cytoplasm is filled with an array of granule droplets usually of two to three types. The secretory cells in the case of accessory glands are typified by the presence of microvilli on their apical cell surfaces and numerous mitochondria in their cytoplasm. SDS-PAGE profile when performed depicted a similarity in most bands of the secretions from both testes and accessory glands, except for four proteins of which two were present only in testes, while the other two only appeared in accessory glands. Their molecular weights were 117 and 55 kDa for testes and 196 and 30 kDa for accessory glands, respectively. This study gives new insights into the intriguing features of male internal reproductive system and it especially constitutes the first report of its kind about the secretion properties of these organs in C. vestalis.     

Silk fibers and silk‐producing organs of Harpactea rubicunda (C. L. Koch 1838) (Araneae,Dysderidae)

Scanning electron microscopy and atomic force microscopy were used to study the silk spinning apparatus and silks of Harpactea rubicunda spiders. Three types of silk secretions that are produced by three kinds of silk spinning glands (ampullate, piriform, and pseudaciniform) and released through three types of spigots, were confirmed for both adult and juvenile spiders. Silk secretions for the construction of spider webs for shelter or retreat are produced by the pseudaciniform silk glands. Silk secretions that are released from spigots in the course of web construction are not processed by the legs during the subsequent process of hardening. Pairs of nanofibril bundles seemed to be part of the basic microarchitecture of the web silk fibers as revealed by AFM. These fiber bundles frequently not only overlap one another, but occasionally also interweave. This structural variability may strengthen the spider web. High‐resolution AFM scans of individual nanofibrils show a distinctly segmented nanostructure. Each globular segment is ～30–40 nm long along the longitudinal axis of the fiber, and resembles a nanosegment of artificial fibroin described by Perez‐Rigueiro et al. (2007). Microsc. Res. Tech., 2013. © 2012 Wiley Periodicals, Inc.     

Ovary of Matsucoccus pini (Insecta,Hemiptera, Coccinea: Matsucoccidae): Morphology,ultrastructure, and phylogenetic implications

The structure of ovary in a representative of the scale insect family Matsucoccidae, Matsucoccus pini, is described at the ultrastructural level. The paired ovaries of M. pini are composed of about 50 ovarioles of telotrophic type that develop asynchronously. An individual ovariole consists of an anterior tropharium (trophic chamber) and posterior vitellarium. The tropharium encloses trophocytes (nurse cells) and early previtellogenic oocytes termed arrested oocytes. In the vitellarium from 1 to 6, linearly arranged oocytes may develop. Analysis of serial sections has shown that each ovariole contains 32 germ cells (trophocytes, arrested oocytes, and developing oocytes). In the cytoplasm of all these cells, small rod‐shaped bacteria are present. In the early vitellogenic oocytes, accessory nuclei arise. As vitellogenesis progresses, these nuclei migrate toward the cortical ooplasm. The obtained results are discussed in a phylogenetic context. Microsc. Res. Tech. 77:327–334, 2014. © 2014 Wiley Periodicals, Inc.     

Olfactory sensilla on antennae and maxillary palps of Fannia hirticeps (Stein, 1892) (Diptera: Fanniidae)

The Fanniidae is one of four families in the superfamily Muscoidea (Diptera), including some important medical and hygienic flies. There is a paucity of reports on the ultrastructure of olfactory sensilla for the fanniid species. To provide more information on the morphology of the antennal and palpal sensilla of fanniid fly, Fannia hirticeps (Stein, 1892) has been studied using scanning electron microscopy. The first two antennal segments, scape and pedicel, are covered by microtrichiae and several chaetic sensilla. Six distinct morphological types of sensilla are recorded on the antennal funiculus, including one trichoid, two basiconic, two coeloconic sensilla, and one clavate sensilla. The measurement and density of each sensilla type are also provided. The trichoid sensilla tend to be longer and denser toward the apex of antennal funiculus. Basiconic sensilla spread all over the funicular surface. F. hirticeps bears two types of coeloconic sensilla, type 2 coeloconic sensilla distributed on the distal part of the anterior surface, whereas type 1 distributed on the rest of the funiculus. Clavate sensilla are found on the base of antennal funiculus. Only one large sensory pit is located on the posterior surface. Maxillary palps bear one type of basiconic sensilla. These results are compared with eight other muscid flies. Our findings provide a morphological basis for future investigations on olfactory‐mediated behavior of this group. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Ultrastructure of dermal denticles in sharpnose shark (Rhizoprionodon lalandii) (Elasmobranchii,Carcharhinidae)

The presence of denticles in the external surface, oral cavity, fins, and clasper of Elasmobranchii has been widely reported. These structures, called body denticles, may be observed on the body surface of sharks. Dermal and oral denticles are made up by a basal plate that is embedded in the dermis, forming a peduncle that grows from the base to the crown. These denticles may protect the skin against abrasion, and improve hydrodynamics and gill arches function. Rhizoprionodon lalandii is a widely distributed and very common species in Brazilian coastal areas. The aims of this study was to compare the morphology of oral and body denticles of R. lalandii to understand the implications of these structures in the behavior of these animals. Morphological analysis showed that there are differences between dermal and oral denticles, which are related to their role in different body regions. Body denticles have three cusps, and well‐defined crests and ridges, and literature data suggest that suggest that hydrodynamics is their main function. Most of the oral denticles have only one cusp, and their morphology and distribution showed that their main functions are preparing food to be swallowed and protecting the oral cavity against abrasion. Microsc. Res. Tech. 78:859–864, 2015. © 2015 Wiley Periodicals, Inc.     

Light and transmission electron microscopy study of the peripheral olfactory organ of the guppy, Poecilia reticulata (Teleostei, Poecilidae)

A study of the peripheral olfactory organ, with special attention to the olfactory epithelium, has been carried out in the guppy (Poecilia reticulata). Guppy is well known to have a vision-based sexual behavior. The olfactory chamber caudally opens directly in an accessory nasal sac, which is bent medially and gives rise to two recesses that can be considered secondary accessory nasal sacs, antero-medial and postero-medial, respectively. The sensory epithelium, which lines only the medial wall of the nasal cavity, is basically flat rising in a very low lamella only in the posterior part. The olfactory receptors are not evenly distributed in the olfactory mucosa, but aggregate in shallow folds separated by epithelial cells with evident microridges. Ciliated olfactory sensory neurons and microvillous olfactory sensory neurons are clearly identified by transmission electron microscopy (TEM). Scarce crypt olfactory neurons are found throughout the sensory folds. The nasal sacs indicates the capacity to regulate the flow of odorant molecules over the sensory epithelium, possibly through a pump-like mechanism associated with gill ventilation. The organization of the olfactory organ in guppy is simple and reminds what is found in early posthatching stages of fish which at the adult state have a well developed olfactory organ. This simple organization supports the idea that the guppy rely on olfaction less than other fish species provided with more extended olfactory receptorial surface.     

Ultrastructural characteristics of the lung of <i>Melanophryniscus stelzneri stelzneri</i> (Weyenberg, 1875) (Anura,Bufonidae)

The lung of the toad, Melanophryniscus stelzneri stelzneri was studied using scanning and transmission electron microscopy. In M.s.stelzneri the parenchyma forms a polygonal network arrangement, therefore the parenchyma is edicular. These spaces are delimited by the interconnection of third order septa which are covered by respiratory epithelium. Small patches of ciliated epithelium without goblet cells appear irregularly distributed on the septa. The respiratory epithelium consists of one type of pneumocyte, which shows characteristics of both type I and type II alveolar cells of higher vertebrates. The pneumocytes are irregular in shape and possess attenuated cytoplasmic processes, which spread around the capillaries to form the outer layer of the air-blood barrier. These cells contain different types of cytoplasmic bodies: electron dense bodies, multivesicular bodies and lamellar bodies. Dense bodies are probably the precursors of lamellar bodies and the multivesicular bodies are incorporated into the latter. Neuroepithelial bodies appear randomly distributed over the septa. These bodies are separated from the lumen of the lung by thin cytoplasmic processes of neighbouring pneumocytes. The air-blood barrier consists of three layers: epithelium, interstitial space and endothelium.
The relatively simple pulmonary structure of M.s.stelzneri is due to a lower degree of partitioning of the pulmonary lumen in comparison to the lung of other bufonid anurans, could be correlated with a well developed cutaneous and buccopharingeal respiration. The testing of this hypothesis awaits further studies.     

Ultrastructure of immature stages of the blowfly Chrysomya putoria (Wiedemann, 1818) (Diptera: Calliphoridae)

Forensic entomology is an area of science that serves as a tool in crime scene investigations. Usually, flies are the first insects to reach a carcass and can oviposit just a few hours after arrival. Therefore, the knowledge of immature stages is essential for correct identification of the species found on corpses. Scanning electron microscopy (SEM) gives detailed information about morphological characters helping to identify the immature forms of flies. Chrysomya putoria (Wiedemann) is a very important fly for forensic entomology, because it has high population densities and is easily found in colonizing carcasses, moreover, it is also a possible causative agent for secondary myiasis. The aim of this study is to identify larvae and puparia of C. putoria using SEM. The first instar larvae were composed of 12 smooth segments separated by spines. Antennae and maxillary palps were visible. Anterior spiracle was absent and only one spiracular opening could be seen at the posterior spiracle. Second and third larval instars were similar to first instar, except for the presence of anterior spiracle that is composed by 11-12 spiracular ramifications. At the anal segment, two spiracular openings were found in second instars and three openings in third instar larvae. Puparia showed a retracted cephalic region and none of the head structures were visible.     

Rhyncophorus palmarum L. (Linnaeus, 1758): A morphological and histological study of the female reproductive system

Rhyncophorus palmarum (Coleoptera) is a pest of great economic importance because of the damage caused in the plants of the Palmae family. This study showed that the female reproductive system of this beetle is composed of two ovaries of the telotrophic meroistic type invariably containing two ovarioles per ovary. Each ovariole is home to about 50 oocytes at different stages of maturation. The proximal region of the ovary is rather peculiar because it houses inside it a large quantity of bacilliform structures immersed in a glycolipoprotein substance, which has never before been described in the literature. The function of these structures is still unknown. However, it is suggested that they can function as a method of neutralizing the resistance of the plant because these insects are responsible for the transmission of the red ring disease to the palm plants. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.     

Inhibitory action of neem aqueous extract (Azadirachta indica A. Juss) on the vitellogenesis of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) ticks

The present study revealed unheard of data about the action of aqueous extracts of neem leaves (Azadirachta indica) on the vitellogenesis of Rhipicephalus sanguineus ticks, proving that these extracts in 10 and 20% concentrations do not have the potential to kill the females; however, in lower concentrations (10%) provokes great morphological alterations in germinative cells such as the emergence of extended cytoplasmic vacuolization areas as well as the fragmentation of the germinal vesicle, even in those oocytes which were in initial stages of development (I-III), showing that neem is a potent agent which acts impeding one of the main metabolic stages of the ticks, i.e., the reproduction. In oocytes in final stages of development (IV-V) azadirachtin (neem's active principle) caused significant reduction in the size and quantity of proteic granules of the yolk and the inversion of their localization where the smaller granules before inside the cell (normal oocyte) were posteriorly observed in the periphery, and the bigger ones in the central region. Thus, the study showed that the alterations found both in the oocytes and in the pedicel cells indicated that azadirachtin acts on the process of tick's reproduction and signalizes that this plant can be used in the future to control ticks with the advantage of not being aggressive to nontarget organism or the environment. Furthermore, data here obtained showed that the most significant efficiency of the aqueous extract of neem is related to the concentration of 10%, proving that higher doses would not be so efficient.     

Egg envelopes of Armadillidium vulgare (Latreille, 1804) (Crustacea,Isopoda Oniscidea): Ultrastructure and lectins binding

The ultrastructural study carried out on (a) oocytes of Armadillidium vulgare during vitellogenesis, (b) mature eggs taken from the ovaries during the parturial moult of the posterior half of the body, and (c) fertilized eggs collected within a few hours of their release into the brood pouch, has clearly demonstrated that before the fertilization the chorion is the only envelope present in the egg of oniscidean isopods. In the mature eggs, the chorion appears as a uniformly electron‐dense lamina, about 0.4–0.5 µm thick, which does not show any specialized area. A second envelope, described by other authors as vitelline envelope, is formed above the oolemma only right after fertilization and appears separated from the chorion by a space full of liquid. The ways in which the genesis of this envelope is realized are not yet clear; it could be interpreted rather as a fertilization membrane. The investigations carried out with the aid of a battery of FITC‐lectins have highlighted the presence at the chorion surface of unfertilized eggs of various saccharide residues distributed in uniform way. No significant change was observed in the pattern of lectins binding to the chorion of eggs taken from the brood pouch, thus demonstrating how, after the fertilization, no significant rearrangement in the distribution of saccharide residues present on the egg surface occurs in A. vulgare. The ways in which, therefore, the recognition, the binding and the entry of the peculiar sperm of oniscidean isopods into the egg occur, still remain all to be deciphered. Microsc. Res. Tech. 79:792–798, 2016. © 2016 Wiley Periodicals, Inc.     

Ultrastructure of the bullfrog (Lithobates catesbeianus‐Shaw, 1802) oviduct in different seasons

The bullfrog (Lithobates catesbeianus) is an exotic animal that adapted perfectly to the climatic conditions of Brazil after introduction in the country. These climatic conditions favor the reproduction and fattening of bullfrogs. However, the development of the bullfrog is significantly reduced in colder regions because low temperatures affect the secretion of gonadotropins and change the sensitivity of the germinal epithelium to gonadotropic hormones. Thus, the temperature and photoperiod are abiotic factors that influence the life cycle of this species, especially with regard to reproduction. Samples from different regions of the oviducts (the infundibulum, magnum, and isthmus) of 9 bullfrogs were collected in the different seasons of the year. The samples were fixed and processed for analysis with scanning and transmission electron microscopy. A comparison of the morphology of the oviducts collected during different seasonal periods suggests that the frogs have greater reproductive potential in the spring due to the development of the structures related to the oviduct, presence of developed cilia, and electron‐dense granules. However, seasonal changes were noted in the animal throughout the year in preparation for reproduction during the spring and summer. Microsc. Res. Tech. 76:523–532, 2013. © 2013 Wiley Periodicals, Inc.     

Antennal morphology and sensilla ultrastructure of three tomicus species (coleoptera: Curculionidae,scolytinae)

The antennal morphology and sensilla ultrastructure of Tomicus yunnanensis, T. minor, and T. brevipilosus were studied by scanning electron microscopy and transmission electron microscopy. Eight common sensilla types were recorded: (1) sensilla trichodea (S.tr.) types 1 and 2 were located on the club and were innervated by five and eight dendrites, respectively; (2) sensilla chaetica (S.ch.) types 1 and 2 had no dendrites in the sensilla lymph lumen; (3) sensilla basiconica (S.b.), top‐protuberated S.b. and fluted cones (Fl.c.) occurred on the club; and Böhm bristles (B.b.) occurred on the funicle. S.b. were the most abundant and were innervated by 10–14 dendritic branches. Top‐protuberated S.b., a new sensilla type, were innervated by one dendrite. Fl.c. were innervated by five dendrites. Only three sensilla furcatea were visible on the antennae of female T. yunnanensis. The possible functions of these sensilla are discussed in relation to their morphology and ultrastructure. No statistical differences between sexes were found in the size and numbers of each sensilla type. Although the three species had similar antennal morphology and sensilla type, sensilla on sub‐segments 3 and 4 of the antennal club of T. minor were much sparser than those of T. yunnanensis or T. brevipilosus. Concerning the antennae of T. yunnanensis, there were more S.tr. type 2, S.ch. type 2 and S.b. and the size of S.tr. type 1 and S.b. were significantly greater than those of T. minor. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.