Ultrastructure of the ovariole sheath in <i>Diatraea saccharalis</i> (Lepidoptera: Pyralidae)

The ultrastructure of the ovariole sheath along the Diatraea saccharalis ovariole was studied by scanning and transmission electron microscopy. Each ovariole is surrounded by an epithelial sheath, a tunica propria and scattered lumen cells. These three components of the ovariole sheath show different ultrastructural features along the ovariole, in the germarium or in the vitellarium; these differences are more evident in the epithelial sheath cells. The epithelial sheath is composed by two layers of cells, the external one running longitudinally and the internal one running circularly in the ovariole. These cells, in vitellarium, present cytoplasmic bundles of myofilaments that are arranged parallel to the long axis of the cells; these myofilaments are apparently related to the contraction movements of the follicles within the ovariole. The acellular tunica propria, composed of finely filamentous material, is attached to the adjacent follicle cells by adhesive dense plates. Between the epithelial sheath and the tunica propria there is a population of lumen cells, with morphological features of secretory activity     

Ultrastructural observations on antennal sensilla of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)

The morphology and ultrastructure of the antennal sensilla of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) was studied by scanning and transmission electron microscopic techniques. Eight morphological types of sensillum were recorded in both sexes: sensilla trichodea (S. trichodea), sensilla basiconc (S. basiconc), sensilla coeloconica (S. coeloconica), sensilla styloconica (S. styloconica), sensilla squamous (S. squamous), sensilla auricillica (S. auricillica), B?hm bristles (B. bristles), and sensilla cavity (S. cavity). S. trichodea were the most abundant sensilla and were distributed over the entire antennal surface. Four different types of S. trichodea and S. basiconc were observed. The number of S. basiconc and S. coeloconica of males were greater than those of females of C. medinalis. S. squamous formed on the dorsal part of the antenna, as a cluster in females and as in a line in males. Higher magnification revealed that S. basiconc had an olfactory function, while the character of longer length of these sensilla suggested that they also played a rolein sensing mechanical or other chemical stimuli. Transmission electron microscopy (TEM) of S. squamous revealed nonporous walls suggesting a non-olfactory function. S. coeloconica, S. styloconica, and S. cavity may be involved in the perception of humidity, temperature, heat, and CO?. Because of their particular location, we infer that B. bristles may function in sensing the position and movements of the antennae, while the function of S. auricillica is as yet unknown. The results obtained provide direct morphological evidence that antennae possess structures that can play a role in finding mates and locating host plants.     

Ultrastructure of antennal sensilla of Erannis ankeraria Staudinger (Lepidoptera: Geometridae)

Erannis ankeraria Staudinger, 1861 (Lepidoptera: Geometridae) is one of the major pests causing serious damages on Larix spp., Quercus L., and Picea Mill. To investigate the conceivable functions of antennal sensilla associated with pheromone detection, we observed the ultrastructure of antennae in female and male of E. ankeraria moths by scanning electron microscopy. Six types (including two subtypes and a new type) of sensilla were recorded and characterized, including Sensilla trichodea (ST I and ST II), Böhm bristles, Sensilla squamiformia, Sensilla chaetica, Sensilla auricillic (SAU) and newly observed serrate‐like sensilla. ST I and SAU were abundant on male antennae, displaying a sexual dismorphism. Serrate‐like sensilla were also peculiar to male antennae. In summary of reported functions of corresponding sensilla, we presumed putative functions of the recorded sensilla in E. ankeraria, providing the morphological basis for sensory mechanisms related to pest management.     

Ultrastructure of larvae and puparia of the blowfly Chrysomya megacephala (Diptera: Calliphoridae)

Chrysomya megacephala is a forensic important fly, and its immature forms also cause myiasis. The adults are the first insects to reach a carcass and can oviposit just a few hours after arrival. Therefore, the knowledge of immature stages of flies is essential for correct identification of the species found on corpses. Scanning electron microscopy (SEM) gives detailed information on the morphological characters that can help identify the immature forms of flies. The aim of this study is to identify larvae and puparia of C. megacephala using SEM. The larval instar body of C. megacephala is similar at all instars. The integument is smooth with small spines located at the limit of all segments. The cephalic region has a group of robust spines with one or two tips. The puparia are very similar to third instar larvae, except for the cephalic structures that are retracted. The integument shows the wrinkles from the third instar larvae and posterior spiracle disc with three spiracular openings localized on the top of an elevation. In conclusion, SEM provides some characteristics to distinguish among Chrysomya species that could help entomologists to identify immature found on corpses.     

Nitric oxide synthase activity in tissues of the blowfly Chrysomya megacephala (Fabricius, 1794).

Although insects lack the adaptive immune response of the mammalians, they manifest effective innate immune responses, which include both cellular and humoral components. Cellular responses are mediated by hemocytes, and humoral responses include the activation of proteolytic cascades that initiate many events, including NO production. In mammals, nitric oxide synthases (NOSs) are also present in the endothelium, the brain, the adrenal glands, and the platelets. Studies on the distribution of NO-producing systems in invertebrates have revealed functional similarities between NOS in this group and vertebrates. We attempted to localize NOS activity in tissues of na?ve (UIL), yeast-injected (YIL), and saline-injected (SIL) larvae of the blowfly Chrysomya megacephala, using the NADPH diaphorase technique. Our findings revealed similar levels of NOS activity in muscle, fat body, Malpighian tubule, gut, and brain, suggesting that NO synthesis may not be involved in the immune response of these larval systems. These results were compared to many studies that recorded the involvement of NO in various physiological functions of insects.     

Protein detection in spermatids and spermatozoa of the butterfly Euptoieta hegesia (Lepidoptera).

This study was undertaken to detect protein components in both sperm types of the butterfly Euptoieta hegesia. These spermatozoa possess complex extracellular structures for which the composition and functional significance are still unclear. In the apyrene sperm head, the proteic cap presented an external ring and an internal dense content; basic proteins were detected only in external portions. In the tail, the paracrystalline core of mitochondrial derivatives and the axoneme are rich in proteins. The extratesticular spermatozoa are covered by a proteic coat, which presented two distinct layers. In eupyrene spermatozoa, acrosome and nucleus were negatively stained, probably because of their high compaction. In the tail, there is no paracrystalline core and the axoneme presented a very specific reaction for basic proteins. The lacinate and reticular appendages are composed of cylindrical sub-units and presented a light reaction to E-PTA and a strong reaction to tannic acid. A complex proteic coat also covers the extratesticular spermatozoa. We found similarities between both extratesticular coats, indicating a possible common origin. Both spermatozoon types are rich in proteins, especially the eupyrene appendages and the extratesticular coats. We believe that both coats are related to the sperm maturation and capacitation processes.     

Ultrastructure of submandibular salivary glands of mouse: TEM and HRSEM observations

The fine structure of submandibular glands of mouse were analyzed using light microscopy (LM), high resolution scanning electron microscopy (HRSEM), and transmission electron microscopy (TEM) methods. For LM, the specimens were embedded in Spurr resin, stained by toluidin blue solutions. For TEM, the tissues of submandibular salivary glands were fixed with modified Karnovsky solution and postfixed with osmium tetroxide. For HRSEM, the tissues were fixed with 2% osmium tetroxide solution in 1/15M sodium phosphate buffer (pH 7.4). The samples were immersed successively in dymethylsulphoxide and freeze cracked. The maceration was made in diluted osmium tetroxide for 24-48 h. The samples were examined by high resolution scanning electron microscopy. The intracellular components of acinar and ductal cells revealed clearly the Golgi apparatus, rough endoplasmic reticulum, secretory granules, and mitochondria. The end bulbs of Golgi lamellae and flattened cisterns of rough endoplasmic reticulum showed the luminal surface. A few mitochondria were identified intermingling between the rough endoplasmic reticulum and the mitochondriales cristae in three-dimensional HRSEM images. Secretory granules were numerous and presented different sizes. Small granules of ribosomes were attached on cistern surface, measuring 20-25 nm in diameter. Numerous arranged microvilli were found on the luminal surface of secretory canaliculus. The contact surfaces of acinar cells revealed complicated interdigitations by cytoplasmic processes. The mitochondria of duct cells were disposed vertically and surrounded by basal infoldings of plasma membranes. Basement membrane showed a spongy-like structure having an irregular surface with various strands and meshes of fine collagen fibrils.     

Age-related changes in the avian primary lymphoid organs (thymus and bursa of Fabricius)

The avian primary lymphoid organs, the thymus and the bursa of Fabricius, undergo age-dependent changes leading in some cases to the complete atrophy of the organ. Nevertheless, the timetable of the involutive process as well as the consequences in the structure and functionality of the organs vary largely in the time frame and structural changes among species. On the other hand, and in contrast with the large body of literature reporting the structural and functional changes in mammalian primary lymphoid organs, the age-dependent changes in avian thymus and bursa of Fabricius are scarce, fragmentary, and heterogeneous. This article reviews the current literature on this topic, and focuses primarily on the involution of the bursa of Fabricius.     

Foreword: the Cosslett festschrift (Ellis Cosslett).

A method for the detection and characterization of clusters of particles observed in section with the electron microscope is presented. Cluster analysis is performed by the division method described by Berthet et al. (1976). Starting from a single cluster, profiles from each electron micrograph are successively classified in sets containing an increasing number of clusters. The decrease in the mean free distance, Λ, between profiles in the clusters, is used for terminating the subdivision procedure. The function relating the mean free distance with the number of clusters is evaluated in each subdivision set. The actual number of clusters is selected on the basis of the slope of that function, at a point where Λ has a value close to the average profile diameter. The method assumes a convex shape for the clusters; the salient feature is that it provides a physical delineation of clusters in the section. Hence, an evaluation of some characteristics of clusters in the three-dimensional sample may be obtained by using standard stereological procedures. Characterization of the volume to which the individual particles of a population are eventually restricted can as a result be performed. Practical problems in the acquisition of the data needed for cluster analysis are discussed and a system using for that purpose a Quantimet 720 image analyser in a basic configuration, connected on line with a PDP 11/10 minicomputer, is presented. Application of the method is illustrated by the analysis of lysosomes in cultured hepatoma (HTC) cells, at the end of mitosis and during the S phase. Cluster analysis shows that in mitotic cells, lysosomes are confined within 3·7% of the total cellular volume, while in cells actively synthesizing DNA they are grouped in clusters representing 5·7% of the cellular volume. Moreover, the average number of particles per cluster falls from a minimum of thirteen at mitosis to only six at the S phase.     

Morphology and distribution of antennal sensilla of female Phyllotreta striolata (Fabricius) (Coleoptera: Chrysomelidae)

Phyllotreta striolata (Fabricius) is an important pest of Brassicaceae in Southeast Asia and North America. Using scanning electron microscopy, we observed the external structure, number, and distribution of the antennal sensilla in P. striolata females to discuss the putative function of these sensilla in host location and oviposition behaviors. The antenna of female P. striolata is filiform, composed of a scape, a pedicel, and a flagellum with 9 flagellomeres. Five types of sensilla were identified, including sensilla cheaetica, sensilla trichodea, Böhm bristles, sensilla auricillica, and sensilla basiconica (five subtypes, SB1–SB2). External structure and distribution of antennal sensilla are compared with data from other insect species. In addition, we discuss the possible functions of antennal sensilla based on their characteristics. Microsc. Res. Tech. 79:219–226, 2016. © 2016 Wiley Periodicals, Inc.     

Epidermal glands in the abdomen of a basal ant Dinoponera lucida (Formicidae: Ponerinae)

The basal ant Dinoponera (Hymenoptera: Ponerinae) has lost the morphologic queen caste so that all females may be potential reproductive individuals, and the nestmate recognition results from cuticular hydrocarbons cues. However, data about the origin of that substance in Ponerinae ants are scarce. This study reports the occurrence of epidermal glands in the abdomen of the ant Dinoponera lucida. In this ant, the epidermis of the abdominal sternites has tall cells with well-developed nucleus contrasting with flattened and collapsed epidermis in the tergites, suggesting a glandular function in the epidermis of the sternites. The possible role of the glandular epidermis in the synthesis of cuticular hydrocarbons for the nestmate recognition is discussed.     

Sensilla and secretory glands in the antennae of a primitive ant: Dinoponera lucida (Formicidae: Ponerinae)

Morphology of the antennae of the female workers of the ponerine ant Dinoponera lucida was examined by light and scanning electron microscopy. In several antennomers, we found secretory gland cells of class I and III. Class III gland cells release their secretion through single pores in the antennal surface, whereas class I secretory cells are seen as tall epidermal cells close to the cuticle. Both gland types have weak reaction for total proteins and neutral polysaccharides. Six distinct sensilla types were observed: trichodea, chaetica, campaniform, basiconica, placodea, and coeloconica. The possible sensory functions of these sensilla and the gland functions are discussed.     

Fluorescence microscopy study on the cytoskeletal displacements during sperm differentiation in the bush‐cricket Tylopsis liliifolia (Fabricius) (Orthoptera: Tettigoniidae)

A study by fluorescence microscopy has been carried out on male gametes from testicular follicles, seminal vesicles, spermatophores, and seminal receptacles of the bush‐cricket Tylopsis liliifolia, focusing the attention on localization and movements of F‐actin and α‐tubulin during sperm differentiation, since data in this respect are lacking in the Orthoptera. F‐actin and α‐tubulin positivity was detected in the testicular follicles, in particular at the bridges connecting spermatids of a same clone and around their nucleus, during the first differentiation stages. During the following differentiation stages in the testes, F‐actin was found at one of the spermatid poles and then, during nucleus elongation, at the whole acrosomal region. A peculiar F‐actin‐positivity was found at the flagellum, more markedly immediately posterior to the nucleus, at the basal body region of the gametes from the testicular follicles and from the other examined districts. Other interesting data from our investigations concerns the α‐tubulin displacements during the differentiation stages of the spermatid and a constant absence of α‐tubulin‐positivity where the centrioles are located. No positivity was also found for both α‐tubulin and nuclear markers at the anterior region of the gamete, where the acrosomal wings are localized. Our results, compared with what is so far known in literature for the insects, lead us to assert that microfilaments and microtubules undergo gradual displacements, markedly in the testicular follicles, during the morphogenesis of the male gamete of T. liliifolia aimed to its organization and motility and probably also to its interaction with the female gamete. Microsc. Res. Tech. 79:81–88, 2016. © 2015 Wiley Periodicals, Inc.     

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.     

Ultrastructure of proteoglycans in the tectorial membrane

The ultrastructure of proteoglycans (PGs) in the tectorial membrane (TM) of the mature chinchilla cochlea was investigated using the cationic dye Cuprolinic blue. When used at a high critical electrolyte concentration, Cuprolinic blue has been shown specifically to bind to the glycosaminoglycan residues of sulfated PGs. After Cuprolinic blue treatment, PGs were observed in the TM which were represented as rod-shaped, electron-dense structures. A perifibrillar, primarily orthogonal, array of PGs was associated with the type A protofibrils. These PGs were distributed in 50 nm intervals along the length of the type A protofibrils. A less common orientation was parallel to the axis of the type A protofibrils. PGs did not appear to be associated with the type B protofibrils. Based upon previous results by other investigators, the TM contains types II and IX collagen, and it appears likely that the type A protofibrils are composed of collagen type II. PGs visualized in the TM in this study thus may represent the glycosaminoglycan residue of type IX collagen which is associated with the type II collagen fibrils. Alternatively, the TM PGs may be small dermatan or chondroitin sulfate PGs.     

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.     

Structural study of the salivary glands of Anocentor nitens (Acari: Ixodidae) during the feeding cycle

The salivary glands of Anocentor nitens (Neumann, 1897 ) occur in pairs and are located in the anterolateral region of the general cavity, with milky white color and approximately equal sizes. They consist of a secretory portion and an excretion duct. In some glandular acini, all the cells had a basophilic appearance they were stained by hematoxylin, whereas others presented cells with different staining affinities. In this work, we describe the variations observed in these glands during the feeding cycle of ticks [after feeding (0 h) and successively at 24, 48, 72, 96, 120, and 144 h]. The cells stained by hematoxylin were shown to be more reactive to Alcian blue, thus demonstrating the presence of acid glycosaminoglycans, whereas those stained using eosin presented weak or no reaction. A strong reaction was found by the use of the periodic acid‐Schiff (PAS) technique, thereby suggesting the presence of glycogen and/or glycoconjugates containing hexose, confirmed by using salivary amylase before PAS, with partial destaining of the slides. Continuing presence of residual staining in these cells suggests the presence of glycoconjugates containing hexose. Cells with nuclei of circular outline and few granules (of different sizes) were found in type II acini, 72 h after collection. Type I acini presented wide lumina and walls composed of larger numbers of cells of cubic to cylindrical shape. The pronounced degranulation shown in this study over the course of the feeding cycle was associated with the release of substances for oviposition. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.     

Ultrastructure of sensilla on the maxillary and labial palps of the adult Xylotrechus grayii (Coleoptera: Cerambycidae)

The longhorn beetle, Xylotrechus grayii (White, 1855), is a serious woodborer pest of many economic and medicinal plants. The maxillary and labial palps of X. grayii adults are strongly involved in mating and host selection behaviors. To further develop control strategies of the pest, sensilla on the palps were studied with scanning and transmission electron microscopy. Five types of sensilla (nine subtypes) were identified: sensilla styloconica (four subtypes, Sty1‐4), sensilla chaetica (2 subtypes, SC1‐2), sensilla digitiformia (Di), sensilla basiconica (Ba), and Böhm sensilla (Bm). Sty and Ba have dendrites and wall pores. SC2 has a tubular body. Di exist only on the terminal segment of the maxillary palp. Sty numbers are the highest in both palps, followed by SC. The number of sensilla in the maxillary palp is significantly higher than that in the labial palp. There are no significant sexual differences in sensilla types, characteristics, distribution, and quantity in the maxillary or labial palps, except for the number of SC sensilla on the maxillary palp. Sensilla functions were compared with correlative sensilla in previous studies. Sty may be sense gustation and olfaction stimulation, while SC are mechanoreceptors. Ba are likely thermo‐ and hygroreceptors. The phylliform depression area formed by Di may exert multiple functions. Bm may sense palp position and movement. These results contribute to the study of the electrophysiology and behavior mechanisms of X. grayii.     

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.