Effects of sublethal doses of imidacloprid in malpighian tubules of africanized Apis mellifera (Hymenoptera,Apidae)

In Brazil, imidacloprid is a widely used insecticide on agriculture and can harm bees, which are important pollinators. The active ingredient imidacloprid has action on the nervous system of the insects. However, little has been studied about the actions of the insecticide on nontarget organs of insects, such as the Malpighian tubules that make up the excretory and osmoregulatory system. Hence, in this study, we evaluated the effects of chronic exposure to sublethal doses of imidacloprid in Malpighian tubules of Africanized Apis mellifera. In the tubules of treated bees, we found an increase in the number of cells with picnotic nuclei, the lost of part of the cell into the lumen, and a homogenization of coloring cytoplasm. Furthermore, we observed the presence of cytoplasmic vacuolization. We confirmed the increased occurrence of picnotic nuclei by using the Feulgan reaction, which showed the chromatin compaction was more intense in the tubules of bees exposed to the insecticide. We observed an intensification of the staining of the nucleus with Xylidine Ponceau, further verifying the cytoplasmic negative regions that may indicate autophagic activity. Additionally, immunocytochemistry experiments showed TUNEL positive nuclei in exposed bees, implicating increased cell apoptosis after chronic imidacloprid exposure. In conclusion, our results indicate that very low concentrations of imidacloprid lead to cytotoxic activity in the Malpighian tubules of exposed bees at all tested times for exposure and imply that this insecticide can alter honey bee physiology. Microsc. Res. Tech. 76:552–558, 2013. © 2013 Wiley Periodicals, Inc.     

Structural and ultrastructural features of the Malpighian tubules of Dolycoris baccarum (Linnaeus 1758), (Heteroptera: Pentatomidae)

The morphology and ultrastructure of the Malpighian tubules of Dolycoris baccarum were analyzed by scanning (SEM) and transmission (TEM) electron microscopy in order to determine their functional organization. The Malpighian tubules are compared with similar structures of other insects based on cell structure and functional organization. The Malpighian tubules of D. baccarum extend from the midgut–hindgut region of the digestive tract. The Malpighian tubules are divided into two regions: the proximal segment is short and flattened and the distal segment is long, stringy in shape and free in hemolymph. The tubules are generally long and narrow. There is a large number of trachea around the tubules. They consist of a single layer of epithelial cells. It is observed in the TEM observation that the epithelial cells have numerous microvilli at the apical side of the cells. At the basal side of the cells, there is a great number of membrane foldings and mitochondria among them. Besides some spherites, mitochondria, lysosome‐like bodies, and large or small granules can be distinguished in the cells. With this study, we aimed to demonstrate the ultrastructure of the Malpighian tubules of D. baccarum and differences or similarities with other species.     

Morphological characterization of the nymphs Rhipicephalus sanguineus ticks (Latreille, 1806) (Acari: Ixodidae). Description of the testes, integument, Malpighian tubules, and midgut on the detachment day

This study presents the morpho-histological and histochemical characterization of the testes, integument, Malpighian tubules, and midgut of engorged Rhipicephalus sanguineus nymphs on the detachment day, showing the morphological and physiological characteristics to this phase in the life cycle of these individuals. The testis is constituted by germinative cells (only spermatogonia) with large, round-shaped and strongly stained nuclei which are organized into cysts by a thin layer of somatic cells. The integument consists of a cuticle subdivided into epicuticle (lipoprotein) and procuticle (glycoproteic), and a layer of epithelial cells which present glycolipoprotein elements. The procuticle presents two distinct regions: the exocuticle (next to the epicuticle) and the endocuticle (next to the epithelial layer). The Malpighian tubules present a simple epithelium with small flat and/or cubic cells, which form its wall and delimitates a lumen full of lipoprotein material. The midgut consists of an epithelial wall formed by two types of digestive cells, spent cells and empty digest cells, and by generative cells supported by a basal lamina and a thin layer of muscular tissue. This study described the main organs of engorged nymphs of R. sanguineus, to generate information that can help researchers to better understand the biology of these ectoparasites; which is fundamental for the development of compounds that are less aggressive to the environment. In addition, if the immature stages of the ticks are controlled, the number of adult ticks able to cause damages to the animals--and to the man as well--is also under control.     

Influence of the insecticide pyriproxyfen on the flight muscle differentiation of Apis mellifera (Hymenoptera, Apidae)

The Brazilian africanized Apis mellifera is currently considered as one of the most important pollinators threatened by the use of insecticides due to its frequent exposition to their toxic action while foraging in the crops it pollinated. Among the insecticides, the most used in the control of insect pragues has as active agent the pyriproxyfen, analogous to the juvenile hormone (JH). Unfortunately the insecticides used in agriculture affect not only the target insects but also beneficial nontarget ones as bees compromising therefore, the growth rate of their colonies at the boundaries of crop fields. Workers that forage for provisions in contaminated areas can introduce contaminated pollen or/and nectar inside the beehives. As analogous to JH the insecticide pyriproxyfen acts in the bee's larval growth and differentiation during pupation or metamorphosis timing. The flighty muscle is not present in the larvae wingless organisms, but differentiates during pupation/metamorphosis. This work aimed to investigate the effect of pyriproxyfen insecticide on differentiation of such musculature in workers of Brazilian africanized honey bees fed with artificial diet containing the pesticide. The results show that the bees fed with contaminated diet, independent of the insecticide concentration used, show a delay in flight muscle differentiation when compared to the control.     

Embryonic development of the alimentary canal of the scorpionfly Panorpa obtusa Cheng (Mecoptera: Panorpidae)

The embryonic origin of the alimentary canal, especially the midgut, is a controversial problem in insects, and it has not been satisfactorily resolved to date. The organogenesis of the digestive system in the embryonic development was observed in the scorpionfly Panorpa obtusa Cheng using light, transmission, and scanning electron microscopy. The embryonic development lasts about 150–160 h at 24°C. The stomodaeum is formed from an invagination in the medioposterior portion of the protocephalon mid‐ventrally posterior to the labral segment at 76 h after oviposition. The proctodaeum arises as an invagination from the caudal end of the abdomen at 78 h. Four anal forks are formed from within the opening of proctodaeum. Three pairs of proctodaeal evaginations are formed from the anterior part of the proctodaeum, and eventually developing into Malpighian tubules, thus are of ectodermal origin. The cardiac and pyloric valves develop from stomodaeum and proctodaeum, respectively, and also of ectodermal origin. The midgut epithelium originates from anterior and posterior midgut rudiments in blind ends of the stomodaeum and proctodaeum, and it is of endodermal origin. The two cell‐bands (rudiments) cover the yolk ventrally and then dorsally, elongate to each other, and eventually fuse to form the midgut. The midgut formation pattern is briefly discussed in different insects. Microsc. Res. Tech. 76:457–466, 2013. © 2013 Wiley Periodicals, Inc.     

Ultrastructural and developmental toxicity of potato and tomato leaf extracts to beet armyworm,Spodoptera exigua (lepidoptera: noctuidae)

Beet Armyworm, Spodoptera exigua is a herbivorous moth and a serious pest of many economically important plants, which are used as food sources. Because of rigorous standards of food quality, usage of synthetic insecticides in crop protection, against pests, is limited. Solanaceae plant extracts may be a relatively cheap source of efficient natural insecticides that can limit usage of synthetic substances. Their biological activity is not fully known. In particular, ultrastructural studies, using transmission electron microscopy, are not usual. In the present article we describe the effects of sublethal concentrations of tomato and potato leaf extracts against S. exigua. Acute lethal effects were not observed. Both extracts exerted similar effects within midgut and fat body cells. Midgut cells were not significantly altered while fat body cells showed prominent swelling of nuclear envelope and endoplasmic reticulum, vacuolization of mitochondria and fusion of fat droplets. These changes were much more intensive within groups exposed to potato than tomato extracts at highest concentration at least. Light microscopy was used to observe and document developmental alterations of S. exigua exposed to potato and tomato leaf extracts. Potato leaf extracts significantly decreased hatching success and caused morphological malformations of imagoes. Among them, malformations of wings were the most prominent. Interestingly, these effects were not observed within populations exposed to tomato extracts at highest concentration at least.     

Sewage sludge toxicity in edaphic organism: analysis of midgut responses in the diplopod Rhinocricus padbergi

Diplopods are agile saprophagous invertebrates with the ability to colonize several soil layers. They feed on waste and decaying organic matter, making the soil in which they are present better aerated and mineral‐enriched. Because of their habits, diplopods have been suggested to be used in studies to analyze possible toxic effects of impacted soils in edafic fauna. In that respect, this study aimed at checking the feasibility of using morphological analysis from millipede's midgut as a biomarker for soils exposed to sewage sludge from sewage treatment plants. Histological and histochemical techniques, applied to the species Rhinocricus padbergi's midgut, were used. Two sewage sludge samples were used; they were collected in two small towns in the countryside of São Paulo State. Both cities are part of the Piracicaba–Capivari–Jundiaí basin (PCJ‐1 and PCJ‐3). For 7 and 15‐day periods, the animals were exposed to 10% concentrations and crude sludge for the PCJ‐1sample and to 1, 10, and 50% concentrations and sludge for the PCJ‐3 sample. The material exposed to the PCJ‐1 sample showed an increase in the presence of cytoplasmic granules in both concentrations in all periods and a slight increase in the rate of epithelial renewal. The material of PCJ‐3 sample presented the same alterations observed in PCJ‐1 sample in higher frequence and over that high quantity of neutral polysaccharides in the hepatic cells layer. We can conclude that with the increase of sewage sludge concentrations tested, there is an increase in morphophysiologicals alterations frequencies found in the diplopods' midgut. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Huge clusters of embryonic stem cells in human embryos: A morphologic study

Background: Nothing is known about huge clusters (HC) of embryonic stem cells (ESC) in human fetal organs (HFO). Aim: To know the status of HC‐ESC in HFO. Methods: Morphology and immunohistochemistry (IHC) in 32 HFO of 7–40 gestational weeks (GW). Results: HC‐ESC were seen in many HFO including central nervous system, spinal cords, spine, soft tissue, bone, skin, thyroid, lung, liver, pancreas, gall bladder, extrahepatic bile duct, adrenal, kidney, bladder, foregut, midgut, hindgut, female and male genital organs, and neurons. HC‐ESC's were composed of two populations depending on constituting cells. One were large cells with ample acidophilic cytoplasms with vesicular nuclei and nucleoli. The other were small cells with scant cytoplasm with hyperchromatic nuclei without nucleoli, resembling lymphocytes. The HC‐ESC were frequently showed neuronal differentiation. HC‐ESC were positive for NCAM, synaptophysin, NSE, chromogranin, PDGFRA, AFP, ErbB2, bcl‐2, KIT, MET. They were negative for CD45, CD3, CD20, EMA, CEA, CA19‐9, cytokeratin (CK) 7, CK8, CK18, CK19, MUC1, MUC2, MUC5AC, and MUC6. The mean Ki‐67 labeling index (LI) was 13% ± 7%. HC‐ESC showed a little glycogen but lacked mucins. These HC‐ESC were seen in 7–25 GW, and they were rarely seen in 26–40 GW. Conclusions: The morphology, IHC, and ontogeny of HC‐ESC were described. Microsc. Res. Tech. 77:825–831, 2014. © 2014 Wiley Periodicals, Inc.     

Localization of metals in cells of pterygote insects

Insects maintain the equilibrium of their internal milieu by diffusion mechanisms, but more frequently by a substantial storage of metals in the cells of numerous organs: digestive tract, Malpighian tubules, fat body, integument, and genital organs. This storage implies the precipitation, of numerous cations (Ca, Mg, K, Mn, Fe, Zn, Cu) in a structure called the spherocrystal, which originates from the endoplasmic reticulum-Golgi complex: elements precipitate on a glycosaminoglycan nucleus in thin peripheral strata. Some spherocrystals contain exclusively mineral compounds, frequently phosphates, whereas others may contain organic compounds such as urates. In some species mineralized lysosomes store Ca, Fe, Zn, and Cu. When fed additional metals found in the environment, insects such as cockroach and ant are able to stay alive and to trap the metals (Cd or Pb, for example) in the peripheral strata of spherocrystals; the cytoplasm is not altered. It seems that these insects are able to resist exposures to high levels of toxic metals. The lysosomes are able to retain toxic heavy metals (Cd or Hg, for example) within metallothionein-like proteins.     

Penetrability of AH plus and MTA fillapex after endodontic treatment and retreatment: A confocal laser scanning microscopy study

The aim of the study was to assess the penetrability of two endodontic sealers (AH Plus and MTA Fillapex) into dentinal tubules, submitted to endodontic treatment and subsequently to endodontic retreatment. Thirty ex vivo incisors were prepared using ProTaper rotary system up to F3 instrument and divided in three groups according to the endodontic sealer used for root canal filling: AH Plus (AHP), MTA Fillapex (MTAF), and control group (CG) without using EDTA previously to the root canal filling. Rhodamine B dye (red) was incorporated to the sealers in order to provide the fluorescence which will enable confocal laser scanning microscopy (CLSM) assessment. All specimens were filled with gutta‐percha cones using the lateral compaction technique. The specimens were submitted to endodontic retreatment using ProTaper Retreatment system, re‐prepared up to F5 instruments and filled with gutta‐percha cones and the same sealer used during endodontic retreatment. Fluorescein dye (green) was incorporated to the sealer in order to distinguish from the first filling. The roots were sectioned 2 mm from the apex and assessed by CLSM. No difference was found between the two experimental groups (P > 0.05). On the other hand, in the control group the sealers were not capable to penetrate into dentinal tubules after endodontic treatment (P > 0.05). In retreatment cases, none of the sealers were able to penetrate into dentin tubules. It can be concluded that sealer penetrability is high during endodontic treatment. However, MTA Fillapex and AH Plus do not penetrate into dentinal tubules after endodontic retreatment. Microsc. Res. Tech. 77:467–471, 2014. © 2014 Wiley Periodicals, Inc.     

20(R)-ginsenoside Rg3, a product of high-efficiency thermal deglycosylation of ginsenoside Rd,exerts protective effects against scrotal heat-induced spermatogenic damage in mice

Heat stress (HS) reaction can lead to serious physiological dysfunction associated with cardiovascular and various organ diseases. Ginsenoside Rg3 (G-Rg3) is a representative component of ginseng rare saponin and can protect against multiple organs, also used as functional food to adjust the balance of the human body, but the therapeutic effect and molecular mechanism of G-Rg3 on male diseases under HS are underexplored. The aim of the present study, G-Rg3 was prepared through the efficient conversion of ginsenoside Rd and investigate the contribution of G-Rg3 to testicular injury induced exposure to HS. All mice were divided into four groups as follows: normal group, HS group, and HS+G-Rg3 (5 and 10 mg/kg) groups. G-Rg3 was administered orally for 14 days, then exposed to a single scrotal heat treatment (43°C, 18min) on the 7th day. After HS treatment, the morphology of testis and epididymis changes, and caused a significant loss of multinucleated giant cells, desquamation of germ cells in destructive seminiferous tubules, and degenerative Leydig cells, further destroying the production of sperm. After administration G-Rg3 (5 and 10 mg/kg/day) for 2 weeks, the spermatogenic-related indexes of testosterone levels and superoxide dismutase (SOD) activity, glutathione (GSH) content significantly (p < 0.01) increase compared with the HS group. Moreover, G-Rg3 treatment effectively ameliorated the production of malondialdehyde (MDA) (p < 0.05 or p < 0.01). Importantly, G-Rg3 exhibited the protective potential against HS-induced injury not only suppressing the protein levels of heme oxygenase-1 (HO-1), hypoxia-inducible factor-1α (HIF-1α), and heat shock protein 70 (HSP70) but also modulating the Bcl-2 family (p < 0.01 or p < 0.001) and activation of mitogen-activated protein kinase (MAPK) signaling pathways (p < 0.01). For most of the parameters tested, the HS+G-Rg3 (10 mg/kg) group exhibited potent effects compared with those exhibited by the low dose (5 mg/kg) group. In conclusion, the present study demonstrated that G-Rg3 exerted protective effects against HS-induced testicular dysfunction via inhibiting the MAPK-mediated oxidative stress and apoptosis in mice.     

Post-embryonic development of the antennal sensilla in Melipona quadrifasciata anthidioides (Hymenoptera: Meliponini)

The sensilla are sensory organs formed by cuticular and cellular structures specialized in reception of chemical and physical stimuli from the environment and transmission to the insect's central nervous system. In function of the great concentration of sensilla, the antennae are the main organs for interaction between bees and with the environment. This work studied the presence of antennal sensilla in the different phases of pupal development of the stingless bee Melipona quadrifasciata anthidioides by means of scanning electron microscopy and light microscopy. The results showed that antennal sensilla begin their development in the transition of the prepupae to the white-eyed pupae and finish it in the pigmented-body pupae phase. The antennal sensilla were exposed to the environment in the black-eyed pupae when the old cuticle is completely digested, suggesting that only in the final pupal phases can these bees perceive the environmental stimuli.     

Histopathological lesions and toxicity in common carp (Cyprinus carpio L. 1758) induced by copper nanoparticles

Different types of metal oxide nanoparticles (NPs) have been suggested for various applications such as water treatment and construction of agricultural pesticides; however, there are concerns about the potential toxicity of these compounds for the nontarget organism especially aquatic organisms. The aims of this study were assessing toxicity and histopathological effects of copper oxide NPs (NPs‐CuO) on common carp (Cyprinus carpio) as a model organism. For this purpose 150 common carp with an average weight 7 ± 1 g were exposed to 0, 10, 20, 30, 40, 60, 80, 100, 150, and 200 mg/l of CuO‐NPs (10 treatment with three replicates) for 96 hrs. After 24, 48, 72, and 96 hrs exposures, mortality rates recorded and gill samples were collected. Statistical analysis showed significant differences in carp survival between control and treatment groups (p < 0.05); regression between fish mortality rate and NPs‐CuO concentration was also revealed (p < 0.01). The LC₅₀ 96h of NPs‐CuO for common carp was estimated as 124.9 mg/l in this study. Various tissue damages were observed in gill of treatments; such as, hypertrophy, hyperplasia, lamellar fusions, erythrocyte infiltration, epithelial lifting; also, there was significant correlation between intensity of tissue lesions and concentration of NPs‐CuO (p < 0.01). The findings of the present study demonstrate that sublethal concentration of NPs‐CuO can lead to serious tissue lesions. Whats more, concentrations above 30 ml/l of NPs‐CuO can lead to some clinical signs; such as skin darkening and death with open mouth as well as definite fish death.     

Development beyond the gastrula stage and digestive organogenesis in the apple-snail <i>Pomacea canaliculata</i> (Architaenioglossa,Ampullariidae)

Development of Pomacea canaliculata from the gastrula stage until the first day after hatching is described. Trochophore embryos are developed after gastrulation, showing the prototroch as a crown of ciliated orange-brownish cells. However, no true veliger embryos are formed, since the prototroch does not fully develop into a velum. Afterward, the connection between the fore- and midgut is permeated and the midgut becomes full of the pink-reddish albumen, which is stored into a central archenteron’s lake, from where it is accumulated into the large cells forming the midgut wall (“giant cells”). Electron microscopy of giant cells in late embryos showed that albumen is engulfed by large endocytic vesicles formed between the irregular microvilli at the top of these cells. By the end of intracapsular development, giant cells become gradually replaced by two new epithelial cell types which are similar to those found in the adult midgut gland: the pre-columnar and the pre-pyramidal cells. Pre-columnar cells have inconspicuous basal nuclei and are crowned by stereocilia, between which small endocytic vesicles are formed. Pre-pyramidal cells have large nuclei with 2-3 nucleoli and show a striking development of the rough endoplasmic reticulum. The genesis of the three cell lineages (giant, pre-columnar and pre-pyramidal cells) is hypothetically attributed to epithelial streaks that occur at both sides of the midgut since early stages of development.     

Enamel erosion and prevention efficacy characterized by confocal laser scanning microscope

The aim of this study was to evaluate the erosion‐inhibiting effect of two toothpastes on the development of erosion‐like lesions, by a confocal laser scanning microscope (CLSM). Forty human enamel blocks were divided into five groups (n = 8), in accordance to evaluate the GC MI Paste Plus and Oral B with stannous fluoride, applied as slurries and associated with toothbrush. Specimens were submitted to an erosion challenge from citric acid (0.5%, pH = 2.8), for 5 min, six times a day, alternating in artificial saliva immersions. Reference group was not exposed to treatment. Part of specimens (Groups 02 and 03) was exposed twice daily just to slurries, for 2 min, therefore specimens from Groups 04 and 05 were also abraded, for 30 s. The enamel surfaces were morphological characterized using CLSM images, with mineral loss being measured using the resulting 3D images referenced to an un‐challenged portion of the sample. Step values were compared using the one‐way ANOVA test. CLSM was shown to be a viable, noncontact, and simple technique to characterize eroded surfaces. The statistical difference in the step size was significant between the groups (P = 0.001) and using multiple comparisons a statistically significant protective effect of toothpastes was shown when these were applied as slurries. Although groups submitted to tooth brush showed mineral loss similar to reference control group, due to the damages of abrasion associated. Microsc. Res. Tech. 77:439–445, 2014. © 2014 Wiley Periodicals, Inc.     

Caries removal with Er:YAG laser followed by dentin biomodification with carbodiimide and chitosan: Wettability and surface morphology analysis

This study aimed to investigate dentin wettability and surface morphology after selective removal of carious lesion by erbium‐doped yttrium aluminum garnet (Er:YAG) laser, followed by dentin biomodification with carbodiimide (EDC) and chitosan (CHI). Seventy‐eight bovine dentin specimens were submitted to caries induction. Specimens were distributed according to methods of carious removal (n = 39): bur at low‐speed (40,000 rpm) or Er:YAG laser (noncontact mode, 250 mJ/pulse and 4Hz). All specimens were etched with 35% phosphoric acid, and subdivided according to dentin biomodification (n = 13): Control (no biomodification), EDC or CHI. The contact angle (n = 10) between adhesive system (3M ESPE) and dentin surface was measured by a goniometer. Eighteen specimens (n = 3) were analyzed by scanning electron microscopy. Data were analyzed by two‐way ANOVA and Tukey's test (α = .05). The method used to remove carious lesion did not influence the wettability of dentinal surface (p = .748). The angles produced on the remaining dentin after biomodification were influenced (p = .007). CHI promoted higher contact angles (p = .007) and EDC did not differ from the control group (p = .586). In the bur‐treated group, most tubules were open, regardless of which biomodifier was used. Laser modified the organic matrix layer. CHI promoted partially closed tubules in some areas while EDC exposed dentinal tubules. Regardless of which method was used for selective removal of carious lesion, biomodification with EDC did not affect the dentin wettability, whereas CHI changed the wettability of remaining dentin. Both biomodifiers promoted a slight change on dentin morphology.     

Toxicity of trifluralin herbicide in a representative of the edaphic fauna: Histopathology of the midgut of Rhinocricus padbergi (Diplopoda)

The aim of this study was to use the diplopod Rhinocricus padbergi as test organism to verify the possible toxic effects of the herbicide trifluralin in a representative of the edaphic fauna. Histological and histochemical techniques were applied in the midgut of the animals after exposure to different concentrations of trifluralin for 7 and 90 days (acute and subchronic exposure, respectively). The individuals from the control group did not present alterations to any of the exposures. In the acute exposure, the group exposed to a quarter and half of the field dose presented an accumulation of cytoplasmic granules in the hepatic cells; the field dose group presented an increase in the rate of epithelial renewal and the group exposed to double the field concentration presented an increase in the release of secretory vesicles. The subchronic response was obtained with the field dose and double field dose because the animals from a quarter and half groups did not survive until the end of the experiment. In both groups, the animals presented disruption of the epithelium and higher occurrence of hemocytes among the hepatic cells. The group exposed to double the field dose presented sites of cytoplasmic vacuolization, dilatation of the intercellular space, increase in the epithelial renewal, and release of secretory vesicles. The results alert for a careful use of this herbicide, since it promoted alterations in the midgut cells of the test organism used. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Morphological alterations of radicular dentine pretreated with different irrigating solutions and irradiated with 980-nm diode laser

Background: The topographical features of intraradicular dentine pretreated with sodium hypochlorite (NaOCl) or ethylenediamine tetraacetic acid (EDTA) followed by diode laser irradiation have not yet been determined. Purpose: To evaluate the alterations of dentine irradiated with 980‐nm diode laser at different parameters after the surface treatment with NaOCl and EDTA. Study design: Roots of 60 canines were biomechanically prepared and irrigated with NaOCl or EDTA. Groups were divided according to the laser parameters: 1.5 W/CW; 1.5 W/100 Hz; 3.0 W/CW; 3.0 W/100 Hz and no irradiation (control). The roots were splited longitudinally and analyzed by scanning electron microscopy (SEM) in a quali‐quatitative way. The scores were submitted to two‐way Kruskal–Wallis and Dunn's tests. Results: The statistical analysis demonstrated that the specimens treated only with NaOCl or EDTA (control groups) were statistically different (P < 0.05) from the laser‐irradiated specimens, regardless of the parameter setting. The specimens treated with NaOCl showed a laser‐modified surface with smear layer, fissures, and no visible tubules. Those treated with EDTA and irradiated by laser presented absence of smear layer, tubules partially exposed and melting areas. Conclusions: The tested parameters of 980‐nm diode laser promoted similar alterations on dentine morphology, dependent to the type of surface pretreatment. Microsc. Res. Tech., 2009. © 2008 Wiley‐Liss, Inc.     

Anatomical and histological studies of the alimentary canal of adult maize leaf weevil,Tanymecusdilaticollis Gyllenhal, 1834 (Coleoptera: Curculionidae)

In this study, ananatomical and histological study was conducted on the alimentary canal of Tanymecusdilaticollis (Coleoptera: Curculionidae), which is an economic polyphagous pest species, to study the relationship between the structure of the alimentary canal and the feeding habit. Therefore, the structure of the alimentary canal of T. dilaticollis was examined using light and electron microscopies. Results have shown that the alimentary canal in T. dilaticollis is consisted of three separate regions as foregut, midgut, and hindgut structurally between the mouth and the anus, which pass from head, thorax, and abdomen. The foregut consists of pharynx, esophagus, crop and proventriculus and in the crop part, expansion is seen compared to other foregut parts. Midgut of T. dilaticollis is the largest part of digestion system. The anterior region of midgut is twofolds wider than the posterior region. The posterior midgut extends tubularly and it is connected to eightgastric caeca. The hindgut of T. dilaticollis consists of fourparts as pylorus, ileum, colon, and rectum. Well‐developed muscle layers are found near the rectum and genital chamber. These results contribute to further studies on the ecology and biological control agents of Coleoptera and to provide a broad comparison of alimentary canal of Coleoptera species.     

Embryonic development of endoderm in chicken (Gallus gallus domesticus)

The poultry industry is a sector of agribusiness which represents an important role in the country's agricultural exports. Therefore, the study about embryogenesis of the domestic chicken (Gallus gallus domesticus) has a great economic importance. The aim of this study was to evaluate embryonic development of the endoderm in chicken (Gallus gallus domesticus). Forty fertilized eggs of domestic chickens, starting from the 1st day of gestation and so on until the 19 days of the incubation were collected from the Granja São José (Amparo, SP, Brazil). Embryos and fetus were fixed in 10% formaldehyde solution, identified, weighed, measured, and subjected to light and scanning electron microscopy. The endoderm originates the internal lining epithelium of the digestive, immune, respiratory systems, and the organs can be visualized from the second day (48 h) when the liver is formed. The formation of the digestive system was complete in the 12th day. Respiratory system organs begin at the fourth day as a disorganized tissue and undifferentiated. Their complete differentiation was observed at the 10 days of incubation, however, until the 19 days the syrinx was not observed. The formation of immune system at 10th day was observed with observation of the spleen, thymus, and cloacal bursa. The study of the organogenesis of the chicken based on germ layers is very complex and underexplored, and the study of chicken embryology is very important due the economic importance and growth of the use of this animal model studies such as genetic studies. Microsc. Res. Tech. 76:803–810, 2013. © 2013 Wiley Periodicals, Inc.