Morphophysiological analysis of the salivary glands of Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) exposed to ozonated water: A control strategy

The tick Rhipicephalus sanguineus sensu lato has great medical and veterinary importance, mainly because the ability to transmit many diseases, causing harm to pets but also risks to public health. The blood spoliation and transmission of pathogens occur because of the immunosuppressive action of these ticks' saliva, a potent mixture of bioactive substances that is secreted by the salivary glands, one of the organs responsible for their biological success, and hence the target of studies for their control. Ozone has promise for use as an alternative acaricide, due to its proven efficiency in controlling agricultural and food pests, besides posing no risk of environmental contamination or to animal and human health. Therefore, this study evaluated the acaricidal potential of exposure of females of R. sanguineus s.l. to ozonated water at many concentrations and analysed the morphophysiological alterations of the salivary glands, employing histological and light microscopic techniques. The results demonstrated that the ozonated water at the concentrations investigated caused severe alterations in the salivary glands, bringing a new perspective for control of R. sanguineus s.l., through an ecologically correct method due to the absence of harm to non‐target organisms and the environment.     

Morphology of the salivary glands of three Squamata species: Podarcis sicula sicula, Tarentola mauritanica and Coluber viridiflavus

The histology, histochemistry and ultrastructure of the salivary glands of three species of squamata, Podarcis sicula sicula (mandibular glands), Tarentola mauritanica (sublingual gland) and Coluber viridiflavus (supra- and infralabial glands) were studied. Each gland contained acidic cells, positive for both periodic acid Schiff and Alcian blue reactions. These cells can be distinguished as seromucous and mucous types based on the different electron density of their granules. α-Amylase, until now detected only in mammalian salivary glands, was not found in any of the salivary glands examined. The ultrastructural study revealed that the salivary glandular cells of T. mauritanica lack intercellular canaliculi, which by contrast, are present between the seromucous cells in the salivary glands of C. viridiflavus and P. s. sicula . Comparable variation is also seen when the ultrastructural features of the secretory granules in salivary glands of the three Squamata species are compared. The salivary granules of T. mauritanica and C. viridiflavus are more or less dense but structureless, while the mucous granules of P. s. sicula have a distinctive and characteristic substructure. Therefore, this study, designed to obtain comparative data on the histology, histochemistry and ultrastructure of the salivary glands of three representative, but hitherto unstudied, species of Squamata, reveals great variation in the structure of these glands within the Squamata lineage, even when compared to previously documented species.     

Morphologic diversity of the minor salivary glands of the rat: fertile ground for studies in gene function and proteomics

In this article the locations and histologic and ultrastructural features of all of the minor salivary glands of the rat are presented; similarities and differences among them are highlighted. These glands are almost as diverse morphologically as the major salivary glands of the rat. The acini of von Ebner's glands are serous; those of the anterior and posterior buccal glands and minor sublingual glands are mucous; and those of the glossopalatal, palatal, and Weber's glands are mucous with serous demilunes. The anterior buccal, minor sublingual and von Ebner's glands have striated and stratified columnar ducts, while only the minor sublingual and von Ebner's glands have intercalated ducts. The glossopalatal, palatal, posterior buccal and Weber's glands have none of these ducts; the tubulo-acini drain abruptly into short terminal ducts composed of stratified squamous epithelium. All of the mucous acini react with an antibody to a mucin (Muc19) of the rat major sublingual gland, but in some of the glands the reaction varies in intensity among the acinar cells. Ultrastructurally, the mucous secretory granules of the anterior buccal, glossopalatal, palatal and Weber's glands are biphasic, while those of the minor sublingual and posterior buccal glands are monophasic. Although there is a considerable body of literature concerning the development, innervation, physiology and proteomics of von Ebner's glands, investigation of the other minor salivary glands of the rat ranges from modest to nearly nonexistent.     

Ultrastructural and histochemical study of the salivary glands of Aplysia depilans (Mollusca, Opisthobranchia)

The digestive system of the sea hare, Aplysia depilans , includes a pair of ribbon-shaped salivary glands. A central duct and a large blood vessel run close to each other along the length of these glands and both are surrounded by a layer of muscle cells. Three cell types form the glandular epithelium: granular cells, vacuolated cells and mucocytes. The granular cells possess cilia and spherical secretion granules, located primarily in the apical region. The granules of immature cells have a low electron density and are mainly formed by neutral polysaccharides with small amounts of proteins. The granules of mature cells are larger, have a high electron density and are mainly formed by proteins with lower amounts of neutral polysaccharides. Transition stages between immature and mature granular cells are observed. The vacuolated cells are large and frequently pyramidal in shape, but after the application of histochemical techniques almost all vacuoles remain uncoloured. The numerous vacuoles contain flocculent material in a clear background and the mitochondria possess large crystalline structures in the matrix. A pyramidal shape is also typical of the mucocytes, which are filled with vesicles containing granular masses surrounded by a network of secretion material. These large cells are strongly stained by Alcian blue, revealing the presence of acidic mucopolysaccharides. This is the first ultrastructural study of the salivary glands in opisthobranch gastropods.     

斑透翅蝉唾液腺细菌群落组成研究

【目的】为明晰蝉类昆虫唾液腺中细菌的组成及其中是否存在内共生菌Candidatus Sulcia muelleri。【方法】以斑透翅蝉Hyalessa maculaticollis (Motschulsky)为材料,采用16S rRNA限制性内切酶片段长度多态性(RFLP)对其唾液腺细菌群落组成进行分析。【结果】斑透翅蝉唾液腺中共存在7种细菌,分别属于变形菌门和厚壁菌门;其中绿脓假单胞杆菌Pseudomonas aeruginosa和肠杆菌Enterobacter sp.为优势细菌,分别占克隆总数的48.7%;另外5种细菌(反硝化细菌热单胞菌Thermomonas brevis、鞘胺醇单胞菌Sphingomonas sp.、芽孢杆菌Bacillus sp.、厌氧球菌Anaerococcus sp.和Methyloversatilis sp.)总共占克隆文库的2.05%。【结论】首次采用分子生物学方法明晰斑透翅蝉唾液腺的细菌群落;其细菌群落组成相对简单,且两种细菌占主导地位;此外,头喙亚目昆虫体内特有的内共生菌Candidatus Sulcia muelleri未在斑透翅蝉唾液腺中检测出,表明该共细菌可能仅在腹部的贮菌体中分布;斑透翅蝉唾液腺中的细菌是否普遍存在于蝉科昆虫唾液腺中以及在取食韧皮部汁液过程中的功能有待进一步研究。     

Histology and ultrastructure of the salivary glands and salivary pumps in the scorpionfly Panorpa obtusa (Mecoptera: Panorpidae)

Liu, S. and Hua, B. 2009. Histology and ultrastructure of the salivary glands and salivary pumps in the scorpionfly Panorpa obtusa (Mecoptera: Panorpidae). —Acta Zoologica (Stockholm) 91 : 457–465. The morphology, histology and ultrastructure of the salivary glands and salivary pumps in the scorpionfly Panorpa obtusa Cheng 1949 were investigated using light microscopy and scanning and transmission electron microscopy. The salivary glands display a distinct sexual dimorphism. The female has only two small sac‐like glands located in the prothorax, while the male possesses six long tubular glands extending into the sixth abdominal segment. The male salivary glands can be divided into five distinct regions. The apical long, thin secretory region possesses numerous secretory cells containing large secretory vesicles; the salivary reservoir expands in diameter, accumulating and temporarily storing the saliva in addition to secreting saliva; the constricted region contains prismatic cells with complex infolded plasma membrane; the sac has an internal brush border to absorb water and ions; the common salivary duct contains longitudinal muscles in the male, but not in the female. The salivary pump possesses independent strong dorsal muscles and abundant internal palm spines near its orifice. The anatomy and ultrastructure of the salivary glands and the salivary pump of scorpionflies as well as their possible functions are briefly discussed.     

Morphology of the anterior digestive system of tonnoideans (Gastropoda: Caenogastropoda) with an emphasis on the foregut glands

ABSTRACT

Tonnoideans are marine carnivorous caenogastropods that prey on different invertebrates, namely polychaetes, sipunculids, bivalve and gastropod molluscs, and echinoderms. The morphology of the digestive system of 20 species from five families of the Tonnoidea was examined (for most of these species for the first time), and the salivary glands of six of them were studied using serial histological sections. Most of the studied families are rather similar anatomically, except Personidae (Distorsio), which differs both in proboscis morphology and the structure of the salivary glands. In most tonnoideans the salivary glands are split morphologically and functionally into anterior and posterior lobes, the latter synthesising strong sulfuric acid. The ducts of the posterior lobes are lined with non-ciliated epithelium and receive usually paired ciliated ducts from the anterior lobes to form a non-ciliated common duct, opening into the buccal cavity. In Personidae, the salivary glands are not separated into lobes, but are instead composed of ramifying tubules that are histologically different in the proximal and distal parts. Radulae of Tonnoidea are rather variable, with different patterns of interlocking teeth, both in the transverse and longitudinal rows, which may be related to particular feeding mechanisms. Due to the peculiarities of Personidae, the close relationship between that family and the rest of the Tonnoidea is questioned.     

Programmed cell death in Bradysia hygida (Diptera, Sciaridae) salivary glands presents apoptotic features

In this work, we present biochemical and morphological evidence that the final steps of programmed cell death (PCD) in the salivary glands of the inferior Diptera, Bradysia hygida, present apoptotic characteristics. In B. hygida, elimination of salivary glands is preceded by the establishment of a typical pattern of protein synthesis; increase in caspase activity; decrease in cell volume; nuclear pyknosis; nuclear DNA breakage; changes in the actin cytoskeleton; and most importantly, destruction of giant cells via formation of apoptotic bodies containing broken DNA or cytoplasm remains. Thus, elimination of B. hygida salivary glands by this process suggests that such mode of PCD is also involved in the destruction of entire organs in insects and, therefore, adds more complexity to the regulation of tissue elimination during development.     

Mechanisms of programmed cell death in the midgut and salivary glands from Bradysia hygida (Diptera: Sciaridae) during pupal–adult metamorphosis

Programmed cell death is involved with the degeneration/remodeling of larval tissues and organs during holometabolous development. The midgut is a model to study the types of programmed cell death associated with metamorphosis because its structure while degenerating is a substrate for the formation of the adult organ. Another model is the salivary glands from dipteran because their elimination involves different cell death modes. This study aimed to investigate the models of programmed cell death operating during midgut replacement and salivary gland histolysis in Bradysia hygida. We carried out experiments of real‐time observations, morphological analysis, glycogen detection, filamentous‐actin localization, and nuclear acridine orange staining. Our findings allow us to establish that an intact actin cytoskeleton is required for midgut replacement in B. hygida and nuclear condensation and acridine orange staining precede the death of the larval cells. Salivary glands in histolysis present cytoplasmic blebbing, nuclear retraction, and acridine orange staining. This process can be partially reproduced in vitro. We propose that the larval midgut death involves autophagic and apoptotic features and apoptosis is a mechanism involved with salivary gland histolysis.     

Serial cultivation of epithelial cells from human and macaque salivary glands

Summary To study the regulation of human salivary-type gene expression we developed cell culture systems to support the growth and serial cultivation of salivary gland epithelial and fibroblastic cell types. We have established 22 independent salivary gland epithelial cell strains from parotid or submandibular glands of human or macaque origin. Nineteen strains were derived from normal tissues and three from human parotid gland tumors. Both the normal and the tumor-derived salivary gland epithelial cells could be serially cultivated with the aid of a 3T3 fibroblast feeder layer in a mixture of Ham’s F12 and Dulbecco’s modified Eagle’s media supplemented with fetal bovine serum, calcium, cholera toxin, hydrocortisone, insulin, and epidermal growth factor. Salivary gland epithelial cells cultured under these conditions continued to express the genes for at least two acinar-cell-specific markers at early passages. Amylase enzyme activity was detected in conditioned media from cultured rhesus parotid epithelial cells as late as Passage 5. Proline-rich-protein-specific RNAs were detected in primary cultures of both rhesus and human parotid epithelial cells. Neither amylase enzyme activity nor PRP-specific RNAs were detected in fibroblasts isolated from the same tissues. In addition, salivary gland epithelial cells cultured under our conditions retain the capacity to undergo dramatic morphologic changes in response to different substrata. The cultured salivary gland epithelial cells we have established will be important tools for the study of salivary gland differentiation and the tissue-specific regulation of salivary-type gene expression.     

Vasculature of the parotoid glands of four species of toads (bufonidae: bufo)

The parotoid glands of toads (Bufonidae) consist of large aggregations of granular glands located between the otic region of the skull and the scapular region. To determine the circulatory pattern of these glands, we perfused the vascular systems of Bufo alvarius, B. marinus, B. terrestris, and B. valliceps with either India ink or Microfil, a fine latex. The perfused glands were studied by gross dissection, microscopic examination, and histology. The vascular patterns of the parotoid glands were compared to the arrangement of vessels in the dorsal skin of Rana sphenocephala (Ranidae), a frog that lacks parotoid glands. The parotoid glands of the four species of toads are supplied with blood by the lateral and dorsal cutaneous arteries and are drained by one or more branches of the internal jugular vein. The dorsal cutaneous artery supplies most of the blood to the parotoid glands in B. terrestris and B. valliceps. In B. alvarius and B. marinus, both the lateral and dorsal cutaneous arteries serve major roles in the blood supply of the glands. These patterns of blood flow have not been described previously for parotoid glands and conflict with earlier accounts for B. alvarius and B. marinus. The arteries and veins associated with the parotoid glands of toads are present in R. sphenocephala, but are arranged differently. In R. sphenocephala, the lateral cutaneous artery supplies the dorsal and lateral skin posterior to the shoulder region, whereas the dorsal cutaneous artery supplies the skin of the shoulder region. In toads, both the lateral and dorsal cutaneous arteries supply the skin of the shoulder region and ramify into subcutaneous capillaries that surround the secretory units of the parotoid glands. Extensive vasculature presumably is important for delivering cholesterol and other precursor molecules to the parotoid glands, where those compounds are converted into toxins.     

棉铃虫幼虫唾液腺cDNA文库的构建及EST分析

棉铃虫Helicoverpa armigera (Hübner)幼虫唾液中的各种酶类及各种生化组分在棉铃虫与植物相互作用及协同进化中起到重要作用; 唾液腺是棉铃虫唾液成分的合成器官。本研究通过构建棉铃虫幼虫唾液腺全长cDNA文库, 测序得到1 502条EST序列, 聚类分析后获得821个unigenes, 为筛选棉铃虫与寄主互作信号因子提供基因信息资源。使用Blast2 GO软件对821个unigenes进行了比对和功能注释, 初步获得棉铃虫幼虫唾液腺中mRNA的构成特征。结果显示, 在棉铃虫唾液腺ESTs文库中, 鉴定得到脂类相关消化酶基因17个, 糖类相关消化酶基因5个, 半胱氨酸蛋白酶基因1个, 丝氨酸蛋白酶基因20个（其中16个为新发现）, 提示唾液腺的主要功能是分泌消化酶进行预消化; 还发现在棉铃虫幼虫唾液腺中存在表皮蛋白、 气味结合蛋白和化学感受蛋白基因。结果为研究棉铃虫预消化系统打下基础。     

Cyclic nucleotide crosstalk in salivary glands from partially fed Dermacentor variabilis (Say)

Enzyme immunosorbent assays were used to measure cyclic nucleotide concentrations in homogenates of salivary glands from partially fed female Dermacentor variabilis. The adenylyl cyclase activator forskolin (100 μM) increased homogenate cGMP concentrations greater than three-fold over controls. Competitive inhibition of nitric oxide synthase with 1 mM l-NMMA, an l-arginine analog, demonstrated that crosstalk occurs downstream of nitric oxide synthesis. Forskolin-stimulated synthesis of cGMP was diminished 58% by the soluble guanylyl cyclase inhibitor ODQ (2 μM). The protein kinase A selective inhibitor Rp-cAMPS (50 μM) inhibited forskolin-stimulated cGMP by 49%. Whole glands treated with 10 μM dopamine increased cGMP levels two-fold in the presence of 1 mM IBMX. Treatment of whole salivary glands with equimolar concentrations of 8-Br-cAMP and 8-Br-cGMP produced no greater fluid uptake than in glands treated with 8-Br-cGMP alone, suggesting that cAMP and cGMP share a downstream target. The protein kinase G-selective inhibitor Rp-8-pCPT-cGMPS (100 μM) impeded 10 mM 8-Bromo-cGMP-stimulated gland weight increases. Pretreatment with verapamil, a Ca²⁺ channel blocker, attenuated cyclic nucleotide-stimulated fluid uptake indicating that whole gland fluid changes are dependent on extracellular Ca²⁺. Together, our data suggest that cGMP production is mediated in part by cAMP-dependent activation of soluble guanylyl cyclase. Experiments measuring changes in whole salivary gland weight support the hypothesis that cAMP and cGMP signaling cascades have a common target and that cyclic nucleotide-stimulated fluid movement is dependent on Ca²⁺ influx.     

Energy expenditure and protein turnover in three species of wallabies (Marsupialia: Macropodidae)

Summary Relationships between basal and fed metabolic rates and whole-body protein turnover rates were examined in three species of wallabies, the red-necked pademelon (Thylogale thetis), parma wallaby (Macropus parma) and tammar wallaby (M. eugenii).There were no significant differences among wallaby species in basal metabolic rate (BMR) which was 30% below eutherian mammals. However, the fed metabolic rate of the tammar was lower than that of the other two species (P<0.05), as was the protein turnover rate (P<0.01) which is consistent with its lower voluntary feed intake and with its lower maintenance nitrogen requirement.Protein turnover rates in the wallabies were 23–47% lower than in eutherian mammals. Similarly, protein synthesis made a lower contribution to fed metabolic rates in the wallabies (7–8%) than in eutherians (17–25%).Thus, compared with several eutherian species, macropodid marsupials have low rates of both energy and protein metabolism, but within the macropodids there is not necessarily a close link between basal metabolic rate and whole-body protein turnover.Abbreviations BMR basal metabolic rate - DEE daily energy expenditure - EE energy expenditure - LSD least significant difference - RQ respiratory quotient     

Ultrastructural investigation of a salivary gland in a centipede: structure and origin of the maxilla I-gland of Scutigera coleoptrata (Chilopoda, Notostigmophora)

The maxilla I-gland of Scutigera coleoptrata was investigated using light and electron microscopy methods. This is the first ultrastructural investigation of a salivary gland in Chilopoda. The paired gland opens via the hypopharynx into the foregut and extends up to the third trunk segment. The gland is of irregular shape and consists of numerous acini consisting of several gland units. The secretion is released into an arborescent duct system. Each acinus consists of multiple of glandular units. The units are composed of three cell types: secretory cells, a single intermediary cell, and canal cells. The pear-shaped secretory cell is invaginated distally, forming an extracellular reservoir lined with microvilli, into which the secretion is released. The intermediary cell forms a conducting canal and connects the secretory cell with the canal cell. Proximally, the intermediary cell bears microvilli, whereas the distal part is covered with a distinct cuticle. The cuticle is a continuation of the cuticle of the canal cells. This investigation shows that the structure of the glandular units of the salivary maxilla I-gland is comparable to that of the glandular units of epidermal glands. Thus, it is likely that in Chilopoda salivary glands and epidermal glands share the same ground pattern. It is likely that in compound acinar glands a multiplication of secretory and duct cells has taken place, whereas the number of intermediary cells remains constant. The increase in the number of salivary acini leads to a shifting of the secretory elements away from the epidermis, deep into the head. Comparative investigations of the different head glands provide important characters for the reconstruction of myriapod phylogeny and the relationships of Myriapoda and Hexapoda.     

Breeding and age structure of the population of Macropus parma on Kawau Island,New Zealand

The parma wallaby, Macropus parma, is one of four species of wallaby introduced on to Kawau Island, New Zealand, in ahout 1870. A sample of sixty-four parmas was shot on the island in February 1973 to provide information on breeding activity and age structure of the population. Incidental data were also collected from the tammar wallaby, Macropus eugenii, which is the other common species on the island. Neither the sex ratio of the shot sample of parmas (thirty-seven males to twenty-seven females) nor that of the tammars (eleven males to eleven females) differed significantly from parity. Onset of sexual maturity of female parmas on Kawau I. was delayed when compared with that of captive females. One female was estimated to have bred at 19 months old but most females in the sample were not mature until 2 years old and a few not until 3 years old. In contrast, female tammars became sexually mature at about 12 months. There did not appear to be any delay in the onset of sexual maturity of male parma wallabies. In the shot sample, none of sixteen female parmas capable of having a young were carrying a pouch young whereas nine of ten female tammars had a pouch young. The mean date of birth of these latter young was 30 January (range 18 January - 11 February) which is consistent with the breeding season of tammars in Australia. Among the mature female parmas two had recently mated, six were in pro-oestrus and six were in anoestrus, indicating that the breeding season in 1973 had just commenced. Estimated months of birth for all parmas under 3 years of age suggested that breeding was continuous in 1970–71 but that there had been a defined breeding season in 1972 with births occurring between March and July. There was an excess of 1– and 2–year-old parmas in the sample. This was the result of continuous breeding in 1970–71, presumably due to the provision of pasture on farmland being developed on the island. A sample of thirty-three tammar skulls resulting from a shoot in November 1972 did not show a nmilar excess of 1– and 2–year-old animals as this species has a rigidly defined breeding pattern and the females can produce only one young a year.     

The salivary glands of the cockroach Nauphoeta cinerea (Olivier)

The innervation of the salivary gland of the cockroach Nauphoeta cinerea (Olivier) has been investigated with the use of light and scanning electron microscopy. Light microscopy of methylene blue stained glands reveals the presence of a dual innervation arising from the ventral nerve cord and the stomodeal nervous system; the principal innervation is that from the ventral nerve cord which passes to the gland via the reservoir ducts. Branches of these nerves form a plexus on the acinar surface, the axons of which exhibit swelling at irregular intervals. The presence of this surface plexus and the axonal swellings was confirmed by scanning electron microscopy both in normal glands and in those in which the basal lamina had been removed by means of an HCl-collagenase digestion method. No acinar plexus was seen to be formed by branches of the stomatogastric nerve that were associated with the gland. However, other branches of this nerve were clearly connected with a complex network of multipolar neurones on the surfaces of the anterior regions of both salivary reservoirs.     

Morphology and histology of the larynx of the common toad Rhinella arenarum (Hensel, 1867) (Anura,Bufonidae)

The structure of the larynx of the toad Rhinella arenarum was exhaustively studied. The laryngeal skeleton consists of three bilaterally symmetrical cartilages: the cricoid and two arytenoids. Internally, each half‐larynx has an anterior and a posterior chamber. The first chamber is delimited by the epithelium covering the arytenoid cartilage and the anterior membrane. The latter consists of fibro‐elastic tissue and contains blood capillaries that, judging by their location and distribution, might serve to maintain vocal cord turgidity. At the level of the cricoid cartilage, two structures are reported here for the first time: the posterodorsal and the anteroventral processes. Both processes are associated with the insertion of the posterior membrane. A cartilaginous rod is located at the free margin of the posterior membrane. This rod appears to support the membrane when the air flows. The distal portion of the larynx communicates with the proximal region of the lung. The epithelium of the laryngeal mucosa contains ciliated cells, goblet cells, secretory cells with short microvilli and neuroendocrine cells immunopositive to PGP 9.5. The results obtained in this study provide new information about the internal organization of the larynx in anurans, which could serve as additional morphological characters for phylogenetic relationships.