Failure of the Amblyomma cajennense nymph to become infected by Theileria equi after feeding on acute or chronically infected horses

Tick-borne diseases in horses are caused by the intraerythrocytic protozoan parasites Theileria equi and Babesia caballi. Although T. equi is highly endemic in Latin America, the New World vector of this important parasite is controversial. The aim of this study was to test the ability of nymph Amblyomma cajennense ticks acquire infection by T. equi following feeding on infected horses. Three experiments were performed: tick acquisition of T. equi from an experimentally infected horse, tick acquisition of T. equi from naturally infected foals and tick acquisition of T. equi from a chronically infected horse. A. cajennense adults were dissected and salivary glands were collected in aliquots. Methyl green pyronin staining of the salivary glands did not show the presence of hypertrophy of acini or cell nuclei normally suggestive of Theileria spp. infection. The pools of salivary glands were negative for Theileria DNA in nested PCR assays. Histopathological analysis failed to detect sporoblast and sporozoites of T. equi in salivary gland acini. This study was not able to observe infection of the A. cajennense by T. equi.     

Ultrastructure of the salivary glands of the planthopper,peregrinus maidis (ashmead) (Homoptera : Delphacidae)

The principal salivary gland of the planthopper, Peregrinus maidis (Ashmead) (Homoptera : Delphacidae), comprises 8 acini of only 6 ultrastructurally different acinar types. In these acini, secretory cells contain elongated vacuoles partly lined by microvilli and by microtubule bundles. These vacuoles are apparently connected with extracellular canaliculi deeply invaginated into secretory cells. Canaliculi of each acinus lead to a ductule lumen, which is lined with spiral cuticular intima, surrounded by duct cells. Striated muscle fibers, supplied with small nerve axons and tracheoles, are found in various acini of the principal gland, usually around secretory and duct cells.In the accessory salivary gland, the 2 large secretory cells contain no elongated vacuoles or canaliculi invaginations. However, in their central region, apically, these cells border a large microvilli-lined canal with its own canal cells. This canal is apparently connected with the cuticle-lined accessory duct, formed by duct cells. Nerve axons, but no muscle fibers, are found in the accessory gland and its duct. It is suggested that the system for transporting secretory material within acini of the principal gland, is basically different from that within the accessory gland.     

Effects of Ricinus communis oil esters on salivary glands of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae)

This study showed the interference of esters extracted from Ricinus communis in the secretory cycle of salivary glands of Rhipicephalus sanguineus ticks, which consequently caused collateral effects on their feeding process. Ticks attached on hosts which were fed with commercial feed containing different concentrations of R. communis oil esters suffered damages such as cytoplasmic changes in their salivary glands, notably in the acinar cells, impairing the functioning of the acini and accelerating the organs degeneration as a whole. It was found that esters interfered with the activity of cellular secretion by changing the glycoprotein of salivar composition especially in acini II cells. It was also shown that the damages caused by esters in the salivary glands cells of these ectoparasites increased in higher concentrations of the product and degenerative glandular changes were more pronounced.     

The effects of dopamine receptor agonists and antagonists on the secretory rate of cockroach (Periplaneta americana) salivary glands

The acinar salivary glands of the cockroach, Periplaneta americana, are innervated by dopaminergic and serotonergic nerve fibers. Serotonin stimulates the secretion of protein-rich saliva, whereas dopamine causes the production of protein-free saliva. This suggests that dopamine acts selectively on ion-transporting peripheral cells within the acini and the duct cells, and that serotonin acts on the protein-producing central cells of the acini. We have investigated the pharmacology of the dopamine-induced secretory activity of the salivary gland of Periplaneta americana by testing several dopamine receptor agonists and antagonists. The effects of dopamine can be mimicked by the non-selective dopamine receptor agonist 6,7-ADTN and, less effectively, by the vertebrate D1 receptor-selective agonist chloro-APB. The vertebrate D1 receptor-selective agonist SKF 38393 and vertebrate D2 receptor-selective agonist R(-)-TNPA were ineffective. R(+)-Lisuride induces a secretory response with a slower onset and a lower maximal response compared with dopamine-induced secretion. However, lisuride-stimulated glands continue secreting saliva, even after lisuride-washout. Dopamine-induced secretions can be blocked by the vertebrate dopamine receptor antagonists cis(Z)-flupenthixol, chlorpromazine, and S(+)-butaclamol. Our pharmacological data do not unequivocally indicate whether the dopamine receptors on the Periplaneta salivary glands belong to the D1 or D2 subfamily of dopamine receptors, but we can confirm that the pharmacology of invertebrate dopamine receptors is remarkably different from that of their vertebrate counterparts.     

Attachment of the Flavescence doree pathogen (MLO) to leafhopper vectors and other insects

Flavescence doree (FD) is an important yellows disease of grapevine, caused by mycoplasma-like-organism (MLO) and is transmitted in the field by the leafhopper Scaphoideus titanus Ball. It can be transmitted in the laboratory between Vicia faba test plants by the leafhopper, Euscelidius variegatus Kbm. A technique to identify a specific attachment system between the MLO and the leafhopper vectors was developed. In this method, called “Double Dot”, extracts of macerated healthy whole insects or organs applied to a support membrane or cryosections of healthy whole leafhoppers, are incubated with a MLO-enriched extract from FD-infected V. faba or FD-infected E. variegatus. Attached MLO cells were identified by immunolabelling using FD-MLO specific monoclonal antibodies. Attachment of MLO cells was obtained on extracts of healthy S. titanus and E. variegatus and on tissues such as salivary glands, hemolymph and alimentary tract. On cryosections, MLO attachment was obtained on acini IV and V of the salivary glands and on some acini III, on the ventriculus of the alimentary tract, and on the abdomen fat bodies. “Double dot” experiments were done using other insect species, and MLO cells attachment was obtained on most MLO-vector insects but also on insects from a few non-vector species.     

The sympathetic and parasympathetic nature of neuropeptide Y-immunoreactive nerve fibres in the major salivary glands of the rat

Summary The distribution and origin of neuropeptide Y in the major salivary glands of the rat was studied by indirect immunofluorescence technique. Numerous nerve fibres immunoreactive for the peptide were seen in the parotid and sublingual glands. Most of the fibres were located around blood vessels and salivary acini. In the submandibular gland the number of immunoreactive nerve fibres around the acini was lower in comparison with that in the parotid and sublingual glands. Some immunoreactive nerve fibres were also found around or along intra- and interlobular ducts in all major salivary glands.A large number of the neuropeptide-containing neuronal cell bodies and nerve fibres were detected in the sympathetic superior cervical ganglion. Sympathetic postganglionic nerve trunks of this ganglion contained numerous immunoreactive nerve fibres as well. A subpopulation of the neuronal cell bodies in the submandibular ganglion were immunoreactive to neuropeptide Y.Both uni- and bilateral superior cervical ganglionectomies caused a significant decrease in the number of immunoreactive nerve fibres around the blood vessels in all the major salivary glands. However, these denervations did not affect the density of nerve fibres around the acini and ducts. On the contrary, unilateral parasympathetic denervation by sectioning the auriculotemporal nerve reduced the fibres around the secretory acini in the parotid gland remarkably, while only a minor reduction in the density of immunoreactive fibres associated with the blood vessels of the gland was detected. Unilateral electrocoagulation of the trigeminal nerve branches caused no detectable change in the density of immunoreactive nerve fibres in any of the major salivary glands.On the basis of the present findings it is concluded that neuropeptide Y-reactive nerve fibres present in all major salivary glands around the blood vessels seem to be mainly sympathetic, whereas those around the acini and ducts seems to be of parasympathetic origin.     

Survival of Theileria parva-infected adult Rhipicephalus appendiculatus under laboratory and quasi-natural conditions

Adult Rhipicephalus appendiculatus Muguga, having high or low intensities of Theileria parva Muguga infection in their salivary glands, were exposed to 20 °C and 85% relative humidity in the laboratory or quasi-natural conditions. Survival of the ticks and T. parva infections in their salivary glands was then monitored over a two year period. Ticks, having an average infection level of 2 infected acini per female, survived for up to 70 or 106 weeks after moulting under the laboratory or quasi-natural conditions respectively. Those having an infection level of 26 infected acini per female, survived for a similar duration except that those under quasi-natural conditions survived for a slightly shorter duration (102 weeks). Similarly, T. parva parasites survived for much longer periods under quasi-natural conditions than under the laboratory conditions. They survived for up to 38 or 78 weeks post salivary gland infection under the laboratory or quasi-natural conditions respectively in both categories of infection levels. There was apparently a density dependent relationship in T. parva survival, with a dramatic fall in infection occurring in ticks with high levels of infection between weeks 10 and 18 or weeks 38 and 46 post salivary gland infection in those exposed to laboratory or quasi-natural conditions before levelling off. This revised version was published online in July 2006 with corrections to the Cover Date.     

The expression of water channel proteins during human salivary gland development: a topographic study of aquaporins 1, 3 and 5

Some members of aquaporin family (AQP) plays crucial functions in salivary synthesis and secretion. These proteins expression has already been reported during salivary gland formation, however no previous studies in human developing glands have been performed. We evaluated AQP1, 3 and 5 expression through the stages of human salivary gland morphogenesis and discuss the possible role of AQP for glandular maturation. Human salivary glands derived from foetuses aged between 14 and 25 weeks were submitted to immunohistochemistry. At the bud stage, membrane expression of AQP1, 3 and 5 were observed within the epithelial bud cells presenting a similar apicolateral pattern, also found at the pseudoglandular stage, present within the terminal portions of future acini, while AQP5 was also particularly strong at the apical membrane of pre-acinar and pre-ductal cells. AQP5 was co-localised with Cytokeratin 7. Similar AQP1, 3 and 5 expression were observed at the following canalicular stage, where distinct and strongly luminal and acinar AQP5 expression is present. During the final terminal bud stage, AQP1 was only identified in serous acini, myoepithelial and endothelial cells, while differentiated mucous acinar cells and ducts were negative. AQP3 was detected at apicolateral membranes of both mucous and serous acini. AQP5 also showed a diffuse expression in mucous and serous acini, in addition to strong apical membrane expression within lumen of intercalated ductal cells. This topographic analysis of AQP1, 3 and 5 revealed differences in the expression pattern throughout salivary gland developmental stages, suggesting different roles for each protein in human glandular maturation.     

The innervation of the salivary gland in the american cockroach: Light and electron microscopic observations

The salivary gland of Periplaneta americana (L.) is innervated from both the stomatogastric nervous system (SNS) and subesophageal ganglion (SEG). Methylene-blue preparations, histological sections and electron microscopy revealed a pair of nerves from the SEG, each of which contains two axons 5–7 μ in diameter, and these are accompanied by several smaller ones. The nerves going to the salivary glands from the SNS contain a dozen or more axons, each less than 2 μ thick. Axons from two sources innervate the efferent salivary ducts, the acini, the anterior ends of the salivary reservoirs, and the reservoir suspensory muscles. A nerve which has reached an acinus forms a plexus upon its surface. Electron micrographs disclose penetration of axons with or without glial wrappings, into the intercellular spaces between gland cells. Axons without glial wrappings have been observed in intimate contact with gland-cell membranes, and several areas which resemble synaptic junctions have been seen.     

Receptors for the neuropeptides,myoinhibitory peptide and SIFamide,in control of the salivary glands of the blacklegged tick Ixodes scapularis

Tick salivary glands are important organs that enable the hematophagous feeding of the tick. We previously described the innervation of the salivary gland acini types II and III by a pair of protocerebral salivary gland neurons that produce both myoinhibitory peptide (MIP) and SIFamide (?imo et al., 2009b). In this study we identified authentic receptors expressed in the salivary glands for these neuropeptides. Homology-based searches for these receptors in the Ixodes scapularis genome sequence were followed by gene cloning and functional expression of the receptors. Both receptors were activated by low nanomolar concentrations of their respective ligands. The temporal expression patterns of the two ligands and their respective receptors suggest that the SIFamide signaling system pre-exists in unfed salivary glands, while the MIP system is activated upon initiation of feeding. Immunoreactivity for the SIFamide receptor in the salivary gland was detected in acini types II and III, surrounding the acinar valve and extending to the basal region of the acinar lumen. The location of the SIFamide receptor in the salivary glands suggests three potential target cell types and their probable functions: myoepithelial cell that may function in the contraction of the acini and/or the control of the valve; large, basally located dopaminergic granular cells for regulation of paracrine dopamine; and neck cells that may be involved in the control of the acinar duct and its valve.     

Autoactive molluscan neuron: Reflex function and synaptic modulation during feeding in the terrestrial slug,Limax maximus

Summary The salivary burster (SB) is an autoactive motoneuron to the salivary duct of the terrestrial slugLimax maximus. The SB is electrically coupled to protractor motoneurons and inhibited by the metacerebral giant cell. During feeding these synaptic inputs cause SB activity to be phase-locked to the protraction-retraction cycle. The SB can be used as a clear and reliable monitor of feeding motorprogram activation.We thank Dr. Joseph Jin Chang for invaluable assistance with some of this work. Support was provided by NSF grant BMS74-15217 to D.J.P., NSF grant BMS74-03572 to A.G., and a grant from the Spencer Foundation.     

Salivary glands of <Emphasis Type="Italic">Amblyomma cajennense</Emphasis> (Acari: Ixodidae): a histological and an ultrastructural overview

The present study on the salivary glands of semi-engorged Amblyomma cajennense females has identified the various cell types present in this tissue and allowed its morphohistochemical characterization. Marking techniques were applied to detect polysaccharides (PAS), proteins (bromophenol blue), lipids (Nile blue) and calcium (von Kossa), as well as those of scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The results obtained by TEM showed and confirmed that these individuals’ glands are also formed by round acini that are connected to the common excretory duct through acinar and intermediate ducts. Histological data as well as ultrastructural ones showed that the glands are formed by types I, II and III acini. In this study with salivary glands polysaccharides, proteins, lipids and calcium were observed in the cytoplasm and/or cell secretion granules—both free or forming complexes, as the intensity of the marking varied according to the cell as well as the type of acini analyzed, showing the structural and functional complexity of the tick salivary glands, characteristics that give the multifunctional character to this organ.     

Evidence that developmental changes in type III acini in the tickAmblyomma hebraeum (Acari: Ixodidae) are initiated by a hemolymph-borne factor

During feeding, certain cells in the salivary gland type III acini of the ixodid tickAmblyomma hebraeum Koch undergo major developmental changes. We induced many of these changes in the ablumenal interstitial cells (AbIC), adlumenal interstitial cell (AdIC), and f-cells of type III acini, by transplanting the salivary gland of the unfed female to the hemocoel of a feeding female. In transplants, AbICs enlarged and formed a labyrinth of extracellular spaces. Extensions of AbICs pushed into the AdIC. Autophagic vacuoles were common in AbICs. The f-cells also enlarged and developed autophagic vacuoles. Complex interdigitation occurred between the f-cells and the AbIC. In transplants, the labyrinth was not as extensive as that of fed unoperated females or of operated females. The AdIC, AbIC, and f-cells did not undergo as extensive a development in unoperated fed males as the same cells did in unoperated fed females. In males AbICs did not develop an extensive labyrinth, and the f-cells did not develop beyond a secretory phase. No autophagic vacuoles were observed in any of these cells. When male salivary glands were transplanted into feeding females, AdIC, AbICs and f-cells developed an ultrastructure similar to the same cells in female transplants. Cells from salivary glands of unfed females cultured for 2 days in TC medium 199 resembled the same cells from control unfed salivary glands. The selectivity of these changes supports the conclusion that a hemolymph-borne salivary gland development factor initiated this development.     

Cytotoxic effects of permethrin in salivary glands of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged females

Because of the medical and veterinary importance of ticks and the wide use of synthetic chemical substances such as permethrin (active ingredient of Advantage® Max3 – Bayer)for their control, this study evaluated the effects of different concentrations (206, 1031 and 2062 ppm) of the acaricide on the salivary glands of Rhipicephalus sanguineus semi-engorged females. Results showed that permethrin is a potent substance that acts morpho-physiologically in the tick glandular tissue, causing changes in the acini shape intense vacuolation in acinar cells, and disruption of the tissue by cell death process, with subsequent formation of apoptotic bodies, especially at higher concentrations, thus precluding the accurate identification of different types of acini. Importantly, it is demonstrated that permethrin acts on salivary gland tissue, as well as affecting the nervous system, accelerating the process of glandular degeneration, and interfering with the engorgement process of female ticks, preventing them from completing the feeding process.     

Effect of long-term treatment of norethisterone (a progestogen-only contraceptive) on salivary gland activity of the cycling albino rat

Effect of long-term treatment of norethisterone (a progestogen-only contraceptive) on the salivary glands of the cycling albino rats was evaluated from histomorphic and karyokinetic standpoints. Norethisterone increased concentration of zymogen granules in the serous acini and also of mucoid material in the mucous-containing acini in the submaxillary gland. In the parotid gland, the acini were hypertrophied, accompanied by cytoplasmic degranulation. In addition, the mitotic cells were seen in both the glands after the treatment. It is suggested that norethisterone perhaps stimulates the salivary gland activity of the rats.     

Morphology of the salivary glands of Mastotermes darwiniensis Froggatt (Isoptera : Mastotermitidae)

The salivary glands of Mastotermes darwiniensis (Isoptera : Mastotermitidae) workers consist of acini made up of peripherally and centrally placed cells with storage cells. A network of small efferent ductules is associated with each acinus. Intracellular ductules of the central and peripheral cells are filled with protruding microvilli. Central cells contain an abundance of rough endoplasmic reticulum and vacuoles. Continua of rough endoplasmic reticulum and vacuoles are present. Pinocytosis does not appear to be involved in secretion by the central cells. Peripheral cells are compartmental, having electron-lucent vesicle accumulations away from intracellular ductules and the basal plasmalemma invaginations. Peripheral cell morphology is similar to cells which are associated with fluid movement. Storage cells contain a large vacuole and exhibit micropinocytosis. Two axon profile types are present near and on the acini and salivary ducts, one of which appears associated with neuronal end plates.     

Expression and localization of immunoreactive-sialomucin complex (Muc4) in salivary glands

Sialomucin Complex (SMC; Muc4) is a heterodimeric glycoprotein consisting of two subunits, the mucin component ASGP-1 and the transmembrane subunit ASGP-2. Northern blot and immunoblot analyses demonstrated the presence of SMC/Muc4 in submaxillary, sublingual and parotid salivary glands of the rat. Immunocytochemical staining of SMC using monoclonal antisera raised against ASGP-2 and glycosylated ASGP-1 on paraffin-embedded sections of parotid, submaxillary and sublingual tissues was performed to examine the localization of the mucin in the major rat salivary glands. Histological and immunocytochemical staining of cell markers showed that the salivary glands consisted of varying numbers of serous and mucous acini which are drained by ducts. Parotid glands were composed almost entirely of serous acini, sublingual glands were mainly mucous in composition and a mixture of serous and mucous acini were present in submaxillary glands. Since immunoreactive (ir)-SMC was specifically localized to the serous cells, staining was most abundant in parotid glands, intermediate levels in submaxillary glands and least in sublingual glands. Ir-SMC in sublingual glands was localized to caps of cells around mucous acini, known as serous demilunes, which are also present in submaxillary glands. Immunocytochemical staining of SMC in human parotid glands was localized to epithelial cells of serous acini and ducts. However, the staining pattern of epithelial cells was heterogeneous, with ir-SMC present in some acinar and ductal epithelial cells but not in others. This report provides a map of normal ir-SMC/Muc4 distribution in parotid, submaxillary and sublingual glands which can be used for the study of SMC/Muc4 expression in salivary gland tumors.