Sequential changes in salivary gland structure during attachment and feeding of the cattle tick, Boophilus microplus

A study of the morphology and histochemistry of the salivary glands of the parasitic stages of Boophilus microplus has been made, glands of feeding females being studied in greatest detail. Of 9 granular cell types present in the female and 10 in the male, 3 probably secrete attachment cement and 4 others glycoproteins and enzymes, possible functions of which are discussed. Two cell types, c₄ and g (the latter being present only in males) are of unknown function. The most likely functions of non-granular epithelial cells and those forming acinus I are in osmoregulation.     

Programmed cell death in the larval salivary glands of <Emphasis Type="Italic">Apis mellifera</Emphasis> (Hymenoptera,Apidae)

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy. Tissue sections were subsequently stained with haematoxylin-eosin, bromophenol blue, silver, or a variant of the critical electrolyte concentration (CEC) method. Ultrathin sections were contrasted with uranyl acetate and lead citrate. Glands were processed for the histochemical and cytochemical localization of acid phosphatase, as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed. The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and, additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen. The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed. The nuclei were pyknotic, with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.     

Ultrastructural Studies on the Trypanosomatid Phytomonas serpens in the Salivary Glands of a Phytophagous Hemipteran

Electron microscopy of salivary glands of the phytophagous hemipteran Phihia picta infected with Phytomonas serpens revealed the presence of flagellates in the gland periphery beneath the gland envelope, in the gland central lumen, between gland cells in the intercellular space and inside the gland cells. In the latter case, flagellates were found in the cytoplasm whether or not it was surrounded by a vacuolar membrane. Flagellates were always of the promastigote type, sometimes displaying a large twisted body. Morphological peculiarities of flagellates in different gland locations are recorded.     

The morphology and ultrastructure of salivary glands of Zoraptera (Insecta)

The salivary glands of two species of Zoraptera, Zorotypus caudelli and Zorotypus hubbardi, were examined and documented mainly using transmission electron microscopy (TEM). The results obtained for males and females of the two species are compared and functional aspects related to ultrastructural features are discussed. The salivary glands are divided into two regions: the secretory cell region and the long efferent duct, the latter with its distal end opening in the salivarium below the hypopharyngeal base. The secretory region consists of a complex of secretory cells provided with microvillated cavities connected by short ectodermal ducts to large ones, which are connected with the long efferent duct. The secretory cell cytoplasm contains a large system of rough endoplasmic reticulum and Golgi apparatus producing numerous dense secretions. The cells of the efferent duct, characterized by reduced cytoplasm and the presence of long membrane infoldings associated with mitochondria, are possibly involved in fluid uptaking from the duct lumen.     

Structural study and expression of the androgen receptors during the reproductive cycle in the Harderian gland of the male Meriones libycus

The goal of this study was to evaluate for the first time the expression of the androgen receptors (AR) in Harderian glands (HG) of the male Meriones lybicus in relation to the reproductive cycle. Six male Harderian glands of the resting period and 6 of the breeding period were collected. The animals were trapped in the desert of Béni Abbès (Algeria). The morphology of the Harderian glands was studied by light microscopy and morphometry, whereas the expression of the androgen receptors was assessed and quantified based on immunohistochemistry techniques. We have shown that the Harderian glands of Meriones libycus are tubuloalveolar glands with wide lumen. The glandular epithelium is composed of two types of cells (types I and II) in the resting season and three types of cells (types I, II and III) in the breeding season. These three types of cells differ in size and shape. Type-I cells have a prismatic shape, an acidophilic cytoplasm, and small lipidic vacuoles, whereas type-II ones are pyramidal in shape, with basophilic cytoplasm. Type-III cells resemble those of type I, and so they are prismatic in shape and have an acidophilic cytoplasm with larger lipidic vacuoles. The immunoreactivity of type-I and type-III cells was mainly cytoplasmic and the intensity of the immunostaining was significantly higher during the breeding season. Among other functions, the Harderian gland seems to be involved in the production of pheromones under the effect of androgens.     

Ultrastructure of male reproductive accessory glands and ejaculatory duct in the Queensland fruit fly,Bactrocera tryoni (Diptera: Tephritidae)

Ultrastructure of male reproductive accessory glands and ejaculatory duct in the Queensland fruit fly (Q-fly), Bactrocera tryoni, were investigated and compared with those of other tephritid flies. Male accessory glands were found to comprise one pair of mesodermic glands and three pairs of ectodermic glands. The mesodermic accessory glands consist of muscle-lined, binucleate epithelial cells, which are highly microvillated and extrude electron-dense secretions by means of macroapocrine transport into a central lumen. The ectodermic accessory glands consist of muscle-lined epithelial cells which have wide subcuticular cavities, lined with microvilli. The electron-transparent secretions from these glands are first extruded into the cavities and then forced out through small pores of the cuticle into the gland lumen. Secretions from the two types of accessory glands then flow into the ejaculatory duct, which is highly muscular, with epithelial cells rich in rough endoplasmic reticulum and lined with a thick, deeply invaginated cuticle. While there are some notable differences, reproductive accessory glands of male Q-flies generally resemble those of the olive fruitfly, Bactrocera oleae, and to a lesser extent the Mediterranean fruit fly, Ceratitis capitata.     

Infectivity of Leucocytozoon caulleryi sporozoites developed in vitro and in vivo

Morii T., Matsui T., Iijima T. and Fiotnaoa F. 1984. Infectivity of Leucocytozoon caulleryi sporozoites developed in vitro and in vivo. International Journal for Parasitology14: 135–139. Infectivity of Leucocytozoon caulleryi sporozoites isolated from various sites in Culicoides arakawae and from the midguts and the salivary glands which had been cultured in vitro after the infective blood meals was studied. Sporozoites isolated from the midguts, the abdominal and thoracic hemocoel and the salivary glands of biting midges on the 2nd day after feeding did not show infectivity to any of the chickens inoculated. Sporozoites obtained from the salivary glands on the 3rd day after feeding caused infection in all the inoculated chickens. The results indicated that sporozoites which had been just released from oocysts or had just reached the salivary glands cannot induce infection in chickens. Sporozoites were produced in the midguts which had been cultured in vitro in Medium 199 or Grace's medium after the infective blood meals, but they showed lower infectivity than those isolated from the salivary glands which had been cultured by the same methods as the midgut cultivation. The development of infectivity of L. caulleryi sporozoites seems to be site-dependent rather than time-dependent. High infectivity of sporozoites develops during their residence in the salivary glands of biting midges.     

Silk formation mechanisms in the larval salivary glands of<Emphasis Type="Italic">Apis mellifera</Emphasis> (Hymenoptera: Apidae)

The mechanism of silk formation inApis mellifera salivary glands, during the 5th instar, was studied. Larval salivary glands were dissected and prepared for light and polarized light microscopy, as well as for scanning and transmission electron microscopy. The results showed that silk formation starts at the middle of the 5th instar and finishes at the end of the same instar. This process begins in the distal secretory portion of the gland, going towards the proximal secretory portion; and from the periphery to the center of the gland lumen. The silk proteins are released from the secretory cells as a homogeneous substance that polymerizes in the lumen to form compact birefringent tactoids. Secondly, the water absorption from the lumen secretion, carried out by secretory and duct cells, promotes aggregation of the tactoids that form a spiral-shape filament with a zigzag pattern. This pattern is also the results of the silk compression in the gland lumen and represents a high concentration of macromolecularly well-oriented silk proteins.     

Distribution of specific proteins in the salivary gland lobes of Culicidae and their relation to age and blood sucking

Specific proteins are produced in different parts of the salivary glands of female Anopheles and Culex. In both species there are concentrated low molecular weight proteins in the median lobes and proteins in the mol. wt range 25,000–60,000 in the distal parts of the lateral lobes. These proteins are not identical in the two species. In the proximal parts of the lateral lobe there are only small amounts of protein in addition to an unknown high molecular weight substance. At the time of blood-sucking 15–35% of soluble gland proteins are injected with the saliva into the host animal, the proteins from the median lobes and the proximal part of the lateral lobes being released in greater quantities than the proteins from the distal parts of the lateral lobes. The typical gland proteins are only extractable in low concentrations from the salivary glands of the newly-hatched adult mosquito, they increase up to the 7th day of the adult development and are stored in the glands.     

Developmental changes in morphology and toxin content of the salivary gland of the australian paralysis tick Ixodes holocyclus

Binnington K.C. and Stone B.F. 1981. Developmental changes in morphology and toxin content of the salivary gland of the Australian paralysis tick Ixodes holocyclus.International Journal for Parasitology11: 343–351. Histological study of the salivary gland of female I. holocyclus has shown that 2 of the 4 cell types present are richest in granules in the unfed stage but have discharged these granules after an attachment period of 24 h. The presence of a toxin in homogenates of salivary glands from unfed females and its absence after 24 h of attachment may be associated with the loss of granules from these 2 cell types shortly after attachment. Another cell type shows a gradual increase in granule content throughout feeding and a fourth, a peak in granule content after an attachment period of 120 h. The latter cell type may produce the paralysing toxin since ticks do not paralyse the host until they have fed for about this time and homogenised glands are most toxic at 120–144 h.     

Genetic Diversity and Population Structure of Trypanosoma brucei in Uganda: Implications for the Epidemiology of Sleeping Sickness and Nagana

Background

While Human African Trypanosomiasis (HAT) is in decline on the continent of Africa, the disease still remains a major health problem in Uganda. There are recurrent sporadic outbreaks in the traditionally endemic areas in south-east Uganda, and continued spread to new unaffected areas in central Uganda. We evaluated the evolutionary dynamics underpinning the origin of new foci and the impact of host species on parasite genetic diversity in Uganda. We genotyped 269 Trypanosoma brucei isolates collected from different regions in Uganda and southwestern Kenya at 17 microsatellite loci, and checked for the presence of the SRA gene that confers human infectivity to T. b. rhodesiense.

Results

Both Bayesian clustering methods and Discriminant Analysis of Principal Components partition Trypanosoma brucei isolates obtained from Uganda and southwestern Kenya into three distinct genetic clusters. Clusters 1 and 3 include isolates from central and southern Uganda, while cluster 2 contains mostly isolates from southwestern Kenya. These three clusters are not sorted by subspecies designation (T. b. brucei vs T. b. rhodesiense), host or date of collection. The analyses also show evidence of genetic admixture among the three genetic clusters and long-range dispersal, suggesting recent and possibly on-going gene flow between them.

Conclusions

Our results show that the expansion of the disease to the new foci in central Uganda occurred from the northward spread of T. b. rhodesiense (Tbr). They also confirm the emergence of the human infective strains (Tbr) from non-infective T. b. brucei (Tbb) strains of different genetic backgrounds, and the importance of cattle as Tbr reservoir, as confounders that shape the epidemiology of sleeping sickness in the region.     

Role of Centrins 2 and 3 in Organelle Segregation and Cytokinesis in Trypanosoma brucei

Centrins are calcium binding proteins involved in cell division in eukaryotes. Previously, we have shown that depletion of centrin1 in Trypanosoma brucei (T. brucei) displayed arrested organelle segregation resulting in loss of cytokinesis. In this study we analyzed the role of T. brucei centrin2 (TbCen2) and T. brucei 3 (TbCen3) in the early events of T. brucei procyclic cell cycle. Both the immunofluorescence assay and electron microscopy showed that TbCen2 and 3-deficient cells were enlarged in size with duplicated basal bodies, multinuclei and new flagella that are detached along the length of the cell body. In both TbCen2 and TbCen3 depleted cells segregation of the organelles i.e. basal bodies, kinetoplast and nucleus was disrupted. Further analysis of the cells with defective organelle segregation identified three different sub configurations of organelle mis-segregations (Type 1–3). In addition, in majority of the TbCen2 depleted cells and in nearly half of the TbCen3 depleted cells, the kinetoplasts were enlarged and undivided. The abnormal segregations ultimately led to aborted cytokinesis and hence affected growth in these cells. Therefore, both centrin2 and 3 are involved in organelle segregation similar to centrin1 as was previously observed. In addition, we identified their role in kinetoplast division which may be also linked to overall mis-segregation.     

Ultrastructure of salivary glands of the chicken red mite <Emphasis Type="Italic">Dermanyssus gallinae</Emphasis> (Acarina,Gamasina, Dermanyssidae)

Structure of salivary glands of the chicken red mite, Dermanyssus gallinae (De Geer, 1778) was studied using transmission electron microscopy. Structure of the glands and their ducts is described. The cell composition, ultrastructural characteristics of the secretion, and peculiarities of its release from cell are revealed.     

The ultrastructural investigation of the midgut in the quill mite Syringophilopsis fringilla (Acari,Trombidiformes: Syringophilidae)

The midgut of the females of Syringophilopsis fringilla (Fritsch) composed of anterior midgut and excretory organ (=posterior midgut) was investigated by means of light and transmission electron microscopy. The anterior midgut includes the ventriculus and two pairs of midgut caeca. These organs are lined by a similar epithelium except for the region adjacent to the coxal glands. Four cell subtypes were distinguished in the epithelium of the anterior midgut. All of them evidently represent physiological states of a single cell type. The digestive cells are most abundant. These cells are rich in rough endoplasmic reticulum and participate both in secretion and intracellular digestion. They form macropinocytotic vesicles in the apical region and a lot of secondary lysosomes in the central cytoplasm. After accumulating various residual bodies and spherites, the digestive cells transform into the excretory cells. The latter can be either extruded into the gut lumen or bud off their apical region and enter a new digestive cycle. The secretory cells were not found in all specimens examined. They are characterized by the presence of dense membrane-bounded granules, 2–4 μm in diameter, as well as by an extensive rough endoplasmic reticulum and Golgi bodies. The ventricular wall adjacent to the coxal glands demonstrates features of transporting epithelia. The cells are characterized by irregularly branched apical processes and a high concentration of mitochondria. The main function of the excretory organ (posterior midgut) is the elimination of nitrogenous waste. Formation of guanine-containing granules in the cytoplasm of the epithelial cells was shown to be associated with Golgi activity. The excretory granules are released into the gut lumen by means of eccrine or apocrine secretion. Evacuation of the fecal masses occurs periodically. Mitotic figures have been observed occasionally in the epithelial cells of the anterior midgut.     

Tripartite parasitic and symbiotic interactions as a possible mechanism of horizontal gene transfer

Herbivory is a highly sophisticated feeding behavior that requires abilities of plant defense suppression, phytochemical detoxification, and plant macromolecule digestion. For plant‐sucking insects, salivary glands (SGs) play important roles in herbivory by secreting and injecting proteins into plant tissues to facilitate feeding. Little is known on how insects evolved secretory SG proteins for such specialized functions. Here, we investigated the composition and evolution of secretory SG proteins in the brown marmorated stink bug (Halyomorpha halys) and identified a group of secretory SG phospholipase C (PLC) genes with highest sequence similarity to the bacterial homologs. Further analyses demonstrated that they were most closely related to PLCs of Xenorhabdus, a genus of Gammaproteobacteria living in symbiosis with insect‐parasitizing nematodes. These suggested that H. halys might acquire these PLCs from Xenorhabdus through the mechanism of horizontal gene transfer (HGT), likely mediated by a nematode during its parasitizing an insect host. We also showed that the original HGT event was followed by gene duplication and expansion, leading to functional diversification of the bacterial‐origin PLC genes in H. halys. Thus, this study suggested that an herbivore might enhance adaptation through gaining genes from an endosymbiont of its parasite in the tripartite parasitic and symbiotic interactions.     

Ultrastructural changes of Drosophila larval and prepupal salivary glands cultured in vitro with ecdysone

Summary Alterations in the ultrastructure of in vitro cultured larval salivary glands of Drosophila melanogaster in response to the steroid hormone ecdysone were studied in relation to complex changes in puffing patterns. We found that the changes in the fine structure of cultured glands reflected progression of the puffing pattern, and they paralleled those seen in vivo. We observed that glue secretion by exocytosis, the main function of salivary glands, took place between puff stage 5 (PS5) and PS7. Glue could not be expectorated under culture conditions but was slowly released from the lumen through a duct into the medium. After the cultured glands reached PS13/PS14, further progress of puffing and fine structural alterations required that the ecdysteroid titer be transiently extremely low or absent. Under in vitro conditions we did not observe the putative new secretory program(s) described for glands in vivo after PS12. However, ultrastructural changes which unambiguously indicated that an autohistolytic process had begun in vitro started to appear after PS17. Many salivary gland cells developed numerous features of progressive self-degradation between PS18 and PS21. Actual degradation of salivary glands in vivo seemed to be rapid, but in vitro degradation was never completed, probably due to a lack of exogenous factors from the hemolymph. Manipulations of ecdysone titer in vitro in the culture medium, known during the larval puffing cycle to cause premature induction of developmentally specific puffing patterns, did not affect the normal development of ultrastructural features of the cytoplasm and nucleus.     

A histological and ultrastructural comparison of the male accessory reproductive glands of diapausing and non-diapausing adults in Bruchidius atrolineatus (Pic) (Coleoptera : Bruchidae)

Cytological variations of the median and the 2 lateral accessory glands of Bruchidius atrolineatus Pic (Coleoptera : Bruchidae) were examined as a function of age and the reproduction of the male. In sexually active virgin males, the secretory epithelium is columnar at emergence, but progressively flattens, and the secretions formed and stored by its cells are expelled by exocytosis into the glandular lumen. After 10 days, the male accessory glands exhibit a stage of repletion, characteristic of glands temporarily storing their secretions in their lumen. In diapausing males, the genital tract is relatively undeveloped and the accessory glands are reduced to tubules, whose lumen, surrounded by an epithelium composed of narrow cells, contains little secreted material. The presence of secretion aggregates in the secretory epithelial cells, the abundance of rough endoplasmic reticulum in them, and the release of a part of their secretions into the glandular lumen, indicate that reproductive diapause in B. atrolineatus is characterized by a decrease in the reproductive function. and not its total arrest.     

Salivary gland of the tick vector of east coast fever. IV. Cell type selectivity and host cell responses to Theileria parva

Responses of cells in the tick salivary gland to parasitism by Theileria parva were studied by electron microscopy. The gland is composed of three distinct types of acini (I, II, III) which together include ten or more different cell types. Of some 30 infected cells observed in the present study, all were E-cells of acinus III. The parasite thus exhibits a high degree of selectivity for acinus and cell type. The glandular cell invaded undergoes massive hypertrophy and accumulates glycogen deposits in its cytoplasm which may serve as an energy source for the growing intracellular parasite. As synthesis of its secretory material declines the product is packaged in progressively smaller secretory granules. The extensive arrays of endoplasmic reticulum are dismantled and eliminated in autophagic vacuoles. Excess secretory granules are also broken down by crinophagy. After 4 days, sporogony is completed and the host cell contains 30,000–50,000 sporozoites in an electron-lucent cytoplasm largely devoid of cytomembranes and secretory granules. Mitochondria are still present and normal in appearance. The loss of basophilia and secretory granules observed heretofore by light microscopy have been attributed to ingestion and destruction of host organelles by the parasite. The pallid appearance of the cytoplasm has been interpreted as a sign of impending degeneration of the host cell. In electron micrographs no ingestion of organelles by the parasite or degenerative changes were found. The host cell clearly remains viable and metabolically active throughout sporogony. The striking changes in its ultrastructure result from active elimination of organelles and inclusions by the host cell itself in response to parasitism.