Developmental stages of Trypanosoma (Megatrypanum) freitasi Rego, Magalh?es & Siqueira, 1957 in the opossum Didelphis marsupialis (Marsupialia, Didelphidae)

Trypanosoma (Megatrypanum) freitasi, a parasite of marsupials of the genus Didelphis, has been found to undergo in the lumen of the scent (anal) glands of its vertebrate host, a cycle such as usually occurs in the intestinal tract of the insect vectors of trypanosomatids and similar to what has been reported for Trypanosoma (Schizotrypanum) cruzi. The invertebrate host of Trypanosoma freitasi is still unknown. Developmental stages of the trypanosome in its mammalian host, especially the dividing epimastigotes, multinucleate plasmodial forms and rosettes found in the lumen of the scent glands of a naturally infected Didelphis marsupialis are described and illustrated.     

Continuous Growth and Differentiation of Trypanosoma (Megatrypanum) freitasi Rego, Magalhaes & Siqueira, 1957, In Vitro

Trypanosoma (Megatrypanum) freitasi, a parasite of didelphid opossum, was known to be very difficult to cultivate in conventional media. Co-cultivation with L929 cell line in Baltz's medium at 27.5° C resulted in luxuriant growth of the trypanosome with the production of epimastigote colonies that adhered to the surface of culture flasks or tubes, and transformation into metacylics. Further transformation was stimulated by raising the incubation temperature. At 37° C the population was of the bloodstream type and resistant to lysis by complement.     

Scent glands in legume flowers

Scent glands, or osmophores, are predominantly floral secretory structures that secrete volatile substances during anthesis, and therefore act in interactions with pollinators. The Leguminosae family, despite being the third largest angiosperm family, with a wide geographical distribution and diversity of habits, morphology and pollinators, has been ignored with respect to these glands. Thus, we localised and characterised the sites of fragrance production and release in flowers of legumes, in which scent plays an important role in pollination, and also tested whether there are relationships between the structure of the scent gland and the pollinator habit: diurnal or nocturnal. Flowers in pre‐anthesis and anthesis of 12 legume species were collected and analysed using immersion in neutral red, olfactory tests and anatomical studies (light and scanning electron microscopy). The main production site of floral scent is the perianth, especially the petals. The scent glands are distributed in a restricted way in Caesalpinia pulcherrima, Anadenanthera peregrina, Inga edulis and Parkia pendula, constituting mesophilic osmophores, and in a diffuse way in Bauhinia rufa, Hymenaea courbaril, Erythrostemon gilliesii, Poincianella pluviosa, Pterodon pubescens, Platycyamus regnellii, Mucuna urens and Tipuana tipu. The glands are comprised of cells of the epidermis and mesophyll that secrete mainly terpenes, nitrogen compounds and phenols. Relationships between the presence of osmophores and type of anthesis (diurnal and nocturnal) and the pollinator were not found. Our data on scent glands in Leguminosae are original and detail the type of diffuse release, which has been very poorly studied.     

From a mono to a digenetic life-cycle: how was the jump for flagellates of the family Trypanosomatidae?

It has been found that species of the digenetic genus Trypanosoma as well as species of monogenetic trypanosomatids of insects can grow in the lumen of the scent glands of opossums reproducing the cycle they perform in the intestinal tract of their insect hosts. Based on these findings, speculations are made on the incompletely known cycles of many mammalian trypanosomes and on the evolution of the family Trypanosomatidae.     

Pollinator attraction to volatiles from virgin and pollinated host flowers in a yucca/moth obligate mutualism

The classic obligate pollination–seed consumption mutualism between yuccas and yucca moths has been thought to be mediated by chemical cues, but empirical data on pollinator attraction to host floral volatiles in this association have been lacking. Here we show that the scent from virgin flowers of the host Yucca glauca is sufficient to attract its obligate pollinator Tegeticula yuccasella in Y‐tube olfactometer tests. Interestingly, both sexes of moths were attracted to the scent stimulus. Because yucca moths mate inside host flowers, the attraction of both females and males to host floral volatiles is likely to increase encounter rates. In a second test, female moths did not discriminate between virgin and hand‐pollinated flowers, indicating no post‐pollination change in scent production by the host that would lead to a reduction in pollinator attraction and thereby limit exploitation of the available seeds in host flowers. However, other mechanisms that could stabilise the mutualism between T. yuccasella and its yucca hosts have already been documented, i.e. selective abortion of heavily infested flowers, and a female‐derived host‐marking pheromone. Headspace collection and GC–MS were used to identify the blend of floral volatiles emitted by Y. glauca, which was found to be very similar to those of two other allopatric capsular‐fruited species, Y. elata and Y. filamentosa, revealing strong conservation of this trait within Yucca section Chaenocarpa.     

Morphological,Molecular, and Phylogenetic Characterization of Nosema ceranae,a Microsporidian Parasite Isolated from the European Honey Bee,Apis mellifera1

ABSTRACT. Nosema ceranae, a microsporidian parasite originally described from Apis cerana, has been found to infect Apis melllifera and is highly pathogenic to its new host. In the present study, data on the ultrastructure of N. ceranae, presence of N. ceranae-specific nucleic acid in host tissues, and phylogenetic relationships with other microsporidia species are described. The ultrastructural features indicate that N. ceranae possesses all of the characteristics of the genus Nosema. Spores of N. ceranae measured approximately 4.4 × 2.2 μm on fresh smears. The number of coils of the polar filament inside spores was 18–21. Polymerase chain reaction (PCR) signals specific for N. ceranae were detected not only in the primary infection site, the midgut, but also in the tissues of hypopharyngeal glands, salivary glands, Malpighian tubules, and fat body. The detection rate and intensity of PCR signals in the fat body were relatively low compared with other examined tissues. Maximum parsimony analysis of the small subunit rRNA gene sequences showed that N. ceranae appeared to be more closely related to the wasp parasite, Nosema vespula, than to N. apis, a parasite infecting the same host.     

Trypanosoma cruzi in the scent glands of Didelphis marsupialis: the kinetics of colonization

This study examined the dynamics of colonization of Trypanosoma cruzi in the scent glands of the opossum Didelphis marsupialis following direct inoculation with 10(5) epimastigotes of isolate G-49 (an opossum-derived strain). One, three, and five days, 1 month, and 1 year after inoculation, scent glands were fixed for analysis using brightfield and electron microscopies. One day after inoculation the parasites, mainly as epimastigotes, were randomly distributed into the lumen. From the third day on, the parasites still in the form of epimastigotes tended to concentrate closer to the epithelium. The flagellates reached the definitive distribution pattern on the fifth day, when they formed huge clusters deep into the foveae. In samples collected 1 month and 1 year after inoculation, the ratio of epimastigotes:trypomastigotes was 1:1, with epimastigotes predominating near the epithelium and trypomastigotes far from it. Our observations suggest that T. cruzi grows continuously in the scent glands and does not depend on adhesion to promote metacyclogenesis. Metacyclogenesis far from the epithelium seems to be an important selective advantage to both host and parasite, since it assures the elimination of the infective forms of the parasite when the host expels the glands' contents, which occurs in frightening situations or at times of stress. The morphological characteristics of infected and noninfected scent glands using transmission and scanning electron microscopies were also described.     

External thoracic scent efferent system of Scutelleridae (Hemiptera: Heteroptera)

The metathoracic scent glands in the Heteroptera produce defence secretions which are spread outside the body through and by using the thoracic external scent efferent system. That complex system was studied in 18 species from 11 genera of four subfamilies, Elvisurinae, Eurygastrinae, Hoteinae and Scutellerinae of the family Scutelleridae (Pentatomoidea). The results have been compared with published data. The pattern of that system is more consistent at the level of genus, mostly very similar in the congeneric species, but mostly variable within higher taxonomic levels, tribes and subfamilies. Five types of the external scent efferent system are recognized within the family Scutelleridae, basic two of them in studied species: (i) peritreme well developed, covering large part of metapleuron, evaporatorium small, developed only on metapleuron, (ii) evaporatorium large, more conspicuous than moderate-sized to small peritreme, extending to mesopleuron as large structure. The results do not support a hypothesis that the system of structures associated externally with metathoracic scent glands is in correlation with type of a habitat. However, these structures are well usable as diagnostic characters for scutellerid genera (e.g. Cantao, Hyperonchus, Scutellera and Solenosthedium).     

Comparative ultrastructure of coxal glands in unfed larvae of Leptotrombidium orientale (Schluger, 1948) (Trombiculidae) and Hydryphantes ruber (de Geer, 1778) (Hydryphantidae)

Coxal glands of unfed larvae Leptotrombidium orientale (Schluger, 1948) (Trombiculidae), a terrestrial mite parasitizing vertebrates, and Hydryphantes ruber (de Geer, 1778) (Hydryphantidae), a water mite parasitizing insects were studied using transmission electron microscopy. In both species, the coxal glands are represented by a paired tubular organ extending on the sides of the brain from the mouthparts to the frontal midgut wall and are formed of the cells arranged around the central lumen. As in other Parasitengona, the coxal glands are devoid of a proximal sacculus. The excretory duct, joining with ducts of the prosomal salivary glands constitutes the common podocephalic duct, opening into the subcheliceral space. The coxal glands of L. orientale are composed of a distal tubule with a basal labyrinth, an intermediate segment without labyrinth, and a proximal tubule bearing tight microvilli on the apical cell surface and coiled around the intermediate segment. The coxal glands of H. ruber mainly consist of the uniformly organized proximal tubule with apical microvilli of the cells lacking the basal labyrinth. This tubule shows several loops running backward and forward in a vertical plane on the side of the brain. In contrast to L. orientale , larvae of H. ruber reveal a terminal cuticular sac/bladder for accumulation of secreted fluids. Organization of the coxal glands depends on the ecological conditions of mites. Larvae of terrestrial L. orientale possess distal tubule functioning in re‐absorption of ions and water. Conversely, water mite larvae H. ruber need to evacuate of the water excess, so the filtrating proximal tubule is prominent.     

Intraspecific Variation of Floral Scent Chemistry in Magnolia kobus DC. (Magnoliaceae)

Magnolia kobus was examined at 32 sites in Japan (109 female-stage flowers from 52 plants) by GC-MS. Major chemical compounds (a total of 36 chemicals) emitted from the flowers were: linalool (and its oxides), limonene, cis- and trans-β-Ocimene, benzaldehyde, benzyl alcohol, benzyl cyanide, and 2-aminobenzaldehyde. Linalool and its oxides were the most abundant components of floral scents in 21 individuals. The rate at which chemical volatiles were emitted ranged from 0.002 to 0.929 μg/flower/hour (average 0.211). High quantitative and qualitative variation in floral scent chemistry among individuals was found throughout the range of M. kobus, especially in central Honshu. The high variability in floral scent chemistry may be due to the importance of visual cues in the reproductive biology of M. kobus which flowers in early spring, resulting in decreased selection for specific floral scent profiles. Alternatively, different scent compounds or chemical profiles may be equally effective in attracting pollinators. Received 25 June 2001/ Accepted in revised form 25 August 2001     

Histological examinations of facial glands in <Emphasis Type="Italic">Saccopteryx</Emphasis><Emphasis Type="Italic">bilineata</Emphasis> (Chiroptera,Emballonuridae), and their potential use in territorial marking

Scent marking is widespread among individuals of Mammalia species, especially in resource defence social systems. Apart from urine and faeces that are used for claiming resource ownership, specialised scent glands are the main source of secretions in scent marking individuals. Most previous studies have described secretory epithelia macroscopically, since many glands are conspicuous. But macroscopically inconspicuous scent glands or morphological structures might then be overlooked. In Saccopteryx bilineata (greater sac-winged bat), behavioural observations suggest that both sexes have, apart from the conspicuous gular glands of males, specialised facial glands to display territorial marking. We investigated the facial glands of two males and one female S. bilineata histologically and found, first, that both sexes possess a bilateral symmetrically intermandibular gland, which is composed of a bed of modified apocrine sudoriferous cells. Second, we found lip glands consisting of modified apocrine sudoriferous cell units with pigmented ducts around the upper and the lower lip. Both gland types are probably involved during territorial marking.     

Floral scent composition in early diverging taxa of Asclepiadoideae,and Secamonoideae (Apocynaceae)

The Apocynaceae–Asclepiadoideae are well known for their specialized floral morphologies and pollination systems and many species have distinct floral aromas. However, our knowledge on the chemistry of floral volatiles in this plant family is relatively limited although it has been suspected that floral scent plays a key function for pollinator attraction. This is the third paper in a series of papers reporting on the floral odours of Asclepiadoideae. Floral odours of eleven species from seven genera (Cibirhiza, Fockea, Gymnema, Hoya, Marsdenia, Stephanotis and Telosma) of early diverging taxa of Apocynaceae–Asclepiadoideae, and two species of Secamone (Apocynaceae–Secamonoideae) were collected using headspace sampling and then analyzed via GC–MS. We detected 151 compounds, of which 103 were identified. The vast majority of chemicals identified are common components in flower odour bouquets of angiosperms. However, striking was the high relative amount of acetoin (97.6%) in the flower scent of Cibirhiza albersiana. This compound has rarely been reported as a flower scent component and is more commonly found in fermentation odours. Bray–Curtis similarities and Nonmetric-Multidiminsional Scaling (NMDS) analyses showed that each of the species has a distinct odour pattern. This is mostly due to only twelve compounds which singly or in different combinations dominated the scent of the species: the benzenoids benzyl acetate, benzaldehyde, methyl benzoate, and 2-phenylethyl alcohol; the monoterpenoids (E)-ocimene, (Z)-ocimene, linalool, and eucalyptol; and the aliphatic compounds acetoin, and (E,Z)-2,6-nonadienal. The floral scent compositions are discussed in relation to tribal affiliations and their potential role for pollinator attraction, and are compared with the scent data available from other Asclepiadoideae species.     

Hormonal control of scent marking in the Indian musk shrew, Suncus murinus viridescens (Blyth)

The musk shrew, Suncus murinus viridescens, marks objects in the environment employing secretions of specialized skin glands and by urination and defecation. Diverse types of scent marking exhibited a significant decline after castration. Both methyltestosterone and ethinyloestradiol have been shown to reactivate the scent marking of castrate male shrews. Ethinyloestradiol effected an immediate elevation of marking pattern of castrate males.     

Fine structure of the male reproductive system in two species ofHaplognathia sterrer (Gnathostomulida,Filospermoidea)

Summary The structure of the male reproductive systems of two species ofHaplognathia cf.lyra andH. cf.rosacea was described. The structure of the testes and the anterior portions of the sperm ducts in both species was found to be similar. However, considerable species differences were found between the structures of the glands and muscles associated with the reproductive systems. These were more elaborate inH. cf.lyra than inH. cf.rosacea. The former species possessed an H-shaped sperm duct gland, three distinct groups of penis muscles and a penis with two cell types and with a lumen. The latter species had paired sperm duct glands, no specialized penis muscles and a penis with only one cell type and without a detectable lumen. No open gonopore was observed in either species. The sperm presumably exit through a ventral tissue connection observed connecting the penis and the ventral epidermis. These findings were discussed in the light of Mainitz's (1977) theory concerning the primitive penis type within the Gnathostomulida.Abbreviations ap anterior-posterior penis muscles - bm basement membrane - csd common sperm duct - dl dorsal lumen of the penis - dp dorsal gland cells of the penis - dv dorsoventral muscles anterior to the penis - dw sperm duct wall cell - e epidermis - ex exit cell - g intestine - gl gut lumen - n nerve - p penis - sd sperm duct - sdg sperm duct gland - tw testes wall cell - vl ventral lumen of the penis - vp ventral gland cells of the penis This project was supported by NSF grant #GB 42211 (R.M. Rieger P.I.). The line drawings have been executed after our design by Ms. Linda McVay     

Structures related to pheromone storage in alar androconia and the female abdominal scent gland of Heliconius erato phyllis,Heliconius ethilla narcaea,and Heliconius besckei (Lepidoptera: Nymphalidae: Heliconiinae)

We describe the morphology of alar androconia and the female abdominal scent gland of Heliconius erato phyllis, Heliconius ethilla narcaea, and Heliconius besckei. Androconial scales of Heliconius, which are arranged in overlapping wing bands, release pheromones during courtship, probably through vibratory movements of male wings over the female to induce her to mate. An antiaphrodisiac is produced by glands located in the valves of the male and is transferred during copulation to the yellow dorsal abdominal sac present in the virgin female, causing this sac to emit a scent that reduces the attractiveness of the female for courtship with other males. Stereomicroscopy, SEM, and TEM analyses were conducted to describe the morphology of the internal and external scales and the external abdominal scent sac. The findings revealed different sizes of external androconial scales and an internal group of porous structural vesicles that are probably related to the preservation of internal space, reception and storage of secretions, and elimination of volatiles when the male is actively involved in courtship. Translucent projections on the female abdominal scent sac create open reservoirs for the reception, storage, and emission of antiaphrodisiac volatiles along with stink clubs. Male valve denticles vary in form and probably attach securely to the female sac during mating, thus ensuring secretion transfer. These features are discussed in the context of a comparative analysis.     

Observations on the metathoracic scent gland system of the backswimmer,Notonecta glauca L. (Heteroptera: Notonectidae)

The closing apparatus in the metathoracic scent gland system of Notonecta glauca is described, and notes on the distribution of metathoracic scent glands in the family Notonectidae are included.     

Floral scent composition predicts bee pollination system in five butterfly bush (Buddleja,Scrophulariaceae) species

Traditionally, plant–pollinator interactions have been interpreted as pollination syndrome. However, the validity of pollination syndrome has been widely doubted in modern studies of pollination ecology. The pollination ecology of five Asian Buddleja species, B. asiatica, B. crispa, B. forrestii, B. macrostachya and B. myriantha, in the Sino‐Himalayan region in Asia, flowering in different local seasons, with scented inflorescences were investigated during 2011 and 2012. These five species exhibited diverse floral traits, with narrow and long corolla tubes and concealed nectar. According to their floral morphology, larger bees and Lepidoptera were expected to be the major pollinators. However, field observations showed that only larger bees (honeybee/bumblebee) were the primary pollinators, ranging from 77.95% to 97.90% of total visits. In this study, floral scents of each species were also analysed using coupled gas chromatography and mass spectrometry (GC‐MS). Although the five Buddleja species emitted differentiated floral scent compositions, our results showed that floral scents of the five species are dominated by substances that can serve as attractive signals to bees, including species‐specific scent compounds and principal compounds with larger relative amounts. This suggests that floral scent compositions are closely associated with the principal pollinator assemblages in these five species. Therefore, we conclude that floral scent compositions rather than floral morphology traits should be used to interpret plant–pollinator interactions in these Asian Buddleja species.     

The development of amphids and amphidial glands in adult Syngamus trachea (Nematoda: Syngamidae)

Nematode amphids are a pair of lateral cephalic sense organs, each comprising a group of sensory endings terminating in a cuticle-lined pit. In Syngamus trachea, a parasite of birds, each amphid is surrounded by two non-nervous supporting elements, a large gland cell basally and a smaller supporting cell anteriorly. The amphidial glands display high levels of secretory activity from five to six days postinfection. Secretory material is discharged through the lumen of the sense organ onto host tissue. The ultrastructure of amphids and amphidial glands has been investigated in newly moulted, immature and mature adults to trace the development of glandular activity and its effect on amphid-amphidial gland relationships. In newly moulted adults, the glands have very low levels of secretory activity and appear to act only as supporting cells to the amphids. As secretory activity increases, the gland cell membrane surrounding the sensory endings is elaborated into a reticulum which probably forms the secretory surface. In mature adults the amphid pit is swollen and filled with secretion; the sensory endings are relegated to the periphery of the lumen. It is suggested that amphidial glands develop from typical supporting cells, but acquire a new role possibly associated with parasite attachment.     

First report of filarial nematodes in free-living pitheciid primates

Although little studied, infections with nematodes of the Onchocercidae Leiper, 1911, predominated by the genera Dipetalonema Diesing, 1861 and Mansonella Faust, 1929, are frequent in wild primates and human populations in the Neotropical forest areas. This study reports natural infections with Dipetalonema freitasi Bain, Diagne & Muller, 1987 and D. gracile (Rudolphi, 1809) in two free-living species of pitheciid primates, extending the known geographical distribution of these species to the forest of the Peruvian Amazon. Adult worms were recovered from the thoracic and abdominal cavities of two species of monkeys, Pithecia monachus monachus (É. Geoffroy Saint-Hilaire) and Cacajao calvus ucayalii (Thomas) (Primates: Pitheciidae), collected along the Yavari-Mirin River basin and analysed via light and scanning electron microscopy. Both host species represent new host records for D. freitasi and D. gracile. Morphometric data are also presented for the sampled filarial worms in addition to morphological details obtained through light and electron microscopy examination of D. freitasi specimens.

     

Functional anatomy of scent glands in Paranemastoma quadripunctatum (Opiliones,Dyspnoi, Nemastomatidae)

The morphological organization and functional anatomy of prosomal defensive (scent) glands in Paranemastoma quadripunctatum, a representative of the dyspnoid harvestmen, was investigated by means of histological semithin sections, software‐based 3D‐reconstruction and scanning electron microscopy. Scent glands comprise large, hollow sacs on either side of the prosoma, each of these opening to the outside via one orifice (ozopore) immediately above coxa I. In contrast to the situation known from laniatorean, cyphophthalmid and some eupnoid Opiliones, ozopores are not exposed but hidden in a depression (atrium), formed by a dorsal integumental fold of the carapace and the dorsal parts of coxae I. Glandular sacs are connected to ozopores via a short duct which is equipped with a specific closing mechanism in its distal part: A layer of modified epidermal cells forms a kind of pad‐like tissue, surrounding the duct like a valve. Several muscles attached to the anterior parts of the glandular reservoir and to the epithelial pad may be associated with ozopore‐opening. The actual mechanism of secretion discharge seems to be highly unusual and may be hypothesized on the basis of corroborating data from behavioral observations, scent gland anatomy and secretion chemistry as follows: Enteric fluid is considered to be directed towards the ozopores via cuticular grooves in the surface of the coxapophyses of legs I. Then, the fluid is sucked into the anterior part of the scent gland reservoirs by the action of dorsal dilator muscles that widen the reservoir and produce a short‐term negative pressure. After dilution/solution of the naphthoquinone‐rich scent gland contents, a secretion‐loaded fluid is thought to be discharged with the help of transversal compressor muscles. This is the first detailed study on the functional anatomy of scent glands and the mechanisms of secretion discharge in the Dyspnoi. J. Morphol. 2011. © 2011 Wiley‐Liss, Inc.