Salivary gland ultrastructure of the unfed adult and feeding female of Haemaphysalis (Rhipistoma) leachi (Ixodoidea: Ixodidae)

The paired salivary glands of unfed adult Haemaphysalis (Rhipistoma) leachi contain one type of agranular and three types of granular alveoli connected to a salivary duct system. Type I agranular alveoli consist of one large, central cell surrounded by peripheral cells with numerous basal membrane infoldings indicative of epithelia involved in fluid transport. Glycogen particles, lipid-like droplets, and the parallel pattern of infolded membranes disappeared from the peripheral cells during feeding. Types II, III, and IV granular alveoli contain some agranular interstitial epithelial cells, cap cells, and fundus cells, but are predominantly composed of structurally different granular cell types a, b, c, d, e, and f. Agranular cells develop during the early stages of feeding. Granular a, c, e, and f cells release their granules directly after attachment to the host and possibly are involved in cement secretion required for firm attachment to it. The b cell granules are replaced by b₁ filamentous granules during feeding. Golgi bodies and rough endoplasmic reticulum (RER) participate in the formation of most types of granules. The d cells contain lamella-like structures and condensing vacuoles, probably responsible for lysosome formation. The main salivary duct and all types of alveoli are innervated by neurosecretory axons.     

Ultrastructural changes in the salivary alveoli ofArgas (Persicargas) persicus (Ixodoidea: Argasidae) during and after feeding

Two types of salivary alveoli are present in adultArgas (Persicargas) persicus: agranular type I and granular type II alveoli. Type I alveoli consist of a large central cell surrounded by a constrictor cell and peripheral cells with numerous infoldings of the basal membrane similar to epithelia involved in active transport. The basal infoldings form a previously undescribed successively convoluted membranous pattern which may increase the capability of peripheral cells for active transport. Type II alveoli consist of three granular cells (a, b, c) and two agranular (adlumenal and ablumenal interstitial) cells. Golgi bodies and rough endoplasmic reticulum are probably involved in the granule formation. The granules are discharged within 5–10 min after feeding commences, and presumably contain anticoagulant substances and pharmacologically active agents that promote the blood flow of the host during tick feeding. Although the adlumenal cells are not structurally affected by feeding, ablumenal cells develop into transporting epithelia.     

Ultrastructure of salivary glands of Ornithodoros (Ornithodoros) moubata (Ixodoidea: Argasidae)

The paired salivary glands of unfed adult Ornithodoros (Ornithodoros) moubata are composed of type I (agranular) and type II (granular) alveoli. Type I alveoli consis of one large central cell surrounded by peripheral cells having the morphology of fluid-transporting epithelia. Type II alveoli contain granular and agranular cells; the former are comprised of morphologically distinct types of cells (a, b, and c) containing granules of different structures and chemical composition with respect to polysaccharide and protein. The agranular cells are the interstitial and cap cells. Golgi bodies and rough endoplasmic reticulum (RER) are found in all granular cells and apparently are involved in granule formation. No appreciable structural changes were observed in type I alveoli during or after feeding. Type c cell granules are released before granules from types a and b cells and may contain anticoagulant substances that promote the blood flow of the host during the tick feeding. Although the cap cells are not structurally affected by feeding, interstitial cells are developed into transporting epithelia.     

Immunohistochemical localization of adenosine 3':5'-cyclic monophosphate in female ixodid tick Amblyomma americanum (L.) salivary glands

Localization of adenosine 3':5'-cyclic monophosphate (cyclic AMP) in alveoli of salivary glands of female Amblyomma americanum (L.) was accomplished with an indirect immunofluorescent technique. Little cyclic AMP fluorescence was seen in Type I alveoli in glands of unfed females but considerable fluorescence was seen in Type I alveoli of glands obtained from females that had fed. The most intense cyclic AMP fluorescence was observed in complex granular cells of Type II and III alveoli in glands of unfed females and glands from females in early stages of tick feeding. In the latter stages of tick feeding an increase in fluorescence in Type III alveoli was observed in cells near the lumen, possibly adluminal interstitial or transformed granular cells.     

Morphofunctional changes in the midgut of the Ixodes females (Acari: Ixodidae) during the immunizing feedings

The changes of the midgut in the females of the tick species Ixodes persulcatus and I. ricinus during the second and third immunizing feeding on rabbits were studied by the histological technics. The alternation of one generation of the digestive cells of nymphal stage and two generations of the digestive cells of adult stage was observed. The generation of secretory cells is absent. The tick completes feeding and drop off when the last generation of the digestive cells is on the initial activity stages. The amount of the blood consumed is not enough for the rhythmical functioning of the midgut. The feeding of tick is broken on the second phase and full satiation does not take place. It is apparently an effect of the interruption of the blood entrance into the midgut cavity of feeding tick as a probable result of anti-ticks resistance of unnatural hosts.     

Effect of forest fires on the population count and spatial structure of the taiga tick (Ixodidae)

Results of two large-scale surveys of Ixodes persulcatus distribution were compared. Surveys were conducted in a south taiga forest area in the Khabarovsk region two and ten years after a great fire. The direct inhibiting effect of the fire on the population of I. persulcatus was not continuous though apparent. By the moment of pyrogenic small forests formation on slash fires, i.e. during the period not exceeding 2-3 complete developmental cycles of I. persulcatus, the abundance and spatial structure of its population are restored completely.     

The infection of ixodid ticks collected from humans with the tick-borne encephalitis virus in Tomsk city and its suburbs

The ticks Ixodes persulcatus and I. pavlovskyi collected from people visited gardens and suburban forests have been examined by the IFA methods on the presence of tick-borne encephalitis virus (TEV). It was established, that the most part of ticks collected from humans belongs to I. persulcatus, despite the fact that I. pavlovskyi dominates on the territory of the city and its suburbs. TEV infection was registered more often in fed ticks, in comparison with those without signs of preceding feeding. Infected specimens constituted 9.4% of 53 I. persulcatus individuals examined. In I. pavlovskyi 3.7% of 27 examined specimens were infected. In fed ticks (41 specimens of I. persulcatus and 20 I. pavlovskvi) the rates of infected specimens were higher and constituted 48.8 and 35%, respectively. In suburban forests, as well as in the city, I. persulcatus was found to attack humans more often than I. pavlovskyi. TEV infection was recorded in 12.7% of 220 unfed I. persulcatus specimens examined and in 41.6% of 142 fed individuals. In I. pavlovskyi, 6.6% of unfed and 25% of fed specimens was infected (33 unfed and 24 fed specimens were examined). Thus, in all territories examined, percent of infected specimens was significantly lower in I. pavlovskyi, and in fed ticks of both species, TEV was recorded more often.     

Histopathological changes in the skin of small mammals in the area of feeding of Ixodes trianguliceps, I. Persulcatus i I. Ricinus (Ixodidae) ticks

Histopathological changes in the places of attachment and feeding of the Ixodes trianguliceps, I. persulcatus, and I. ricinus on natural hosts are similar. They pass through the stages of skin inflammation: perforation, exudation and proliferation. The mouthparts of tick contact with the host tissues. The fibrin deposit and callogen capsule surround the mouthparts. These species of ticks do not form the cement. Formation of the feeding cavity is caused by the salivation of ticks and neutrophil infiltration into surrounding tissues. Unnatural hosts (mice, rabbits) do not form the callogen capsule in the places of feeding, the callogen fibres are disposed irregular. Mast cells accumulate in the tick attachment site in the skin of natural host and degranulate in response to the tick salivary antigens releasing the histamine. Mediators involve cutaneous hypersensitivity reactions.     

The taxonomic aspects of the transmission of the causative agent of Lyme disease

According to world literature data 17 species of ixodid ticks have been studied for natural infection with the Lyme disease agent. Analysis of the data on the level of the infection, transovarial and transphase transmission has shown that main biological vectors of Borrelia burgdorferi are the species of the subgenus Ixodes s. str. - I. ricinus, I. persulcatus (Eurasia), I. dammini, I. pacificus (North America). Potential vectors are I. scapularis, I. dentatus, Amblyomma americanum, Dermacentor variabilis. Single isolations were registered for I. neotomae, Haemaphysalis leporispalustris, D. occidentalis. Nonidentified spirocheta was isolated from A. americanum, D. variabilis, D. parumapertum, Rhipicephalus sanguineus. No agent was isolated from I. cookey, D. albipictus, R. reticulatus, H. concinna. On the basis of comparative and ontogenetic data the species from a group of main vectors: I. ricinus, I. persulcatus, I. pacificus had been attributed by me to the phyletic group persulcatus before Lyme disease was discovered and its causative agent isolated. The question whether I. scapularis belongs to the group persulcatus was also discussed at that time but left open due to somewhat aberrant structure of gnathosoma at preimaginal phases (Filippova, 1969, 1971, 1973). 6 Palaearctic, 2 Indomalayan and 3 Nearctic species were referred to the group persulcatus at the time. I. dammini was described later, in 1979. Gnathosoma of its preimaginal phases has an intermediate structure between I. scapularis and other species of the group persulcatus. Sexually mature phase and nymph of I. dentatus have much in common with Palaearctic members of the group, I. pavlovskyi, I. kazakstani, I. kashmicus. Preimaginal phases of I. scapularis and nymph of I. dentatus were studied by me on the collection material. Thus, it is possible to speak of the belonging of main vectors of B. burgdorferi to a common phyletic group within the subgenus Ixodes s. str. and, therefore, of common origin of ecological medium for the agent. At the same time each species of the vector is an evolutionally developed difference of ecological medium for B. burgdorferi. Roots of the group persulcatus could originate as far as in Paleocene before the land connection between North America and Europe disappeared. Conditions for the existence of recent species, however, appeared considerably later and their flourishing is dated by Pliocene. The main epidemiological role belongs now to I. ricinus, I. persulcatus, I. dammini, I. pacificus.(ABSTRACT TRUNCATED AT 400 WORDS)     

Fetal mouse lung in circumfusion system cultures.

Fetal mouse lungs were cultivated, using the dual-rotary circumfusion system for tissue culture, and their histotypic development was surveyed for 75 days by phase-contrast and electron microscopy. Alveoli, terminal bronchioles and alveolar macrophages were photographed periodically with still and time-lapse phase-contrast microscopy. Their histotypic appearance was confirmed by electron micrographs of the 1- and 2 1/2-month-old specimens. These revealed typical alveoli surrounded by a basal lamina and composed of types I and II pneumocytes containing various lamellar-body forms within the type II cells, the alveolar lumen, and the alveolar macrophages. There was a shift from almost all type II cells in the 1-month-old alveoli to the presence of frequent type I cells as constituents of the alveoli in the 2 1/2-month-old cultures. The terminal bronchioles were tubules consisting of ciliated cells with Clara cells interspersed between them. The ciliated cells contained as many as 30 cilia or basal bodies per section and numerous microvilli. They were attached to each other and to the Clara cells by junctional complexes and accessory desmosomes which were generally in the apical ends of the cells. The Clara cells typically had glycogen granules interspersed between lamellae of the endoplasmic reticulum, contained numerous well dispersed mitochondria, occasional lysosome-like granules and crystalloid bodies which appeared to be tubular. Some Clara cells presented a moderatley dense secretory granule in the center of the whorl of the endoplasmic reticulum.     

The effect of level of feeding, genetic merit, body condition score and age on biological parameters of a mammary gland model

An evolutionary algorithm was applied to a mechanistic model of the mammary gland to find the parameter values that minimised the difference between predicted and actual lactation curves of milk yields in New Zealand Jersey cattle managed at different feeding levels. The effect of feeding level, genetic merit, body condition score at parturition and age on total lactation yields of milk, fat and protein, days in milk, live weight and evolutionary algorithm derived mammary gland parameters was then determined using a multiple regression model. The mechanistic model of the mammary gland was able to fit lactation curves that corresponded to actual lactation curves with a high degree of accuracy. The senescence rate of quiescent (inactive) alveoli was highest at the very low feeding level. The active alveoli population at peak lactation was highest at very low feeding levels, but lower nutritional status at this feeding level prevented high milk yields from being achieved. Genetic merit had a significant linear effect on the active alveoli population at peak and mid to late lactation, with higher values in animals, which had higher breeding values for milk yields. A type of genetic merit × feeding level scaling effect was observed for total yields of milk and fat, and total number of alveoli produced from conception until the end of lactation with the benefits of increases in genetic merit being greater at high feeding levels. A genetic merit × age scaling effect was observed for total lactation protein yields. Initial rates of differentiation of progenitor cells declined with age. Production levels of alveoli from conception to the end of lactation were lowest in 5- to 8-year-old animals; however, in these older animals, quiescent alveoli were reactivated more frequently. The active alveoli population at peak lactation and rates of active alveoli proceeding to quiescence were highest in animals of intermediate body condition scores of 4.0 to 5.0. The results illustrate the potential uses of a mechanistic model of the mammary gland to fit a lactation curve and to quantify the effects of feeding level, genetic merit, body condition score, and age on mammary gland dynamics throughout lactation.     

Detection of two zoonotic Babesia microti lineages, the Hobetsu and U.S. lineages, in two sympatric tick species, ixodes ovatus and Ixodes persulcatus, respectively, in Japan

The species Babesia microti, commonly found in rodents, demonstrates a high degree of genetic diversity. Three lineages, U.S., Kobe, and Hobetsu, are known to have zoonotic potential, but their tick vector(s) in Japan remains to be elucidated. We conducted a field investigation at Nemuro on Hokkaido Island and at Sumoto on Awaji Island, where up to two of the three lineages occur with similar frequencies in reservoirs. By flagging vegetation at these spots and surrounding areas, 4,010 ticks, comprising six species, were collected. A nested PCR that detects the 18S rRNA gene of Babesia species revealed that Ixodes ovatus and I. persulcatus alone were positive. Lineage-specific PCR for rRNA-positive samples demonstrated that I. ovatus and I. persulcatus carried, respectively, the Hobetsu and U.S. parasites. No Kobe-specific DNA was detected. Infected I. ovatus ticks were found at multiple sites, including Nemuro and Sumoto, with minimum infection rates (MIR) of ～12.3%. However, all I. persulcatus ticks collected within the same regions, a total of 535, were negative for the Hobetsu lineage, indicating that I. ovatus, but not I. persulcatus, was the vector for the lineage. At Nemuro, U.S. lineage was detected in 2 of 139 adult I. persulcatus ticks (MIR, 1.4%), for the first time, while 48 of I. ovatus ticks were negative for that lineage. Laboratory experiments confirmed the transmission of Hobetsu and U.S. parasites to hamsters via I. ovatus and I. persulcatus, respectively. Differences in vector capacity shown by MIRs at Nemuro, where the two species were equally likely to acquire either lineage of parasite, may explain the difference in distribution of Hobetsu throughout Japan and U.S. taxa in Nemuro. These findings are of importance in the assessment of the regional risk for babesiosis in humans.     

The subgenus Persicargas (Ixodoidea: Argasidae: Argas). 23. Fine structure of the salivary glands of unfed A. (P.) Arboreus Kaiser, Hoogstraal, and Kohls.

Salivary glands of the unfed adult Argas (Persicargas) arboreus (family Argasidae) contain 2 types of alveoli, one nongranular and one granule-secreting. The fine structure of the nongranular alveolus is similar to that of the family Ixodidae. In the granule-secreting alveolus, the presence of 3 types of secretory cells, each with morphologically distinct granular inclusions, confirms histological and histochemical observations on argasid salivary glands. Epithelial cells with numerous membranous infoldings, mitochondria, microtubules, and a complex canalicular system probably concerned with fluid regulation and secretion are located between granule-secreting cells and form caps over their basal regions. The luminal border of both secretory and epithelial cells is microvillate. The alveolar lumen leads into the chitinous alveolar duct which lacks the complex valvular structure of ixodid alveoli. Axons containing neurosecretory material occur in both nongranular and granule-secreting alveoli and probably control salivary secretion.     

Multistage mechanism of reproductive sympatric isolation of the closely-related species, Ixodes persulcatus and I. Pavloskyi (Ixodidae)

The closely related species Ixodes persulcatus and I. pavlovskyi are vectors of agents tick-born encephalitis and Lyme disease. These species have two great disjunctive sympatric distribution areas (fig. 1), where both species inhabit the same biotopes, and during the same season. In these conditions there are prerequisites of contacts of opposite sexes of the different species during ambushing the hosts on vegetation. Sequence of "switching on" of the morphological, physiological, and hostparasite factors of reproductive isolation has been established. The features and the range of manifestation of these factors at different combinations of sexes and species of the partners have been revealed. The studied factors, such as precopulative and postcopulative components of the mechanism of reproductive isolation of I. persulcatus and I. pavlovskyi were considered. A hypothesis for sympatric origin of the species as a result of specialization the adults of I. persulcatus to mammals, and I. pavlovskyi to birds is proposed.     

Ultrastructural aspects of saliva production by the granule-secreting acini of the tick Ixodes persulcatus females

Fine structure of salivary glands was investigated in the tick Ixodes persulcatus females. Ultrastructure of II and III acini in unfed and feeding females is described. Changes in size and structure of the secretory products during feeding are demonstrated.     

Morphofunctional changes in the midgut of virgin tick females of the genus ixodes (Acarina: Ixodidae) during and after feeding

The midgut epithelium of the virgin females in Ixodes ricinus, L. pacificus, and I. persulcatus is represented by the digestive cells of nymphal phase and stem cells. Digestion of feed is performed by the one generation of digestive cells of nymphal phase and two generations of digestive cells of adult females. The generation of secretory cells and two generations of digestive cells are absent. The feeding of virgin females is not completed but interrupted in the second phase during the period of slow feeding and preparation of the intestine for the third phase.     

Peritrophic matrix in the midgut of tick females of the genus Ixodes (Acari: Ixodidae)

The formation of the peritrophic matrix in the midgut of females of 5 ixodid tick species (Ixodes pacificus, I. pavlovskyi, I. persulcatus, I. ricinus and I. scapularis) was studied by means of light and electron microscopy in different periods of the feeding and after detachment. The formation of the peritrophic matrix started when the first food portions came into the gut lumen, 9-12 hours after the attachment. Renovation of the peritrophic matrix took place during the whole feeding period; every new generation of midgut cells synthesized their own matrix. It was deposited on the apical surface of every midgut cell in the beginning of differentiation, and was functioning during the life of the cell. The peritrophic matrix separates spaces of the cavitary and cytozoic digestions.     

Developmental changes in salivary glands of nymphs and adults of the spinose ear tick Otobius megnini

Otobius megnini has an autogenous 1 host life cycle, where larvae and nymphs stay attached inside the ear canal for long periods, but the adult tick is free living and can lay several egg batches without feeding. In order to obtain information about anatomical structures involved in this particular life cycle, nymphs and adults of O. megnini were dissected and salivary gland images were obtained in situ with the use of scanning electronic microscopy. Measurements of salivary alveoli were obtained with the use of ImageJ 1.40 g software. In the nymphs, the Type I alveoli are relatively small (mean diameter: 19.6 μm) compared with those of the adults (mean: 43.4 μm) and other soft ticks in the literature. Type II alveoli in nymphs are similar (mean: 82.6 μm) to previously described alveoli in adult soft ticks. In contrast, the adults of O. megnini Type II alveoli are smaller (mean: 36.8 μm) and have a wrinkled surface. These findings provide more evidence that Type I alveoli take part in absorption of moisture during the free-living tick stages.     

The meadow tick Dermacentor reticulatus in natural foci of tick-borne encephalitis in Udmurtia

In southern Udmurtia, where occur both Dermacentor reticulatus and Ixodes persulcatus, in the years of high abundance of small mammals feeding periods of larvae and nymphs of these ticks coincide completely. The duration of simultaneous parasitism of preimago of the above ixodid species on animals is of epidemiological significance as far as within this period the infection of young stages of D. reticulatus with the virus of tick-borne encephalitis occurs. This is confirmed by the isolation of I strain of the virus from D. reticulatus collected at the state of engorged nymphs on hosts common with I. persulcatus and moulted in the laboratory.     

Morphological barrier in mechanisms of reproductive isolation acting in areas of sympatry of closely related species Ixodes persulcatus-I.pavlovskyi and I. persulcatus-I. ricinus (Ixodidae)

A morphometric analysis of different male and female organs used in the process of copulation, including an attachment of the spermatophore to the genital opening of female and placing the spermatophore into the vestibular part of vagina, was carried out for the pairs of ixodid tick species Ixodes persulcatus--I. pavlovskyi and I. persulcatus--I. ricinus in areas of their sympatry. In two disjunctive areas of sympatry of I. persulcatus--I. pavlovskyi (South-East Altai and Southern Primorye), the morphological barrier is displayed in the precopulation mechanism of reproductive isolation and does act only in the pair I. pavlovskyi female--I. persulcatus male. It is proved by the reliable prevalence of the hypostome diameter (in widest part) of I. persulcatus male over the genital opening length of I. pavlovkyi female. Because of this reason, the copulation between these partners cannot take place. On the other hand, there are no any morphological obstacles for a copulation within each species examined and in the combination I. persulcatus female--I. pavlovskyi male, and in any combination of the partners I. persulcatus--I. ricinus in sympatry areas (samples from two locations in the north-west of the Eastern-European Plane have been examined). Morphological peculiarities causing differences in host preference of tick species, that in turn realizes the isolation mechanisms between species, are discussed. Postcopulation non-genetic components of the reproductive isolation mechanism have been found in examined species.