Three new species of the genus Aporcelaimoides Heyns, 1965 from Vietnam (Nematoda,Dorylaimida, Aporcelaimidae), with an updated taxonomy of the genus

Three new species of Aporcelaimoides from natural habitats in Vietnam are studied, described and illustrated, including line drawings, LM and/or SEM pictures. Aporcelaimoides brevistylum sp. n. is characterized by its body 1.95–2.90 mm long, lip region offset by deep constriction and 17–18 µm broad, ventral side of mural odontostyle 11–14 µm long with aperture occupying 62–71% of its length, neck 663–767 µm long, pharyngeal expansion occupying 58–66% of total neck length, uterus a simple tube 85–182 µm long, pars refringens vaginae absent, V = 55–63, tail short and rounded (34–46 µm, c = 49–76, c’ = 0.6–0.8), spicules 67–86 µm long, and one ventromedian supplement out the range of spicules. Aporcelaimoides minor sp. n. is distinguished in having body 2.09–2.61 mm long, lip region offset by deep constriction and 19–20 µm broad, mural odontostyle 14–16 µm long at its ventral side with aperture occupying 73–84% of its length, neck 579–649 µm long, pharyngeal expansion occupying 57–66% of total neck length, uterus a simple tube 44–69 µm long, pars refringens vaginae well developed, V = 48–56, female tail very short, rounded conoid or truncate (14–26 µm, c = 90–146, c’ = 0.3–0.6), and male unknown. Aporcelaimoides silvaticum sp. n. is characterized by its body 2.09–2.60 mm long, lip region offset by depression and 17–18 µm broad, mural odontostyle 11–12 µm long at its ventral side with aperture occupying 60–66% of its length, neck 597–720 µm long, pharyngeal expansion occupying 58–64% of total neck length, uterus a simple tube 128–243 µm long, pars refringens vaginae well developed, V = 58–60, tail short and rounded (27–37 µm, c = 67–94, c’ = 0.6–0.7), spicules 64–75 µm long, and two or three widely spaced ventromedian supplements bearing hiatus. The genus Aporcelaimoides is restored, its diagnosis emended, and three species of Sectonema, namely Sectonema amazonicum, Sectonema haguei and Sectonema moderatum, transferred to it. An updated list of its species, a key to their identification and a tabular compendium with the most important morphometric features are also presented.     

Paurodontella parapitica n. sp. (Nematoda: Hexatylina,Sphaerularioidea) from Kermanshah Province,Western Iran

Paurodontella parapitica n. sp., collected from the rhizosphere of an apple tree in Kermanshah province, western Iran, is described. The new species is characterized by a body length of 505 to 723 µm (females) and 480 to 600 µm (males), lip region continuous by depression; 7 to 8 μm broad, 3 to 4 µm high, stylet length 7 to 9 µm or 1 to 1.3 times the lip region diameter, short postuterine sac of 4 to 6 μm long, lateral fields with five to six incisures; outer incisures crenated and inner incisures weakly crenated, excretory pore situated 90 to 100 µm from anterior end; functional males common in the population, with spicules 24 to 26 μm long. Tail of both sexes similar, almost straight and elongate-conoid. The new species resembles in morphology and morphometrics to four known species of the genus, namely P. apitica, P. minuta, P. myceliophaga, and P. sohailai. The results of phylogenetic analyses based on sequences of D2/D3 expansion region of 28S rRNA gene revealed this genus is polyphyletic in four different clades in Tylenchid.     

Calcaridorylaimus castaneae sp. n. (Nematoda,Dorylaimidae) from Bulgaria with an identification key to the species of the genus

An unknown species belonging to the genusCalcaridorylaimus Andrássy, 1986 was collected from the litter of broadleaf forests dominated by Castanea sativa Mill. and mixed with Quercus daleshampii Ten. and Fagus sylvatica L. on Belasitsa Mountain, south-western Bulgaria. Calcaridorylaimus castaneae sp. n. is characterised by its long body (1.4–2.1 mm), lip region practically not offset, vulva transverse, short odontostyle (14.5–16 μm) and tail (75.5–110.5 μm, c=14.7–23.6; c’=2.9–4.4) in females and 38–46 μm long spicules with small spur before their distant end in males. It is most similar to C. andrassyi Ahmad & Shaheen, 2004, but differs in having transverse vs pore-like vulva and shorter spicules (38–46 μm vs 52–57 μm). An identification key to the species of the genus Calcaridorylaimus is proposed. Phylogenetic analyses were performed on 18S and D2-D3 expansion domains of 28S rRNA genes by Neighbor-Joining, Maximum Likelihood and Bayesian Inference methods. The phylograms inferred from 18S sequences showed closest relationships of the new species with some species belonging to the genus Mesodorylaimus. However, insufficient molecular data for members of both genera do not allow the phylogenetic relationships of Calcaridorylaimus and the new species described herein to be elucidated.     

Margollus bokanicus n. sp. from Iran,the Fourth Species of a Rare Nematode Genus (Dorylaimida,Tylencholaimellidae)

Margollus bokanicus n. sp., collected from natural habitats in Khorasaneh district, Bokan, West Azarbaijan province, Iran, is described. Morphological and morphometric data are provided as well as drawings and light microscopy illustrations. The new species is characterized by a medium size body length (0.60 to 0.73 mm), labial and postlabial sclerotizations, lip region 7-μm wide, offset by constriction and long neck (167 to 207 μm), long pharyngeal basal bulb (27 to 36 μm) or 16% to 17% of total neck length, female genital system monodelphic–opisthodelphic, anterior branch reduced to a uterine sac (26–29 μm) or 1.1 to 1.3 times the body diameter, long posterior uterus (25–28 μm) or 1.1 to 1.3 times the body diameter, V = 40 to 47, cylindroid female tail (17 to 24 μm, c = 31 to 38, c’ = 1.1 to 1.4), and males unknown. This taxon is easily distinguishable from other Margollus species by its smaller general size and more posterior vulva. A compendium of Margollus species is also presented.     

First Report of Korean Cyst Nematode,Heterodera koreana,Parasitic on Bamboo,Phyllostachys nigra,from Iran

Bamboo is grown sporadically in the north of Iran and is confined to very limited areas. The history of growing bamboo was to some extent simultaneous with the entrance, commencement, and growth of the tea industry in the north about a century ago. The bamboo was used for making baskets to transfer the harvested tea foliage from farm to the factory and other linked functions. A main area allocated for bamboo growing is located in Lahidjan Agricultural Research Station (LARS) in the north of Iran, where several species of bamboo were cultivated in an area of 5 ha. The species include five species of Phyllostachys (viz., P. aurea, P. bambusoides, P. decora, P. nigra, P. vivax) and one species of Arundinaria gigantean, Pleioblastus fortune, and Semiarundinaria fastuosa; however, only P. aurea and P. nigra have been precisely identified. A survey on plant parasitic nematodes associated with bamboo mainly on P. nigra in LARS revealed second-stage juveniles of cyst forming nematode in soil samples. Further analysis of root and soil samples led to recovery of a cyst nematode belonging to the genus Heterodera and the Afenestrata group. Cysts, vulval cone, and second-stage juveniles were studied for morphological and morphometric features. The classical identification was followed by amplification of the ribosomal RNA-ITS region and the D2-D3 expansion segments of 28S large-subunit rRNA gene; the amplified fragments were sequenced, edited, and compared with those of the corresponding published gene sequences. New D2-D3 and rRNA-ITS gene sequences were deposited in the GenBank database under the accession numbers {"type":"entrez-nucleotide","attrs":{"text":"KR818910","term_id":"924272655","term_text":"KR818910"}}KR818910 and {"type":"entrez-nucleotide","attrs":{"text":"KR818911","term_id":"924272662","term_text":"KR818911"}}KR818911, respectively. Based on the morphological and molecular data, the species of the cyst-forming nematode was identified as H. koreana (Vovlas et al., 1992; Mundo-Ocampo et al., 2008). The body contour of cysts was mainly subspherical, vey often with irregular shape (Fig. 1A), yellowish to light brown, thin cuticle with fine zigzag pattern, without fenestration, lacking bulla, and underbridge. Vulval lips protruded, cuticular pattern of vulval cone with a tuberculate area (Fig. 2B), and vagina embedded into vulval lips. The second-stage juveniles cylindrical and slender, hemispherical cephalic framework, with three lines in lateral field, well-developed rounded stylet knobs, tail conoid tapring to fine rounded terminus, phasmids posterior to anus. The cyst measurements were (n = 21) length 502 ± 70 (420 to 640) µm; width = 408 ± 60 (320 to 520) µm; length/width = 1.23 ± 0.09 (1.07 to 1.5) µm. The morphometric characters of vulval cone were measured (n = 7): fenestral length = 62.4 ± 6.5 (51 to 71) µm; fenestral width = 50.7 ± 3.2 (45 to 54) µm; vulval slit = 51.9 ± 4.3 (46 to 59) µm; distance from vulva to anus = 51.3 ± 4.4 (43 to 56) µm. Second-stage juveniles showed the following morphometric characters (n = 14): L = 455 ± 11.3 (437 to 472) µm; a = 29.9 ± 0.9 (28.3 to 31.5); b΄ = 2.7 ± 0.4 (2.2 to 3.5); c = 7.4 ± 0.9 (6 to 8.9); ć = 6.1 ± 0.4 (5.1 to 6.7); lip region height = 3 µm; lip region width = 7.5 ± 0.5 (7 to 8) µm; stylet length = 18.1 ± 0.5 (17 to 19) µm; anterior end to median bulb = 72.2 ± 1.7 (70 to 75) µm; anterior end to secretory-excretory pore = 99.7 ± 2.5 (96 to 103) µm; maximum body width = 15.2 ± 0.4 (15 to 16) µm; body width at anus = 10.1 ± 1 (8 to 11) µm; tail length = 62.0 ± 6.9 (51 to 74) µm; hyaline part of tail = 44.0 ± 1.8 (40 to 47) µm. The egg measurements for 11 individuals were length = 102.5 ± 7.9 (93 to 119) µm; width = 39.3 ± 4.2 (33 to 46) µm; length/width = 2.6 ± 0.3 (2.0 to 3.1). The morphology, morphometric characters and molecular data of the population of H. koreana isolated from bamboo in Iran are in agreement with those previously reported for this species (Vovlas et al., 1992; Mundo-Ocampo et al., 2008). At present, five species of Heterodera belonging to the Cyperi and Afenestrata groups were reported from bamboo, H. bamboosi (Kaushal and Swarup, 1988; Wouts and Baldwin, 1998) on Bambusa sp. from India; H. koreana on P. pubescence, P. aurea, and P. nigra from South Korea and the United States; and H. hainanensis (Zhuo et al., 2013), H. fengi (Wang et al., 2013), and H. guangdongensis (Zhuo et al., 2014) on P. pubescence from China; thus showing host suitability of bamboo for at least five species of cyst-forming nematodes. A greenhouse test performed by planting rice seed cv. Hashemi in soil containing H. koreana showed successful multiplication of Korean cyst nematode on rice seedlings after 2 mon. The exact date of the establishment of bamboo plantation in LARS is not precisely clear, but it indicates that the Korean cyst nematode was most likely brought with the imported bamboo seedlings from unknown origin several decades ago. According to our best knowledge, this is the first report of occurrence of H. koreana from Iran. So far the Korean cyst nematode was reported from South Korea, Thailand, and the United States, Florida (from nurseries); this study includes the distribution of this cyst-forming nematode in Iran and expands the information of the occurrence of H. koreana for the world.     

Taxonomic Status of Berntsenus brachycephalus (Nematoda: Aphelenchina)

Berntsenus brachycephalus is redescribed from original material collected by Thorne in Utah, and hololectotype and paralectotypes are designated. This species is characterized by an expanded lip region, offset head, heavily sclerotized stylet conus, and two rows of oogonia and spermatogonia. The hololectotype male is 720 µm long, and paralectotype males are 864 to 952 µm long. Paralectotype females are 840 µm long. Tails of both sexes are amucronate.     

Further Studies on Soil Nematode Fauna in North Western Iran with the Description of One New Species

Heterodorus youbertghostai n. sp. is described and illustrated based on morphological, morphometric, and molecular data. The new species was found in two geographically distant points in northwestern Iran and is characterized by having angular lip region, separated from the rest body by a constriction, body length of 1,432.5 to 1,751.3 µm, odontostyle length of 24 to 28 µm, rod-like odontophore, 37.0 to 42.5 µm long, lacking flanges at base, double guiding ring at 14 to 16 µm distance from anterior end, pharyngeal bulb comprising 40% to 48% of pharynx, intestine usually containing green material, female reproductive system amphidelphic with less divided short uterus, specific structure of pars distalis vaginae, bluntly conical tail, dorsally convex and ventrally flat, with rounded tip and saccate bodies in ventral side. The new species comes more close to H. conicaudatus and H. irregularis by its morphology and morphometric characters. Compared to former, it has remarkable difference in vulva position and tail characters, and compared to the latter, it could be separated by shorter body, posteriorly located vulva, wider lip region, and longer tail. In phylogenetic analyses using partial sequences of 28S rDNA D2-D3, the new species formed a fully supported clade with several isolates of H. brevidentatus, prevalent in Iran. The other nordiid taxon, Enchodorus dolichurus, already reported from Iran, was also sequenced for the same genomic region and included in phylogenetic analyses.     

Description of Meloidoderita salina sp. n. (Nematoda,Sphaeronematidae) from a micro-tidal salt marsh at?Mont-Saint-Michel Bay in France

Meloidoderita salina sp. n. is described and illustrated from the halophytic plant Atriplex portulacoides L. (sea purslane) growing in a micro-tidal salt marsh in the Mont-Saint-Michel Bay in France. This new species is the first member of Meloidoderita Poghossian, 1966 collected from a saline environment, and is characterized by the following features: sedentary mature females having a small swollen body with a clear posterior protuberance; slightly dorsally curved stylet, 19.9 µm long, with posteriorly sloping knobs; neck region irregular in shape and twisted; well developed secretory-excretory (S–E) pore, with markedly sclerotized S-E duct running posteriorly; prominent uterus bordered by a thick hyaline wall and filled with eggs. The adult female transforms into a cystoid. Eggs are deposited in both egg-mass and cystoid. Cystoids of Meloidoderita salina sp. n. display a unique sub-cuticular hexagonal beaded pattern. Male without stylet, pharyngeal region degenerated, S-E duct prominent, deirids small, developed testis 97.5 µm long, spicules 18.4 µm long, cloacal opening ventrally protruded, small phasmids posterior to cloaca opening and situated at 5.9 (3.2–7.7) µm from tail end, and conical tail ending in a rounded terminus marked with one (rarely two) ventrally positioned mucro. Additionally, some young malesof the new species were observed enveloped in the last J2 cuticle. Second-stage juvenile body 470 µm long, with a 16.4 µm long stylet, prominent rounded knobs set off from the shaft, hemizonid anterior and adjacent to S-E pore, small deirids located just above S-E pore level, genital primordium located at 68–77% of body length, phasmids small and located at about 19 µm from tail tip, and tail 38.7 µm long, tapering to finely pointed terminus with a finger-like projection. Phylogenetic analyses based on the nearly full length small subunit ribosomal DNA sequences of Meloidoderita salina sp. n. revealed a close relationship of the new species with Sphaeronema alni Turkina & Chizhov, 1986 and placed these two species sister to the rest of Criconematina.     

Two New Species of Actinolaimidae Thorne, 1939 (Nemata: Dorylaimida) from China

Two new species of Actinolaimidae are described from China. Trachactinolaimus brevicaudatus n. sp. is 3.4-4.4 mm long; a = 40-60, b = 3.5-5.2, c = 20-21 in female and 34-39 in male; odontostyle length is 29-33 µm; spicules are 67-77 µm long; and the stoma has four onchia with numerous mural denticles. The female has a longitudinal vulva, and the male has 20 to 23 contiguous ventromedian supplements. Egtitus sinensis n. sp. is 1.7-2.2 mm long; a = 24-33, b = 3.1-3.9, c = 16-19 in female and 0.7-0.9 in male; odontostyle length is 25-29 µm; spicules are 55-56 µm long; and the prerectum 53-77 µm long. The cardia is short and blunt conoid, 13-19 µm long. The male has 12 to 13 ventromedian supplements at intervals of 2-3 µm.     

Descriptions of Chitwoodius brasiliensis n.sp., Chitwoodius rusticulus n.sp., and Vancterlindia venata n.sp. (Dorylaimida: Tylencholaimidae: Vanderlindiinae)

Chitwoodius brasiliensis n.sp. is described from soil around plant roots in Vicosa, MG State, Brazil. It has a body 1.9 mm (1.6-2.5) long, odontostyle and odontophore 33 μm (31-37) and 34 μm (31-36) long, respectively, and males with spicules 59-61 μm long. Chitwoodius rusticulus n.sp. from Colombian rain forest differs from other species of the genus in having a weakly muscular anterior part of the oesophagus, a pore-like vulva, and an unsclerotized vagina. Vanderlindia venata n.sp. from lucerne soil in South Kilimanjaro, Tanzania, differs from the type and only other known species of its genus in having a smaller and less slender body (L = 3.34 mm [3.15-3.71], a = 50 [46-53]), odontostyle 2.3-2.5 times lip region width long, and stylet guiding ring located at 1.2-1.4 times lip region width from anterior end.     

Morphological and Molecular Characterization of Gracilacus wuae n. sp. (Nematoda: Criconematoidea) Associated with Cow Parsnip (Heracleum maximum) in Ontario,Canada

Gracilacus wuae n. sp. from soil associated with cow parsnip in Ontario, Canada is described and illustrated. Morphologically, females have a long stylet ranging from 80 to 93 µm long, the lip region not offset from the body contour, without lateral lips but with large and flat submedian lobes, the mouth opening slit-like elongated laterally and surrounded by lateral flaps, the excretory pore is anterior to the knobs of the stylet; males without stylet and the pharynx degenerated. The fourth-stage juveniles lack a stylet, the pharynx degenerated, and can be differentiated into preadult females and males based on the position of the genital primordia. The third-stage juveniles are similar to females but smaller. Phylogenetic studies using the rDNA small subunit 18S, large subunit 28S D2/D3, and internal transcribed spacer (ITS) sequences collectively provide evidence of a grouping with other Gracilacus and some species of Paratylenchus with stylet length of females longer than 41 µm deposited in GenBank.     

Particle Deposition in a Child Respiratory Tract Model: In Vivo Regional Deposition of Fine and Ultrafine Aerosols in Baboons

To relate exposure to adverse health effects, it is necessary to know where particles in the submicron range deposit in the respiratory tract. The possibly higher vulnerability of children requires specific inhalation studies. However, radio-aerosol deposition experiments involving children are rare because of ethical restrictions related to radiation exposure. Thus, an in vivo study was conducted using three baboons as a child respiratory tract model to assess regional deposition patterns (thoracic region vs. extrathoracic region) of radioactive polydisperse aerosols ([d16–d84], equal to [0.15 µm–0.5 µm], [0.25 µm–1 µm], or [1 µm–9 µm]). Results clearly demonstrated that aerosol deposition within the thoracic region and the extrathoraic region varied substantially according to particle size. High deposition in the extrathoracic region was observed for the [1 µm–9 µm] aerosol (72%±17%). The [0.15 µm–0.5 µm] aerosol was associated almost exclusively with thoracic region deposition (84%±4%). Airborne particles in the range of [0.25 µm–1 µm] showed an intermediate deposition pattern, with 49%±8% in the extrathoracic region and 51%±8% in the thoracic region. Finally, comparison of baboon and human inhalation experiments for the [1 µm–9 µm] aerosol showed similar regional deposition, leading to the conclusion that regional deposition is species-independent for this airborne particle sizes.     

Hemicycliophora hellenica n. sp. (Nemata: Criconematidae) from Greece

Hemicycliophora hellenica n. sp. is described and illustrated from a bisexual population found in Filippias, Epirus, Greece, in the rhizosphere of giant reed (Arundo donax) and unidentified aquatic plants along the edge of irrigation canals. Hemicycliophora hellenica n. sp. is characterized by a long stylet (more than 120 µm), body length (L = 1,078-1,634 µm; R = 303-362), and lateral field marked by three lateral lines in females and four in males. The lip region is distinctive in that the female labial disc is oval and offset from the first lip region annule, and protrudes markedly; the male labial disc is very large, offset and rectangular, and protrudes distinctly. Hemicycliophora hellenica n. sp. resembles H. megalodiscus Loof by possession of a strongly protruding labial disc, but differs in having a longer female body, more body annules, longer stylet, and dorsally and ventrally indented head annules. The male labial plate is larger than in H. megalodiscus, and the male has four incisures in the lateral field vs. three in H. megalodiscus.     

EXCITABLE MEMBRANE ULTRASTRUCTURE : I. Freeze Fracture of Crayfish Axons

Cross-sectioned and cross-fractured crayfish axons display regions in which axon and Schwann cell surface membranes are regularly curved and project into the axoplasm. At these regions (projections) the two membranes run precisely parallel, separated by a gap of 130–140 Å. Longitudinal fractures through the axons expose the inner fractured surface of either the internal (face A) or the external (face B) leaflet of axon and adjacent Schwann cell surface membranes. On both membranes the projections appear as elongated structures oriented with the long axis parallel to the long axis of the nerve fiber. On face A of the axon surface membrane they are seen as elongated indentations 0.5–1.2-µm long, 0.12–0.15-µm wide. The indentations contain parallel chains of globules. The chains repeat every 120–125 Å and are oriented obliquely in such a way that if one looks at the axon surface from the extracellular space, the axis of the chains is skewed counterclockwise to the long axis of the indentations by an acute angle (most often 55–60°). The globules repeat along the chain every 80–85 Å. Globules of adjacent chains are in register in such a way that the axis on which globules of neighboring chains are aligned forms an angle of 75–85° with the axis of the chains. The complex structure can be defined as a globular array with a rhomboidal unit cell of 80–85 x 120–125 Å. On face B of the axon surface membrane the complementary image of these structures is seen. The projections of the Schwann cell surface membrane also contain groupings of globules; however, these differ from those in the axonal projections in size, pattern of aggregation, and fracture properties. Several possible interpretations of the meaning of these membrane specializations could be proposed. They could be: (a) structures involved in the mechanism of excitation, (b) regions of presumed metabolic couplings, and (c) areas of cell-to-cell adhesion.     

Meloidogyne haplanaria n. sp. (Nematoda: Meloidogynidae), a Root-knot Nematode Parasitizing Peanut in Texas

Meloidogyne haplanaria n. sp. is described and illustrated from specimens parasitizing peanut in Texas. The perineal pattern of the female is rounded to oval with a dorsal arch that is high and rounded except for striae near the vulva, which are low with rounded shoulders. The striae are distinctly forked in the lateral field, and punctations often occur as a small group near the tail tip and singly within the whole perineal pattern. The female stylet is 13-16 µm long and has broad, distinctly set-off knobs. The excretory pore opens 40-118 µm from the head, approximately halfway between the anterior end and the metacorpus. Males are 1.2-2.4 µm in length and have a high, wide head cap that slopes posteriorly. The labial disc and medial lips are partially fused to form an elongated lip structure. In some specimens the labial disk is distinctly separated from the lips by a groove. The stylet is 17-22 µm long and has wide knobs that are rounded and distinctly set off from the shaft. Mean second-stage juvenile length is 419 µm. The head region is not annulated, and the large labial disc and crescent-shaped medial lips are fused to form a dumbbell-shaped head cap. The stylet is 9-12 µm long and has rounded, posteriorly sloping knobs. The slender tail, 58-74 µm long, has a distinct, inflated rectum and a slightly rounded tip. The hyaline tail terminus is 11-16 µm long. The isozyme phenotypes for esterase and malic dehydrogenase do not correspond to any other recognized Meloidogyne species. Tomato and peanut are good hosts; corn and wheat are very poor hosts; and cotton, tobacco, pepper, and watermelon are nonhosts.     

Water transport in the midrib tissue of maize leaves : direct measurement of the propagation of changes in cell turgor across a plant tissue

Water movement across plant tissues occurs along two paths: from cell-to-cell and in the apoplasm. We examined the contribution of these two paths to the kinetics of water transport across the parenchymatous midrib tissue of the maize (Zea mays L.) leaf. Water relations parameters (hydraulic conductivity, Lp; cell elastic coefficient, ε; half-time of water exchange for individual cells, T_½) of individual parenchyma cells determined with the pressure probe varied in different regions of the midrib. In the adaxial region, Lp = (0.3 ± 0.3)·10⁻⁵ centimeters per second per bar, ε = 103 ± 72 bar, and T_½ = 7.9 ± 4.8 seconds (n = seven cells); whereas, in the abaxial region, Lp = (2.5 ± 0.9)·10⁻⁵ centimeters per second per bar, ε = 41 ± 9 bar, and T_½ = 1.3 ± 0.5 seconds (n = 7). This zonal variation in Lp, ε, and T_½ indicates that tissue inhomogeneities exist for these parameters and could have an effect on the kinetics of water transport across the tissue.

The diffusivity of the tissue to water (D_t) obtained from the sorption kinetics of rehydrating tissue was D_t = (1.1 ± 0.4)·10⁻⁶ square centimeters per second (n = 6). The diffusivity of the cell-to-cell path (D_c) calculated from pressure probe data ranged from D_c = 0.4·10⁻⁶ square centimeters per second in the adaxial region to D_c = 6.1·10⁻⁶ square centimeters per second in the abaxial region of the tissue. D_t D_c suggests substantial cell-to-cell transport of water occurred during rehydration. However, the tissue diffusivity calculated from the kinetics of pressure-propagation across the tissue (D_t′) was D_t′ = (33.1 ± 8.0)·10⁻⁶ square centimeters per second (n = 8) and more than 1 order of magnitude larger than D_t. Also, the hydraulic conductance of the midrib tissue (Lp_m per square centimeter of surface) estimated from pressure-induced flows across several parenchyma cell layers was Lp_m = (8.9 ± 5.6)·10⁻⁵ centimeters per second per bar (n = 5) and much larger than Lp.

These results indicate that the preferential path for water transport across the midrib tissue depends on the nature of the driving forces present within the tissue. Under osmotic conditions, the cell-to-cell path dominates, whereas under hydrostatic conditions water moves primarily in the apoplasm.

     

Thermus parvatiensis RLT sp. nov., Isolated from a Hot Water Spring,Located Atop the Himalayan Ranges at Manikaran,India

A Gram negative, yellow pigmented, rod shaped bacterium designated as RL^T was isolated from a hot water spring (90–98 °C) located at Manikaran in Northern India. The isolate grows at 60–80 °C (optimum, 70 °C) and at pH 7.0–9.0 (optimum pH 7.2). Phylogenetic analysis of 16S rRNA gene sequences and levels of DNA–DNA relatedness together indicate that the new isolate represents a novel species of the genus Thermus with closest affinity to Thermus thermophilus HB8^T (99.5 %) followed by Thermus arciformis (96.4 %). A comparative analysis of partial sequences of housekeeping genes (HKG) further revealed that strain RL^T is a novel species belonging to the genus Thermus. The melting G+C content of strain RL^T was calculated as 68.7 mol%. The DNA–DNA relatedness value of strain RL^T with its nearest neighbours (>97 %) was found to be less than 70 % indicating that strain RL^T represents a novel species of the genus Thermus. MK-8 was the predominant respiratory quinone. The presence of characteristic phospholipid and glycolipid further confirmed that strain RL^T belongs to the genus Thermus. The predominant fatty acids of strain RL^T were iso-C_17:0 (23.67 %) and iso-C_15:0 (24.50 %). The results obtained after DNA–DNA hybridization, biochemical and physiological tests clearly distinguished strain RL^T from its closely related species. Thus, strain RL^T represents a novel species of the genus Thermus for which the name Thermus parvatiensis is proposed (=DSM 21745^T= MTCC 8932^T).

Electronic supplementary material

The online version of this article (doi:10.1007/s12088-015-0538-4) contains supplementary material, which is available to authorized users.     

First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea,Decapoda)

For the first time, a morphological study of haemocytes from the crab Carcinus aestuarii was carried out by means of light microscopy and differing cytochemical assays. Analysis of haemocyte size frequency distribution (performed by means of a Coulter Counter) revealed the presence of two distinct haemocyte fractions in C. aestuarii haemolymph, depending on cell size. The first fraction was of about 3–5 µm in diameter and 30–50 fL in volume, the second was of about 6–12 µm in diameter and over 200 fL in volume. Mean cell diameter and volume were 8.20±1.7 µm and 272.30±143.5 fL, respectively. Haemocytes observed under light microscope were distinguished in three cell types: granulocytes (28%; 11.94±1.43 µm in diameter) with evident cytoplasmic granules, semigranulocytes (27%; 12.38±1.76 µm in diameter) with less granules than granulocytes, and hyalinocytes (44%; 7.88±1.6 µm in diameter) without granules. In addition, a peculiar cell type was occasionally found (about 1%): it was 25–30 µm in diameter and had a great vacuole and a peripheral cytoplasm with granules. Granulocyte and semigranulocyte granules stained in vivo with Neutral Red, indicating that they were lysosomes. Giemsa’s dye confirmed that granulocytes and semigranulocytes were larger than hyalinocytes. Pappenheim’s panoptical staining and Ehrlich’s triacid mixture allowed to distinguish granule-containing cells (including semigranulocytes) in acidophils (64%), basophils (35%) and neutrophils (1%). Hyalinocytes showed always a basophilic cytoplasm. Haemocytes were positive to the PAS reaction for carbohydrates, even if cytoplasm carbohydrate distribution varied among cell types. Lastly, lipids were found on cell membrane and in cytoplasm of all haemocyte types in the form of black spots produced after Sudan Black B staining. The morphological characterisation of C. aestuarii haemocytes by light microscopy was necessary before performing both ultrastructural and functional studies of circulating cells.Key words: Carcinus aestuarii, crab, haemocytes, light microscopy, cytochemical assays, morphological characterisation.