The rate of preimaginal development of the blowfly, Calliphora vicina R.-D. (Diptera, Calliphoridae) under field and laboratory conditions

Open field experiments with the blowfly, Calliphora vicina originating from the environs of St. Petersburg showed that the correlation between the rate of its preimaginal development under the natural conditions and the mean daily temperature could be very closely approximated by linear regression. The sum of effective temperatures required for development from the egg to the puparium constituted ca 140 degree-days and from the egg to the adult, ca 320 degree-days with the lower thresholds of 5.8 and 4.8°C, respectively. The minimum duration of development (6–8 days from the egg to the puparium and 16–20 days from the egg to the adult) was recorded during the period of the highest mean temperatures (22–23°C) from July 8 to August 15 of 2010 and 2011. The rate of development under natural thermorhythms was not siggnificantly different from that under the laboratory conditions at constant temperatures from 12 to 23°C.     

Morphology of the egg shell and the developing embryo of the Red Palm Weevil,Rhynchophorus ferrugineus (Oliver)

The harvested eggs of Rhynchophorus ferrugineus are ovo-cylindrical shaped, averaged 1.09 mm in length and 0.43 mm in width, with ratio of L\W 4.42. The chorionic layer of electron dense material is seen covering the exochorion structure of the eggs. The egg main body chorion exhibits a polygonal pattern and architecture surface of the egg is supported by a system of irregular interconnecting grooves. The micropylar apparatus of the eggs of the Red Palm Weevil, R. ferrugineus is described in the present study for the first time. Two micropylar openings are found closed to the center of the posterior wide pole of the egg. Each micropylar opening presents a single small orifice and its surrounding chorion is porous and densely set with tiny projections allowing the spermatozoa to penetrate the egg. Respiratory aeropyles are distributed on the borders of reticulations in the area chorionic surface of egg capsule. The hatching region is detected on the anterior part at the opposite side of the egg. Changes in the appearance and shape of R. ferrugineus eggs as well as the incidence of embryonic development are observed.     

Development of eggs,larvae and juveniles of the labrid fish,Halichoeres poecilopterus,reared in the laboratory

Embryonic, larval and juvenile development of the labrid fish,Halichoeres poecilopterus, is described using a laboratory-reared series. The eggs, measuring 0.60–0.72 mm in diameter, were pelagic and spherical with a single oil globule (0.12–0.16 mm in diameter). Hatching occurred 18 h 48 min after spawning. The newly-hatched larvae, measuring 1.46–1.70 mm TL, had 8–114 + 16–18 myomeres. A conspicuous melanophore appeared on the dorsal finfold 8 h after hatching, at ca. 2 mm TL. The yolk was completely absorbed 3 days after hatching, at 2.52–2.72 mm TL. Flexion of the notochord started at ca. 6 mm TL and was finished at ca. 8 mm TL. Aggregate numbers of all fin rays were completed at ca. 14 mm TL. Squamation was almost completed at ca. 20 mm TL.     

Reproduction of the sandpaper skate,Bathyraja kincaidii (Garman 1908) in the eastern North Pacific

The sandpaper skate, Bathyraja kincaidii, is commonly taken as bycatch in trawl fisheries off central California. The impact of fisheries exploitation on this population is unknown, however little is known about its biology, including its reproductive life history. To determine reproductive aspects of B. kincaidii, 506 individuals were collected from commercial trawls, museum collections, and monthly trawl and longline surveys conducted by the National Marine Fisheries Service. Males ranged from 112 to 635 mm TL, whereas females ranged from 113 to 610 mm TL. Males reach first maturity at 440 mm TL and at ~3 years, whereas females reach first maturity at 450 mm TL, approximately 1 year later. Size and estimated age at 50 % maturity were similar between sexes: 492 mm TL and 7.5 years for males and 467 mm TL and 7.1 years for females. Gonadosomatic index (GSI) and hepatosomatic index (HSI) values indicate a continuous reproductive cycle, with a seasonal peak in the summer and fall, and a resting period following egg case deposition.     

Oviparous reproduction of Vestia ranojevici moravica confirms taxonomic value of life history characters in carpathian clausiliids (Gastropoda: Pulmonata)

Based on five-year observation of 36 snails collected in the ?eladná valley (Moravia, Czech Republic) and their progeny reared in laboratory, it was found that Vestia ranojevici moravica (Brabenec, 1952) is oviparous. The oviparity in V. ranojevici is in line with the previous assumption of its reproductive strategy based on anatomical studies alone. It supports the separate position of the species in the genus Vestiaa (subgenus Brabenecia). V. ranojevici was kept at room temperature (18–25°C) between March and October; in winter snails were stored at 3°C. The egg-laying period started in late March, ca. 1 month after raising the temperature, and lasted till September, with one maximum in spring. In the laboratory the snails laid eggs in batches (usually 4–6 eggs, max 11) or singly. The number of batches per pair per season was 1–8; the corresponding number of eggs — 3–40. The eggs were gelatinous, with separate calcium carbonate crystals in the external envelope. The average egg size was 1.80 × 1.51 mm, but actual size and shape were highly variable: larger and more elongated eggs were produced mainly by laboratory reared animals. Freshly deposited eggs did not contain shelled embryos nor the reproductive tract of dissected individuals contained eggs. At room temperature the eggs hatched after ca. 14–16 days, then the shell growth lasted 20–29 weeks; the snails laid eggs 30 weeks after completion of shell growth indicating one-year life cycle under laboratory conditions. Adult individuals collected in the field reproduced during 4–5 consecutive years. Comparison of V. ranojevici reproduction to the results of breeding of other Carpathian clausiliids in the same laboratory shows that similar reproductive strategies may have evolved independently in different lineages of the group: both the oviparous and egg retaining species belong to the genera Vestia and Balea.     

Production,maturity, reproductive cycle and fecundity of small-spotted catshark,Scyliorhinus canicula (Chondrichthyes: Scyliorhinidae) from the northern coast of Tunisia (Central Mediterranean)

Small-spotted catshark Scyliorhinus canicula (Linnaeus, 1758) is confronted since 2000 to a drastic decline of captures in Tunisian northern areas where it formerly was quantitatively and qualitatively very abundant. A study conducted in the area showed that males and females sexually matured between 400–478 mm and 364–471 mm total length (TL), respectively. The largest male and female were 521 mm and 531 mm TL, respectively and weighed 449 and 445.5 g, respectively. There was a not significant relationship total mass vs. TL between males and females. Conversely, the relationship of liver mass vs. TL significantly differed between sexes. Both HSI and GSI values did not present significant differences between males and females. There was a not significant relationship total mass vs. TL between males and females. Both male and female HSI reached high values in adults, significantly higher than those of sub-adults. Both male and female GSI values increased with TL of specimens; additionally, they showed significant differences between juveniles and sub-adults and also between sub-adults and adults. The OSI values were significantly different between juvenile and sub-adult females, and also between sub-adult and adult females. Monthly mean values of adult male and female HSI did not significantly vary throughout the year. Significant monthly changes were recorded in mean values of adult male however no significant changes were observed in monthly mean of GSI and OSI for adult females. Vitellogenic activity and production of egg cases permanently occurred throughout the year. The diameter of largest yolky oocytes ranged from 20.1 to 23.0 (mean = 21.43 ± 1.07) and weighed from 0.64 to 0.90 g (mean = 0.78 ± 0.09). Some measurements were taken on egg cases: length with horns ranged 42.5-55.7 mm with mean = 48.9 ± 2.9, width ranged 13.6–19.7 mm with mean = 16.50 ± 0.9, while the total mass of complete egg case ranged between 1.8 and 4.6 g, with mean = 2.4 ± 0.4. An annual fecundity estimation based on production of egg cases and oocytes during one year counted in adult females, enabled us to consider it between 40 and 240.     

Development ofDiatraea Saccharalis [Lep.: Pyralidae] at constant temperatures

At constant temperatures between 15.6 and 32°C the incubation time of eggs ofDiatraea saccharalis (F.) was reduced by each increase in temperature. At 34°C the time decreased. Highest (98.6%) and lowest (9.9%) egg hatch occurred at 26 and 34°C, respectively. Larvae completed development at temperatures ranging from 22 to 34°C; however, only 4.4% of the larvae pupated at 34°C. Duration of the larval stage at 30°C (♂=18.1 days; ♀=19.1 days) was ca. 14 days shorter than at 22°C. Maximum rate of development in the pupal stage occurred at 28°C (ca. 6.8 days), and a higher temperature increased developmental time and mortality. Adult longevity and egg production generally were reduced with increasing temperatures and egg production was highest at 24°C (729.8 eggs/ moth). As many as 7 larval stages occurred; but most larvae completed development in 5 stages, and none completed development in less than 5 stages. The female larval stage was ca. 1 day longer than that of males, and this difference occurred primarily in the 5th stage.     

Hatching Response of Meloidogyne incognita acrita to Electric Shock

The influence of electric shock on hatch of Meloidogyne incognita acrita from egg masses taken from roots of ''Acala SJ-I'' cotton (Gossypium hirsutum L.) was studied. Egg masses in tap water were individually placed between the tips of needle electrodes 1 mm apart and exposed to potentials of l, 10, 20, and 60 vdc/mm at 1, 1, 1, and 86 milliamperes dc, respectively, for periods of 2 and 60 seconds. Hatched larvae were counted at five-day intervals for 60 days. Of the eight treatment combinations used, six gave a greater egg hatch than the control. The largest hatch, 520 percent greater than the control, resulted from exposure to 1 vdc/mm for 60 seconds; 60 vdc/mm for 2 and 60 sec decreased egg hatch 11 and 94 percent of the untreated control. Hatched larvae from all treatments except the 60 vdc/mm, 60-second exposure were infective and reproduced on young cotton plants in a glasshouse.     

Development of the bitterling,Tanakia tanago (cyprinidae), with a note on minute tubercles on the skin surface

The development of eggs and larvae and minute tubercles on the skin surface in larvae ofTanakia tanago were observed. The eggs began to hatch approximately 52 hours after insemination and the larvae reached free-swimming stage 19 days after hatching at water temperature of 22±1°C. The egg and larval development and minute tubercles on the skin surface in larvae of this species were similar to those ofAcheilognathus lanceolata andA. limbata. However,T. tanago was distinguishable in egg and larval development fromA. lanceolata andA. limbata by the following characters: the perivitelline space was narrower, embryonic and larval development was faster, and minute tubercles on the skin surface of the anteriormost parts of the yolk sac, and of the body and head were hemispheric in shape. From these characters,T. tanago is considered to be more specialized thanA. lanceolata andA. limbata.     

Seed characteristics and testa textures of Pratensis, Orobon, Lathyrus, Orobastrum and Cicercula sections from Lathyrus (Fabaceae) in Turkey

The seed morphologies and testa textures of 23 taxa belonging to the Pratensis, Orobon, Lathyrus, Orobastrum and Cicercula sections of Lathyrus that can widely be found in Turkey were analysed. The findings obtained in this study and previous findings (34 taxa in total) were compared and interpreted at the level of the sections. Morphological properties including seed size, general shape, surface shape, colour, hilum length and width were measured under stereomicroscopy. The seeds were of spheroidal, subprolate and prolate (P/E = 0.90–1.58) types and medium in size. The smallest seeds belonged to Lathyrus inconspicuus var. inconspicuus (P = 2.19 ± 0.25 mm, E = 2.08 ± 0.16 mm) and the largest to L. sativus (P = 5.88 ± 0.74 mm, E = 5.36 ± 0.57 mm). The smallest hilum belonged to L. inconspicuus var. stenophyllus (0.38 ± 0.04 mm) and the largest to L. sylvestris (4.50 ± 0.58 mm). Testa textures such as papillae shape, dense, ribbing and presence or absence of a waxy layer were examined using scanning electron microscopy (SEM). In addition, some photographs included in this study were taken via stereomicroscopy and SEM.     

The plasma-membrance IMP pattern as related to animal/vegetal polarity in the amphibian egg

Freeze-fracture electron microscopy of the plasma membrane of the fertilized, uncleaved Xenopus egg shows that intramembranous particles (IMPs) range in size from ca. 50 to 200 Å and that more IMPs are attached to the E-face than to the P-face. The overall IMP densities of the animal and the vegetal hemisphere do not differ significantly. IMP-free regions (?, ca. 0.1 μm) on the tips of surface protrusions were irregularly distributed in the animal and the vegetal half (E-face) occupying ca. 8.5 and 2%, respectively of the free area. The relative densities for 16 different IMP sizes have been compared, on the basis of seven animal and seven vegetal halves, counting (E-faces only) ca. 10,000 IMPs in each hemisphere. For IMP sizes of ≤81 Å, a significant difference (P < 0.0005) was found, more small IMPs being present in the animal half. Some evidence for IMP-associated thin elements was found. These findings are discussed in relation to plasma membrane anisotropy and the morphogenetic role of the egg cortex.     

Ontogenetic shift to dependence on salmon-derived nutrients in Dolly Varden char from the Iliamna River,Alaska

The upstream migration, spawning, and death of anadromous, semelparous Pacific salmon brings nutrients to terrestrial and aquatic communities around the Pacific Rim. Many fishes use these resources but the relationship between fish body size and the reliance on salmon-derived nutrients might follow one of several patterns related to the onset of egg consumption with body size as fish grow, and possible shifts to alternative prey such as fishes as they grow larger still. In this study, these size-dependent hypotheses of marine subsidy use by resident Dolly Varden, Salvelinus malma, were tested using diet and stable isotope analyses. S. malma did not shift abruptly to a reliance on salmon eggs after they became large enough to eat eggs (i.e., no gape limitation). Rather, fish large enough to eat eggs but < 150 mm showed diets that blended salmon nutrients with aquatic insects, likely because they were spatially segregated from the highest concentration of spawning sockeye salmon, Oncorhynchus nerka. From intermediate through the largest sizes observed (150 to > 600 mm long) S. malma received ca. 80 % of their nutrients from salmon (eggs, flesh, and maggots that had scavenged dead salmon) based on diet analysis and stable isotope ratios despite being large enough to consume fish, as many similarly-sized salmonids do in other ecosystems. The few fish sampled in June, prior to the availability of salmon subsidies, had stable isotope signatures that also reflected heavy (ca. 90 %) reliance on marine sources, likely because they had eaten little since the end of the salmon run the previous fall. This apparent avoidance of piscivory in favor a rich yet pulsed marine subsidy highlights the importance of healthy salmon runs for the sake of not only the salmon but resident fishes that consume them.     

Towards the conservation of a critically endangered species, <Emphasis Type="Italic">Aphanius farsicus</Emphasis>: embryogenesis and development

Human-induced disturbance or anthropogenic activities, natural disturbance and also limited distribution, have forced Aphanius farsicus populations to the edge of extinction, during the past few years. As knowledge of fish ontogeny is fundamental for accurate stock assessment, and reproductive potential determination, thus for future conservation and management of the critically endangered species, we describe the first detailed normal embryonic development of this cyprinodontid fish from fertilization to post-hatching. Aphanius farsicus spontaneously spawned at 24°C. The newly-laid eggs were hyaline and spherical in shape (1.40 ± 0.02 mm, n = 11), sticky and full of egg yolk (telolecithal/macrolecithal) with oil droplets. We documented the main developmental stages at 24°C to zygote (0 h), cleavage (2 h), blastula (2 h), gastrula (17 h), segmentation, (34 h), pharyngula (100 h) and hatching (162 h).     

Bemerkungen zur Biologie sowie Haltung und Vermehrung des Sokotra-Riesengeckos, Haemodracon riebeckii (Peters, 1882)

The Socotra Giant Gecko, Haemodracon riebeckii, is the largest species of lizard on Socotra Island. The nocturnal, arboreal and rupiculous living geckos are omnivorous. Two pairs were kept in terrariums and were fed with various insects (crickets, locusts, cockroaches), sweet fruits and other feeding stuff (such as meat, fish). Temporarily H. riebeckii was kept together with other lizards (Eublepharis macularius, Trachylepis socotrana), without any signs of aggressive behaviour. Juveniles and adults of both sexes are able to produce a sound. These acoustic signals seem to be related to predators, because never any intraspecific function could be observed. Within seven years of captive breeding two females produced 253 eggs. Usually two white and sticky soft-shelled eggs were laid within one clutch, more rarely a single egg was laid. The two eggs of a clutch were always laid on the same day. H. riebeckii belongs to the geckos that bury their eggs and practice some brood care, but no special parental care. The female is able to proof with her hind legs the deep and shape of a hollow in the substrate to bury the eggs, which were buried in a sticky and soft-shelled condition. They are oval in shape (egg length 16.4-19.8 mm, egg width 12.4-17.8 mm, quotient EL:EW 1.22±0.05) and have in the beginning a weight ranging from 1.7100 to 2.5201 g. As typical for geckos with hard-shelled eggs the egg weight decreases during the incubation period. The loss can be between 5.59 to 30.29%. The development of eggs up to hatching of young depends upon temperature and the germinal stage in the laid egg. The time difference between the hatching of the young within one clutch of two eggs was usually 1 to 5 days. In some cases there were, however, longer differences (up to 61 days), which are probably caused by different developmental stages of the embryos during the time of egg laying. The shortest incubation period recorded during our investigations was 83 days for eggs incubated at constant temperature of 28 to 29.5 °C and the longest 359 days at 26 to 26.5 °C. Constant high incubation temperatures caused a premature hatching of young. In normal hatched young were the yolk sac retracted and the navel closed. In premature hatched young were the yolk not resorbed and the mortality within the first three month comparatively high. The snout-vent length (SVL) of newly hatched young is from 27 to 39 mm and the tail length (TL) from 25 to 38 mm (SVL:TL index 0.90-1.27), the weight is from 0.7688 to 1.5366 g. Young specimens are distinguished from adults by the brown/white striped lower jaw and the white-banded tail. Young which hatched in the terrarium were eaten by the adults. A loss of young can be avoided if they are raised individually.     

Axone,an Ethnic Probiotic Containing Food,Reduces Age of Sexual Maturity and Increases Poultry Production

Axone (Akhuni) is a homemade synbiotic (Nagamese fermented soybean product) served as side dish in North Eastern India. In this study, effects of Axone feeding on growth, weight gain, sexual maturity and egg production on Vanaraja birds (a strain of poultry bird developed at PDP Hyderabad for villages and backyard poultry) were evaluated. Axone incorporation in commercial poultry feed at the rate of 5 % (W/W) significantly improved growth rate (weight gain) both in male (p 0.001) and female (p 0.05) chicks, reduced age by 13 days at first egg laying (p 0.01), increased egg production (p ≤ 0.001) and improved egg weight (p ≤ 0.01). Microbiological analysis of Axone sample revealed that the major bacteria in Axone samples were Bacillus coagulans, well known for their probiotic value.     

Spawning, egg development and early ontogenesis in rock cod Patagonotothen ramsayi (Regan, 1913) caught on the Patagonian Shelf and maintained in captivity

Rock cod Patagonotothen ramsayi (Regan, 1913) is one of the most abundant fish of the family Nototheniidae inhabiting the Patagonian Shelf and upper Slope in the southwest Atlantic. Recently, P. ramsayi became an important commercial species around the Falkland Islands with annual catch of 60,000–75,000 t. The present study aimed to reveal previously unknown aspects of reproductive biology of P. ramsayi during the first successful maintenance of adults for more than a year in an aquaculture facility with running seawater. The fish spawned at the end of austral winter. During spawning, males changed their coloration dramatically, occupied artificial shelters on the bottom and showed aggressive territorial behaviour. Egg masses were light-yellow to light-orange irregular spongiform. They were negatively buoyant, but located outside shelters and were ignored by males. Egg diameters varied between 2.1 and 2.3 mm, and the number of eggs per egg mass ranged from 26,800 to 123,400. Embryogenesis lasted 28–32 days. Total lengths of newly hatched larvae ranged from 6.2 to 6.7 mm. The yolk sac feeding period lasted approximately 11 days, during which the larvae showed negative phototaxis. One-month-old larvae attained 8.8–9.0 mm in length. This study confirms that P. ramsayi exhibit the reproductive strategy typical for nototheniid species occupying low-latitude peripheries of their distributional range, characterised by a combination of r-features (small eggs and larvae, high fecundity) and K-features (territorial behaviour and possible nest guarding).     

Pelagic eggs and larvae of Coryphaenoides marginatus (Gadiformes: Macrouridae) collected from Suruga Bay, Japan

The pelagic eggs, yolk-sac and pelagic larvae of the macrourid fish, Coryphaenoides marginatus, from Suruga Bay in southern Japan, are described. The identification of the pelagic eggs based on 16S rRNA gene nucleotide sequences agreed with that obtained from morphological analyses. The spherical eggs, 1.14–1.30 mm in diameter, contained a single oil globule 0.30–0.38 mm in diameter, and had hexagonally patterned ornamentation on the chorion, 0.025–0.033 mm in width. Many melanophores were present on the anterodorsal region of the embryo after the caudal end had detached from the yolk. Within a day after hatching, each of the yolk-sac larvae had a body axis that was bent slightly at the anterior trunk region, many dorsal and lateral melanophores on the trunk plus several on the gut, and small irregular wrinkles on the dorsal and anal fin membranes. The pelagic larvae had a short caudal region in comparison to other known congeners (length 2.0–3.2+ times head length vs. 4–7, respectively), a short stalked pectoral fin base, and no elongate first dorsal and pelvic fin rays. They were further characterized by the presence of numerous very dense melanophores from just behind the eye to the anterior part of the caudal region at 5.1 mm head length (25.8+ mm total length). The significant difference in vertical distribution between the pelagic eggs and larvae (dominant depths ca. 200–350 m vs. ca. 10–100 m, respectively), with no subsequent collection of pelagic larvae with greater than 6 mm head length, indicate two stages (rising and falling) of ontogenic vertical migration.     

Temperature-based Prediction of Egg-Mass Production by Meloidogyne incognita

A mturation-rate relationship for Meloidogyne incognita on Lycopersicon esculentum ''Rutgers'' was derived and used to estimate harvest dates for maximmn egg hatch from laboratory cultures at ambient temperatures. Daily maturation increments were totaled (nematode maturation total, NMT) and correlated with hatch from isolated white, yellow, and amber egg masses. Hatch per mass fluctuated periodically from ca. 1.0 NMT, when egg masses were first visible, to 2.5 NMT by which time plants showed stress. Maximum yields from white and yellow masses occurred, with a shorter than expected periodicity, at 1.5-1.8 and 2.1-2.2 NMT. White masses decreased from 90% of the total masses at 1.0 NMT to 5% at 2.5 NMT, as the proportion of yellow and amber masses increased concomitantly. Harvested masses per gram of root varied from 97 to 276; hatch per gram of root, 11,000 to 86,000.     

Development of the bitterling,Paracheilognathus himantegus (Cyprinidae), with a note on minute tubercles on the skin surface

The development of the eggs and larvae and minute tubercles on the skin surface ofParacheilognathus himantegus larvae were observed. The egg began to hatch approximately 68 hours after insemination and the larvae reached the free-swimming stage 23 days after hatching at water temperature of 22±1°C. The larval development and minute tubercles on the skin surface of this species were similar to those ofAcheilognathus lanceolata, A. limbata, A. signifer andTanakia tanago. However, the shape of the ripe eggs ofP. himantegus differed from those of the four species. As regards the shape of eggs, there was a common characteristic amongP. himantegus, Rhodeus uyekii andA. limbata from Korea. As regards larval development,P. himantegus had two characters also found inRhodeus. These facts seem to suggest thatP. himantegus is closely related toA. lanceolata, A. limbata, A. signifer andT. tanago but is more specialized than these four species, except forA. limbata from Korea.     

Development of the cottid fish,Pseudoblennius percoides,reared in the laboratory,with brief descriptions of juvenileP. marmoratus andP. zonostigma

Embryonic, larval and juvenile development of the cottid fish,Pseudoblennius percoides were described on the basis of a series of laboratory-reared specimens. The eggs were demersal, adhesive, almost spherical in shape, measuring 1.66–1.82 mm in diameter, and with numerous various-sized oil globules. Neighboring eggs adhered to each other to form an egg mass. Hatching occurred between 13 and 16 days after spawning at a water temperature of 15.4 to I6.5°C. Newly hatched larvae measured from 6.5 to 7.3 mm, averaging 6.9 mm TL, and possessed 40 myomeres. Absorption of the yolk was completed at about 7.5 mm TL. Flexion of the notochord started and finished at about l0 mm TL and about 14 mm TL, respectively. Aggregate numbers of all fin rays were completed at over 16 mm TL, when the larvae reached the juvenile stage. The pigment pattern became the same as that of adults in juveniles longer than 25 mm TL. Lateral lines were completed at over 44 mm TL, when the juveniles attained to the young stage. The early stages of this species were clearly distinguished from those ofP. cottoides, and the juveniles of fourPseudoblennius species, i.e.P. percoides, P. cottoides, P. marmoratus andP. zonostigma, could be identified mainly by their pigment patterns.