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The production of haploids and doubled haploids (DHs) on unfertilized ovule culture was examined in 19 wild species of gentians (Gentiana ssp.) classified into eight sections. Of the 19 species including 35 strains, embryo-like structures (ELSs) were obtained from unfertilized ovule culture in 15 species, and regenerated plants were produced in 11 species. ELS production has varied greatly among the15 species, i.e., 0.5–79.2% frequency of responding flower buds and 0.01–1.99 ELSs per flower bud. Of the ELS-producing species, almost all were classified into the sects. Pneumonanthe or Cruciata. Species in sect. Pneumonanthe showed higher responses than those in sect. Cruciata. In examining the effect of flower bud stage on ELS production, more than twice as many ELSs were observed at the anther-dehiscent stage than that at the anther-indehiscent stage. Ploidy level was determined in 117 randomly selected regenerated plantlets, which suggests that most were diploid (46.2%) and haploid (32.5%). When 12 diploid plants were examined using simple sequence repeat (SSR) markers, 8 (66.7%) were DH. This study revealed that unfertilized ovule culture can be applied not only on cultivated gentian species but also on a number of wild species. The production of haploids and DHs in wild gentians provides novel prospects for ornamental and/or medicinal gentian breeding. This is the first report of a successful haploid and doubled haploid production by unfertilized ovule culture of various wild species of gentians (Gentiana spp.).

Analysis of the distribution of ostracods from the Armkhi Formation (Callovian) of the Tsudakhar section (Central Dagestan) enabled the identification of a stratigraphic sequence of six assemblages determined by paleoecological conditions and the first ostracod-based biostratigraphic zonation of the Callovian of Dagestan. Beds with Pleurocythere khapissovi and the Neurocythere cruciata intermedia Zone of the European ostracod scales were identified in the middle Callovian; this facilitated the interregional correlation for ostracods. Paleoecological analysis demonstrated the formation of the lower part of the section (beds with P. khapissovi and the N. cruciata intermedia Zone) in the coastal zone of the upper subtidal with normal marine salinity, good aeration, and low sedimentation rates. Sediments of the upper part of the section (paleoecological intervals with Schuleridea translucida–Cytherella perennis and with C. perennis) accumulated in the lower zone of the upper subtidal with a high sedimentation rate, high (and constantly increasing) eutrophication, and lowered pH values. Analysis of the paleogeographic distribution of the species studied revealed the connections of the Caucasus Sea to Western and Eastern Europe and to Central Asia.

The Genji firefly, Luciola cruciata, is widely distributed throughout the major Japanese islands (Honshu, Shikoku, and Kyushu) and distinguished into two ecological types on the basis of the flash interval of the mate-seeking males (4-sec slow-flash or 2-sec fast-flash intervals). The boundary of the ecological types corresponds to the Fossa Magna, a great rupture zone that separates eastern and western Japan. Although the degree of genetic differentiation of the two types has been evaluated using allozyme and mitochondrial DNA sequence data, it has not been evaluated using genome-wide data. Based on the genome-wide data obtained using single-end restriction-site-associated DNA (RAD-Seq), principal component, gene-level phylogenetic tree, admixture, and Wright’s fixation index analyses, we identified three phylogenetic groups in L. cruciata: East-Honshu, West-Honshu, and Kyushu. This grouping corresponds to the ecological types: East-Honshu to the slow-flash type and West-Honshu and Kyushu to the fast-flash type. Although introgression was exceptionally observed around adjacent or artificially transplanted areas, gene flow among the groups was almost absent in the natural populations. The phylogenetic tree under the coalescent model also evaluated differentiation among the East-Honshu, West-Honshu and Kyushu groups. Furthermore, because the distribution patterns of the three groups are consistent with the geological history of Japanese islands, a vicariant speciation scenario of L. cruciata is concluded. In addition, we identified genetic markers that can be used to distinguish the three genetic groups for genetic management of firefly transplantation in nature conservation and regeneration.

Background Study of phytochemicals and pharmacological properties of bryophytes has been neglected for a long time because of the three main reasons i.e. (i) difficulty in collection in large amount for analysis; (ii) their availablility only in particular season and (iii) their restricted geographic distribution. So, the aim of this work was to propagate Lunularia cruciata under in vitro condition for comparing its pharmacological properties and phytocmecial constituents with naturally grown counterparts. Method Axenic culture of L. cruciata was established by propagating gemmae under in vitro condition. Appropriate culture conditions, media, and the effect of hormones on growth and development were studied. The phytochemical composition was determined by GC-MS analysis and pharmacological activity was evaluated by assessing the antioxidant and anti-diabetic activities. For the antioxidant activity ABTS+ [2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)], DPPH• (2,2-diphenyl-1-picrylhydrazyl) and metal chelating assays were done and for evaluation of the in vitro anti-diabetic activity α-glucosidase and α-amylase inhibitory activities were done. Result Growth of L. cruciata was recorded in half strength MS media. Benzylaminopurine (BAP: 2 mg/L) and 1-Naphtheleneacetic acid (NAA: 0.5 mg/L) were the successful hormonal combination. GC-MS analysis revealed the existence of nine key compounds in both in vitro and naturally grown L. cruciata. Result of antioxidant and anti-diabetic activity showed that in vitro grown L. cruciata has a more or less similar antioxidant and anti-diabetic activities as naturally grown ones. This result confirms the possibility of using in vitro grown plants in place of naturally grown plants for research and clinical purposes.

Lutzomyia cruciata (Diptera: Phlebotominae) is an important Leishmania vector with wide distribution in America. However, geographical patterns of morphological variation of populations along its longitudinal and latitudinal distribution are still unknown. In this work, we quantified morphological variation and disparity in 243 specimens (females) from nine sample groups of Lu. cruciata from two biogeographical provinces in Mexico: Veracruzan Province (five groups) and Yucatan Peninsula Province (four groups). We registered two linear morphometric data of a reproductive structure (spermatheca) and one landmark configuration for wing shape. Our questions were if intraspecific morphological variation and disparity are related to contrasting environmental factors in these two provinces. First, our results from canonical variates analyses of the spermatheca and wing shape revealed more differences among groups of specimens within a biogeographic province than between. Second, our regression analyses revealed that morphometric variation is weakly related to 20 environmental variables considered for the two biogeographic provinces. Levels of morphological disparity in the spermatheca and wing shape within the two provinces are the same regardless of environmental homogeneity in the Yucatan Province or heterogeneity in the Veracruzan Province. These patterns of morphological variation and disparity weakly correlated with environmental variables suggest that intraspecific variation in Lu. cruciata is not clinal within a province. Subsequent investigations will be required to elucidate suspected correlations of local discontinuous phenotypes with genetic divergence within a biogeographical province.

The complete plastid genome of Gentiana leucomelaena Maxim., belonging to the most species-rich section Chondrophyllae in Gentiana, was determined and analyzed in this study. It has a circular-mapping molecular with the length of 131,856 bp, the shortest one among all available Gentiana plastomes. Gentiana leucomelaena has gene mutation, for example ndh and rpl2 intron, and reversed SSC region comparing with the available species in sections Cruciata, Frigida, Kudoa, Isomeria and Microsperma. Phylogenetic analysis showed that G. leucomelaena clustered together with section Cruciata with a long branch. The plastome provides in this work will contribute to elucidate the phylogenetics and evolution in Gentiana.

The presented checklist and red list include 231 liverwort (Marchantiophyta) and two hornwort species (Anthocerotophyta). Overall, 22.9% of liverworts are evaluated as threatened (CR – 4.3%, EN – 7.8%, VU – 10.8%), while 17 of species (7.4%) are categorized as NT, 11.6% as DD and 3.0% as RE. A total of 128 species (55.4% of the liverwort flora of Slovakia) were assessed as LC. For the liverwort Lunularia cruciata the criteria were not applicable (NA), since this species is not native to Slovakia. Currently, only two hornworts are known in Slovakia, of which Anthoceros agrestis is assessed as LC and Phaeoceros carolinianus as NT. The main factors of the liverwort and hornwort endangerment and retreat are anthropogenic changes in natural conditions and degradation of the ecosystems caused by agriculture, forestry, aquaculture, climatic changes, industry, and traffic pollution.

Volatile oils (VOs) obtained from the aerial parts of species from the Disynaphiinae subtribe (genera Acanthostyles, Campovassouria, Disynaphia, Grazielia, Raulinoreitzia, and Symphyopappus) of Eupatorieae (Asteraceae) collected in Rio Grande do Sul, southern Brazil, were characterized using GC–MS. The yield of VOs relative to fresh material ranged from 0.2 to 1.7% between the Disynaphiinae subtribe samples (Acanthostyles buniifolius, Campovassouria cruciata, Disynaphia ericoides, D. ligulifolia, D. spathulata, Symphyopappus casarettoi, S. reticulatus, S. itatiayensis, Grazielia gaudichaudieana, G. intermedia, G. nummularia, G. serrata, Raulinoreitzia crenulata, and R. tremula). The VOs in this subtribe were mainly composed of terpene compounds. Compositional multivariate analysis demonstrated clustering between the samples of Grazielia and Raulinoreitzia on one side, which contain relatively higher proportions of monoterpenes, and Disynaphia and Campovassouria, which contain more oxygenated sesquiterpenes. The samples of Symphyopappus and Acanthostyles, which contain more sesquiterpene hydrocarbons, were located between these two main groups. Multivariate analysis accounting for the recent proposals of subtribal recircumscription of Eupatorieae was also performed. The odd chemistry of S. itatiayensis was in accordance with the recent attempts to exclude it from the genus Symphyopappus.

The Qinghai-Tibetan Plateau (QTP) and adjacent areas are centres of diversity for several alpine groups. Although it is known that the QTP acted as a source area for diversification of the alpine genus Gentiana, the evolutionary processes underlying diversity in this genus, especially the formation of narrow endemics, are still poorly understood. Hybridization has been proposed as a driver of plant endemism in the QTP but few cases have been documented with genetic data. Here, we describe a new endemic species in Gentiana section Cruciata as G. hoae sp. nov., and explore its evolutionary history with complete plastid genomes and nuclear ribosomal internal transcribed spacer sequence data. Genetic divergence within G. hoae ~3 million years ago was followed by postglacial expansion on the QTP, suggesting Pleistocene glaciations as a key factor shaping the population history of G. hoae. Furthermore, a mismatch between plastid and nuclear data suggest that G. hoae participated in historical hybridization, while population sequencing show this species continues to hybridize with the co-occurring congener G. straminea in three locations. Our results indicate that hybridization may be a common process in the evolution of Gentiana and may be widespread among recently diverged taxa of the QTP., Here, we describe a new species, Gentiana hoae from the Qinghai-Tibetan Plateau (QTP), and explore its evolutionary history in this region. Pleistocene glaciations are suggested as a key factor shaping the population history of G. hoae. Phylogenetic analysis suggests that G. hoae participated in historical hybridization, while population sequencing show this species continues to hybridize with the co-occurring congener G. straminea in three locations. Our results indicate that hybridization may be a common process in the evolution of Gentiana and may be widespread among recently diverged taxa of the QTP.

The maculosa species-group of the genus Promalactis Meyrick, 1908 is proposed based on the features of the forewing pattern. Members of this group have a white speckle in the forewing cell and usually with more than eight white markings distributed mostly at or above the fold and along the dorsal margin. Eighteen new species are described: P. coloristigmosa sp. nov., P. cruciata sp. nov., P. lateridentalis sp. nov., P. latispinata sp. nov., P. lonchodes sp. nov., P. magnispina sp. nov., P. plicata sp. nov., P. bomiensis sp. nov., P. fasciserrata sp. nov., P. brevipalpa sp. nov., P. libona sp. nov., P. medilargissima sp. nov., P. ramivalvata sp. nov., P. aculeiformis sp. nov., P. apicitriangula sp. nov., P. curvispinosa sp. nov., P. medimacularis sp. nov., and P. serraticostalis sp. nov. Photographs of adults and genitalia of the new species are provided. A total of 50 species of the maculosa species-group are listed, including 32 previously known species.

A collection of microcaddisflies from sites on the Cuito, Cuanavale, Cuembo, Cuando, Cubango, and Lungue Bungo Rivers in Angola has yielded seven new species and led to new records for a further five species. The new species include Hydroptila cuembica sp. nov., Hydroptila moxica sp. nov., Oxyethira palisada sp. nov., Orthotrichia ferreirae sp. nov., Or. meyi sp. nov., and Or. mlamboi sp. nov., and another species for which we erect a new genus, Aenigmatrichia asymmetrica gen. et sp. nov. The new records are for Catoxyethira kunenica Mey & de Moor 2019, Hydroptila brigittae Gibon 1987b, Hydroptila cruciata Ulmer 1912, Hydroptila maoae Gibon, Guenda, & Coulibaly 1994, Oxyethira sechellensis Malicky 1993, and a larva identified as that of a species of Tricholeiochiton. Beyond the known Angolan microcaddisfly fauna we resolve several taxonomic problems: We refer to Orthotrichia a species described by Jacquemart in Hydroptila, to become Orthotrichia trifurcata (Jacquemart 1962), comb. nov., with Orthotrichia hydroptiloides Wells & Andersen 1995 (from Tanzania) as a junior synonym; and we recognise Orthotrichia kivuensis Jacquemart 1956 (from Bukana, on Lake Kivu) as a junior synonym of Or. sanya Mosely 1948a from the Orange and Fish rivers. A list is given of the 16 microcaddisfly species now known from Angola: Representation is probably typical of sand and silt-based streams and pools, with the exception of Catoxyethira and Orthotrichia.

In this study, we examine 500 specimens of Anaplecta collected from China, of which 26 samples were used for COI sequencing. We confirm eight new species, i.e., Anaplecta corneola Deng & Che sp. nov., Anaplecta staminiformis Deng & Che sp. nov., Anaplecta arcuata Deng & Che sp. nov., Anaplecta strigata Deng & Che sp. nov., Anaplecta furcata Deng & Che sp. nov., Anaplecta cruciata Deng & Che sp. nov., Anaplecta nigra Deng & Che sp. nov. and Anaplecta bicolor Deng & Che sp. nov. based on morphological and molecular data using ABGD and GMYC analyses. The results of ABGD and GMYC were basically consistent with the morphospecies of Anaplecta. The intraspecific and interspecific genetic distances of Anaplecta ranged from 0 to 6.6% and 16.8% to 31.8%, respectively. We found the male genitalia of Anaplecta to exhibit intraspecific variation, especially in the phallomeres.

The chemical composition of the volatile fraction from Galium verum L. (leaves and flowers) and Cruciata laevipes Opiz (whole plant), Rubiaceae, was investigated. Samples from these two plant species were collected at full bloom in Val di Susa (Western Alps, Turin, Italy), distilled in a Clevenger-type apparatus, and analyzed by GC/FID and GC/MS. A total of more than 70 compounds were identified, making up 92%&ndash, 98% of the total oil. Chemical investigation of their essential oils indicated a quite different composition between G. verum and C. laevipes, both in terms of the major constituents and the dominant chemical classes of the specialized metabolites. The most abundant compounds identified in the essential oils from G. verum were 2-methylbenzaldheyde (26.27%, corresponding to 11.59 &mu, g/g of fresh plant material) in the leaves and germacrene D (27.70%, 61.63 &mu, g/g) in the flowers. C. laevipes essential oils were instead characterized by two sesquiterpenes, namely &beta, caryophyllene (19.90%, 15.68 &mu, g/g) and trans-muurola-4(15),5-diene (7.60%, 5.99 &mu, g/g), two phenylpropanoids, benzyl alcohol (8.30%, 6.71 &mu, g/g), and phenylacetaldehyde (7.74%, 6.26 &mu, and the green-leaf alcohol cis-3-hexen-1-ol (9.69%, 7.84 &mu, g/g). The ecological significance of the presence of such compounds is discussed.

Wide hybridization, which is a powerful tool to broaden genetic variation, has been used in breeding of many crops. However, in ornamental gentian few wide hybridizations have been reported. Interspecific hybridizations between two gentian cultivated species (Gentiana scabra and G. triflora) and 11 wild species, which were classified in five sections, were carried out using ovule culture. When G. scabra was used as a female parent, normal seedlings and hybrid plants were obtained from eight and five interspecific combinations, respectively. The yield of seedling produced from ovule culture depended on interspecific combinations, ranging from 0.3 to 427.7 normal seedling per flower. In the hybridization of G. triflora with five wild species, normal seedlings and plants were produced in five and four interspecific combinations, respectively. The yield of normal seedling ranging from 0.4 to 228.3 was different between not only interspecific combinations but also reciprocal crosses. Two cultivated species are classified in sect. Pneumonanthe, and successful production of hybrids was obtained from the hybridization with species classified in sections Pneumonanthe or Cruciata. The hybrid nature of the produced plants was confirmed by molecular marker and morphology. The production of interspecific hybrids opens a novel prospect in ornamental gentian breeding.

Until recently, fertile colonies of the unisexual Mediterranean liverwort Lunularia cruciata were considered to have a restricted distribution in Britain, due in particular to damaging effects of cold winters. Records of male-expressing plants and sporophytes were almost restricted to southern parts of England and Wales, while female-fertile plants had a somewhat wider range. A different picture has emerged from a more recent survey and, together with findings of other recorders, fertile plants of both genders of L. cruciata have been found in more northerly localities. Sporophytes have also been recorded somewhat more widely, but are likely to be limited by both separation of the sexes and a female-biased sex ratio. The British distribution of fertile plants is compared from records before and after 1990 since when there has been an increase in seasonal and annual temperature. It is suggested that warmer conditions have promoted an increase in fertility. This appears to be the first proposed example of r...

Lunularia cruciata (L.) Dumort. ex Lindb., the only species of the monotypic liverwort family Lunulariaceae (Lunulariales), is reported for the first time for the bryoflora of Kashmir Himalaya. It is easily recognizable by crescent-shaped gemma cups on the surface of the thalli.

Medial medullary syndrome (MMS) consists of ipsilateral lingual paresis, contralateral hemiparesis and lemniscal sensory loss. Hemiplegia cruciata (HC) consists of ipsilateral arm paresis and contralateral leg paresis. Both syndromes develop as a result of pathologies affecting V4 segment of the vertebral artery and anterior spinal artery. A 54-year-old man was admitted with left sided weakness. Neurological examination revealed left hemiparesis and right lingual paresis. Acute diffusion defect of anterior part of medulla oblongata was observed on Diffusion Weighted Imaging (DWI) and the patient was diagnosed with medial medullary syndrome. A month later, he developed another stroke causing a newly-emerging right arm paresis and progression of left leg paresis. DWI examination was repeated, which revealed an acute diffusion defect of craniocervical junction extending till third cervical vertebra. The patient was diagnosed with HC following MMS. We presented these two extremely rare syndromes which developed consecutively in a patient with stroke recurrence.

Gentiana section Cruciata (Gentianaceae) is mainly found on alpine mountains across Eurasia, with the greatest species diversity occurring on the Qinghai–Tibet Plateau. Gentiana crassicaulis Duthie ex Burk. and the closely related species Gentiana robusta King ex Hook. f. and Gentiana tibetica King ex Hook. f. in sect. Cruciata show extremely high morphological similarity, which could be one of the main reasons for their misidentification. This study targets the authentication of the three closely related species in section Cruciata using morphometric analysis of flower shape. We collected 130 individuals in 14 populations of G. crassicaulis, 38 individuals in 12 populations of G. tibetica, and 45 individuals in 13 populations of G. robusta throughout the respective geographical ranges of the species. We used the parameters for bract length (l1), bract width (w1), inflorescence circumference (c), inflorescence width (w2), calyx length (l2), and corolla length (l3) to quantify the flower shape. The analyses showed that the parameters Rb (l1/w1), Ri (w1/w2), and l3 were significantly different between two of the species: G. crassicaulis and G. tibetica could be identified through corolla length, and although very similar to G. crassicaulis and G. tibetica, G. robusta is distinguishable using the parameters Rb and Ri.

Cruciata articulata (L.) Ehrend. is a herbaceous species distributed in parts of Western Asia and the Mediterranean region. While research on other species in the Cruciata genus has revealed the presence of a range of flavonoids and terpenoids, few such studies have been conducted on C. articulata. Thus, in the current study, a phytochemical investigation of C. articulata was carried out. Molecular networking identified a large cluster of compounds sharing distinctive MS-MS fragmentation patterns that were targeted for isolation, leading to the isolation of five undescribed monoterpenoid glycosides, cruciasides C-G, along with two known monoterpenoid glycosides. The structures of these compounds were elucidated by using chemical and spectroscopic analyses, including 1D and 2D NMR, and MS-MS fragmentation. Structures for the ions observed in the MS-MS were proposed, and based on these fragmentation patterns, structures for several of the minor components observed in the molecular network were also proposed. All isolated compounds were tested for cytotoxic, anti-inflammatory, antimicrobial, and α-glucosidase inhibitory properties, but did not display any activity.

Somatic hybridization by protoplast fusion is used in breeding programs to obtain plant material that has inherited valuable traits from two different species, and in order to broaden plant genetic diversity. Somatic hybrids of the genus Gentiana could provide a useful source of new ornamental cultivars and of secondary metabolites. However, in order to evaluate its further usefulness, detailed characterization of the newly created hybrid is essential. Here, genome composition and stability of interspecific gentian somatic hybrids obtained following electrofusion of cell suspension-derived protoplasts of diploid Gentiana kurroo Royle with leaf mesophyll-derived protoplasts of tetraploid G. cruciata L. were characterized using various molecular markers and flow cytometry. AFLP and ISSR analyses revealed that all 21 hybrid plants and 3 lines of hybrid callus were genetically closer to G. cruciata than to G. kurroo. According to the results of chloroplast DNA analysis with the use of CAPS markers, all somatic hybrids inherited chloroplasts from “mesophyll” fusion partner G. cruciata. Flow cytometry revealed that polyploidization occurred, and probably it took place at the early stage of post-fusion culture. In consequence, gradual elimination of nuclear DNA, mixoploidy, and high genetic instability were observed in most hybrid plants and calli during the subsequent 4 years of in vitro culture.

The liverwort Lunularia cruciata, known for being a species tolerant to pollution able to colonize urban areas, was collected in the town of Acerra (South Italy) to investigate the biological effects of air pollution in one of the three vertices of the so-called Italian Triangle of Death. The ultrastructural damages observed by transmission electron microscopy in specimens collected in Acerra were compared with samples collected in the city center of Naples and in a small rural site far from sources of air pollution (Riccia, Molise, Southern Italy). The biological response chain to air pollution was investigated considering vitality, photosynthetic efficiency, heat shock protein 70 (Hsp70) induction and gene expression levels, and chlorophyll degradation and related ultrastructural alterations. Particularly, a significant increment in Hsp70 expression and occurrence, and modifications in the chloroplasts' ultrastructure can be strictly related to the environmental pollution conditions in the three sites. The results could be interpreted in relation to the use of these parameters as biomarkers for environmental pollution.

The horse mackerels of the genus Trachurus (Pisces: Carangidae) are commercially important pelagic fish resources worldwide. There have been several studies of the parasites infecting fishes of this genus, the first one concerning the occurrence of the coccidian Goussia cruciata Thelohan, 1892. Horse mackerels have been targeted in recent projects, in a number of countries that exploit this fish resource (Argentina, Chile, European and Mediterranean countries, North America), with a view of using parasites to define population structure, in conjunction with other methods and techniques, to achieve a better understanding of fish stocks and to minimize risks of localized extinctions. However, information on the occurrence of parasites in this fish genus is scattered through a substantial number of scientific journals, some of them of difficult access. The present review aims to compile the up-to-date knowledge on the occurrence of parasites of horse mackerels, from protozoans to metazoans, their dis...

Background Species belonging to the genus Lernaea are cosmopolitan parasites that can infect many different freshwater fish hosts. Due to a high degree of morphological intraspecific variability and high levels of interspecific similarities, their classification is extremely difficult and controversial. Although the suitability of the shape of cephalic horns has been questioned decades ago by some experimental infection studies, this character still plays the central role in the identification of Lernaea spp. Methods We used the nominal species Lernaea cyprinacea and Lernaea cruciata to test the hypothesis that the shape of the anchor can exhibit host-induced morphological variability, and that the two taxa may be synonymous. Results We examined 517 wild or farmed specimens of five host fish species (four cyprinids and a mosquitofish), and found that all 16 parasite specimens collected from mosquitofish could be morphologically identified as L. cruciata, whereas the remaining 25 parasite specimens were all identified as L. cyprinacea. We experimentally infected goldfish and mosquitofish specimens with offspring (copepodids) of a single L. cyprinacea specimen: the adult parasites from goldfish were morphologically identified as L. cyprinacea, and those from mosquitofish as L. cruciata. We then used molecular data to corroborate that all these specimens are conspecific. Conclusions Our results suggest that L. cyprinacea and L. cruciata may be synonyms, misidentified as different species as a result of host-induced morphological variation. Given the current shortage of molecular data for the genus Lernaea, in order to resolve the taxonomy of this genus (determine the exact number of species), future studies should aim to sequence as much molecular data as possible, and conduct further experimental infections.

Early work on the photoperiodic responses of liverworts had shown a very clear induction of dormancy by long days in Lunularia cruciata, especially in the Mediterranean strains, while in short-day conditions growth of this species continued rapidly without interruption. An obvious requirement for an attempt is to devise a suitable bioassay method which will demonstrate the presence or absence of suspected inhibitory or other substance. For this purpose the gemmae were chosen as the most suitable material. Thalli which have been arrested in their growth by long-day treatment were found to exhibit resistance to drought. Lunularia thalli growing actively on a moist medium will desiccate and die if removed from the moist substrate and exposed to atmospheres of relative humidity less than 90%; i.e., their drought resistance is negligible. While there are clearly quantitative and probably also qualitative differences between species which so far have not been detected, the generalizations below must be regarded as almost certainly too sweeping.

Seven new bisbibenzyls (1-7) were isolated from the methanol extract of the liverwort Lunularia cruciata along with one previously known bibenzyl and five known bisbibenzyls. The structures of compounds 1-7 were elucidated on the basis of the spectroscopic data. These newly isolated bisbibenzyls may be divided into two groups, the acyclic bisbibenzyls, perrottetins (1-3), and the cyclic analogues, riccardins (4-7). Besides standard perrottetin and riccardin structures (1 and 4, respectively), they contain phenanthrene (3 and 5), dihydrophenanthrene (2), and quinone moieties (6 and 7), rarely found in natural products. The new compounds 3 and 5, as well as the known riccardin G, exhibited cytotoxic activity against the A549 lung cancer cell line with IC 50 values of 5.0, 5.0, and 2.5 μM, respectively. Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3097]

In this paper, we show the results of research that can inform conservation measures elsewhere in Europe for the endangered butterfly Euphydryas aurinia . A five year field study was undertaken to identify the host plant preference of larvae of Euphydryas aurinia provincialis in the Mediterranean and which signals are used by females to lay their eggs. The females oviposit on Gentiana cruciata , Scabiosa columbaria and Cephalaria leucantha ; the larvae feed on all these plants and additionally on Lonicera caprifoliumin the wild and on Gentiana lutea in the laboratory. The females do not show any preference for a specific host plant and the larvae move from one species of plant to another without any difficulty. The most important factors in determining the female oviposition are the visibility, accessibility and sun-exposure of the host plants. The vegetative state of host plants is the key factor in larval use of plants during the pre- and post-diapause period. The large-sized host plants, G. cruciata and C. leucantha , are optimal for the growth and survival of the pre-diapause I-III larval instar, while they are unavailable to the larvae in Spring because of their delayed vegetative growth. The post-diapause larvae preferentially feed on plants of S. columbaria , and to lesser degree L. caprifolium , as they provide and abundant food source compared with G. cruciata and C. leucantha. The results also suggest that, there is an evolutionary advantage in large numbers of caterpillars feeding together, with the females of E. aurinia provincialis preferring to lay their eggs nearby or above egg batches laid previously by another female, and selecting large plants for oviposition. Despite the competition for food among caterpillars, the oviposition behaviour of females is advantageous and increases the larval survival rate on large plants. The gregarious larval behaviour provides several benefits during both pre-diapause period (avoiding starvation) and post-diapause period (efficiency in thermoregulation).

Aim We investigated the historical biogeography and diversification of Gentiana L. (Gentianaceae). Our study depicts the origin and dispersal routes of this alpine genus, and the role of the uplift of the Qinghai–Tibet Plateau (QTP) and past climate changes as triggers for its diversification. Location Tibeto-Himalayan region and world-wide mountain habitats. Methods Our sampling represents more than 50% of the extant Gentiana species, including all sections across their entire geographical ranges. We investigated the evolutionary history of Gentiana using phylogenetic reconstructions (maximum likelihood and Bayesian inference) of ITS, atpB–rbcL and trnL–trnF sequences, as well as molecular dating with beast. We tested two approaches of ancestral area reconstructions (DEC, DIVA) in BioGeoBEARS and investigated diversification rates using BAMM. Results The common ancestor of Gentiana and subtribe Gentianinae lived in the QTP region at around 34 (25–45) million years ago (Ma), and 40 (29–52) Ma respectively. From the surroundings of the QTP, Gentiana lineages dispersed to eastern China, Taiwan, Europe, North and South America, Australia and New Guinea, from mid-Miocene onward (c. 15 Ma–present), with only one older dispersal event to Europe (c. 37–21 Ma). Diversification rates gradually increased over time, and two switches of diversification rates were identified in Gentianinae (c. 7 Ma, simultaneously in the Pneumonanthe/Cruciata lineage and in Tripterospermum). Main conclusions Gentiana existed in the QTP region throughout most of its uplift history following the India-Asia collision. This region acted as the primary source area for dispersals to many areas of the world. Because steady increase in diversification rates coincides with the extension of the QTP, we argue that the museum theory rather than the explosive radiation theory prevails for gentians in this region, although rare shifts of diversification rates are associated with niche shifts across the alpine/subalpine ecotone.

The Rubiaceae is characterized by considerable diversity in floral features. To shed light on the variation of floral developmental patterns in Rubiaceae, a comparative study was carried out on pentamerous flowers of Rubia tinctorum L. and tetramerous flowers of Cruciata (Cruciata laevipes Opiz.; Cruciata glabra (L.) Ehrend.), two herbaceous genera of the tribe Rubieae. We aim to study the mechanism responsible for meristic variation and to discuss the expression of andromonoecy in C. laevipes. Developmental peculiarities include the time overlap in the initiation of petal and stamen whorls in Cruciata, the lack of calyx, and a very weak sympetaly in all species examined. Our results show that changes in initial meristem size can play a significant role in meristic variation. The late expression of gender differentiation in C. laevipes may be an explanation for the plasticity of andromonoecy in response to the environmental changes.

The liverwort Lunularia cruciata was collected from the town of Acerra, in the heart of the so-called ‘Land of Fires’ a large area in the eastern part of Campania region of Italy affected by burning of waste and fraudulent dumping and one of the vertices of the “Italian Triangle of Death” so said for the high incidence and mortality from tumors. The data obtained from these samples were compared with samples collected in two other sites representing two different environmental conditions. The soil below the samples, and gametophytes, were collected and analyzed for the concentration of Al, As, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se, V. DNA damage, Reactive Oxygen Species production and localization, activity of antioxidant enzymes and presence of chelating molecules were investigated. All biomarkers provided an answer closely related to the pollution conditions at the 3 sites. We discuss the data considering the possibility of using these biological changes as environmental pollution biomarkers. Finally, it is underlined the importance of phytochelatins due to of their specificity for metal pollution.

The Rubiaceae tribe Rubieae has a world-wide distribution with up to 1,000 species. These collectively exhibit an enormous ecological and morphological diversity, making Rubieae an excellent group for macro- and microevolutionary studies. Previous molecular phylogenetic analyses used only a limited sampling within the tribe or missed lineages crucial for understanding character evolution in this group. Here, we analyze sequences from two plastid spacer regions as well as morphological and biogeographic data from an extensive and evenly distributed sampling to establish a sound phylogenetic framework. This framework serves as a basis for our investigation of the evolution of important morphological characters and the biogeographic history of the Rubieae. The tribe includes three major clades, the Kelloggiinae Clade (Kelloggia), the Rubiinae Clade (Didymaea, Rubia) and the most species-rich Galiinae Clade (Asperula, Callipeltis, Crucianella, Cruciata, Galium, Mericarpaea, Phuopsis, Sherardia, Valantia). Within the Galiinae Clade, the largest genera Galium and Asperula are para- and polyphyletic, respectively. Smaller clades, however, usually correspond to currently recognized taxa (small genera or sections within genera), which may be used as starting points for a refined classification in this clade. Life-form (perennial versus annual), flower shape (long versus short corolla tube) and fruit characters (dry versus fleshy, with or without uncinate hairs) are highly homoplasious and have changed multiple times independently. Inference on the evolution of leaf whorls, a characteristic feature of the tribe, is sensitive to model choice. Multi-parted leaf whorls appear to have originated from opposite leaves with two small interpetiolar stipules that are subsequently enlarged and increased in number. Early diversification of Rubieae probably started during the Miocene in western Eurasia. Disjunctions between the Old and the New World possibly are due to connections via a North Atlantic land bridge. Diversification of the Galiineae Clade started later in the Miocene, probably in the Mediterranean, from where lineages reached, often multiple times, Africa, eastern Asia and further on the Americas and Australia.

Gentiana section Cruciata is widely distributed across Eurasia at high altitudes, and some species in this section are used as traditional Chinese medicine. Accurate identification of these species is important for their utilization and conservation. Due to similar morphological and chemical characteristics, correct discrimination of these species still remains problematic. Here, we sequenced three complete chloroplast (cp) genomes (G. dahurica, G. siphonantha and G. officinalis). We further compared them with the previously published plastomes from sect. Cruciata and developed highly polymorphic molecular markers for species authentication. The eight cp genomes shared the highly conserved structure and contained 112 unique genes arranged in the same order, including 78 protein-coding genes, 30 tRNAs, and 4 rRNAs. We analyzed the repeats and nucleotide substitutions in these plastomes and detected several highly variable regions. We found that four genes (accD, clpP, matK and ycf1) were subject to positive selection, and sixteen InDel-variable loci with high discriminatory powers were selected as candidate barcodes. Our phylogenetic analyses based on plastomes further confirmed the monophyly of sect. Cruciata and primarily elucidated the phylogeny of Gentianales. This study indicated that cp genomes can provide more integrated information for better elucidating the phylogenetic pattern and improving discriminatory power during species authentication.

Cruciata pedemontana (Bellardi) Erhend. var. pedemontana – Tenda, Grimaseta-Poggiale [42°34’N 9°13’E], fruticée naine, 1470 m, 27.6.2006, Gamisans, J. 184 (G); Secteur Rotondo, commune d’Evisa, col de Vergio, sur la crête à env. 400 m au SE du col, chaos rocheux et fruticée, 1540 m, une vingtaine de plantes, 27.6.2016, Delage, A. s.n. (Hb. CBNC), pointage GPS et photos; Secteur Ospedale-Cagna, commune de Lévie, Montagne de Cagna, Pianu d’Ovace, pelouse mésophile dans la partie N du plateau, 1230 m, plusieurs dizaines de plantes, 30.6.2016, Delage, A. & T. Fernez s.n. (Hb. CBNC), pointage GPS. Trois nouvelles stations pour ce taxon très rare dans l’île. Si l’espèce était déjà connue du col de Vergio, seul le var. procumbens (Ascherson) Natali & Dutartre y avait été récolté (voir NATALI & DUTARTRE, Candollea 50: 578-583, 1995). Elle est en revanche nouvelle pour le secteur Ospedale-Cagna. Chez cette dernière population les échantillons récoltés ne présentent aucune pilosité étalée ou elle est limitée à quelques poils. Les individus prélevés ou photographiés au col de Vergio montrent quant à eux une pilosité étalée très nette bien que peu abondante, alors que la plante de Tenda a une pilosité nette, rétrorse et relativement abondante. Le taxon passe au statut R (rare).

Lutzomyia cruciata Coquillet (Diptera: Psychodidae: Phlebotominae) is a potential vector of Leishmania sp.; its geographical distribution in Mexico is widespread, but its life history is unknown. The present study gives relevant information on the life cycle, morphology, survival and reproduction of Lu. cruciata observed over successive generations under laboratory conditions. Seven successive generations were produced. A total of 975 adults were obtained in a sexual proportion of 1.1 : 1 (female : male). Each Lu. cruciata female produced 20.7 eggs and 1.9 adults, approximately, with a proportion of eggs per female of 2.7% (first generation) and 21.3% (second generation). The life cycle of Lu. cruciata, from egg to adult, occurred in 52.7 ± 0.52 days. The largest percentage of mortality occurred during the egg stage (48.5%) and the first larval instar (26.5%), whereas in the pupal stage mortality was the lowest (9.1%). Lutzomyia cruciata exhibits sexual dimorphism based on size, which is exhibited as of the second larval instar, males being smaller than females. The maximum survival of females and males was 10 and 15 days, respectively. An overview of the immature stages of the species made with an electronic scanning microscope is included. This paper contributes basic information on aspects of Lu. cruciata that were previously unknown related to its life history.

In 2006, ca. 60 individuals of Gentiana cruciata were found in the Strzyzowskie Foothills. Until recently, only one locality of the species in the area has been reported from Kolaczyce in the Wisloka Riv- er valley by Knapp in the 19th century. The new locality is situated in the Wielopolka River valley on an S-SW-W slope with varied inclination. In total, 16 phytosociological releves were taken with the use of the Braun-Blanquet method in patches comprising Gentiana cruciata. The patches were characterised by high proportions of species from the classes Molinio-Arrhenathereta, Festuco-Brometea, and Trifolio-Geranietea. Given the dominance of meadow species, the community was classified as a thermophilic fresh meadow from the alliance Arrhenatherion. An upward trend was noted in the analysed population, as the number of the indi- viduals increased to ca. 160 in 2014.

The Mediterranean liverwort flora of the surroundings of Santiago, Central Chile, has been little studied. This paper reports 21 species from the area, including one species new to Central Chile (Cephaloziella divaricata), 9 new to the Metropolitan region (Cephaloziella divaricata, Clevea spathysii, Fossombronia sp., Frullania pluricarinata, Lejeunea globosiflora, Leptoscyphus expansus, Lethocolea radicosa, Riccardia sp., Targionia hypophylla ) and 6 new to Quillota province (Clasmatocolea vermicularis, Clevea spathysii, Gongylanthus dusenii, Lethocolea radicosa, Lunularia cruciata, Symphyogyna circinata). Taxonomic notes are provided on selected species of particular interest. Most of the species grow terrestrially; Frullania pluricarinata and Lejeunea globosiflora occur epiphytic on bark in dry Mediterranean forest. Riccia is the most species-rich genus in the area. Three Riccia species, R. crystallina, R. nigrella and R. trichocarpa, have characteristic disjunct ranges, occurring in Mediterran...

La leishmaniasis es una enfermedad infecciosa causada por un protozoo del género Leishmania. En Honduras es una importante causa de morbilidad. El propósito del estudio fue caracterizar la eco epidemiología de la leishmaniasis cutánea no ulcerada (LCNU) en Honduras. Se seleccionaron 4 municipios endémicos a LCNU (Amapala, San Francisco de Coray, Reitoca y Alubarén). Para realizar la captura de Lutzomyia se usaron trampas miniatura tipo CDC, los individuos capturados fueron identificados según los criterios de Young & Duncan. Se realizaron búsquedas de pacientes en conjunto con personal de la Secretaría de Salud de Honduras para realizar la caracterización de las especies de Leishmania usando PCR. Se determinó la ingesta alimenticia de las hembras ingurgitadas utilizando la amplificación de ITS de los principales animales domésticos y humanos. Se colectaron 156 muestras de pacientes con sospecha clínica de LCNU 130 (83.33 %), resultaron positivas a Leishmania spp., siendo Leishmania infantum/chagasi la única especie circulante en la zona. Por medio de la utilización de criterios de Young & Duncan, se logró identificar un total de 5,951 Lutzomyia, de la cuales 3,543 (60 %) son machos y 2,404 (40 %) son hembras con o sin ingesta de sangre. De esta población de insectos capturados, se identificaron diez especies de Lutzomyia, siendo Lu. longipalpis, Lu. evansi y Lu. cruciata las de mayor predominio. Los cuatro municipios presentaron una baja diversidad de especies de Lutzomyia (DMg =1.04). Se analizaron 450 hembras ingurgitadas de las especies: Lu. longipalpis, Lu. cruciata, Lu. evansi, Lu. chiapanensis y Lu. sanguinaria. Las principales fuentes alimenticias encontradas fueron: Gallus gallus, Sus scrofa, Canis familiaris. Se encontró una tasa de infección del 0.27 %, encontrándose a Lu. longipalpis infectada con Leishmania infantum/ chagasi. Con este estudio se logró caracterizar la eco epidemiología de la LCNU en Honduras, confirmando a L. chagasi/infantum como el principal agente etiológico y a Lu. longiplapis su principal vector. DOI: http://dx.doi.org/10.5377/rct.v0i14.1799 Revista Ciencia y Tecnología No. 14, junio 2014: 115-128

Qualitative phytochemical analyses of eight species from the genus Galium (G. corrudifolium, G. cruciata, G. divaricatum, G. lucidum, G. mollugo, G. palustre, G. parisiense, and G. verum), followed by spectrophotometric evaluation of their total phenolic, flavonoid and iridoid content, as well as antiradical capacity, were conducted. G. cruciata contained the greatest amount of total phenolics (111.00 mg gallic acid equivalents g–1 dry extract), while G. verum had the greatest amount of flavonoids (23.11 mg quercetin equivalents g–1 dry extract) and iridoids (461.30 mg aucubin equivalents g–1 dry extract). The best antioxidant (antiradical) activity was shown by G. cruciata (IC50 (ABTS) = 30.30 µg mL–1; IC50 (DPPH) = 27.62 µg mL–1), followed by G. divaricatum, G. verum and G. palustre. The same species were rich in various bioactive constituents and would be appropriate for inclusion in further investigations considering their biomedical potential. This work is licensed under a Creative Commons Attribution 4.0 International License.

The sand fly Lutzomyia cruciata has been associated with the transmission of Leishmania mexicana to humans in Mexico. This species has a wide distribution in Mexico occupying different microhabitats and environments. In this work comparisons of the egg exochorion of Lu. cruciata from different physiographic areas are presented. Study sites are from different states of southern Mexico. Exochorion analysis was carried out using scanning electron microscopy (SEM). Results show differences in the exochorionic pattern among samples from Veracruz (AVER), Yucatán (HOYU) and Chiapas (TACH). The morphotype “Chiapas” has a polygonal crest pattern, the morphotype “Veracruz” shows parallel and longitudinal crests with some or few connections, and the morphotype “Yucatán” has weak connections between crest ridges. These morphological differences could be the result of local adaptations or evidence of divergence within the nominal unit Lutzomyia cruciata.

The following new corticolous species are described.Thelopsis cruciataAptroot & M. Cáceres, with an olive-green thallus, immersed red-brown perithecia andc.50–100 cruciate ascospores per ascus, 7–10×4–7 µm, from Brazil.Topelia argentinensisAptroot, L. I. Ferraro & M. Cáceres, with a verrucose, partly almost isidiate, greenish grey thallus, immersed pinkish perithecia and 8 uniseriate muriform ascospores per ascus, 12–17×7–11 µm, from Argentina.Topelia tetrasporaAptroot & M. Cáceres, with ac.0·1–0·6 mm thick layer of coralloid to usually flattened, irregularly palmately branched isidiac.0·05 mm diam., ascospores 15–19×2–6-septate, muriform, ellipsoid, 39–50×11–16 µm, only up to 4 per ascus maturing, from Brazil.Thelopsis inordinatais reported for the first time from South America, from Guyana. A key is given to all currently known species ofTopeliaandThelopsis. The distinction between these genera has become arbitrary with the recent addition of more or less intermediate species.

Surirella cruciata A. Schmidt and S. terryi Terry are rare and poorly known diatom species originally described from North America and previously illustrated only by drawings. The goal of this study was to examine and document the morphology and distribution of these species. First photographs designating the lectotype of S. terryi and the neo- and epitypes of S. cruciata are presented. Our revision of the distribution records shows that both species are found mostly along the east coast of the USA with a main occurrence in the New England states, while records from outside North America were not confirmed. Surirella terryi and S. cruciata sometimes co-occur, although S. terryi is a strictly freshwater species, while S. cruciata tolerates wider range of salinities from strictly fresh to brackish waters.

The Alcon Blue Maculinea alcon and the Mountain Alcon Blue Maculinea 'rebeli' are obligate ant-parasitic butterflies with no distinct morpho- logical and genetic differences but clear ecological separation. 2. The two butterflies generally lay their eggs on distinct host plant species: M. alcon on Gentiana pneumonanthe and M. 'rebeli 'o nGentiana cruciata. The egg-laying behaviour of these two ecotypes has previously only been investi- gated in sympatric, but not in syntopic populations. We studied the egg-laying preferences of M. alcon and M. 'rebeli' in a unique area where they co-occur, and where they use different host plants. 3. The distance to the nearest other host plant does not seem to influence the decision of female butterflies as to whether to lay eggs on a specific host plant. 4. The two butterflies laid their eggs on different parts of their host plants: M. alcon preferred the sepals of G. pneumonanthe flower buds, while M. 'rebeli' laid most eggs on the leaves of G. cruciata. 5. The major factor correlated with the number of eggs laid on the host plant seemed to be plant quality (number of flowers, stem length, etc.) for both butterflies. 6. The present study reveals behavioural differences between the two Alcon Blue forms under unique syntopic conditions, a hidden intraspecific diversity that is worth conserving.

The assemblages of fossil charophyte fructifications from the La Huerguina Formation at the La Huerguina stratotype and the Las Hoyas section, along with some other samples from isolated outcrops are composed of Atopochara trivolvis var. triquetra, Globator maillardii var. trochiliscoides, Globator maillardii var. biutricularis var. nov., Clavator harrisii var. reyi, Ascidiella cruciata and Mesochara harrisii. Some reworked utricles of Ascidiella iberica var. iberica were also found. The in situ assemblage belongs to the Cruciata-Paucibracteatus Biozone, which is Late Barremian–Early Aptian in age. These results confirm that the entire deposition of the La Huerguina Formation took place within this biozone and not earlier, as previously thought. The top of the unit can be limited to the uppermost Barremian with biostratigraphic data from ostracods. Globator maillardii var. biutricularis var. nov. is defined as the end form for the Globator lineage according to present knowledge. It is characterised by its unique utricle morphology, showing a basal ring representing a second, external utricular layer. This layer shows a primitive structure, reminiscent of G. maillardii var. mutabilis, in contrast to the internal utricle, which is more derived and similar to G. maillardii var. trochiliscoides. In comparison with other non-marine formations of the same age in the Iberian Chain and in Europe, the charophyte assemblages from the La Huerguina Formation appear to be relatively poor and monotonous, suggesting that some of the species found elsewhere never reached this part of the basin owing to the brief development of non-marine facies there. The main differences in composition between the samples studied are indicative of the palaeoecological conditions. Atopochara trivolvis triquetra was found to be dominant in shallow lacustrine facies and Globator maillardii var. trochiliscoides and var. biutricularis are associated with temporary lakes from a well-drained palustrine area. Clavator harrisii and Mesochara harrisii thrived on floodplains whereas Ascidiella cruciata grew in palustrine environments with significant edaphic activity.

In order to evaluate the use of parasites as biological tags to identify populations of the oceanic horse mackerel, Trachurus picturatus , two samples of 100 fish each, caught off the Madeira and Canary Islands from January to June 2009, were examined for the presence of anisakids, trypanorhynchs, acanthocephalans and liver coccidians. In T. picturatus from Madeira, prevalence of the coccidian Goussia cruciata reached 82%, whereas prevalence of the nematode Anisakis sp., the acanthocephalan Rhadinorhynchus cadenati and the trypanorhynch Nybelinia lingualis were 12.0%, 8.9% and 7.0%, respectively. In samples from the Canary Islands, prevalence of G. cruciata was 8.0%, Anisakis sp. and R. cadenati reached 5.0% and 42.0% respectively, and N. lingualis was not recovered. The absence of N. lingualis and the lower prevalence of Anisakis sp. appear to be related to the smaller size of the oceanic horse mackerels examined from the Canary Islands. Although significant differences in prevalence of two parasites, G. cruciata and R. cadenati , were found between the two regions investigated (χ 2 =125.13, df=1, p=0.000 and χ 2 =40.77, df=1, p=0.000), only G. cruciata was considered useful as a biological tag for the identification of populations of T. picturatus . In order to reach sound conclusions, an expansion of the temporal and spatial sampling strategy is recommended.

The methanol extracts from the underground and aerial part of the two species of Gentiana genus, Gentiana asclepiadea L. and Gentiana cruciata L. from Serbia, were investigated for their antigenotoxic activity against wellestablished mutagenic agent ethyl methanesulfonate (EMS) using the in vivo sexlinked recessive lethal (SLRL) test on Drosophila melanogaster. For this purpose, three days old Canton S males were treated with the potent mutagen EMS in concentration of 0.75 ppm, alone and combined with methanol extracts obtained from underground or aerial part of G. asclepiadea and G. cruciata in concentration of 5%, separately. Although EMS in concentration of 0.75 ppm increased the mutation frequency in all three broods, post-treatments with methanol extracts obtained from the underground and aerial part of G. asclepiadea and G. cruciata in concentration of 5%, respectively, drastically reduced the frequency of sex-linked recessive lethal mutations induced by EMS. Compared to the sucrose, as a negative control, methanol extract obtained from underground part of G. cruciata showed the most potent antigenotoxic activity. Extracts from the underground and aerial part of the two species of Gentiana genus, G. asclepiadea L. and G. cruciata L. from Serbia used in our experiments showed a clear antimutagenic effect, reducing the frequency of mutations induced by a strong mutagen such as EMS. [Projekat Ministarstva nauke Republike Srbije, br. III43004 i br. III41010]

The present study presents morphological and chaetotaxic descriptions of the immature stages of Lutzomyia (Tricholateralis) cruciata (Coquillett), a probable vector of leishmaniasis in Mexico. The egg exochorion is consistent with the species already known as Lu. (Tricholaterialis), but different from the Lu. cruciata egg of Chiapas, Mexico. The fourth instar larva of Lu. (Tricholateralis) cruciata possesses a novel antenna, combining morphological characteristics of categories 2 and 3 for neotropical sandflies. Differences between the chaetotaxy of first and fourth instar larvae of Lu. cruciata with those of Lu. (Lutzomyia) and Lu. (Tricholaterlis) are compared and discussed. This is the first time in which the chaetotaxy and morphology of pupa of a species belonging to Lutzomyia (Tricholateralis) sensu Galati have been described, and we recorded for the first time the anterior prothorax setae, which was previously only considered for Old World species.