Non-invasive method of sex identification of crane chicks by the DNA from capillary vessels of allantois

The non-invasive method of determining the sex of chicks after hatching based on the DNA isolated from capillary vessels of allantois of the egg-shell membranes was demonstrated on four crane species (Gruinae, Aves), which were bred in the Crane Breeding Centre of the Oka Nature Reserve in 2009–2012. Using the EE0.6 molecular marker of sex, the gender of 26 Siberian (Grus leucogeranus), 15 Red-crowned (G. japonensis), 4 Common (G. grus) and 1 Demoiselle (Anthropoides virgo) crane chicks was identified. This method can be recommended for determining the sex of chicks and the ratio of sexes in cranes that reproduce both in captivity and natural populations.     

Genetic passportization and identification of Siberian cranes (Grus leucogeranus Pallas) in captivity

The genetic diversity of the founders of an artificial population of the Siberian crane Grus leucogeranus Pallas (rare species of cranes) was characterized using 10 microsatellite loci. It was established that the allelic diversity (on average, 5.9 alleles per locus) and genic (H _O = 0.739) diversity of the Siberian crane is rather high and comparable with the estimations for natural populations of different crane species. Genetic passportization of the birds (119 individuals) from the register of the Siberian crane International Studbook was carried out at the initial stage. The efficiency of genetic passportization for individual identification, identification of the origin, paternity analysis, and exclusion of inbreeding was demonstrated in Siberian cranes under natural mating and artificial insemination. Cases of natural reproduction in pairs of Siberian cranes imprinted to the human and continuous storage of spermatozoa in the female reproductive ducts were registered.     

Rhythm of honeydew excretion by the tea aphid and its attraction to various natural enemies

The rhythm of honeydew excretion by the tea aphid Toxoptera aurantii (Boyer) and its attraction to following 9 species (or subspecies) of beneficial insects, Aphidius sp., Chrysopa sinica Tjeder, Chrysopa septempunctata Wesmael, Sphaerophoria menthastri L., Coccinella septempunctata L., Leis axyridis (Pallas) ab. bimaculata Hemmelmann, L. axyridis (Pallas) ab. conspicua Faldermaenn, L. axyridis (Pallas) var. spectabilis Faldermaenn and L. axyridis (Pallas) var. novemdecimpunctata Faldermaenn, were investigated. Forty-five wingless virginoparae nymphs, reproduced by the same wingless virginogenia adult within 1 h, were introduced onto tea seedlings with one aphid per seedling. The honeydews excreted from different instars of nymphs and adults were collected under 21°C, 85% RH, 3500 lx and 12 L:12 D photoperiod. It took 32.4 d ±5.8 d for the tested tea aphids to complete the development of their nymph and adult stages, during which 325.6±35.8 droplets (ca. 41.98 μl ±6.14 μl and 45.34 mg ± 8.76 mg) of honeydews were secreted. During the 1st and 4th instars, there was a logistic regression relationship between the amount of honeydews excreted and the time (days). The honeydew secretion during the first two instars was less than that from the later instars. Adult aphids survived for 22.0 d ± 0.0 d, and excreted 176.31 ± 22.38 droplets (ca. 30.38 μl ± 5.32 μl) of honeydews in a rate of 1 drop per ca. 30–50 min for 5–8 h with a pause for 2–5 h before next secretion series. A batch of forty-five virginoparae female adults, reproduced by the same virginoparae female adult within 1 h, were introduced onto the tea seedlings (one aphid/seedling) under 13–21°, 85% RH, 3500 lx and 12 L:12 D photoperiod. Temperature showed a significant effect on the amounts of honeydews excreted within the range of 13–21°. Honeydews excreted by the aphids significantly increased the searching and retention time of the tested 9 species of natural enemies in a positive dose-response fashion. The searching times of Aphidius sp. and S. menthastri were the longest and the shortest, respectively, among all the 9 species, while the searching and retention time of L. axyridis (Pallas) var. spectabilis was the longest among the four varieties of L. axyridis. Tea aphid oneydew is considered as an important contact kairomone for the tested natural enemies.     

Effects of a microsporidium from the convergent lady beetle, Hippodamia convergens Guérin-Méneville (Coleoptera: Coccinellidae), on three non-target coccinellids

A microsporidium from Hippodamia convergens was transmitted horizontally to three non-target coccinellid hosts (Adalia bipunctata L., Coccinella septempunctata L. and Harmonia axyridis Pallas) under laboratory conditions. For all species examined, microsporidia-infected larvae took significantly longer to develop than did uninfected larvae but the microsporidium had no effect on larval mortality. Adult sex ratios of uninfected and microsporidia-infected adults were about 1:1 (♀:♂) and did not differ significantly. At the end of a 90-day trial, microsporidia-infected H. convergens produced significantly fewer eggs and did not live as long as uninfected individuals. Differences in fecundity and longevity were not observed for the three non-target coccinellids that were examined. Mean spore counts from smear preparations of microsporidia-infected A. bipunctata did not differ significantly from H. convergens, suggesting that A. bipunctata (a native coccinellid) is a suitable host for the microsporidium but infection was lighter in C. septempunctata and H. axyridis (introduced species). Vertical transmission of the pathogen was observed during the 90-day trial by examining eggs and larvae that were produced by microsporidia-infected adults. For all species examined, 100% vertical transmission of the pathogen was eventually observed. Three eugregarines were found in two adult A. bipunctata: Gregarine A trophozoites are similar in size to those of Gregarina katherina Watson (described earlier from Coccinella spp.), Gregarine B trophozoites are similar in size to those of Gregarine A but are morphology distinct, and Gregarine C trophozoites are similar in size to G. barbarara Watson (described earlier from A. bipunctata).     

Identification of the sex of a wide range of Carinatae birds by PCR using primer sets selected from chicken EE0.6 and its related sequences

A 0.6 kb EcoRI fragment (EE0.6), cloned from the W chromosome of chickens, is a nonrepetitive sequence and contains an exonlike sequence, ET15, which is likely a part of a pseudogene. The EE0.6 sequence is conserved in all species of birds examined both in Carinatae and Ratitae. A counterpart sequence of EE0.6 is present on the Z chromosome. The extent of diversity between the W- and Z-linked sequences are variable among species. The W- and Z-linked EE0.6 sequences, cloned from 12 different species, were compared and four forward and three reverse primers were selected to amplify parts of the EE0.6 sequence by polymerase chain reaction (PCR). By choosing a suitable combination of primers for EE0.6 and a set of primers for a Z/W-common sequence, as an internal control, the sex of 36 species belonging to 16 different orders of Carinatae could be determined clearly by PCR. The sex of two other species representing different orders could be determined by Southern blot hybridization using ET15 as a probe. For the two Ratitae species, emu and ostrich, EE0.6 sequences on W and Z chromosomes could not be distinguished either by PCR or Southern blotting.     

Analysis of creatine kinase variation in some members of the family Acipenseridae

Creatine kinase (E.C. 2.7.3.2) was examined in stellate sturgeon Acipenser stellatus Pallas, Russian sturgeon A. gueldenstaedtii Brandt, European sterlet A. ruthenus L., Siberian sterlet A. ruthenus marsiglii Brandt, and great sturgeon (beluga) Huso huso L., using polyacrylamide gel electrophoresis. Two loci for creatine kinase were identified: CK-A* in white skeletal muscle and CK-C* in stomach wall muscle. Most species proved to be monomorphic at the CK-A* locus, showing the same phenotype represented by a single band. Heterogeneity and polymorphism in creatine kinase, determined by the CK-A* locus, were found only in Russian sturgeon. Based on the results of densitometric analysis of band staining intensity, we have advanced a hypothesis that synthesis of subunits of the CK-A* product in this species was controlled by eight genes. However, the genotype frequencies in the sample were significantly different from those theoretically expected upon free and independent gene recombination. The results of this study support the hypothesis on the absence of heterodimeric creatine kinase molecules in the skeletal muscle of Russian sturgeon. Locus CK-C* in stellate sturgeon was revealed as a single, intensely stained, rapidly migrating fraction, whereas in Russian sturgeon, the enzyme activity in this zone was very weak. No creatine kinase was found in liver, kidneys, spleen, heart, and intestine mucous tunic.     

Tracking the impact of drought on functionally different woody plants in a Mediterranean scrubland ecosystem

Climate warming is predicted to amplify drought stress. Thus, it is important to understand how coexisting plant species respond to severe droughts. Here we study how seven Mediterranean woody plant species with different evolutionary history and functional characteristics (Pinus halepensis Mill., Juniperus phoenicea L., Pistacia lentiscus L., Rhamnus lycioides L., Rosmarinus officinalis L., Genista scorpius (L.) DC., and Globularia alypum L.) responded to a severe winter drought during 2011–2012 in Spain. The study site is located in the Valcuerna valley, Monegros desert, northeastern Spain. We evaluated how the drought affected the annual growth-ring formation of the species by using dendrochronology and quantified the intensity of drought-induced defoliation and mortality and compared it between species and groups of species with different evolutionary history. Radial growth of all species was strongly reduced by the 2012 drought. The pre-Mediterranean species (P. halepensis, J. phoenicea, P. lentiscus and R. lycioides) reduced growth more than the Mediterranean species (R. officinalis, G. scorpius and G. alypum). Defoliation was significantly higher in pre-Mediterranean than in Mediterranean species. When species were analyzed separately we found that P. halepensis was the species with the highest growth reduction but J. phoenicea was defoliated more severely and showed higher mortality rates as a consequence of drought. In the case of the Mediterranean shrubs, drought-induced mortality was only noticeable in R. officinalis. Drought impacted growth of all species but this did not induce mortality in all of them. Growth reduction was dependent on evolutionary history. However, functional characteristics of the species such as leaf stomatal regulation and root architecture may be more important than evolutionary history on explaining drought-induced mortality. Indeed, species with shallow root systems such as J. phoenicea and R. officinalis were the most adversely affected by the drought.     

Chemical composition studies on some aquatic macrophytes in three scottish lochs. II. Potassium,sodium, calcium,magnesium and iron

The potassium, sodium, calcium, magnesium and iron levels in seven species of freshwater macrophytes (Juncus effusus L., Iris pseudacorus L., Carex rostrata Stokes, Glyceria maxima (Hartm.) Holmberg, Nuphar lutea (L.) Sm., Polygonum amphibium L. and Schoenoplectus lacustris (L.) Pallas) in three Scottish lochs of different trophic levels were studied during 1975 and the results obtained were compared with similar studies in other areas.     

中国植物区系新记录种

报道了采自新疆地区的百花丹科、龙胆科、紫草科的４ 个中国新记录种———塔尔巴哈台彩花（ Ａｃａｎｔｈｏｌｉｍｏｎｔａｒｂａｇａｔａｉｃｕｍ Ｇａｍａｊｕｎ） 、西亚龙胆（ Ｇｅｎｔｉａｎａ ｓｅｐｔｅｍｆｉｄａ Ｐａｌｌａｓ） 、早春龙胆（ Ｇ．ｖｅｒｎａ Ｌ．ｓｕｂｓｐ． ｐｏｎｔｉｃａ Ｍ． Ｓｏｌｔｏｋｏｖｉｃ） Ｈａｙｅｋ 、塔城滨紫草（ Ｍｅｒｔｅｎｓｉａ ｔａｒｂａｇａｔａｉｃａ Ｂ．Ｆｅｄｔｓｃｈ ．） 。     

Influence of larval rearing diet on the intensive searching behaviour ofHarmonia axyridis [Col.: Coccinellidae] Larvae

The capture of prey by last instar larvae of the aphidophagous coccinellidHarmonia axyridis Pallas (Col.,Coccinellidae) modified larval movement so that extensive search was replaced by intensive search. The continuous rearing of this species on eggs ofEphestia kühniella Zell. (Lep.,Pyralidae) or on the aphidAcyrthosiphon pisum Harris (Homopt.,Aphidae) led to conditioning. Only larvae that ate the same prey as they were reared on, adopted intensive searching movements after feeding. Using larvae reared on substitute prey in biological control systems may decrease the efficiency of the released predators.     

Analysis of the mitochondrial DNA variation in pond smelt Hypomesus olidus (Osmeridae)

Pond smelt Hypomesus olidus (Pallas, 1814), one of the five species of the genus Hypomesus, family Osmeridae, was examined for intraspecific variation of the mitochondrial DNA cytb (1062 bp) and COI (567 bp) genes. Among the ten single substitutions discovered, only one, leading to the substitution of isoleucine by valine, was nonsynonymous, while the remaining substitutions were synonymous. The degree of genetic divergence among pooled nucleotide sequences in H. olidus populations examined constituted 0.4% on average, ranging from 0.2 to 0.6%. These values were not higher than the levels of divergence between the individuals within the populations. Phylogenetic analysis of the populations examined did not reveal their subdivision depending of their geographic location, and pointed to the absence of intraspecific differentiation of the species.     

Winners and losers of land use change: A systematic review of interactions between the world’s crane species (Gruidae) and the agricultural sector

While agricultural intensification and expansion are major factors driving loss and degradation of natural habitat and species decline, some wildlife species also benefit from agriculturally managed habitats. This may lead to high population densities with impacts on both human livelihoods and wildlife conservation. Cranes are a group of 15 species worldwide, affected both negatively and positively by agricultural practices. While eleven species face critical population declines, numbers of common cranes (Grus grus) and sandhill cranes (Grus canadensis) have increased drastically in the last 40 years. Their increase is associated with higher incidences of crane foraging on agricultural crops, causing financial losses to farmers. Our aim was to synthesize scientific knowledge on the bilateral effects of land use change and crane populations. We conducted a systematic literature review of peer‐reviewed publications on agriculture‐crane interactions (n = 135) and on the importance of agricultural crops in the diet of cranes (n = 81). Agricultural crops constitute a considerable part of the diet of all crane species (average of 37%, most frequently maize (Zea mays L.) and wheat (Triticum aestivum L.)). Crop damage was identified in only 10% of all agriculture‐crane interactions, although one‐third of interactions included cranes foraging on cropland. Using a conceptual framework analysis, we identified two major pathways in agriculture‐crane interactions: (1) habitat loss with negative effects on crane species dependent on specific habitats, and (2) expanding agricultural habitats with superabundant food availability beneficial for opportunistic crane species. The degree to which crane species can adapt to agricultural land use changes may be an important factor explaining their population response. We conclude that multi‐objective management needs to combine land sparing and land sharing strategies at landscape scale. To support viable crane populations while guaranteeing sustainable agricultural production, it is necessary to include the perspectives of diverse stakeholders and streamline conservation initiatives and agricultural policy accordingly.     

Typification of Pallas’ names in <Emphasis Type="Italic">Salix</Emphasis>

Summary  The original collections of Salix species described by Pallas are discussed. Eight names validated by Pallas and one by Nasarov are typified. The use of six currently accepted names, S. arbutifolia Pall., S. arctica Pall., S. berberifolia Pall., S. caspica Pall., S. divaricata Pall. and S. rhamnifolia Pall., is confirmed. Salix serotina Pall. retains its place as a synonym of S. viminalis L. Salix gmelinii Pall. is resurrected and proposed as the correct name for the taxon previously known as S. dasyclados Wimm. S. burjatica Nasarow and S. jacutica Nasarow, the old synonyms of S. dasyclados, therefore are now also synonyms of S. gmelinii. A new combination S. rosmarinifolia L. subsp. sibirica is proposed.     

Differentiation of two hemp nettle species (<Emphasis Type="Italic">Galeopsis bifida</Emphasis> Boenn. and <Emphasis Type="Italic">G. tetrahit</Emphasis> L.) inferred from morphological characters and DNA markers

Two species of the genus Galeopsis L., G. tetrahit L. and G. bifida Boenn. (family Lamiaceae), are problematic to distinguish often wrongly recognized, and treated by some taxonomists as a single species. Morphological diagnostical characters of these species are variable and partly overlap. Species independence of G. tetrahit and G. bifida was evaluated and their diagnostic characters verified using ISSR and RAPD markers. A total of 57 ISSR and 28 RAPD fragments were obtained providing distinct subdivision of the accessions examined into two groups. Analysis of molecular data using the neighbor-joining method showed that the accessions studied fell into two clades in the same way as demonstrated by the analysis of 20 morphological characters using single linkage method. These results suggest that G. tetrahit and G. bifida are distinct species. The morphological characters were found to be more variable compared to the molecular markers, although the combined of these characters provided differentiation of the species.     

Intersex Occurrence in Rainbow Trout (Oncorhynchus mykiss) Male Fry Chronically Exposed to Ethynylestradiol

This study aimed to investigate the male-to-female morphological and physiological transdifferentiation process in rainbow trout (Oncorhynchus mykiss) exposed to exogenous estrogens. The first objective was to elucidate whether trout develop intersex gonads under exposure to low levels of estrogen. To this end, the gonads of an all-male population of fry exposed chronically (from 60 to 136 days post fertilization – dpf) to several doses (from environmentally relevant 0.01 µg/L to supra-environmental levels: 0.1, 1 and 10 µg/L) of the potent synthetic estrogen ethynylestradiol (EE2) were examined histologically. The morphological evaluations were underpinned by the analysis of gonad steroid (testosterone, estradiol and 11-ketotestosterone) levels and of brain and gonad gene expression, including estrogen-responsive genes and genes involved in sex differentiation in (gonads: cyp19a1a, ER isoforms, vtg, dmrt1, sox9a2; sdY; cyp11b; brain: cyp19a1b, ER isoforms). Intersex gonads were observed from the first concentration used (0.01 µg EE2/L) and sexual inversion could be detected from 0.1 µg EE2/L. This was accompanied by a linear decrease in 11-KT levels, whereas no effect on E2 and T levels was observed. Q-PCR results from the gonads showed downregulation of testicular markers (dmrt1, sox9a2; sdY; cyp11b) with increasing EE2 exposure concentrations, and upregulation of the female vtg gene. No evidence was found for a direct involvement of aromatase in the sex conversion process. The results from this study provide evidence that gonads of male trout respond to estrogen exposure by intersex formation and, with increasing concentration, by morphological and physiological conversion to phenotypic ovaries. However, supra-environmental estrogen concentrations are needed to induce these changes.     

Molecular identification of Kiss/GPR54 and function analysis with mRNA expression profiles exposure to 17α-ethinylestradiol in rare minnow <Emphasis Type="Italic">Gobiocypris rarus</Emphasis>

17α-ethinylestradiol (EE2) is a widely existed endocrine disrupting chemical in water environment. Kisspeptins act as indispensable regulators through GPR54 in the hypothalamic–pituitary–gonadal (HPG) axis. This study aimed to provide further understanding of the effect of EE2 on HPG axis. Molecular cloning and tissue distribution of kiss genes and GPR54s were performed in Gobiocypris rarus. The mRNA expression profiles of kiss1, kiss2, GPR54s and GnRHs were detected in G. rarus brain and/or gonad following 3- and 6-days EE2 (1, 5, 25 and 125 ng/L) exposure. Results showed that kiss genes and GPR54s were highly expressed in brain and gonad. Both kiss1 and kiss2 were increased in female brain and suppressed in male brain following EE2 exposure. GnRHs were inhibited in a concentration-dependent manner in male brain following 3-days EE2 exposure. In gonad, GPR54b was almost suppressed in all of EE2 concentrations. The present findings suggest that EE2 impacts the genes expression of Kiss/GPR54-GnRH system in G. rarus, thereby probably disturbing the neuroendocrine homeostasis.     

Sex Allocation by a Parasitoid Wasp (Hymenoptera: Ichneumonidae) to Different Host Species: A Question for the Mechanism of Host Size Estimation

Sex allocation by the polyphagous solitary pupal parasitoid wasp Pimpla luctuosa Smith to a small host species, Galleria mellonella (L.), and a large host species, Mamestra brassicae L., was investigated to test whether female wasps responded to hosts of different sizes across different host species. In the experiments, both host species were presented to each test female wasp. Primary and secondary sex ratio experiments revealed that female wasps laid more female eggs in larger pupae of each host species, indicating that female wasps recognized size differences within host species. The wasp sex ratio (male ratio) from M. brassicae, however, was much higher than that expected on the basis of the sex ratio curve from different-sized G. mellonella. Larger hosts of each host species yielded larger wasps, indicating that the host size estimation by female wasps across different host species was incomplete or was not simple. These results suggested that P. luctuosa evaluated host size not only by physical measures such as dimension but also by other unknown measures. A possible explanation for the adaptiveness of different sex ratio responses by Pimpla luctuosa to different host species was discussed.