Testing Species Delimitations in Four Italian Sympatric Leuciscine Fishes in the Tiber River: A Combined Morphological and Molecular Approach

Leuciscine fishes represent an important component of freshwater ichthyofauna endemic to northern Mediterranean areas. This lineage shows high intra-specific morphological variability and exhibits high levels of hybridization, two characteristics that contribute to systematic uncertainties, misclassification of taxa and, potentially, the mismanagement of biodiversity. This study focused on brook chub, Squalius lucumonis, an endemic taxon of Central Italy. The taxonomic status of this species has long been questioned, and a hybrid origin from sympatric leusciscines (S. squalus x Rutilus rubilio, or S. squalus x Telestes muticellus) has been hypothesised. A phenotypic (evaluating shape and meristic counts) and genetic (using mitochondrial and nuclear markers) investigation of these four taxa was conducted to test species delimitation in sympatric areas and to evaluate the taxonomic status of S. lucumonis. One hundred and forty-five individuals of all four taxa were collected within streams of the lowest portion of the Tiber River basin and analysed; this region encompasses a large portion of the S. lucumonis distribution. The different morphological and genetic approaches were individually examined, compared, and then combined in a quantitative model to both investigate the limits of each approach and to identify cases of misclassification. The results obtained confirm the cladogenetic non-hybrid origin of S. lucumonis, highlight the need for immediate conservation actions and emphasise the value of an integrated approach in the study of leuciscines evolution.     

Immunohistochemical identification of aquaporin 2 in the kidneys of young beef cattle

Aquaporin 2 (AQP2) is a small, integral tetrameric plasma membrane protein that is expressed in mammalian kidneys. The specific constitution of this protein and its selective permeability to water means that AQP2 plays an important role in hypertonic urine production. Immunolocalization of AQP2 has been studied in humans, monkeys, sheep, dogs, rabbits, rats, mice and adult cattle. We analyzed the expression of AQP2 in kidneys of 7-month-old Polish-Friesian var. black and white male calves. AQP2 was localized in the principal cells of collecting ducts in medullary rays penetrating the renal cortex and in the collecting ducts of renal medulla. AQP2 was expressed most strongly in the apical plasma membrane, but expression was observed also in the intracellular vesicles and basolateral plasma membrane. Our study provides new information concerning the immunolocalization of AQP2 in calf kidneys.     

Comparative analysis of DNA polymorphisms and phylogenetic relationships among Syzygium cumini Skeels based on phenotypic characters and RAPD technique

The Indian black berry (Syzygium cumini Skeels) has a great nutraceutical and medicinal properties. As in other fruit crops, the fruit characteristics are important attributes for differentiation were also determined for different accessions of S. cumini. The fruit weight, length, breadth, length: breadth ratio, pulp weight, pulp content, seed weight and pulp: seed ratio significantly varied in different accessions. Molecular characterization was carried out using PCR based RAPD technique. Out of 80 RAPD primers, only 18 primers produced stable polymorphisms that were used to examine the phylogenetic relationship. A sum of 207 loci were generated out of which 201 loci found polymorphic. The average genetic dissimilarity was 97 per cent among jamun accessions. The phylogenetic relationship was also determined by principal coordinates analysis (PCoA) that explained 46.95 per cent cumulative variance. The two-dimensional PCoA analysis showed grouping of the different accessions that were plotted into four sub-plots, representing clustering of accessions. The UPGMA (r = 0.967) and NJ (r = 0.987) dendrogram constructed based on the dissimilarity matrix revealed a good degree of fit with the cophenetic correlation value. The dendrogram grouped the accessions into three main clusters according to their eco-geographical regions which given useful insight into their phylogenetic relationships.     

不同耕作措施对冬小麦-夏玉米复种连作系统土壤有机碳和水分利用效率的影响

在连续8年田间定位试验的基础上,分析了关中平原冬小麦 夏玉米复种连作系统2008—2009年连续两个生长季期间不同耕作措施(结合秸秆还田和不还田)对土壤有机碳和水分利用率的影响.结果表明: 相对于传统耕作,保护性耕作有利于土壤有机碳、水分利用效率和作物产量的提高,其中在“深松+秸秆还田”耕作模式下的增幅最高,土壤有机碳含量在0～30 cm土层增幅达到19.5%,水分利用效率和作物产量提高了16.9%和20.5%,而免耕模式则有效提高了0～10 cm土层有机碳含量.在该地区土壤和气候条件下,深松结合秸秆粉碎还田是最理想的耕作模式,最有利于土壤有机碳累积,并提高水分利用效率和作物产量.     

Über Blutbildungsherde im Mesenchym menschlicher Embryonen

Zusammenfassung Das lockere Bindegewebe des embryonalen Körpers wurde auf seine blutbildenden Eigenschaften näher untersucht. Es stellte sich heraus, daß das Mesenchym der Muskulatur, der Subcutis und der Nerven zu einer Hämopoese befähigt ist. Hierbei werden hauptsächlich Erythrocyten gebildet. Gelapptkernige Formen waren seltener zu finden und traten hauptsächlich bei den älteren Embryonen auf. Eine Lymphopoese war in den genannten Mesenchymlagern nicht nachzuweisen, außer natürlich im Bereich der kleinen Lymphknoten, die gelegentlich an den Extremitäten angeschnitten wurden, und zwar merkwürdigerweise oft an Stellen, wo man sie später zu finden nicht mehr gewohnt ist. Undifferenzierte Mesenchymzellen erhalten sich am längsten an der Adventitia der kleineren Gefäße, wo sie uns neben kleineren rundkernigen Formen entgegentreten.Trotz des für eine solche Untersuchung noch relativ kleinen Materials konnten zeitliche Schwankungen in dem Auftreten der Blutbildungsherde festgestellt werden. Die Differenzierung des Mesenchyms nach den verschiedensten Richtungen (Blutbildung, Fettorgane) wird durch eine besonders intensive Vascularisation eingeleitet, ein Vorgang, der besonders in der Subcutis recht eindrucksvoll hervortritt. Es entstehen zunächst im Gewebe große runde undifferenzierte Hämocytoblasten, aus denen dann durch Teilung und Differenzierung Erythroblasten und kernhaltige rote Blutkörperchen entstehen. Die Entkernung geht im Gewebe hauptsächlich durch Austritt des Kernes aus der Zelle vor sich.     

Molecular cytogenetic analysis of the Appenine endemic cyprinid fish Squalius lucumonis and three other Italian leuciscines using chromosome banding and FISH with rDNA probes

Karyotype and other chromosomal characteristics of the Appenine endemic cyprinid fish, Toscana stream chub Squalius lucumonis, were analysed using conventional banding and FISH with 45S and 5S rDNA probes. The diploid chromosome number (2n = 50) and karyotype characteristics including pericentromeric heterochromatic blocks and GC-rich CMA₃-positive sites corresponding to both positive Ag-NORs and 45S rDNA loci on the short arms of a single medium-sized submetacentric chromosome pair were consistent with those found in most European leuciscine cyprinids. On other hand, 5S rDNA FISH in the Toscana stream chub and three other Italian leuciscines, S. squalus, Rutilus rubilio and Telestes muticellus, revealed a species-specific hybridization pattern, i.e. signals on four (S. lucumonis), three (S. squalus and R. rubilio) and two (T. muticellus) chromosome pairs. Whereas all the species shared the 5S rDNA loci on the largest subtelocentric chromosome pair, a “leuciscine” cytotaxonomic marker, S. lucumonis showed both classes of rDNA loci tandem aligned on the short arms of chromosome pair No. 12. The present findings suggest that the observed high variability of 5S rDNA loci provides a powerful tool for investigation of karyotype differentiation in karyologically conservative leuciscine fishes.     

Summer diet selection of snowshoe hares: a test of nutritional hypotheses

We examined summer diet selection by snowshoe hares in the southwestern Yukon, Canada, and attempted to explain how plant qualities such as nutritional content (protein, energy, fiber, water) and defensive compounds affect hare food choice. Male hares were placed in enclosures in natural vegetation areas dominated by shrubs and in both open and closed spruce forests. Each site contained a variety of herbaceous plant species. After 24 h, hares were sacrificed, their stomachs collected, and the contents removed and analyzed. Of the 30 plant species recorded in the enclosures only 10 were identified in the stomach contents of the hares. The summer diet is composed predominantly of five plant species; Lupinus arcticus, Salix spp. Shepherdia canadensis, Betula glandulosa and Festuca altaica. The selection of these species changes between sampling times. Protein, energy and water contents were highest in leaf tissues of most species early in the season, and most species had an increase in fiber through the summer. Protein content consistently explains the largest amount of variation in diet selection, although protein selection is modified by extremely high concentrations of defence compounds. It is likely that hares are not protein limited and are selecting for other plant characteristics correlated with protein content such as energy content. It is significant that hares continue to ingest heavily defended species (e.g. Shepherdia canadensis, B. glandulosa and L. arcticus) when many less defended plants are available to be eaten (e.g. F. altaica, Epilobium latifolium and Anemone parviflora). Our results support the idea that hares are selecting dietary items on the basis of energy content. No single plant species can satisfy all of the hares’ nutritional requirements and the selection patterns may reflect the need to balance many conflicting plant qualities.