Geometric morphometrics as a tool to resolve taxonomic problems: the case of Ophioglossum species (ferns)

A modern method, geometric morphometrics, was used to clarify the taxonomic position of the European Ophioglossum species: O. azoricum, O. lusitanicum, and O. vulgatum. The identification of these taxa by traditional methods is rather difficult, due to different taxonomic interpretations. Sterile leaf shapes were investigated using a landmark-based method and the Fourier analysis of outlines. Both methods highlight the shape and the base of the leaf as an important diagnostic character

A comparative study of the seed germination capabilities of Anacamptis palustris (Orchidaceae), a threatened terrestrial orchid, and other more common Anacamptis species, by asymbiotic culture in vitro

The increasing emphasis on terrestrial orchid conservation has led to conservation actions for a wide range of threatened Mediterranean species. Many terrestrial orchids are currently at great risk for extinction as a result of a multiplicity of threatening processes. We focus on orchid seed germination capabilities in vitro, specifically germination capability of a threatened species, Anacamptis palustris, compared to three other more common species (A. laxiflora, A. morio, and A. papilionacea), and also discuss its potential impact on orchid distribution and conservation. Asymbiotic germination tests were performed with mature seeds using BM-1 medium. In vitro seed germination and protocorm developmental stages were evaluated up to 20 weeks after sowing. Significant differences in seedling development were detected among the species, and a correlation was evident between the rarity of the species and their germinability. Thus, the presence of intrinsic, biological factors that affect and limit the distribution of A. palustris may exist

Outline analysis for identifying Limodorum species from seeds

Limodorum trabutianum Batt. is an orchid species of the Italian flora, with a central-western stenomediterranean distribution, that is sporadic in the western part of the distribution area of the more common L. abortivum (L.) Sw., an eurimediterranean species. It occurs in Italy only with a few populations in Tuscany, Latium, Umbria, Sicily and Sardinia [1], often with L. abortivum [2], [3], [4] from which it is easily recognizable only during anthesis for the denser inflorescence spike, the ribbon-like lip without differentiation in epychile and hypochile, and for the spur that is very short or absent [5]. On the contrary, the identification of these two taxa during the fruiting phase is rather difficult or even impossible. The aim of this study is to verify the taxonomic value of Limodorum seeds, particularly of their shape, as highlighted from recent studies for other orchids [6], [7], in order to establish its usefulness for recognizing the two species. We have identified 5 Italian populations of the two taxa: 2 populations of L. trabutianum, one within the Marturanum Regional Park (Barbarano Romano, Viterbo), the other near Cortona (Arezzo), and 3 populations of L. abortivum, near S. Martino al Cimino (Viterbo), in the M. Casoli Reserve (Bomarzo, Viterbo), and in the same site of L. trabutianum within the Marturanum Park. The phenology of these populations was monitored to collect mature seeds from naturally dehiscing capsules. The intra- and interspecific variability of seed shapes was analyzed with the methodology of Elliptic Fourier descriptors [8], which allows to describe in terms of harmonics each two-dimensional shape with a closed outline. For this outline analysis we used the software package SHAPE 1.3 [9]. An average of 100 seeds from each species and from each site was photographed with a NIKON Coolpix 5000 camera mounted on a LEITZ-ARISTOPLAN microscope, obtaining 500 digital images with a resolution of 300 dpi and a size of 800 x 1000 pixels. All images were prepared using Adobe Photoshop 7.0: as a first step, every foreign element was eliminated from the picture, thereby isolating the single seed, then its contrast with the background was maximized, and finally all images were saved in .bps format (24bit). The color images were converted to binary with Chain Coder before tracing the outlines in Chain-code, a coding system that describes the geometrical information of the shapes. Then the Chain-code file was transformed into a Normalized Elliptic Fourier file using Chc2Nef using 20 harmonics. The matrix of the harmonic coefficients underwent a process of data normalization based on the first harmonic, to transform the data into shape variables. Subsequently, a PCA was performed on the variancecovariance matrix of normalized coefficients using PrinComp, which gives a graphical output of the principal components (average shape ± standard deviations). The first results of the outline analysis confirm a low intraspecific variability of seed shape, but show a very high interspecific variability: L. abortivum seeds are very elongated, from fusiform to filiform, while L. trabutianum seeds are much wider and have a very lower length/ width ratio. These results allow to distinguish between these two species even during the fruiting phase, simply using seed shape as a diagnostic character, avoiding the use of traditional morphometric analysis which need microscopic measurements

Massive pyuria as an unusual presentation of giant infected urachal remnant in a child

Urachal remnants (URs) are manifestations of an incomplete regression of the urachus; therefore, there may be different types of remnants such as cyst, sinus tract, diverticulum or patent urachus. The clinical presentation of a urachal anomaly includes umbilical discharge, lower abdominal pain and urinary tract infection, although a UR may also be asymptomatic. We present the case of a 2.5-year-old girl who presented with abdominal pain, stranguria and massive pyuria in which a giant infected UR was found. The diagnosis was made using abdominal MRI. The child was subjected to  laparoscopic-assisted drainage and had an uneventful postoperative course.Keywords: differential diagnosis, infected urachal remnant, laparoscopy, pyuri

Differential effects of increasing temperature on the germination of five wild species with varying range sizes in a Carrara marble quarry

Assessing the performance of spontaneous plants in abandoned quarries under future environmental scenarios is important for successful restoration practices. Air warming is one of the most relevant ongoing climatic changes in the Mediterranean. We tested the effects of increasing temperature on the germination of five species spontaneously colonising the abandoned sectors of a Carrara marble quarry (Tuscany, central Italy). We selected five plant species with different widths of their distribution range, from local endemic to invasive alien: Santolina pinnata, Globularia incanescens, Hypericum coris, Helichrysum italicum, and Buddleja davidii. Seeds were collected in situ and their germination was tested in laboratory conditions at 10, 15, 20, 25, and 30°C. The effects of temperature and species on germination percentage (GP) and mean germination time (MGT) were tested through two-way ANOVA. Increasing temperatures negatively affected the GP of S. pinnata and H. coris. G. incanescens performed better at intermediate temperatures, while H. italicum benefited from increasing temperatures. B. davidii showed a similar high GP under all the treatments. MGT decreased with increasing temperature for all the species, except for S. pinnata, which showed an increase of MGT with increasing temperature. We highlighted that, under future warmer climatic conditions, the two endemic species will be disadvantaged with respect to the species with a wider distribution range and the non-native species in the studied quarry. This evidence is relevant for restoration planning since seeding the two endemic species could be unsuccessful under future environmental scenarios, when H. italicum and B. davidii will be more competitive in the colonisation of the quarry

A comparative study of the seed germination capabilities of Anacamptis palustris (Orchidaceae), a threatened terrestrial orchid, and other more common Anacamptis species, by asymbiotic culture in vitro

The increasing emphasis on terrestrial orchid conservation has led to conservation actions for a wide range of threatened Mediterranean species. Many terrestrial orchids are currently at great risk for extinction as a result of a multiplicity of threatening processes. We focus on orchid seed germination capabilities in vitro, specifically germination capability of a threatened species, Anacamptis palustris, compared to three other more common species (A. laxiflora, A. morio, and A. papilionacea), and also discuss its potential impact on orchid distribution and conservation. Asymbiotic germination tests were performed with mature seeds using BM-1 medium. In vitro seed germination and protocorm developmental stages were evaluated up to 20 weeks after sowing. Significant differences in seedling development were detected among the species, and a correlation was evident between the rarity of the species and their germinability. Thus, the presence of intrinsic, biological factors that affect and limit the distribution of A. palustris may exist

Rapid adaptation of seed germination requirements of the threatened Mediterranean species Malcolmia littorea (Brassicaceae) and implications for its reintroduction

AbstractMalcolmia littorea (Brassicaceae) is a threatened species growing in the coastal sandy dunes of the west-Mediterranean basin. In this study, the seed germination and seedling emergence requirements of this species were investigated in the only remaining native population in Italy. The highest germination percentage was achieved in darkness with scoring under safe green light at 5–10°C. Seedling emergence was highest when seeds were buried between 1 and 10mm in depth. The results suggest that germination and seedling emergence are adapted to Mediterranean coastal habitats by employing a common mechanism of light-inhibited germination and by germinating at cooler temperatures before the onset of the summer drought. Seeds were also collected from plants cultivated at a botanical garden and from plants reintroduced by sowing or by transplanting. For those populations, germination was maximal between 10–25°C, suggesting that the thermal germination behaviour may be affected by the maternal environment of seed production within one generation. It is suggested to use seeds produced in the same environment to which they will be used for the reintroduction of this species

QuantiFERON-TB Gold Plus evaluation for latent tuberculosis infection among Italian healthcare workers: a cross-sectional study

Background: healthcare workers (HCWs) are at higher risk of being exposed to tuberculosis (TB). The aim of this study was to estimate LTBI prevalence and evaluate associated risk factors among Italian HCWs using a new generation IGRA test, Quantiferon Plus (QTF-Plus), providing also an analytical evaluation of this test. Methods: this cross-sectional study was conducted during the annual health surveillance program performed in 2017, evaluating 1470 HCWs of a teaching hospital in Rome, tested with QTF-Plus. A standardized questionnaire was collected for multivariate risk analysis. The independent variables associated with a higher frequency of LTBI were tested in a multivariate logistic regression model. Results: LTBI was diagnosed in 50 out of 1470 (3.4%) of HCWs. The male gender and age>40 years were significantly associated with the risk of LTBI. The levels of interferon-γ were significantly lower in subjects with a recent conversion than in those with an older positivity. Moreover, interferon-γ values exceeding 8 IU/ml were recorded in 8 out of 35 remote subjects but in no recent subjects. The concordance between TB1 and TB2 results was 88%, while in 6 out of 50 subjects we found a discrepancy. Conclusion: the LTBI condition among Italian HCWs was very low. In subjects with discordant outcomes between TB1 and TB2, especially when the test values are close to the cut-off, is recommended a repetition of the test to confirm its positivity. Key words: tuberculosis, latent tuberculosis infection, interferon-gamma release assay, quantiferon plus, healthcare worker