Artificial Germination Activation of Dialium corbisieri by Imitation of Ecological Process

Species of the gender Dialium commonly are trees found in Central African rainforests. They produce tasty sugary fruits, feeding numerous frugivores, but are, despite their valuable nutritional value, rarely exploited by humans. A potential reason for this could be the complexity of symbiotic dependence between trees and pollinators, germination activators, and dispersers causing problems in ancestral and contemporary domestication. We investigated Dialium corbisieri reproduction in the Democratic Republic of the Congo, Bandundu Province. Here we give a key for an artificial activation of germination of these trees ecologically adapted to the digestive system of their ape dispersers: By perforation of the impermeable seed coat protection, water assimilation and subsequent activation of germination becomes possible. By this nicking, pretreatment germination increases from 0 to 96%, representing an inexpensive and simple treatment to be used under natural conditions and in developing countries. The use of this mechanical activation for forest management, conservation, and economic use is discussed.     

Ophiostomatoid fungi associated with three spruce-infesting bark beetles in Slovenia

Context

Ophiostomatoid fungi can severely affect the health and economic value of Norway spruce trees (Picea abies). Although the diversity of ophiostomatoid species and their associations with insects have been well-investigated in central and northern Europe, little is known about the conditions in south-eastern Europe.

Aim

This study aims to study the assemblages of ophiostomatoid fungi associated with three bark beetle species (Ips typographus, Ips amitinus, and Pityogenes chalcographus) that infect Norway spruce in Slovenia.

Methods

Bark beetles were sampled in four phytogeographic regions in Slovenia. The fungi found on the bark beetles were identified based on morphology, DNA sequence comparisons of ITS regions and phylogenetic analysis. The species compositions of the fungal associates of the three insect species were compared and the pairwise associations of the occurrence of the fungal species were analysed.

Results

Thirteen different species were found. The most commonly encountered fungal associates of the beetles were Ophiostoma bicolor, Ophiostoma brunneo-ciliatum, Grosmannia piceiperda, Ophiostoma ainoae, Ceratocystiopsis minuta, and Grosmannia penicillata. The composition of the fungal associates differed among the bark beetle species, but not among the phytogeographic regions.

Conclusions

This study confirms that ophiostomatoid species are common associates of the investigated bark beetle species. Many ophiostomatoid species have strong host associations. I. typographus and P. chalcographus can act as effective vectors for O. bicolor, O. ainoae, G. piceiperda and O. brunneo-ciliatum, whereas I. amitinus often carries G. piceiperda and C. minuta in Slovenian forests.     

Treatment-related toxicities in tumor-bearing cats treated with temozolomide alone or in combination with doxorubicin: a pilot assessment

A retrospective study assessing treatment-related toxicities in tumor-bearing cats treated with temozolomide (TMZ) alone or in combination with doxorubicin was conducted. TMZ was administered orally once a day for 5 days every 3 weeks at a dose of 20 mg/cat. Tumor response was evaluated with standard World Health Organization criteria and toxicity was monitored using veterinary co-operative oncology group-common terminology criteria for adverse events (VCOG--CTCAE) criteria. Ten tumor-bearing cats with various types of malignancies were treated with TMZ-based chemotherapy. Eight cats were evaluable for response. Two cats achieved a complete response, one achieved stable disease and five achieved a partial response. Four grade III and one grade IV hematological toxicities, and one grade IV gastrointestinal toxicity were observed. Four cats were euthanased as a result of apparent toxicity. One cat was euthanased as a result of severe and prolonged myelosuppression with fever. Three were euthanased for grade III pleural and pericardial effusions. Effusion was seen in cats treated with higher cumulative dose of TMZ (P = 0.0046). Planned additional case accrual was discontinued because of unacceptable levels of toxicity despite evidence of efficacy in some of the cats. Additional investigation is needed to elucidate this unexpected apparent cumulative toxicity.     

Cellular-resolution in vivo imaging of the feline retina using adaptive optics: preliminary results

Purpose To perform cellular‐level in vivo imaging of the feline retina using an adaptive optics flood illumination fundus camera (AO FIFC) designed for the human eye. Materials and methods Cellular‐level images were obtained from three eyes of two normal sedated cats. Ocular aberrations were corrected using an AO system based on a 52‐acuator electromagnetic deformable mirror and a 1024 lenslet Hartmann–Shack sensor (both Imagine Eyes, Orsay, France). A square 3°×3° area of the ocular fundus was flood‐illuminated by a pulsed LED emitting at 850 nm and imaged onto a low‐noise, high‐resolution CCD camera. The animal’s pupils were dilated and the effective pupil size was set to 7.5 mm. Conjunctival atraumatic clips were used to avoid eyeball movements and eyelid closure. The cornea was artificially hydrated throughout the experiments. Each acquisition consisted of 20 consecutive images, out of which 10 were numerically averaged to produce an enhanced final image. Results The total amount of ocular aberrations was greatly reduced by the AO correction, from 2.4 to 0.21 microns root mean square on average. The resulting images presented white dots distributed at a density similar to that of cone photoreceptors and they allowed us to visualize small blood vessels and nerve fiber bundles at a higher resolution than classically obtained with conventional fundus photography. Conclusion Retinal imaging with cellular resolution was feasible in cats under sedation using an AO FIFC designed for human eyes without any optical modification. The AO FIFC technology could find new applications in clinical, pharmacological, and toxicological investigations.     

Could fetal fluid and membranes be an alternative source for Mesenchymal Stem Cells (MSCs) in the feline species? A preliminary study

Domestic cats are preferred models for normal physiology and several human diseases. In the present study feline fetal fluids and membranes were evaluated as possible sources of MSCs. Samples were recovered from 4 pregnant queens after ovarian-hysterectomy. Gestational sacs were separated from uterine wall; after allantoic and amniotic fluids aspiration and chorion-allantois and amniotic membranes separation, all cell lineages were cultured into 25 cm(2) flasks, in DMEM/TCM199, in a 5% CO(2) incubator at 38.5 °C. At passage 3, chondrogenic, osteogenic and adipogenic differentiation ability were evaluated by culturing cell monolayers in differentiating media for 21 days. Cellular characterization with CD90, CD44, CD105, CD73, CD34, CD14, CD45, was performed by flow cytometry. In all samples, adherent fibroblastoid spindle-shaped cells were observed. Positive von Kossa and Alizarin Red staining confirmed osteogenesis. Alcian blue staining of matrix glycosaminoglycans illustrated chondrogenesis, and positive Oil Red O lipid droplets within cell cytoplasm suggested adipogenesis. All cell lines isolated were positive for CD90, CD44, CD105 and negative for CD34, CD14 and CD45; as unexpected and different from human cells, feline cells resulted negative for CD73. Based on this preliminary results, fetal fluids and membranes could represent an alternative sources for mesenchymal stem cells in feline species.     

Methicillin-resistant Staphylococcus aureus (MRSA) isolated from small and exotic animals at a university hospital during routine microbiological examinations

Clinical specimens of small animals (n=869) were screened for the occurrence of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA; MRSA) during routine microbiological examinations, and results were confirmed by a multiplex PCR strategy. The genetic relatedness of all mecA-positive S. aureus isolates was further investigated by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), PCR for Panton-Valentine leukocidine genes (PVL) and staphylococcal cassette chromosome mec-typing (SCCmec). A total of 61 S. aureus isolates were found during a 20-month period of investigation, 27 (44.3%) of them harbouring the mecA gene for methicillin-resistance. The majority of MRSA were isolated in specimens from dogs (n=18) and cats (n=4). One guinea pig and one rabbit were found to be positive for an MRSA infected site. Similarly, three exotic animals, a turtle, a bat and a parrot, were found to be infected with MRSA. PFGE and MLST analysis revealed a certain genotype ("A" and "A-1") dominating the isolate collection (23 of 27). Furthermore, one isolate showed homologous PFGE pattern to the German epidemic strain Barnim ("BE") and another one ("BE-1") was considered to be closely related. A third genotype ("B") was detected in two cases. Two different sequence types (ST) were identified among the 27 MRSA isolates. PFGE type "A" and both strains related to the Barnim epidemic strain were assigned to ST22, whereas ST239 was associated to PFGE profile "B". The present data show that certain MRSA genotypes are capable of infecting a wide spectrum of small and exotic animals, especially in clinical facilities.     

Differentiation and structure of an Italian landrace of celery (Apium graveolens L.): inferences for on farm conservation

The “Sedano Nero di Trevi” (Black Celery from Trevi) is a landrace grown in Umbria (Italy) at risk of extinction. A morpho-physiological and molecular characterisation of the landrace and a detailed investigation of its cultural and cultivation context were carried out with the aim of facilitating a request for a quality mark, implement in situ (on farm) conservation schemes that are being developed in Italy and register the landrace in the European common catalogue of ‘conservation varieties’. These actions can help the landrace survive. Six farmer populations of the landrace and four élite varieties were characterised for thirteen morpho-physiological traits and nine AFLP primer combinations. The “Sedano Nero” farmer populations were better performers than élite varieties for important agronomic traits, such as yield, under the local agronomic conditions. Both morpho-physiological traits and molecular markers clearly distinguished the landrace from the élite cultivars. In addition, a structure was detected in the landrace with farmer populations being characterised by specific alleles. These results are discussed in connection with the local seed system and possible in situ conservation strategies for landraces.     

Effects of sensor spatial resolution on landscape structure parameters

We examined the effects of increasing grain size from 20 m to 1100 m on landscape parameters characterizing spatial structure in the northern Wisconsin lake district. We examined whether structural parameters remain relatively constant over this range and whether aggregation algorithms permit extrapolation within this range. Images from three different satellite sensors were employed in this study: (1) the SPOT multispectral high resolution visible (HRV), (2) the Landsat Thematic Mapper (TM), and (3) the NOAA Advanced Very High Resolution Radiometer (AVHRR). Each scene was classified as patches of water in a matrix of land. Spatial structure was quantified using several landscape parameters: percent water, number of lakes (patches), average lake area and perimeter, fractal dimension, and three measures of texture (homogeneity, contrast, and entropy). Results indicate that most measures were sensitive to changes in grain size. As grain size increased from 20 m using HRV image data to 1100 m (AVHRR), the percent water and the number of lakes decreased while the average lake area, perimeter, the fractal dimension, and contrast increased. The other two texture measures were relatively invariant with grain size. Although examination of texture at various angles of adjacency was performed to investigate features which vary systematically with angle, the angle did not have an important effect on the texture parameter values. An aggregation algorithm was used to simulate additional grain sizes. Grain was increased successively by a factor of two from 20 m (the HRV image) to 1280 m. We then calculated landscape parameter values at each grain size. Extrapolated values closely approximated the actual sensor values. Because the grain size has an important effect on most landscape parameters, the choice of satellite sensor must be appropriate for the research question asked. Interpolation between the grain sizes of different satellite sensors is possible with an approach involving aggregation of pixels.     

A GIS-based index for relating landscape characteristics to potential nitrogen leaching to wetlands

We developed a spatially-explicit, quantitative Nitrogen Leaching Index to assess the potential for non-point source subsurface nitrogen pollution to wetlands. The index was based on the leaching potential of the watershed soils, the amount of nitrogen available for leaching, and the spatial position of nitrogen sources in the watershed. A raster or cell-based geographic information system (GIS) was used to estimate the necessary data inputs for calculating the index, such as soil hydrologic group, land use/soil type combination, groundwater residence time, and location of septic systems. The Total and Average Watershed Nitrogen Leaching Index (TWNLI and AWNLI) were calculated by summing and averaging, respectively, individual cell contributions over a watershed.Analysis of nine wetland watersheds in central New York state, USA, with mixed forest and agricultural land uses illustrated the use of the index for identifying and ranking wetlands with potential nitrogen pollution. Results showed that the spatial characteristics of a watershed potentially can effect subsurface nitrogen delivery to groundwater-dominated wetlands. The use of an index based on watershed soils, topography, and land use may be useful for assessing potential nitrogen pollution to wetlands at a regional scale.     

The Development of Adventitious Roots on Hardwood Cuttings of Gooseberries

Changes in percentage dry weight and in carbohydrate levels and distribution were followed in Golden Brown Lockyer onion bulbs for three months at storage temperatures of 4°, 15°, 25° and 37°C. The percentage dry weight within the bulbs increased from the outer to the inner leaf bases. Storage temperature and length of storage did not influence water loss. Sucrose concentrations increased from the outer leaf bases to become highest in the inner leaf bases, and in all leaf bases were higher at higher temperatures. Glucose and fructose levels also tended to increase from outer to inner leaf bases. After eight weeks fructose levels increased rapidly at the lower storage temperatures and this is attributed to low-temperature hydrolysis of fructans. The only fructan detected was the trisaccharide fraction, which was barely detectable in outer leaf bases and maximal in the inner ones. Trisaccharide levels were generally lower at the lower temperatures, thus supporting the suggestion of low-temperature hydrolysis of fructans. No information was obtained regarding storage temperature effects on fructans having a higher degree of polymerization than the trisaccharide. The results are discussed relative to temperature and time and the development of storage rots.