Detection and characterization of phytoplasmas in diseased stone fruits and pear by PCR-RFLP analysis in Turkey

During the late summer-early autumn of 2002, surveys were carried out in Turkey to determine the presence of phytoplasma diseases in fruit trees. Phytoplasmas were detected and characterized by PCR-RFLP analysis and TEM technique in stone fruit and pear trees in the eastern Mediterranean region of the country. Six out of 24 samples, including almond, apricot, peach, pear and plum, gave positive results in PCR assays. RFLP analysis usingSspI andBsaAI enzymes of PCR products obtained with primer pair f01/r01 enabled identification of the phytoplasmas involved in the diseases. Stone fruit trees, including a local apricot variety (‘Sakıt’) and a pear sample, were found to be infected with European stone fruit yellows (ESFY, 16SrX-B) and pear decline (PD, 16SrX-C) phytoplasmas, respectively. This is the first report in Turkey of PD phytoplasma infecting pear and of ESFY phytoplasma infecting almond, apricot, myrobalan plum and peach; ESFY phytoplasma infecting Japanese plum was previously reported. http://www.phytoparasitica.org posting July 21, 2005.     

Validation of a human immunoturbidimetric assay to measure canine albumin in urine and cerebrospinal fluid.

The aim of this study was to validate an automated immunoturbidimetric assay used to quantify human albumin in urine and to accurately measure canine albumin concentrations in both urine and cerebrospinal fluid. The partial homology existing between human and canine albumin limited the accuracy of the human assays in measuring canine albumin without method modifications. Thus, the assay was modified by calibrating the analyzer with calibrators made in the laboratory containing known concentrations of canine albumin. To prepare the set of calibrators, the albumin concentration of pooled sera of healthy dogs was assessed in 5 replicates using the BromocresolGreen assay. Pooled samples were aliquoted and serially diluted to obtain the expected concentrations of albumin (0.5, 1, 5, 13, and 30 mg/dl) for establishing the canine calibration curve. Thereafter, the performance was assessed by analyzing canine urine and CSF The modified assay accurately quantified canine albumin in both specimens, as indicated by the following. Intra- and interassay variability was 0.92% and 2.74%, respectively; recovery was 99.66% and 99.07% in urine and 105.02% in CSF No interference was detected when hemolysate and glucose were added to urine. The test was linear within the verified range (0-225 mg/dl). These results demonstrate that the modified human albumin immunoturbidimetric assay can be a useful tool in the veterinary diagnostic laboratory. It is accurate and tends itself to automatization on chemistry analyzers.     

Use of computer tomography for imaging of Crassicauda grampicola in a Risso's dolphin (Grampus griseus).

A mature male Risso's dolphin (Grampus griseus) stranded along the coasts of Friuli Venezia Giulia, northeast Italy, in May 2001. Parasitic infection with Crassicauda grampicola is often found in the tympanic bullae and pterygoid sinuses in many of the Risso's dolphins examined from the same area. For this reason, it was decided to perform computed tomography of the head to assess this imaging technique for the diagnosis of crassicaudosis in dolphins. A full postmortem examination confirmed the pathologic findings of the computed tomography scan. This technique can be considered a useful adjunct in the diagnosis of cranial crassicaudosis in live dolphins.     

Molecular Identification of a New Member of the Clover Proliferation Phytoplasma Group (16SrVI) Associated with Centaurea Solstitialis Virescence in Italy

In the United States, yellow starthistle (Centaurea solstitialis) is an annual invasive weed with Mediterranean origins. Malformed plants displaying witches' broom, fasciations, abortion of buds and flower virescence symptoms were observed in central Italy. Attempts to transmit the causal agent from the natural yellow starthistle host to periwinkle by grafting, resulted in typical symptoms of a phytoplasma, i.e. yellowing and shortening of internodes. The detection of phytoplasmas was obtained from both symptomatic yellow starthistle and periwinkle by the specific amplification of their 16S-23S rRNA genes. PCR amplification of extracted DNA from symptomatic plant samples gave a product of expected size. Asymptomatic plants did not give positive results. An amplicon obtained by direct PCR with universal primers P1/P7 was cloned and sequenced. The homology search using CLUSTALW program showed more than 99% similarity with Illinois elm yellows (ILEY) phytoplasma from Illinois (United States) and 97% with Brinjal little leaf (BLL) phytoplasma from India. Digestion of the nested-PCR products with restriction enzymes led to restriction fragment length polymorphism patterns referable to those described for phytoplasmas belonging to the clover proliferation (16S-VI) group. Since this is a previously undescribed disease, the name Centaurea solstitialis virescence has been tentatively assigned to it. This is a new phytoplasma with closest relationships to ILEY and BLL, but distinguishable from them on the basis of 16S rDNA homology, the different associated plant hosts and their geographical origin.     

Salinity of nutrient solution influences the shelf-life of fresh-cut lettuce grown in floating system

Quality, microbiological and enzymatic characteristics of fresh-cut lettuce (Lactuca sativa var. longifolia, ‘Duende’), grown in floating system with three electrical conductivities of nutrient solutions (2.8, 3.8 and 4.8 mS cm^?1), were investigated in order to evaluate the effect of salinity on product shelf-life during cold storage (9 d at 4 °C). Pre-harvest salinity of 3.8 and 4.8 mS cm^?1 improved the properties of fresh-cut lettuce, since CO₂ production was reduced with a subsequent control of the decay process. Fresh-cut processing caused an activation of polyphenol oxidase and peroxidase; in all cases the product obtained by salinity treatments was less subject to oxidase activity and browning phenomena during storage. Increased salinity reduced the number of mesophilic bacteria and of moulds and yeasts, assessed by plate counts on different culture media; in contrast, Enterobacteriaceae levels were unaffected by pre-harvest treatments. The research demonstrated that an increase in nutrient solution electrical conductivity, through the use of floating system, affects fresh-cut lettuce characteristics, improving shelf-life of the product.     

Productive and biochemical responses of durum wheat to UV filtration

A UV exclusion experiment was conducted on durum wheat (Triticum durum Desf. var. Claudio) grown in pots. Plants were grown under three different radiative treatments in greenhouses covered with plastic filters: Teflon, transparent to the entire region of natural UV‐visible sunlight (TEF); polyester, transparent above 312 nm (MYL, excluding UVB) and Lee, transparent above 400 nm (LEE, excluding both UVA and UVB). Analyses have been carried out to determine the concentration of photosynthetic pigments in leaves, UV‐absorbing compounds, nitrogen and carbon in leaves, culms and spikes and proteins and gluten in grains. In particular, plants grown under UV exclusion showed a reduction in protein and dry gluten content (consequently without variation in the ratio dry gluten/protein), but at the same time, a significant increase of gluten index, which is a parameter to define the quality of gluten, was observed. The results highlighted the influence played by UV radiation on some biochemical parameters, mainly UV‐absorbing compounds, leaf nitrogen and grain protein characteristics of durum wheat cultivated under Mediterranean conditions. In particular, natural level of UV in the Mediterranean improves the characteristics of durum wheat flour important for pasta production (high dry gluten level), while the UV exclusion could improve characteristics of flour important for bread production (high gluten index).     

Long-term deep sedation using Zoletil and haloperidol for the treatment of streptococcal pneumonia in an orangutan (Pongo pygmaeus)

A 13-year-old Bornean orangutan diagnosed with life threatening Streptococcus pyogenes broncho-pneumonia was kept in a state of deep sedation for 20 days via continuous intra-venous (IV) infusion of zolazepam -tiletamine and IV haloperidol to allow consistent IV administration of ceftazidime and gatifloxacine. The use of long-term deep sedation allowed carrying out a particularly demanding treatment not generally associated with zoological patients. The treatment was ultimately successful.