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.     

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).     

The effect of erector spinae plane block on perioperative analgesic consumption and complications in dogs undergoing hemilaminectomy surgery: a retrospective cohort study

ObjectiveTo compare the perioperative use of analgesics and complication rates in dogs administered an erector spinae plane (ESP) block or a traditional opioid-based (OP) treatment as part of analgesic management during hemilaminectomy.Study designRetrospective cohort study.AnimalsMedical records of 114 client-owned dogs.MethodsGeneral data included demographics, duration of procedure, number of laminae fenestrated, perioperative use of steroid and non-steroidal anti-inflammatory drugs. Intra- and postoperative analgesics used in 48 hours and complications rates were compared between groups. Opioid use was expressed in morphine equivalents [ME (mg kg^?1)]. Continuous data were compared using the Mann–Whitney U test and incidence of events with a Fisher's exact tests. Multiple linear regression was used to evaluate association between perioperative ME consumption (dependent variable) with other independent variables. Data are presented as median (range). Differences were considered significant when p < 0.05.ResultsGroup ESP comprised 42 dogs and group OP 72 dogs. No differences were observed in the general data. Intraoperative ME was 0.65 (0.20–3.74) and 0.79 (0.19–5.60) mg kg^?1 in groups ESP and OP, respectively (p = 0.03). Intraoperative infusion of lidocaine was administered intravenously (IV) to 23.8% and 68% of groups ESP and OP, respectively (p < 0.0001). Intraoperative infusion of ketamine was administered IV to 21% and 40% of groups ESP and OP, respectively (p = 0.04). Regression analysis revealed the ESP block as the only independent variable affecting the perioperative ME consumption. Pharmacological intervention to treat cardiovascular complications was administered to 21.4% and 47.2% of dogs in groups ESP and OP, respectively (p = 0.008). There were no differences in postoperative complication rates.Conclusions and clinical relevanceESP block was associated with reduced perioperative opioid consumption, intraoperative adjuvant analgesic use and incidence of pharmacological interventions to treat cardiovascular complications in dogs undergoing hemilaminectomy.     

FEASIBILITY FOR USING HYPOFRACTIONATED STEREOTACTIC VOLUMETRIC MODULATED ARC RADIOTHERAPY (VMAT) WITH ADAPTIVE PLANNING FOR TREATMENT OF THYMOMA IN RABBITS: 15 CASES

Thymoma is a relatively common tumor in rabbits. Treatment with surgery, radiation therapy, and chemotherapy alone or in combination has been reported with varying outcomes. Stereotactic volumetric modulated arc radiotherapy delivered in a hypofractionated manner allows high doses of radiation to be delivered to the target volume while allowing sparing of adjacent critical structures. This therapy is ideally suited for thymomas in rabbits given their size, the difficulty of multiple anesthesia episodes and the complexity of the radiotherapy plans required due to the tumor's proximity to the heart, lungs, and mediastinal structures. Fifteen rabbits with thymoma were prospectively recruited for this observational, single institution, single arm clinical study. All rabbits were imaged with both computed tomography (CT) and magnetic resonance imaging (MRI). A total dose of 40 Gy in six fractions was delivered using a single arc over an 11‐day period with repeat CT simulation done every other fraction for adaptive planning. Follow‐up evaluation was done through repeat CT and MRI imaging and revealed complete responses using the Response Evaluation Criteria in Solid Tumors (RECIST) criteria. Two rabbits had died at 618 and 718 days, 10 were alive and three were lost to follow‐up. Observed acute and late effects were graded according to the Veterinary Radiation Therapy Oncology Group (VRTOG) criteria and were found to be minimal.     

Use of touch-down polymerase chain reaction to enhance the sensitivity of Mycobacterium bovis detection.

The confirmatory diagnosis of Mycobacterium bovis (M. bovis) in animal samples is carried out by culture in Stonebrink media. However, culture is very slow because of the extremely long duplication time of the bacillus and difficult because of the scarcity of bacilli in diagnostic samples. This study describes the development of a single-tube touch-down polymerase chain reaction (PCR) protocol for the detection of M. bovis using primers that target the IS6110 element. Spiked water and milk as well as routine diagnostic samples (milk and nasal swabs) from M. bovis-positive cattle were tested. This protocol allows the rapid and sensitive detection of M. bovis in bovine samples by enhancing the sensitivity of standard PCR amplification.     

Follicular Dynamics in Heifers during Pre-pubertal and Pubertal Period Kept under Two Levels of Dietary Energy Intake

The objective of this study was to characterize follicular dynamics in pre-pubertal, pubertal and post-pubertal periods, as well as the effect of high-energy intake on follicular development and age at puberty in heifers. Thirty-one Nelore (Bos indicus) heifers, 6 months old, were randomly assigned to receive two different diets: one of low (GI) and other of high dietary energy intake (GII). Animals were evaluated in relation to body weight gain by being weighed every 21 days. Heifers were evaluated every other day by real-time linear ultrasonography to characterize ovarian structures development from weaning to post-pubertal period. Blood samples were collected to determine plasmatic concentrations of progesterone by RIA method. The ovulation was determined when progesterone concentrations were >1 ng/mL in three consecutive samples, and by ultrasound images of corpus luteum; and oestrous behaviour in some animals. Age at puberty differed among heifers of GII (17.00 +/- 0.46 months) compared with heifers of GI (19.87 +/- 0.47 months; p < or = 0.05). Maximum size of the dominant follicles at pre-pubertal period was greater in GII heifers than in GI (10.52 +/- 0.33 and 9.76 +/- 0.15 mm, respectively; p < or = 0.05). As heifers approached first ovulation time, size of dominant follicle increased (11.75 +/- 0.37 mm for GI and 12.52 +/- 0.91 mm for GII; p < or = 0.05). Body weight at puberty was not different in both groups (302.33 +/- 27.31 kg for GI and 326.19 +/- 27.78 kg for GII heifers; p > 0.05). We conclude that animals receiving high dietary energy intake attained the puberty earlier and the development of follicles were different than in low dietary energy intake.     

A new hemolytic assay for bovine serum complement and its application during experimental bovine anaplasmosis

A new hemolytic assay for bovine complement is presented. Using this assay we found a significant reduction in bovine serum complement activity during the acute phase of anaplasmosis, and an increase in the sensitivity of the red blood cells (RBC) to bovine complement lysis in vitro. The new hemolytic test is performed with bovine RBC, rabbit anti-bovine RBC serum and bovine serum complement. An isotonic sucrose Tris-buffered saline solution of ionic strength 0.094 and pH 7.2 was found to be adequate for this test. The titres obtained with this new assay, which uses autologous RBC, are comparable with those obtained using the guinea pig RBC assay. The finding of a reduction in bovine serum complement during anaplasmosis may be suggestive of a mechanism responsible for the pathology of this disease.     

Phenological investigations of different winter-deciduous species growing under Mediterranean conditions

Phenological stages are the result of biorhythms and environmental factors, these last are probably the same ones that caused, during evolution, adjustments of the species to different climate. The present study was carried out in a Phenological Garden located in central Italy (Perugia, Umbria Region) which contains indicator species, common to all International Phenological Gardens. The aim of this study was to determine and analyse the average trends of development of eight plant species and their phenological adjustment to the Mediterranean environment, over a nine-year period (1997–2005). The results of the statistical analyses show a strong relationship between the temperature trends and vegetative seasonal evolutions interpreted by phenological data for all the species considered. Moreover, it was demonstrated that the plants studied may approach or close completely the timing gaps eventually created during the first phenological phases, adjusting thus the beginning of subsequent phenophases.     

Histopathological effects of cadmium on the gills of the freshwater fish, Macropsobrycon uruguayanae Eigenmann (Pisces, Atherinidae)

Evaluations of histopathological lesions in gill tissue were carried out in the freshwater fish Macropsobrycon uruguayanae following 30 and 60 days of exposure to 1.5 mg 1^-1 of cadmium. The study was conducted on both fed and starved animals in order to determine the influence of feeding condition on cadmium toxicity. The main lesions observed and quantified were: (1) hyperplasia of primary lamellar epithelium; (2) hyperplasia of secondary lamellar epithelium; (3) separation of respiratory epithelium; (4) shortening of secondary lamellae; (5) epithelial necrosis; (6) fusion of adjacent secondary lamellae; (7) hypertrophy of respiratory epithelium; (8) lamellar telangiectasis; (9) hyperplasia of chloride cells; (10) mucinous metaplasia; and (11) inflammatory infiltration. Lesions 6, 8, 9 and 11 were only induced by exposure to cadmium, while lesion 4 could be produced only by starvation. Starved fish also showed a reduction in total body weight and length. Lesion 2 was shown to be non-specific, and produced by either cadmium, starvation or even exposure time. The possible mode of action of the experimental factors are discussed in relation to the observed pathologies.