Survey on viroids infecting grapevine in Italy: identification and characterization of Australian grapevine viroid and Grapevine yellow speckle viroid 2     

G. Gambino  B. Navarro  E. M. Torchetti  P. La Notte  A. Schneider  F. Mannini  F. Di Serio 《European journal of plant pathology / European Foundation for Plant Pathology》2014,140(2):199-205

Five viroid species have been reported from grapevine. Hop stunt viroid (HSVd) and Grapevine yellow speckle viroid 1 (GYSVd-1) are distributed worldwide, whereas Grapevine yellow speckle viroid 2 (GYSVd-2), Australian grapevine viroid (AGVd) and Citrus exocortis viroid (CEVd) are found only sporadically. However, the presence of AGVd and GYSVd-2 in several countries, including China, Turkey and Tunisia, suggests a wider dissemination, possibly also in Europe, where AGVd has never been found and GYSVd-2 has been occasionally identified in Italy. Taking advantage of a multiplex RT-PCR assay recently developed for detecting simultaneously these five viroids, vines growing in Italy in commercial vineyards and germplasm collections were surveyed. Besides confirming the widespread presence of HSVd and GYSVd-1 in the field, GYSVd-2 and/or AGVd were identified in two grapevine table cultivars (Sultanina Bianca and Red Globe) from germplasm collections. Tests extended to vines cultivated in southern Italy confirmed the presence of both viroids, which were further characterized. No major sequence divergences between the AGVd and GYSVd-2 variants from Italy and those previously described from other countries were observed. Phylogenetic analysis supported the close relationships among AGVd variants from Italy, Tunisia and Australia. To our knowledge this is the first report of AGVd in Europe and the first molecular characterization of GYSVd-2 isolates from a European country.  相似文献   

Evaluation of recombinant Bhlp29.7 as an ELISA antigen for detecting pig herds with swine dysentery     

La T  Phillips ND  Hampson DJ 《Veterinary microbiology》2009,133(1-2):98-104

Swine dysentery (SD) results from infection of the porcine large intestine with the anaerobic intestinal spirochaete Brachyspira hyodysenteriae. Diagnosis of SD traditionally has relied on detecting the spirochaete in the faeces of acutely affected pigs. To date simple and reliable serological assays that can be applied as a diagnostic tool at the herd level have not been available. In the current study a recombinant histidine tagged 29.7 kDa lipoprotein of B. hyodysenteriae (His6-Bhlp29.7) was used as an ELISA plate-coating antigen. Sera (n=1121) from slaughter-aged pigs on 19 farms were tested in this ELISA. Following optimization of the ELISA conditions using hyperimmune control sera, a set of 464 sera from slaughter-aged pigs from five herds where SD did not occur was tested. From these results a suitable cut-off value for herd negativity was defined as the mean optical density reading plus three standard deviations. Testing of 337 pig sera from six farms with SD then showed that the sensitivity of the test at the herd level was 100%, with all six farms having one or more serum samples exceeding the cut-off value for negativity. Finally, 320 sera from eight herds suspected of having SD were examined. Four of these herds were shown to have pigs with titres consistent with SD. The true health status of the other four herds that were serologically negative could not be confirmed. In conclusion, when used on sets of 40 sera from slaughter-aged pigs the His6-Bhlp29.7 ELISA as established proved to be a useful adjunct to the diagnosis of SD at the herd level.  相似文献   

Multilocus sequence typing as a tool for studying the molecular epidemiology and population structure of Brachyspira hyodysenteriae     

Tom La  Nyree D. Phillips  Belinda L. Harland  Phatthanaphong Wanchanthuek  Matthew I. Bellgard  David J. Hampson   《Veterinary microbiology》2009,138(3-4):330-338

The purpose of this study was to develop and apply a multilocus sequence typing (MLST) scheme to study the molecular epidemiology of Brachyspira hyodysenteriae, the aetiological agent of swine dysentery. Sequences of seven conserved genomic loci were examined in 111 B. hyodysenteriae strains. Fifty-eight of these previously had been analysed by multilocus enzyme electrophoresis (MLEE), and for some the results of pulsed field gel electrophoresis (PFGE), restriction endonuclease analysis (REA) and/or serotyping also were available. The discriminatory power of these methods was compared. The strains were divided into 67 sequence types (STs) and 46 amino acid types (AATs) by MLST. The Index of Association value was significantly different from zero, indication that the population was clonal. Eleven clonal complexes (Cc) comprising between 2 and 10 STs were recognised. A population snapshot based on AATs placed 77.5% of the isolates from 30 of the AATs into one major cluster. The founder type AAT9 included 13 strains from nine STs that were isolated in Australia, Sweden, Germany and Belgium, including one from a mallard. The MLST results were generally comparable to those produced by MLEE. The MLST system had a similar discriminatory power to PFGE, but was more discriminatory than REA, MLEE or serotyping. MLST data provided evidence for likely transmission of strains between farms, but also for the occurrence of temporal “micro-evolution” of strains on individual farms. Overall, the MLST system proved to be a useful new tool for investigating the molecular epidemiology and diversity of B. hyodysenteriae.  相似文献   

Characterisation of Escherichia fergusonii isolates from farm animals using an Escherichia coli virulence gene array and tissue culture adherence assays     

Wragg P  La Ragione RM  Best A  Reichel R  Anjum MF  Mafura M  Woodward MJ 《Research in veterinary science》2009,86(1):27-35

Escherichia fergusonii has been associated with a wide variety of intestinal and extra-intestinal infections in both humans and animals but, despite strong circumstantial evidence, the degree to which the organism is responsible for the pathologies identified remains uncertain. Thirty isolates of E. fergusonii collected between 2003 and 2004 were screened using an Escherichia coli virulence gene array to test for the presence of homologous virulence genes in E. fergusonii. The iss (increased serum survival) gene was present in 13/30 (43%) of the test strains and the prfB (P-related fimbriae regulatory) and ireA (siderophore receptor IreA) genes were also detected jointly in 3/30 (10%) strains. No known virulence genes were detected in 14/30 (47%) of strains. Following confirmatory PCR and sequence analysis, the E. fergusoniiprfB, iss and ireA genes shared a high degree of sequence similarity to their counterparts in E. coli, and a particular resemblance was noted with the E. coli strain APEC O1 pathogenicity island. In tissue culture adherence assays, nine E. fergusonii isolates associated with HEp-2 cells with a ‘localised adherence’ or ‘diffuse adherence’ phenotype, and they proved to be moderately invasive. The E. fergusonii isolates in this study possess both some phenotypic and genotypic features linked to known pathotypes of E. coli, and support existing evidence that strains of E. fergusonii may act as an opportunistic pathogens, although their specific virulence factors may need to be explored.  相似文献   

Structural development of Pacific red snapper Lutjanus peru from hatching to the onset of first feeding          下载免费PDF全文

Iram Zavala‐Leal  Silvie Dumas  Edgar O. López‐Villegas  Renato Peña  Mauricio Contreras‐Olguín  Laura Flores‐Montijo  José De La Cruz‐Agüero 《Aquaculture Research》2015,46(5):1162-1174

Successful rearing of larval fish requires culture conditions and feeding strategies matching the ontogenetic status of larvae. This study describes the external morphology and development of organs and structures involved in the feeding process (i.e. sensorial organs, mouthparts and digestive system) from hatching until first feeding in Pacific red snapper. Hatching occurred 26 h after fertilization at 26°C and total length (TL) was 2.45 ± 0.08 mm. The larvae showed an undifferentiated eye and digestive tract. At 48 hah, TL was 3.31 ± 0.12 mm. Yolk and oil globule were still present. The mouth was still closed, but the Meckel's, quadrate, hyoid and hyomandibular cartilages were present. The retina was formed by 5 layers, and a thin layer of pigment epithelium was observed in the outer nuclear layer (ONL). At 70 hah, TL was 3.44 ± 0.22 mm. A remnant of oil globule was still present. The mouth and anus were open. At 93 hah, the number of cones in the ONL of the retina have increased and there was more pigment in the pigment epithelial layer. A joint between Meckel's and the quadrate cartilage and also a joint between the hyomandibular cartilage and the skull were present. The presence of live feed was detected in the digestive tract of these larvae. Based on these observations, the Pacific red snapper larvae is functional to start ingesting live feed between the 3rd and 4th day after hatching.  相似文献   

Differential expression of molecular rust resistance components have distinctive profiles in <Emphasis Type="Italic">Coffea arabica</Emphasis> - <Emphasis Type="Italic">Hemileia vastatrix</Emphasis> interactions     

Geleta Dugassa Barka  Eveline Teixeira Caixeta  Robson Ferreira de Almeida  Samuel Mazzinghy Alvarenga  Laércio Zambolim 《European journal of plant pathology / European Foundation for Plant Pathology》2017,149(3):543-561

Countering the economic hurdle caused by coffee leaf rust disease is most appealing at this time as it has posed a major threat to coffee production around the world. Establishing differential expression profiles at different times following pathogen invasion in both innate and acquired immunities unlocks the molecular components of resistance and susceptibility. Suppression subtractive hybridization (SSH) was used to identify genes differentially over-expressed and repressed during incompatible and compatible interactions between Coffea arabica and Hemileia vastatrix. From 433 clones of expressed sequence tags (ESTs) sequenced, 352 were annotated and categorized of which the proportion of genes expressed during compatible interaction were relatively smaller. The result showed upregulation and downregulation of various genes at 12 and 24 h after pathogen inoculation in both interactions. The use of four different databases in searching for gene homology resulted in different number of similar sequences. BLASTx against EMBL-EBI (European Molecular Biology Laboratory-European Bioinformatics Institute) database being with the maximum (100%) hits for all the annotated sequences. RT-qPCR analysis of seven resistance-signaling genes showed similar expression patterns for most of the genes in both interactions, indicating these genes are involved in basal (non-specific) defense during which immune reactions are similar. Using SSH, we identified different types of resistance related genes that could be used for further studies towards resistant cultivar development. The potential role of some of the resistance related proteins found were also discussed.  相似文献   

Glaucoma with Descemet's membrane detachment in five horses          下载免费PDF全文

Michala de Linde Henriksen  Noelle La Croix  David A. Wilkie  Mary Lassaline‐Utter  Karen R. Brantman  Gillian L. Beamer  Leandro B. C. Teixeira  Richard R. Dubielzig 《Veterinary ophthalmology》2017,20(3):273-279

  相似文献   

Patient characteristics influencing the variability of distributed parameter‐based models in DCE‐CT kinetic analysis          下载免费PDF全文

M. D. La Fontaine  L. S. McDaniel  L. N. Kubicek  R. J. Chappell  L. J. Forrest  R. Jeraj 《Veterinary and comparative oncology》2017,15(1):105-117

Kinetic parameter variability may be sensitive to kinetic model choice, kinetic model implementation or patient‐specific effects. The purpose of this study was to assess their impact on the variability of dynamic contrast‐enhanced computed tomography (DCE‐CT) kinetic parameters. A total of 11 canine patients with sinonasal tumours received high signal‐to‐noise ratio, test‐double retest DCE‐CT scans. The variability for three distributed parameter (DP)‐based models was assessed by analysis of variance. Mixed‐effects modelling evaluated patient‐specific effects. Inter‐model variability (CV_inter) was comparable to or lower than intra‐model variability (CV_intra) for blood flow (CV_inter:[4–28%], CV_intra:[28–31%]), fractional vascular volume (CV_inter:[3–17%], CV_intra:[16–19%]) and permeability‐surface area product (CV_inter:[5–12%], CV_intra:[14–15%]). The kinetic models were significantly (P<0.05) impacted by patient characteristics for patient size, area underneath the curve of the artery and of the tumour. In conclusion, DP‐based models demonstrated good agreement with similar differences between models and scans. However, high variability in the kinetic parameters and their sensitivity to patient size may limit certain quantitative applications.  相似文献   

Inheritance of coffee leaf rust resistance and identification of AFLP markers linked to the resistance gene     

Giovani Greigh de Brito  Eveline Teixeira Caixeta  Ana Paula Gallina  Eunize Maciel Zambolim  Laércio Zambolim  Valdir Diola  Marcelo Elhers Loureiro 《Euphytica》2010,173(2):255-264

The most important disease of Coffea arabica is coffee leaf rust caused by the fungus Hemileia vastatrix. The purpose of this study was to characterize the inheritance of coffee resistance gene(s) to race II of this pathogen and to identify and map molecular markers linked to this trait. Different populations were used: F₂ (160 plants), BCr (20), and BCs (135), derived from a cross between the resistant genotype Híbrido de Timor UFV 427-15 and the susceptible cultivar Catuaí Amarelo UFV 2143-236 (IAC 30). The segregation analysis showed that the resistance of Híbrido de Timor to race II of the H. vastatrix is conferred by a single dominant gene. The amplification of 176 AFLP (Amplified fragment length polymorphism) primer combinations using bulked segregant analysis (BSA) allowed the identification of three molecular markers linked to the resistance gene. Genetic mapping of these three markers in the F₂ population indicated that they are distributed on both sides, flanking the resistance gene. The markers E.CTC/M.TTT405 and E.CGT/M.TGT300 were found linked to the resistance gene at 8.69 cM (LOD 18.91) and 25.10 cM (LOD 5.37), respectively, while E.CCT/M.TTC230 was localized on the other side of the gene, at 20.50 cM (LOD 6.15). These markers are the first rust resistance markers identified in Híbrido de Timor and can be useful for marker assisted selection in coffee breeding programs.  相似文献   

Carbon dioxide fluxes across the Sierra de Guadarrama, Spain     

R. Inclán  C. Uribe  D. De La Torre  D. M. Sánchez  M. A. Clavero  A. M. Fernández  R. Morante  A. Cardeña  M. Fernández  A. Rubio 《European Journal of Forest Research》2010,129(1):93-100

Understanding the spatial and temporal variation in soil respiration within small geographic areas is essential to accurately assess the carbon budget on a global scale. In this study, we investigated the factors controlling soil respiration in an altitudinal gradient in a southern Mediterranean mixed pine–oak forest ecosystem in the north face of the Sierra de Guadarrama in Spain. Soil respiration was measured in five Pinus sylvestris L. plots over a period of 1 year by means of a closed dynamic system (LI-COR 6400). Soil temperature and water content were measured at the same time as soil respiration. Other soil physico-chemical and microbiological properties were measured during the study. Measured soil respiration ranged from 6.8 to 1.4 μmol m^?2 s^?1, showing the highest values at plots situated at higher elevation. Q ₁₀ values ranged between 1.30 and 2.04, while R ₁₀ values ranged between 2.0 and 3.6. The results indicate that the seasonal variation of soil respiration was mainly controlled by soil temperature and moisture. Among sites, soil carbon and nitrogen stocks regulate soil respiration in addition to soil temperature and moisture. Our results suggest that application of standard models to estimate soil respiration for small geographic areas may not be adequate unless other factors are considered in addition to soil temperature.  相似文献