EFFECT OF DEEP DIGITAL FLEXOR TENDON ORIENTATION ON MAGNETIC RESONANCE IMAGING SIGNAL INTENSITY IN ISOLATED EQUINE LIMBS—THE MAGIC ANGLE EFFECT

Ten normal equine isolated limbs were imaged using a knee coil in a 1.5 Tesla magnetic field, with short echo time sequences (TE < 15 ms). Magnetic resonance imaging was performed on each isolated limb in different positions, with and without extension of the metacarpophalangeal joint. Deep digital flexor tendon orientation ranged from 20 to 60 degrees in relation to the static magnetic field. Increased intratendinous signal intensity was observed when the angle between the deep digital flexor tendon and the constant magnetic field approached 55 degrees ("magic angle"). The increased signal intensity was independent from extension of the metacarpophalangeal joint. Recognition of the magic angle phenomenon is essential for proper evaluation of magnetic resonance imaging studies of the equine foot.     

Fate of feed plant DNA monitored in water buffalo (Bubalus bubalis) and rabbit (Oryctolagus cuniculus)

The effect of the digestion process in the gastro-intestinal tract (GIT) of animal models on the fate and integrity of plant DNA has been widely evaluated since DNA availability and integrity is a key factor for hypothetical horizontal gene transfer of recombinant DNA from GM crop-derived feeds to animal and human gut microflora. In this study, plant DNA sequences from high and low copy number genes were monitored in GIT and tissues of buffaloes and rabbits. Using a real-time PCR approach to track plant DNA in animal samples, we demonstrated the persistence of fragmented plant DNA blood and tissues of buffaloes and rabbits raised with conventional feeding.     

Characterization of the cytochrome b gene fragment of Puccinia species responsible for the binding site of QoI fungicides

The fragment of the cytochrome b (cyt b) gene responsible for the binding site of QoI fungicides was sequenced for different Puccinia species by using DNA and RNA as template for PCR and RT-PCR, respectively. Degenerated primers for the cyt b gene amplified in P. recondita f.sp. tritici a 450 bp fragment, which was cloned and sequenced. At cDNA level, several Thermal Asymmetric InterLaced (TAIL)-PCR cycles were needed to produce a 996 bp long fragment, which corresponded to almost the whole cyt b gene (about 1160-1180 bp, without introns). This fragment was sequenced and specific primers were designed. Amplification with cyt b specific primers using genomic DNA as template revealed the presence of an intron of about 1500 bp length after the codon for glycine at amino acid position 143. By using the same primer pair, the cyt b gene fragment was amplified and sequenced both at cDNA and genomic DNA level also for other rust species, including P. graminis f.sp. tritici (length: 506 bp), P. striiformis f.sp. tritici (755 bp), P. coronata f.sp. avenae (644 bp), P. hordei (660 bp), P. recondita f.sp. secalis (687 bp), P. sorghi (709 bp), and P. horiana (478 bp). At the same position as for P. recondita f.sp. tritici, an intron of about 1500-1600 bp length was detected also in all other Puccinia species. High homologies were observed among all Puccinia species for both the exonic and intronic fragments of the cyt b gene. Specific primers for the cyt b gene of all eight Puccinia species were developed, which easily amplified the fragment of the gene including all possible mutations known to confer resistance to QoIs in several plant pathogens. However, in all tested isolates of the Puccinia species included in this study, the sequence of cyt b gene fragment did not contain any point mutations.     

Immunohistochemical diagnosis of canine ovarian epithelial and granulosa cell tumors.

In humans and canines, the morphology of granulosa cell tumors is extremely variable and causes diagnostic difficulties. In human pathology, immunohistochemistry has been widely used for the diagnosis of granulosa cell tumors, whereas, limited studies are present in canine species. The aim of this study was to investigate the expression of cytokeratins, vimentin, and inhibin-alpha in canine normal ovaries, epithelial ovarian tumors, and granulosa cell tumors to establish an immunohistochemical panel for the differential diagnosis of ovarian tumors. Formalin-fixed, paraffin-embedded tissue sections from 4 normal ovaries, 8 granulosa cell tumors, and 6 epithelial ovarian tumors (2 adenomas and 4 adenocarcinomas) sections were obtained and stained with hematoxylin and eosin and immunohistochemically for cytokeratin AE1/AE3, cytokeratin 7, vimentin, and inhibin-alpha. In normal ovaries, cytokeratin 7, cytokeratin AE1/AE3, and vimentin were expressed in the surface epithelium. Granulosa cells were negative for cytokeratin 7 and displayed variable expression of vimentin, cytokeratin AE1/AE3, and inhibin-alpha toward follicular maturation. Granulosa cell tumors were negative for cytokeratin 7 and positive for inhibin-alpha. Conversely, ovarian epithelial cells tumors were positive for cytokeratin 7 and negative for inhibin-alpha. Both granulosa and epithelial cell tumors displayed variable expression of vimentin. Cytokeratin AE1/AE3 was expressed by all epithelial-derived tumors and 6 of 8 granulosa cell tumors. The results of this study suggest that useful immunohistochemical markers to distinguish epithelial ovarian tumors from granulosa cell tumors are cytokeratin 7 and inhibin-alpha.     

First detection of canine parvovirus type 2c in South America

Since its sudden emergence in the early 1970s, canine parvovirus type-2 (CPV-2) has been evolving through the generation of novel genetic and antigenic variants (CPV-2a/b/c and a number of additional mutations) that are unevenly distributed throughout the world. In order to develop strategies to control the spread of these variants and to understand virus evolution is fundamental to genotype field isolates from different geographic locations. In the present paper we have examined 25 isolates of CPV from clinical samples of Uruguayan dogs collected during year 2006. A fragment of the VP2 gene of the virus was analyzed using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and DNA sequence analysis. Out of the 25 isolates analyzed, only one was characterized as CPV-2a and 24 were characterized as CPV-2c, indicating that this type is currently the prevalent field CPV circulating in Uruguay. This is the first report of CPV-2c in the American continent and it also represents the highest frequency of this type observed in a dog population so far. Its presence in South American supports the assumption that CPV-2c is reaching a worldwide distribution as occurred with 2a/2b antigenic types.     

Wood coated with plasma-polymer for water repellence

The effects of cold HMDSO and HMDSO + SF₆ plasma treatments on spruce, chestnut and poplar wood were investigated in order to obtain a hydrophobic surface evaluated by microscopic, spectroscopic (FT-IR and LIBS) and contact angle analyses. Cold plasma treatments can increase the surface hydro-repellence without modifying the external aspect and bulk properties of different woods.     

Virulence and pCM1 plasmid carriage are related to BOX-PCR fingerprint type in strains of Clavibacter michiganensis subsp. michiganensis that cause bacterial wilt and canker of tomato in Argentina

Clavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial wilt and canker in tomato, producing important economic losses worldwide. Its virulence has been related to several putative virulence factors present on a chromosomal pathogenicity island and on plasmids pCM1 and pCM2, in strain NCPPB382. We genotypically characterized a collection of Cmm isolates from the main greenhouse tomato-producing areas of Argentina by BOX-PCR fingerprinting and screened for the presence of genes and plasmids involved in pathogenicity by PCR. In addition, we evaluated in vitro cellulolytic activity and virulence in planta of selected strains. BOX-PCR fingerprinting clustered strains into four groups. Group II was dominant and included the most virulent strains, while Group III was the smallest and had the least virulent strains. All local strains exhibited similar cellulolytic activity. Most of the examined strains carry two plasmids of similar size to those of NCPPB382, although there were strains with one or three plasmids. By PCR amplification of repA, pCM1 was detected only in strains belonging to Group III, which includes local strains closely related to reference strain NCPPB382. All analysed pathogenicity genes were widespread among strains, and so in strains belonging to Groups I and II, celA found on pCM1 in NCPPB382 could be found in the chromosome or in plasmids other than pCM1. This study contributes to a better understanding of the diversity of Cmm genetic profiles and virulence of strains present in Argentina. Such information could be useful for the selection of strains for screening of host resistance and development of resistant tomato varieties.     

Root-lesion nematodes of potato: Current status of diagnostics,pathogenicity and management

Root-lesion nematodes of the genus Pratylenchus are migratory endoparasites with worldwide economic impact on several important crops including potato, where certain species like P. penetrans, P. neglectus, and P. scribneri reduce the yield and quality of potato tubers. Morphological identification of Pratylenchus spp. is challenging, and recent advancements in molecular techniques provide robust and rapid diagnostics to differentiate species without the need of specialist skills. However, the fact that molecular diagnostics are not available for all Pratylenchus species means that there are limitations in worldwide application. In general, root-lesion nematodes are difficult to manage once introduced into agricultural land and damage can be related to pathogenicity and population densities. In addition, root-lesion nematodes interact with fungi such as Verticillium dahliae, resulting in disease complexes that enhance the damage inflicted on the potato crop. Management interventions are often focused on limiting nematode reproduction before planting crops and include the application of nematicides, and cultural practices such as crop rotation, cover crops, biofumigation, and biological control. Understanding the limitations of the available crop protection strategies is important and there are many gaps for further study. This review discusses the status of the diagnosis, distribution, pathogenicity, and management of the main species of root-lesion nematodes, reported to infect potatoes worldwide, and highlights areas for potential future research.     

Lactobacillus rhamnosus as additive for maize and sorghum ensiling

The effects of Lactobacillus rhamnosus AT195, a potential probiotic microorganism cultured in buffalo "scotta" whey, on chemical and microbiological composition in maize and sorghum ensiling were evaluated. Both crops were harvested, chopped, and treated or not with the selected strain prior to ensiling in fiberglass vertical silos; 90 days after ensiling, silages were sensorially evaluated and sampled. Different chemical components were evaluated both on fresh crops and silages: in particular, the water-soluble carbohydrates content was investigated by high-field NMR spectroscopy and the carbohydrate fermentation profile was performed by GC. Besides phenotypic identification and typing, microbiological studies included Lb. rhamnosus genotype typing by RAPD-PCR. All silages, inoculated or not, were well preserved, as their chemical and microbiological data along with the fermentation profiles showed. The selected strain used as inoculum influenced the lactic acid population of silages and evidenced a good survival performance during the ensiling process of both maize and sorghum. Moreover, the use of Lb. rhamnosus strain efficiently improved the quality of the multifactorial ensiling process by significantly reducing the ammonia nitrogen content of both maize and sorghum silages.     

Effects of Baking Temperature on Crumb-Staling Kinetics

This study evaluated the effects of bread baking temperature on the staling kinetics of crumb. Bread dough was leavened and baked in sealed molds. Cooking trials were performed at various temperatures ranging from 90 to 110°C. The crumb samples were then stored at 20°C at constant moisture, and staling was evaluated by measuring crumb elastic modulus (using an Instron dynamometer) and starch retrogradation degree (using differential scanning calorimetry). Results show that the cooking temperature greatly influences bread staling. The lower the cooking temperature, the lower the staling rate, both in terms of crumb hardening and of starch retrogradation. Starch and protein solubility was evaluated on crumb cooked at 90 and 110°C. An increase in cooking temperature resulted in an increase in protein insolubilization and starch granule disruption.