Subtilase cytotoxin encoding genes are present in human, sheep and deer intimin-negative, Shiga toxin-producing Escherichia coli O128:H2

Shiga toxin-producing Escherichia coli (STEC) O128:H2 is recognised worldwide to be an important non-O157 STEC associated with human illness and in particular with causing haemolytic uraemic syndrome. This serotype is commonly isolated from sheep and is being increasingly isolated from deer. We determined the virulence profile and genetic relationships of one human, six sheep and five deer intimin-negative STEC O128:H2 strains isolated in Spain over a 7-year period. Our goals were to establish the presence of other virulence-associated factors, such as SubAB, in intimin-negative STEC O128:H2 strains involved in human disease and in that case, to determine if sheep and/or deer represent a reservoir of SubAB-positive STEC O128:H2. All the strains lacked the eae gene and carried subtilase cytotoxin (SubAB) encoding genes (subAB) and tia genes, but not saa gene, suggesting the presence of the recently identified new variant of SubAB, encoded on a putative pathogenicity island together with tia. We report for the first time the presence of subtilase cytotoxin encoding genes in intimin-negative STEC O128:H2 strains pathogenic for humans and how this finding might explain their clinical relevance despite neither carrying eae nor stx subtypes associated with severe clinical outcomes, but only stx1c and stx2b. Multilocus sequence typing analysis revealed that STEC O128:H2 strains from sheep and deer belong to the clonal lineage of STEC O128:H2 strains involved in diarrhoeal and haemorrhagic diseases in humans. Our results indicate that sheep and deer represent a reservoir of SubAB-positive STEC O128:H2 strains and thus a potential source of human infection.     

Gastroprotective potential of frutalin, a D-galactose binding lectin, against ethanol-induced gastric lesions

The present study was designed to verify whether frutalin (FTL) affords gastroprotection against the ethanol-induced gastric damage and to examine the underlying mechanism(s). Gastric damage was induced by intragastric administration of 0.2 ml of ethanol (96%). Mice in groups were pretreated with FTL (0.25, 0.5 and 1 mg/kg; i.p.), cimetidine (100 mg/kg; p.o.), or vehicle (0.9% of NaCl, 10 mL/kg; p.o.), 30 min before ethanol administration. They were sacrificed 30 min later, the stomachs excised, and the mucosal lesion area (mm2) measured by planimetry. Gastroprotection was assessed in relation to inhibition of gastric lesion area. To study the gastroprotective mechanism(s), its relations to capsaicin-sensitive fibers, endogenous prostaglandins, nitric oxide, sulphydryls, ATP-sensitive potassium channels, adrenoceptors, opioid receptors and calcium channels were analyzed. Treatments effects on ethanol-associated oxidative stress markers GSH and MDA were measured in gastric tissue. FTL afforded a dose-unrelated gastroprotection against the ethanol damage. However, it failed to prevent the ethanol-induced changes in the levels of GSH and MDA. It was observed that the gastroprotection by FTL was greatly reduced in animals pretreated with capsazepine, indomethacin, L-NAME or glibenclamide. Considering the results, it is suggested that the FTL could probably be a good therapeutic agent for the development of new medicine for the treatment of gastric ulcer.     

Distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens in nervous and non-nervous organs of European seabass (Dicentrarchus labrax) during the course of an experimental challenge

The distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens was examined by immunohistochemistry in the nervous and non-nervous organs of juvenile European seabass (Dicentrarchus labrax) during the course of an intramuscular infection. Histological changes resulting from the infection were evaluated from 3 days to 2 months post-infection. The specific antibody response was also studied 2 months post-challenge. Viral proteins were present throughout the experimental period in the retina (inner nuclear layer, ganglion layer, outer limiting membrane, and outer plexiform layer), brain (cerebellum and tectum opticum), and liver (hepatocytes and endothelial cells). These proteins were also observed in the renal tubular cells, white pulp of spleen, and in fibroblasts and cartilage of caudal fin. This is the first report of RGNNV proteins appearing in these organs, where the immunostaining was only detected at certain sampling times after the onset of mortality. Brain and retina of virus-exposed fish showed high levels of vacuolation, while accumulation of fat vacuoles was observed in the liver. RGNNV infection also induced a specific antibody response as measured by an ELISA. In summary, this is the first study demonstrating the presence of viral proteins in cells of caudal fin, kidney and spleen of European seabass.     

Geospatial variation of physical attributes and sugarcane productivity in cohesive soils

Precision Agriculture - Sugarcane productivity is highly variable across space, and knowledge about the spatial distribution of soil physical attributes is important to guide management practices...     

Response of in vitro obtained oil palm and interspecific OxG hybrids to inoculation with Phytophthora palmivora

Bud rot (BR) disease caused by Phytophthora palmivora is the most devastating disease in oil palm cultivation in America. Oil palms that survived BR epidemics were found in areas highly devastated by the disease; these palms were introduced into Cenipalma's in vitro micropropagation (cloning programme). A severity scale was developed for in vitro palms from five ortets inoculated with two different P. palmivora isolates. Then, eight ortets of Elaeis guineensis and two ortets of the OxG interspecific hybrid were evaluated in two inoculation trials under chamber growth conditions. The clone performance response was consistent with that reported in the field for the corresponding ortets, and two contrasting ortets, one susceptible (ortet 57) and one resistant (ortet 34) were identified. We monitored and compared defence responses to P. palmivora in the contrasting ortets. An increase in reactive oxygen species (ROS) production after inoculation with the pathogen was observed, with higher accumulation of H₂O₂ in the resistant plants. Catalase (CAT) activity in resistant plants increased after inoculation with the pathogen from 24 hr post‐infection (hpi) and remained high during the observation time. In the susceptible ortets, there was a significant increase on catalase activity only at 48 hpi. Peroxidase (POS) activity increased in clones from both susceptible and resistant ortets, but the increase was much greater in the susceptible ones. Phenylalanine ammonia lyase activity increased in response to inoculation, and these increases were greater in clones of the susceptible ortet than in clones of the resistant ortet.     

Targeted metabolite analysis and antioxidant potential of Rumex induratus

Targeted metabolite analysis of aqueous extract of Rumex induratus leaves, in terms of phenolic compounds and organic acids, and the study of its antioxidant activity against the DPPH(*) radical, a reactive oxygen species, hypochlorous acid, and a reactive nitrogen species, nitric oxide ((*)NO), were performed. The samples were collected in several locations, spontaneously occurring or from greenhouse culture, at different stages of development and seasons. The phenolic composition was achieved by high-performance liquid chromatography (HPLC)-diode array detection, and four hydroxycinnamic acid derivatives and 10 flavonoid glycosides (C- and O-heterosides) were determined. Organic acids composition was established by HPLC-UV, revealing five compounds. The total amount of phenolic compounds and organic acids were affected by growing conditions and developmental phase. The aqueous extract exhibited a dose-related activity against all tested reactive species.     

Influence of landuse on soil erosion risk in the Cuyaguateje watershed (Cuba)

Landuse changes may dramatically enhance erosion risk. Besides deforestation, also arable landuse may have an important influence on soil loss. We investigated the erosion risk in a 151 km² subwatershed of the Cuyaguateje watershed (Cuba) using the RUSLE model. It was found that the valleys used for agriculture have the highest erosion risk, with actual erosion surpassing soil loss tolerance. Over the period 1985–2000, about 14 km² of forest has been converted into arable land. As a result, the area with a very high erosion risk increased with 12%. On arable land it was found that the crop management factor C of a “tobacco/maize” rotation was 0.478, compared to 0.245 for a rotation of various crops (sweet potato, beans, maize, cassava and fallow). When maize in the “tobacco/maize” rotation was intercropped with a leguminous crop (hyacinth bean) the C factor decreased to a value of 0.369. Also contouring may halve soil loss on moderate slopes (< 10%) when high ridges are applied, which is in Cuba generally the case for maize, cassava and sweet potato.     

Expression of toll-like receptor 2 (TLR2) in porcine leukocyte subsets and tissues

Toll-like receptors (TLR) are a group of pattern recognition molecules that play a crucial role in innate immunity. TLR2 recognises a variety of microbial components leading to the development of inflammatory and immune responses. To characterise the expression and functional properties of porcine TLR2 (pTLR2), we have raised a panel of monoclonal antibodies (mAb) against this molecule. Mouse 3T3 cell transfectants expressing pTLR2 were used for immunisation of mice. The specificity of these antibodies was confirmed by their reactivity with CHO cells transfected with pTLR2 but not with pTLR4 or with non-transfected cells. Using one of these mAbs, named 1H11, pTLR2 was found on cells of the innate immune system, including monocytes, macrophages, and granulocytes, but not on peripheral blood lymphocytes. Staining of tissue sections showed that pTLR2 is also expressed on epithelial cells lining the tracheobronchial and intestinal tracts, bile ducts in the liver and renal tubules, and on the basal layer of the epidermis. This distribution is consistent with a surveillance function at entry sites, allowing for early detection of microbial invasion.