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.     

A multiplex PCR for the detection of Brucella spp. and Leptospira spp. DNA from aborted bovine fetuses

Bovine brucellosis and leptospirosis are important causes of bovine abortion around the world. Both diseases can be serologically diagnosed, but many factors may cause false positive and negative results. Direct methods based on bacteriological isolation are usually employed, but they are difficult, time consuming and dangerous. Monoplex polymerase chain reaction (PCR) have been successfully described for the detection of Brucella spp. and Leptospira spp. Aiming at improvement in the direct diagnosis, a multiplex PCR (mPCR) for the detection of these agents in aborted bovine fetuses is described. The detection threshold of the mPCR was evaluated in experimentally contaminated bovine clinical samples using a conventional proteinase K/SDS or a boiling-based extraction protocols. The mPCR was applied to two groups of clinical samples: 63 episodes of bovine abortion and eight hamsters experimentally infected with Leptospira interrogans serovar pomona. Adopting microbiological isolation as reference, the test showed a sensitivity of 100% in both groups of clinical samples. Seven samples collected from bovine fetuses were Brucella spp. culture negative but showed positive results in mPCR. Regarding Leptospira spp. detection, similar results were observed in three bovine clinical samples. All hamsters infected with Leptospira were positive in both microbiological culture and mPCR. The boiling extraction protocol showed better results in some clinical samples, probably by the removal of PCR inhibitors by heat treatment. The high sensitivity, simplicity and the possibility of detection of both bacteria in a single tube reaction support the use of the mPCR described in the routine diagnosis.     

<Emphasis Type="Italic">Achromobacter insolitus</Emphasis> and <Emphasis Type="Italic">Zoogloea ramigera</Emphasis> associated with wheat plants (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

This study reports for the first time the presence of diazotrophic bacteria belonging to the genera Achromobacter and Zoogloea associated with wheat plants. These bacterial strains were identified by the analysis of 16S rDNA sequences. The bacterium IAC-AT-8 was identified as Azospirillum brasiliense, whereas isolates IAC-HT-11 and IAC-HT-12 were identified as Achromobacter insolitus and Zoogloea ramigera, respectively. A greenhouse experiment involving a non-sterilized soil was carried out with the aim to study the endophytic feature of these strains. After 40 days from inoculation, all the strains were in the inner of roots, but they were not detected in soil. In order to assess the location inside wheat plants, an experiment was conducted under axenic conditions. Fifteen days after inoculation, preparations of inoculated plants were observed by the scanning electron microscope, using the cryofracture technique, and by the transmission electron microscope. It was observed that all isolates were present on the external part of the roots and in the inner part at the elongation region, in cortex cells, but not in the endodermis or in the vascular bundle region. No colonizing bacterial cells were observed in wheat leaves.     

Characterization of the extracellular matrix in the chorioallantoic membrane of water buffalo (Bubalus bubalis) in early gestation

Placenta is formed by a parenchyma rich in extracellular matrix (ECM), and this structure guarantees the proper development of the embryo and placental functioning. Recently, studies have focused on the characterization of ECM in the placenta and foetal membranes of different species. This work aimed to analyse the composition of the ECM and to quantify the types of collagens in its composition. For this, 33 chorioallantoic membranes were used at different gestational ages, which were grouped into five groups. Subsequently, haematoxylin–eosin staining, Masson trichrome and picrosirius were performed for histological analysis. Through the technique of polarized light, it was possible to quantify the total collagen present in the membranes and finally the immunohistochemical technique was performed to verify the presence of collagens and glycoproteins. It was possible to verify that the chorioallantoic membranes have, in all the gestational periods of the initial third of gestation, the same histological structures, being the most significant difference the membrane thickening that occurs gradually during the gestation. However, we notice the appearance of binucleate cells only from group II. In addition, it was verified that a gradual increase of collagen occurs until the group IV, yet from the group V begins to occur a decrease of this protein. In addition, collagen I, collagen III, fibronectin and laminin were present in all membranes. With this, we concluded that the buffalo chorioallantoic membrane presents ECM in constant remodelling at the beginning of gestation and can be used as biomaterial in works on regenerative biology.     

Genetics of Melon Yellows Virus Resistance Derived from Cucumis melo ssp. agrestis

The Melon yellows virus (MYV), a whitefly-transmitted closterovirus, is one of the major pathogens causing crop losses in protected melons in southeastern Spain. An accession of the wild Asiatic Cucumis melo ssp. agrestis (Cma) shows resistance to MYV infection. Results indicate the participation of two resistance mechanisms in this source: firstly, an antixenotic reaction against Trialeurodes vaporariorum, the disease vector, and secondly, resistance to the virus. The combined effect of these two mechanisms confers Cma a higher level of resistance, expressed as a delayed and milder infection.The genetics of resistance to the Melon yellows closterovirus have been studied in two families derived from Cma. As under natural infection conditions, the effect of antixenosis and virus resistance cannot be distinguished, a biometrical model that permits separation of the two resistance mechanisms operating in the same resistant source, has been proposed to determine genetic control of MYV resistance.The genetic analysis has been conducted by fitting the disease progress curves of each generation to the biometrical model instead of fitting the final disease ratios. The scoring of disease incidence over time allows for the comparison of data from assays conducted in different conditions (2 years/4 transplanting dates), thus reinforcing the analysis.The results agree with a simple control of the resistance to MYV derived from Cma, with incomplete penetrance of the gene and partial dominance of resistance. The effect of antixenosis on the spread of this plant virus is highly significant in Cma, but not in segregant generations.Since there do not exist crossability barriers between this accession and the cultivated melon, Cma could be readily used in breeding programmes to obtain melon varieties resistant to MYV.     

Effect of the density on the growth and survival of the Caribbean scallops Argopecten nucleus and Nodipecten nodosus in suspended culture

Juveniles of the Caribbean scallops Argopecten nucleus and Nodipecten nodosus were suspended in pearl nets at densities of 25%, 30% and 50% coverage of the bottom of the net (BC) during days. Additionally, adults of A. nucleus (>35 mm) were placed in lantern nets at densities of 40%, 50% and 65% BC during days. Stocking density affected the growth of animals, with higher growth rates obtained at the lower densities. Although the survival of the two scallops was not influenced at the densities studied, the 15-day net changes, and culture in water of <28 °C produced better survival of A. nucleus and N. nodosus respectively. Predators of the families Cymatiidae, Grapsidae, Portunidae, Majidae and Xanthidae were found in the culture nets of juvenile A. nucleus , but during the adult phase they were absent. Predators exhibited a dense-independent behaviour in relation to scallops. There was not any association between predator frequency and survival of A. nucleus .     

Evaluation of the Microalgae Paste Viability Produced in a Mollusk Hatchery in Southern Brazil

The present study was conducted to define a methodology to produce and store small‐scale microalgae paste to be used in a mollusk hatchery. Microalgae were cultured in 500 L fiberglass tanks, under temperature of 20 ± 2 C, Guillard f/2 culture medium, and continuous light intensity of 203–226 μmol photons/m²/sec. Cultures were centrifuged at 2000 g at the exponential growth phase. Microalgae cell quality after centrifugation and during storage was determined by analyses with Evan’s blue stain and by counting the number of total marine bacteria. Treatments with and without additive were applied to the microalgae paste produced, which was distributed into 100 mL plastic containers, capped, and stored under refrigeration at 4 ± 1 C. Results indicated that in the Chaetoceros muelleri paste, centrifugation did not damage the cells and the number of total marine bacteria reduced significantly from 2.9 × 10⁶ to 8.3 × 10⁵ colony‐forming units per milliliter. Chaetoceros muelleri and Chaetoceros calcitrans pastes stored with addition of 0.1% ascorbic acid had a shelf life shorter than 2 wk. For the treatment without additive, results with Evan’s blue stain showed that cells (99%) remained viable until the sixth week of storage for C. muelleri and seventh week of storage for Skeletonema sp. and C. calcitrans. The number of bacteria did not increase during storage for C. calcitrans and Skeletonema (P > 0.05). For C. muelleri, an increase in bacteria (P < 0.05) was observed after the sixth week of storage. This study demonstrated the feasibility to produce and store microalgae paste for a period of 2–8 wk, which allows it to be used as food source and also optimizes the use of microalgae cultured in laboratory.     

Influence of pH and calcium on the growth,polysaccharide production and symbiotic association of<Emphasis Type="Italic"> Sinorhizobium meliloti</Emphasis> SEMIA 116 with alfalfa roots

The Sinorhizobium -legume interaction is sensitive to a number of environmental factors, soil acidity being one of the most important. In the typic Hapludoll soil of the central-southern region of Córdoba (Argentine) it was found that the nodulation of alfalfa ( Medicago sativaL.) roots was impaired with a reduction in shoot dry matter under conditions of soil acidity (pH 5.5) Our results showed that the addition of lime as dolomitic limestone at a concentration of 1 t ha ^-1to acid soil caused a significant increase in the nodulation of alfalfa roots inoculated with the strain Sinorhizobium meliloti SEMIA 116 (recommended inoculum for alfalfa) in the greenhouse experiments. The success of the lime treatment may be related not only to an increase in the pH values but also to an increase in the Ca concentration, improving the growth of S. meliloti and its nodulation ability under acidic conditions. In this study, we also demonstrated an increment in the bacterial growth rate as well as in the production of exopolysaccharides and lipopolysaccharides under low pH (5.5) and a high Ca concentration (5 mM) in the culture medium.     

Additive gene action for resistance to Puccinia striiformis F.SP. Tritici in Triticum dicoccoides

Summary Seven single-plant selections of wild emmer, with temperature-sensitive minor-effect genes for stripe rust resistance, were intercrossed in eight combinations. The resulting progenies were studied for a possible additive gene action.The transgressive segregation towards resistance in F₂ observed in all the combinations indicates that additive gene action for resistance indeed occurs in wild emmer. The common occurrence of this phenomenon in random combinations suggests further that several minor-effect genes are involved.Following selection of the most resistant plants in F₂, a marked shift towards resistance was noted in F₃, which demonstrates a positive response to selection. In some instances, additive resistance selected for (in F₂) at the high temperature-profile was expressed (in F₃) also at the low temperature-profile. This kind of resistance, when utilized in breeding programmes, promises therefore to be effective over a range of temperatures.