A PCR technique for the detection of Jaagsiekte sheep retrovirus in the blood suitable for the screening of ovine pulmonary adenocarcinoma in field conditions

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring contagious lung neoplasia caused by jaagsiekte sheep retrovirus (JSRV). Although no specific circulating antibodies against the virus can be detected in infected sheep, JSRV proviral DNA sequences can be found in peripheral blood leukocytes (PBLs) in clinically affected and in a proportion of in contact animals. In this study, existing hemi-nested PCR procedure is compared with a new one-step PCR technique that was developed to minimise potential DNA contamination and reduce sample and reagent handling. Different blood preparations were assessed and the best results were achieved on DNA prepared from buffy coat. The sensitivity of this PCR was lower in JSRV infected sheep without lesions of OPA than in clinically affected sheep, which indicate that this PCR may not be not fully appropriate for screening of individual sheep, but rather to provide results at flock level. This PCR is the only currently available blood test for detection of JSRV infected sheep and may be useful in epidemiological studies and in control programmes of OPA.     

Effects of tension of the girth strap on respiratory system mechanics in horses at rest and during hyperpnea induced by administration of lobeline hydrochloride

OBJECTIVE: To determine whether tension of the girth strap of a saddle would sufficiently affect rib motion and reduce lung volume to alter pulmonary resistance in horses. ANIMALS: 10 healthy adult horses. PROCEDURE: We used classical techniques to measure the effects of tightening a girth strap (15 kg of tension) on pulmonary dynamics during eupnea and hyperpnea in horses. Respiratory impedance was evaluated by use of oscillometry, and resistance and reactance data were partitioned into lung and chest wall components. Rib cage and abdominal contributions to tidal volume and minute ventilation were measured by use of respiratory inductance plethysmography. Effects of strap tension on functional residual capacity (FRC) were measured during eupnea by use of a helium-dilution technique. In a subgroup of 6 horses, we also measured transdiaphragmatic pressures during eupnea and hyperpnea induced by administration of lobeline hydrochloride (0.2 mg/kg, i.v.). RESULTS: Pulmonary resistance measured by use of oscillometry but not by use of classical methods was significantly increased by the tension of the girth strap. However, the increase in pulmonary resistance could not be explained by a decrease in FRC. Motion of the rib cage was significantly reduced during eupnea and hyperpnea. However, ventilatory variables (tidal volume, minute ventilation, and peak flows), FRC, and transdiaphragmatic pressures were unaltered by strap tension. CONCLUSIONS AND CLINICAL RELEVANCE: Although tension of the girth strap caused measurable changes in respiratory mechanics (loss of rib motion and increased pulmonary resistance), there was no evidence that ventilation was limited.     

Sphaerospora dicentrarchi and S. testicularis (Myxozoa: Sphaerosporidae) in farmed European seabass (Dicentrarchus labrax) from Italy

The distribution of Sphaerospora dicentrarchi Sitjà-Bobadilla et Alvarez-Pellitero, 1992 and S. testicularis Sitjà-Bobadilla et Alvarez-Pellitero, 1990, myxozoan parasites of European seabass Dicentrarchus labrax (L.), was investigated in different farming systems in Italy. In total, 1406 fish were examined. High S. dicentrarchi prevalence was observed in all the farming systems involved in this survey (extensive farms: 51.5%; intensive farms: inland 59.6%, inshore floating cages 76.2%, offshore floating cages 41.6%) except for submersible cages (7.4%). S. testicularis was detected only in nine male fish from two intensive farms. The epidemiology and pathological effects of the parasites are discussed.     

Adenosine deaminase activity in serum, erythrocytes and lymphocytes of rats infected with Leptospira icterohaemorrhagiae

Leptospirosis is a systemic disease of humans and domestic animals, mainly dogs, cattle and swine. The course of human leptospirosis varies from mild to severe fatal forms and the most severe form of human leptospirosis is principally caused by Leptospira interrogans serovar icterohaemorrhagiae (L. icterohaemorrhagiae). The enzyme adenosine deaminase (ADA) plays an important role in the production and differentiation of blood cells. The aim of this study was to evaluate the activity of ADA in serum, erythrocytes and lymphocytes of rats infected with L. icterohaemorrhagiae, as compared with non-infected rats. Twenty-four adult rats, divided into two uniform groups (A and B) were used for the enzymatic assays. The animals in Group B were inoculated intraperitoneally with 2×10(8) leptospires/rat, and the rodents in Group A (control) were not-inoculated. Blood collection was performed on days 5 and 15 post-infection (PI) and the blood used to assess the ADA activity. The infection by L.icterohaemorrhagiae altered erythrocyte count, hemoglobin concentration and hematocrit, causing a decrease in all these parameters on day 15 PI. Lymphocytes decreased significantly on day 15 PI, and ADA activity in serum was inhibited in infected rats on days 5 and 15 PI and its activity in erythrocytes were increased on day 5 PI. On day 5 PI, we found an increase in ADA activity in erythrocytes of infected rats. No correlation was observed between hematocrit and erythrocyte ADA activity on days 5 and 15 PI. The ADA activity was inhibited in rats infected on day 15 PI. A positive correlation (r(2)=60) was also observed between the number of lymphocytes and ADA activity in lymphocytes on day 15 PI (P<0.05). In conclusion, our results showed that the ADA activity is altered in serum, lymphocytes and erythrocytes in experimental infection by L.icterohaemorrhagiae in rats, concomitantly with hematological parameters.     

Impact of storage conditions on the apple allergen Mal?d?1

Consumption of fresh apple fruits can induce allergic reactions in humans. The apple allergen Mal?d?1 is responsible for these allergic reactions in humans in Central Europe and North America. Biosynthesis of Mal?d?1 depends on apple cultivar, and its concentration increases with time during fruit storage. However, data on the impact of different fruit storage conditions during long-term storage are scarce. Hence, the Mal?d?1 contents of eight apple cultivars were analyzed for this study during long-term storage in a cold chamber as well as under controlled atmosphere conditions (CA). After harvest, apple fruits were stored for 12, 20, 28 or 36 weeks in a cold-chamber at +?2?°C or as under controlled atmosphere conditions of 1.5?% CO₂, 1.5?% O₂ at +?2?°C. Mal?d?1 content in apple fruit of all eight cultivars examined increased during fruit storage. In most cases, differences between Mal?d?1 of apple fruits stored in the cold chamber and under CA conditions were significant, but inconsistent. In apple cv. ??Elise??, fruits stored in the cold chamber contained more Mal?d?1 compared with those stored in CA, whereas the situation reversed in other varieties like cv. ??Boskoop??. The greatest Mal?d?1 content was measured in fruits of cvs. ??Golden Delicious?? and ??Gala??, whereas the smallest Mal?d?1 level was in cvs. ??Elise?? and ??Pinova?? over the whole storage time. Overall, this experiment showed the complexity of the relationship between the Mal?d?1 content, allergenicity of apple fruits, different cultivars, storage conditions and storage time. Persons allergic to apple fruits should consume the fruits as fresh as possible or only after a limited storage time. Furthermore, they should prefer apple varieties with a low content of allergenic proteins, such as cv. ??Elise?? or cv. ??Boskoop?? as a cultivar known for its large polyphenol content.     

Investigating the persistence of accuracy of genomic predictions over time in broilers

Accuracy of genomic predictions is an important component of the selection response. The objectives of this research were: 1) to investigate trends for prediction accuracies over time in a broiler population of accumulated phenotypes, genotypes, and pedigrees and 2) to test if data from distant generations are useful to maintain prediction accuracies in selection candidates. The data contained 820K phenotypes for a growth trait (GT), 200K for two feed efficiency traits (FE1 and FE2), and 42K for a carcass yield trait (CY). The pedigree included 1,252,619 birds hatched over 7 years, of which 154,318 from the last 4 years were genotyped. Training populations were constructed adding 1 year of data sequentially, persistency of accuracy over time was evaluated using predictions from birds hatched in the three generations following or in the years after the training populations. In the first generation, before genotypes became available for the training populations (first 3 years of data), accuracies remained almost stable with successive additions of phenotypes and pedigree to the accumulated dataset. The inclusion of 1 year of genotypes in addition to 4 years of phenotypes and pedigree in the training population led to increases in accuracy of 54% for GT, 76% for FE1, 110% for CY, and 38% for FE2; on average, 74% of the increase was due to genomics. Prediction accuracies declined faster without than with genomic information in the training populations. When genotypes were unavailable, the average decline in prediction accuracy across traits was 41% from the first to the second generation of validation, and 51% from the second to the third generation of validation. When genotypes were available, the average decline across traits was 14% from the first to the second generation of validation, and 3% from the second to the third generation of validation. Prediction accuracies in the last three generations were the same when the training population included 5 or 2 years of data, and a decrease of ~7% was observed when the training population included only 1 year of data. Training sets including genomic information provided an increase in accuracy and persistence of genomic predictions compared with training sets without genomic data. The two most recent years of pedigree, phenotypic, and genomic data were sufficient to maintain prediction accuracies in selection candidates. Similar conclusions were obtained using validation populations per year.     

Chemical and Nutritional Composition of Tejate, a Traditional Maize and Cacao Beverage from the Central Valleys of Oaxaca, Mexico

Foam-topped cacao and maize beverages have a long history in Mesoamerica. Tejate is such a beverage found primarily in the Zapotec region of the Central Valleys of Oaxaca, Mexico. Historically tejate has been ceremonially important but also as an essential staple, especially during periods of hard fieldwork. However, the nutritional contribution of traditional foods such as tejate has not been investigated. We analyzed tejate samples from three Central Valley communities, vendors in urban Oaxaca markets and one migrant vendor in California, USA for their proximate composition, amino acid content and scores, and mineral and methylxanthine content. Nutritional and chemical variation exists among tejate recipes, however, the beverage is a source of energy, fat, methylxanthines, K, Fe and other minerals although their availability due to presence of phytates remains to be determined. Tejate is a source of protein comparable to an equal serving size of tortillas, with protein quality similarly limited in both. Tejate provides the nutritional benefits of maize, and some additional ones, in a form appealing during hot periods of intense work, and year round because of its cultural significance. Its substitution by sodas and other high glycemic beverages may have negative nutritional, health and cultural consequences.     

Trypanocidal Action of (?)-Elatol Involves an Oxidative Stress Triggered by Mitochondria Dysfunction

Natural compounds have shown good potential for the discovery of new chemotherapeutics for the treatment of Chagas’ disease. Recently, our group reported the effective trypanocidal activity of (−)-elatol, extracted from the red macroalgae Laurencia dendroidea present in the Brazilian coast against Trypanosoma cruzi. However, the mechanism of action of this compound has remained unclear. There are only hypotheses concerning its action on mitochondrial function. Here, we further investigated the mechanisms of action of (−)-elatol on trypomastigotes of T. cruzi. For this, we evaluated some biochemical alterations in trypomastigotes treated with (−)-elatol. Our results show that (−)-elatol induced depolarization of the mitochondrial membrane, an increase in the formation of mitochondrial superoxide anion and loss of cell membrane and DNA integrity. Additionally, (−)-elatol induced formation of autophagic vacuoles and a decrease in cell volume. All together, these results suggest that the trypanocidal action of (−)-elatol involves multiple events and mitochondria might be the initial target organelle. Our hypothesis is that the mitochondrial dysfunction leads to an increase of ROS production through the electron transport chain, which affects cell membrane and DNA integrity leading to different types of parasite death.     

Scouting the application of sourdough to frozen dough bread technology

The use of sourdough, even in combination with cryoprotectant (skim milk, sucrose and trehalose), conventional additives (guar gum, diacetyl tartaric acid esters of monoglycerides, ascorbic acid), honey or fructose and glucose, in frozen dough technology was investigated. After frozen storage, the leavening performance of doughs, and the hardness and texture of breads were compared to those of an unfrozen dough, and to those of a conventional frozen dough. All frozen doughs showed a longer fermentation time and a lower volume increase, with respect to unfrozen dough. When sourdough was combined with cryoprotectant, honey or both, the leavening performance improved compared to the use of sourdough alone. Compared to the conventional frozen dough, higher leavening performance was reached combining sourdough with cryoprotectant alone or together with honey. Sourdough combined with honey, fructose and glucose, honey and cryoprotectant, or conventional additives decreased bread hardness compared to the unfrozen dough bread and to the conventional frozen dough bread. Independently from the use of sourdough, conventional additives allowed to reach a specific volume not significantly different from that of unfrozen dough bread, and breads containing honey were characterized by low values of hardness and by high values of red index.     

Characterization of Phaseolus vulgaris L. Landraces Cultivated in Central Italy

Eight Phaseolus vulgaris L. landraces cultivated on farm in marginal areas of Central Italy (Lazio region) were investigated in order to evaluate chemical composition of storage proteins and secondary metabolites fractions. The total protein content showed some differences among landraces; the maximum value was next to 30 g for 100 g of dry weight. The seed storage proteins were screened by polyacrylamide gel electrophoresis (SDS/PAGE): seven landraces exhibited phaseolin patterns type S, one landrace showed a phaseolin pattern type T. A morphological analysis of cotyledon parenchyma performed by scanning electron microscopy (SEM) revealed differences in size of starch granules. Moreover the polyphenolic composition was investigated using HPLC-APCI; from the methanol extracts a flavonoid, kaempferol, and a coumarin, 5,7-dimethoxycoumarin, were identified. To our knowledge, this is the first time that 5,7-dimethoxycoumarin has been reported in P. vulgaris seeds.