Importance of Some Biochemical Parameters in the Quality and Yield of Rapeseed (Brassica napus L.)

Samples of rapeseed from three Italian growing environments (Bologna, Perugia and Palermo) were analysed for glucose content and dry weight of 1000 seeds every three or four days starting from the end of flowering until complete ripening. In addition, the content of oil, soluble and total proteins, glucosinolates and myrosinase activity was determined in samples of mature seeds. The cultivars used were jet Neuf and Lingot (type 0) and Tandem, Jade and Santana (type 00). From the results it emerged that the point of intersection of the two branches of the linear regression plots for different glucose-consumption kinetics found during seed filling, in addition to being strongly affected by the climate of the test environment, is correlated with quantitative and qualitative production, independently of the genotype.     

Neurotoxic syndrome of sheep due to the ingestion of plants of the Mediterranean scrub: clinical, histopathological, histochemical and ultrastructural observations

A severe neurological disorder was observed during a very dry season, in sheep pasturing on land with mediterranean vegetation. During the crisis, lasting 5-20", ataxia and convulsions were the main clinical signs. Brain congestion, spongy appearance of the liver, interstitial nephritis and hyperpigmentation of visceral lymph nodes were the most important macroscopic lesions. Histopathological, histochemical and ultrastructural findings confirmed liver and kidney lesions and showed considerable neurovisceral storage of lipofuscin. Although further toxicological and experimental studies are needed to elucidate the origin of the disease, the Authors attribute the outbreak of the neurological syndrome to the ingestion of a neurotoxin containing plant.     

Comparative study of three feeding methods for draught horses of the Swiss army

Three feeding methods were compared in 36 4- to 6-year-old Franche-Montagne horses during the military school of St-Luzisteig (GR) of Spring 1992. The horses were separated into 3 groups: a group with the traditional oats-hay ration (OH), a group with a pelleted feed and hay ration (PFH), and a group with the complete diet (CD). Feed analyses were performed and food consumption, eating behavior and digestibility were studied. The horses received their daily amount of feed in 3 portions covering the requirements for a medium work: OH = 8 kg hay and 3 kg oats, PFH = 8 kg hay and 3 kg pelleted feed and CD = 10 kg of the complete diet. For the 3 rations, the amount of digestible crude protein for horses was higher than the reference value for the requirement of a 600 kg horse with a medium work. In the 3 diets, the calcium content was higher than the required 32 g per day (g/d). Not enough sodium (OH: 1.2 g/d; PFH: 7.3 g/d; CD: 9.6 g/d) and too much potassium (OH: 140.3 g/d; PFH 153.0 g/d; CD: 167.5 g/d) were present in the diets, both without consequences for the blood parameters. In 3 meals of 60 minutes, the horses of the group OH, PFH and CD ingested 82%, 89% and 92%, respectively, of the daily ration. The complete diet was ingested more quickly than the hay. The number of mastications per minute was smaller for the complete diet than for the hay. Ingestion times were similar for oats and pelleted feed. However, the number of mastications per minute was smaller for the pelleted feed. The digestibility of nutrients was not influenced by the method of feeding. In conclusion, these results demonstrate that the 3 types of ration studied here are adequate for the swiss army horses if sodium is added to the diet. However, despite the fact that both PFH and CD correct excessive supply or deficiencies of nutrients and despite the fact that these two feeding methods offer nutrients in amounts that are closer to the requirements of the horse, the method PFH was introduced in 1994.     

Real-time TaqMan polymerase chain reaction detection and quantification of cow DNA in pure water buffalo mozzarella cheese: method validation and its application on commercial samples

Mozzarella cheese obtained from buffalo (Bubalus bubalis) milk is a typical Italian product certificated by means of the European Protected Designation of Origin (PDO). Mozzarella cheese can also be obtained from bovine milk or bovine/buffalo milk mixtures, but in this case, it cannot be sold as PDO product, and its label must report the actual ingredients. However, bovine milk in PDO products was frequently detected in the past, suggesting fraudulent addition or accidental contamination. Several methods based on end-point polymerase chain reaction (PCR) have been profitably applied in a large number of tests to detect the presence of undeclared ingredients, also in dairy products. In the present study we report a real-time PCR method able to quantify bovine milk addition to pure buffalo cheese products. We validated a normalized procedure based on two targets: bovine mitochondrial cytochrome b (cyt b) to detect and quantify the bovine DNA and nuclear growth hormone (GH) gene used as a universal reference marker. With the use of this real-time PCR assay, 64 commercial mozzarella di bufala cheese samples purchased at local supermarkets, dairy shops, or directly from cheese manufacturers were analyzed. The results obtained demonstrate that most of the commercial samples were contaminated with bovine milk. Therefore, this assay could be conveniently employed to carry out routine and accurate controls aimed not only to discourage any fraudulent behavior but also to reduce risks for consumer health.     

First Isolation of Rotavirus Associated with Neonatal Diarrhoea in Guanacos (Lama guanicoe) in the Argentinean Patagonia Region

Group A rotavirus (RV) and coronavirus (CV) are common viral pathogens associated with neonatal diarrhoea in numerous animal species. The purpose of this work was to investigate the presence of these viral agents in two farm populations of captured guanacos (Lama guanicoe) in the Argentinean Patagonia region, that developed severe diarrhoea outbreaks. Stool and serum samples were analysed for RV and bovine CV antigen and antibody by enzyme‐linked immunosorbent assay. Rotavirus was detected in faeces from two new‐born guanacos with acute diarrhoea, one in each farm. After electrophoretic analysis, each isolated strain, showed a distinctive long dsRNA electropherotype characteristic of group A rotaviruses (4:2:3:2). In addition, 95% (38 of 40) of the sampled animals were positive for RV antibodies, suggesting a high prevalence of RV infection in the populations tested. No evidence of CV circulation by antigen or antibody analysis was observed. To our knowledge, this is the first report of the detection and isolation of RV associated with neonatal diarrhoea in Lama guanicoe.     

alpha(1)-Acid glycoprotein modulates apoptosis in bovine monocytes

alpha(1)-Acid glycoprotein (AGP, orosomucoid) is a normal constituent of bovine blood. AGP is an immunocalin, a binding protein that can also exert several immunomodulatory functions. In this paper we investigated the effect of bovine alpha(1)-acid glycoprotein (boAGP) on spontaneous and staurosporine-induced apoptosis of blood derived monocytes purified using magnetic cell sorting techniques. Bovine AGP was purified from blood following a chromatographic protocol. The homogeneous protein was used to stimulate the cells as well to raise a polyclonal antibody, that was used throughout all the experiments. When monocytes were incubated with high concentrations of boAGP (0.9 mg/ml), similar to those found in bovine plasma during systemic reaction to inflammation, their spontaneous apoptosis rate was suppressed, as determined by caspase-3/7 enzymatic activity assay. Similar results were obtained when apoptosis was induced by adding staurosporine, a potent protein kinase inhibitor. The apoptosis-modulating activity of boAGP was dependent on its concentration, since physiological concentrations of boAGP (0.3 mg/ml) did not exhibit a statistically significative anti-apoptotic activity. We also investigated whether this apoptosis-modulating activity was dependent on the terminal sialic acid residues exposed on the surface of the protein. Enzymatic treatment with neuraminidase, that cleaves terminal sialic acid residues, completely abolished boAGP's anti-apoptotic activity. These results suggest that the protective effect of AGP is likely mediated by its sialic acid terminal groups.     

A rapid chromatographic method for determining the glucosinolate content in crambe seed

Crambe abyssinica is becoming a reliable source of industrial oil with good technical characteristics because of its high erucic acid content. The possibility of using the industrial defatted meal of crambe as a feed could further promote the interest in this crop, but is prevented by the high glucosinolate content (70‐150 μmol/g). Therefore the possibility of reducing this parameter by selection was evaluated and a search for a new quick analytical procedure begun. When hydrolysed by myrosinase (thioglucoside glucohydrolase EC 3.2.3.1.), at pH 6.5 or higher, epiprogoitrin, the main glucosinolate in crambe seed (>90% of total glucosinolates), gives (5R)‐5‐vinyl‐1,3‐oxazolidine‐2‐thione quantitatively. The latter can be easily estimated by high‐performance liquid chromatography. This is the background of the proposed analytical procedure that permits rapid analysis of about 100 samples per day using about 25 mg of crambe meal (five seeds) for assay. The method can also be used to select for glucosinolate content in rapeseed and, probably with some adjustment, all crops that have a 2‐hydroxy‐glucosinolate as the main glucosinolate.     

High levels of genetic differentiation between Wolbachia-infected and non-infected populations of Folsomia candida (Collembola, Isotomidae)

The Gram-negative -Proteobacterium Wolbachia pipientis has been described as an obligate endosymbiont in many arthropod species, where it induces a variety of reproductive alterations, including parthenogenesis. Recently, this microorganism has also been detected in the parthenogenetic collembolan Folsomia candida. Here, we confirm the occurrence of the endosymbiont also in two Italian parthenogenetic populations of F. candida using ultrastructural (electron microscopy) and molecular (PCR screening on two bacterial genes) evidence. The strain isolated in the Italian populations has almost-identical gene sequences compared to that previously isolated in other populations of F. candida. In addition, we discovered a population of Folsomia cf. candida, which showed the presence of both males and females. This population is not infected by Wolbachia. A screening of two mitochondrial genes (COI and COII) showed that the bisexual population has high levels of genetic divergence in comparison with the parthenogenetic ones, even suggesting the possibility that it belongs to a different species. Furthermore, the remarkably high levels of genetic divergence between the two parthenogenetic populations suggests a possible influence of Wolbachia on inducing such differentiation, and, in the long term, speciation.     

Carboplatin sustained delivery system using injectable microspheres

A controlled carboplatin delivery system using biodegradable polymer has been used in this study. The purpose was to evaluate the local and systemic effects of injectable, biodegradable microspheres containing carboplatin when injected as aqueous suspension subcutaneously in rats. Biocompatibility and toxicity of empty microspheres and microspheres loaded with carboplatin were evaluated by clinical and histological examination. The diffusion of carboplatin in tissues and time of drug release were evaluated by platinum determination in plasma and tissues over the time. The results of the study suggest that microspheres provide a sustained slow release of carboplatin and that multiple inoculations of microspheres containing drug and no evidence of local or systemic toxicity is found. This device may be useful in the treatment of solid tumours.