Structural features of lignin obtained at different alkaline oxidation conditions from sugarcane bagasse

Lignin is a macromolecule frequently obtained as residue during technological processing of biomass. Modifications in chemical structure of lignin generate valuable products, some with particular and unique characteristics. One of the available methods for modification of industrial lignin is oxidation by hydrogen peroxide. In this work, we conducted systematic studies of the oxidation process that were carried out at various pHs and oxidizing agent concentrations. Biophysical, biochemical, structural properties of the oxidized lignin were analyzed by UV spectrophotometry, Fourier transform infrared spectroscopy, scanning electron microscopy and small angle X-ray scattering. Our results reveal that lignin oxidized with 9.1% H₂O₂ (m/v) at pH 13.3 has the highest fragmentation, oxidation degree and stability. Although this processing condition might be considered quite severe, we have concluded that the stability of the obtained oxidized lignin was greatly increased. Therefore, the identified processing conditions of oxidation may be of practical interest for industrial applications.     

Development of molecular markers to assess the level of introgression of Populus tremula into P. alba natural populations

Morphological traits traditionally adopted to discriminate between Populus alba L. and P. tremula L. have frequently led to misclassifi‐cation of their spontaneous hybrid P. × canescens Sm. Moreover, they may not be of any help in cases of spontaneous backcross phenomena. These limitations can be overcome by molecular markers, which are not environmentally influenced nor subjectively assessed. In this study, the effectiveness of amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers in species and hybrid discrimination was evaluated by analysing a set of reference samples of P. alba, P. tremula and P. × canescens. Species‐specific and species‐indicative AFLPs, as well as diagnostic SSR alleles, were recorded in both P. alba and P. tremula reference samples. The results allowed a clear distinction between the two poplar species and their hybrid. Using these diagnostic markers, a natural population of P. alba trees sampled along the Ticino river basin in northern Italy was analysed, and P. × canescens individuals, intermingled with P. alba trees, were detected.     

Estimating clinical chemistry reference values based on an existing data set of unselected animals

In an attempt to standardise the determination of biological reference values, the International Federation of Clinical Chemistry (IFCC) has published a series of recommendations on developing reference intervals. The IFCC recommends the use of an a priori sampling of at least 120 healthy individuals. However, such a high number of samples and laboratory analysis is expensive, time-consuming and not always feasible, especially in veterinary medicine. In this paper, an alternative (a posteriori) method is described and is used to determine reference intervals for biochemical parameters of farm animals using an existing laboratory data set. The method used was based on the detection and removal of outliers to obtain a large sample of animals likely to be healthy from the existing data set. This allowed the estimation of reliable reference intervals for biochemical parameters in Sarda dairy sheep. This method may also be useful for the determination of reference intervals for different species, ages and gender.     

Bovine and rabbit models for the study of a Staphylococcus aureus avirulent mutant strain, RC122

Staphylococcus aureus is the main etiological agent of bovine mastitis. Intramammary infections are difficult to cure and vaccination appears to be an alternative to prevent the disease. Research has focused on the development of mutants affected in the synthesis of pathogenicity determinants. We constructed a mutant strain (RC122) after chemical mutagenesis. In a mouse model, the strain was shown to be 1500 times less virulent, showed similar kinetics of disappearance in the kidney as its parental strain, and a good degree of protection against a challenge from homologous and heterologous strains.

The objective of the present report was to study the avirulent RC122 S. aureus mutant strain in rabbit and bovine infection models. The results clearly show that RC122 was less virulent than its parental strain in a rabbit skin model, and was also correlated with its avirulence as an udder pathogen. These traits make the RC122 mutant strain interesting as a potential strain for an experimental vaccine trial in dairy herds.

     

Incidental captures of sea turtles by swordfish and albacore longlines in the Ionian sea

ABSTRACT:   Incidental catches in pelagic longline fishing pose a serious threat to sea turtle populations throughout the world's seas and oceans. In this work, carried out in the framework of the EC-DG-Fisheries 98/008 project, information on sea turtle catch rates from swordfish and albacore longline fishing activities observed in Italian waters off the Ionian Sea during 1999 and 2000, are reported. In addition, biometric data, health status, and tagging return rate of sea turtles captured are provided. A total of 200 sea turtles were caught (198 loggerhead turtles and 2 green turtles), comprising 0.5–15.7% of the total catch in number of individuals. The estimates of the sea turtles caught by the total fishing effort of both longlines in the whole study area were 1084 specimens in 1999 (95% CI = 667–1502) and 4447 specimens in 2000 (95% CI = 3189–5705). Although all sea turtles were released alive, nearly half of them had hooks that could not be removed and remained deeply embedded in the digestive tract.     

Larval population structure of Engraulis encrasicolus in the Strait of Sicily as revealed by morphometric and genetic analysis

In order to investigate the larval population structure, specimens of Engraulis encrasicolus larvae from five different locations in the Strait of Sicily were analyzed by means of otolith readings, morphometric and genetic techniques. The distribution of age in day, identified by means of the otolith readings and associated with the oceanographic parameters, was useful to identify possible spawning areas and transport dynamics. The presence of more than one spawning area suggested the possibility that two or more sub‐populations may co‐exist in the study area. The morphometric characteristics were more adequate than genetic parameters to discriminate the different larval groups. The most relevant variables for the separation were the mouth length (ML) and the body diameter (BD). The population structure by means of genetic data reported the presence of two phylogroups co‐occurring among samples in each sampling locations specimens. The pattern of genetic divergence among anchovy larvae in the Strait of Sicily was congruent with previous studies conducted on adult populations present in other Mediterranean areas with different molecular markers. The habitat‐specific nature of the morphological variation and the lack of corresponding genetic variation among larvae from the different locations suggested that the observed differences in morphology could be linked to environmental parameters. The body form differences among different larvae samples could reflect the nutritional status of larvae. In fact, these differences were found among anchovy larvae collected in areas with different oxygen and fluorescence, which is an index of primary productivity and is linked to the availability of food for anchovy larvae.     

Biological characterization of bovine herpesvirus 1 recombinants possessing the vaccine glycoprotein E negative phenotype

Intramolecular recombination is a frequent event during the replication cycle of bovine herpesvirus 1 (BoHV-1). Recombinant viruses frequently arise and survive in cattle after concomitant nasal infections with two BoHV-1 mutants. The consequences of this process, related to herpesvirus evolution, have to be assessed in the context of large use of live marker vaccines based on glycoprotein E (gE) gene deletion. In natural conditions, double nasal infections by vaccine and wild-type strains are likely to occur. This situation might generate virulent recombinant viruses inducing a serological response indistinguishable from the vaccine one. This question was addressed by generating in vitro BoHV-1 recombinants deleted in the gE gene from seven wild-type BoHV-1 strains and one mutant strain deleted in the genes encoding gC and gE. In vitro growth properties were assessed by virus production, one step growth kinetics and plaque size assay. Heterogeneity in the biological properties was shown among the investigated recombinant viruses. The results demonstrated that some recombinants, in spite of their gE minus phenotype, have biological characteristics close to wild-type BoHV-1.     

Real-time PCR for Quantification of Toxigenic Fusarium Species in Barley and Malt

A real-time PCR technique was applied for the quantification of trichothecene-producing Fusarium species (TMTRI assay) as well as the highly toxigenic Fusarium graminearum (TMFg12 assay) present in barley grain and malt. PCR results were compared to the amounts of trichothecenes detected in the samples to find out if the PCR assays can be used for trichothecene screening instead of expensive and laborious chemical analyses. DNA was extracted from ground kernels using a commercial DNA extraction kit and analysed in a LightCycler® system using specific primers and fluorogenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium DNA and F. graminearum DNA present in barley grain and malt samples, respectively. Both TaqMan assays were considered to be sensitive and reproducible. Linearity of the assays was 4–5 log units when pure Fusarium DNAs were tested. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay could be used for estimation of the deoxynivalenol (DON) content in barley grain. Furthermore, the TMFg12 assay for F. graminearum gave a good estimation of the DON content in north American barley and malt samples, whilst the correlation was poor among Finnish samples. DON content and the level of F. graminearum DNA were found to be naturally low in Finnish barleys.