Study of Organic Matter Evolution in Citrus Compost by Isoelectrofocusing Technique

Three composts from citrus-processing industry wastes, sampled at prefixed times during the composting process, were studied with the aim to follow the organic matter evolution by using the isoelectrofocusing (IEF) technique. Results indicated that IEF qualitative analyses allowed evaluation of the organic matter transformation during composting process, showing a decrease of IEF peaks focused at lower values of pH (less stabilized organic matter) and a corresponding increase of peaks focused at higher value of pH (more humified material). The parameter A %, defined as the areas sum of IEF peaks focused at pH>4.7, could be considered particularly effective as a “threshold value” to evaluate the level of organic matter evolution for the considered composts.     

Porcine circovirus type 2 detection in in vitro produced porcine blastocysts after virus sperm exposure

This study was aimed at assessing the capability of semen experimentally infected with porcine circovirus type 2 (PCV2) to produce porcine blastocysts PCR positive for PCV2. Embryos were obtained from in vitro maturation (IVM) and in vitro fertilization (IVF) of porcine oocytes or by parthenogenesis. Sperm suspension was exposed to PCV2b and utilized for IVF. PCV2 spiked semen did not reveal any reduction in sperm viability or motility but its ability to produce infected blastocysts was irrelevant as only one out of 15 blastocysts obtained by IVF were PCV2b; however two blastocysts were PCV2a positive. Furthermore, the presence of PCV2 was demonstrated also in embryos obtained by parthenogenesis (one out of 17 was PCV2b and one PCV2a positive). Even if PCV2 firmly attaches to the surface of spermatozoa, experimentally spiked sperm were not effective in infecting oocytes during IVF and in producing PCR positive embryos. The infected blastocysts we obtained derived most probably from infected oocytes recovered at the abattoir.     

Serum Protein Fraction in Mature Horses and Relationship With Metabolic and Hematological Parameters

This study was conducted to determine the effect of age on serum protein fractions and their relationship with metabolic and hematological profiles in mature horses. Twenty-five mature Italian Saddle horses (mean age 13.6 ± 4.8 years) fed the same diet (grass hay and concentrate) were stratified according to age as first maturity, M1 (≤10 years old); second maturity, M2 (>10 and <15 years old); and old, O (>15 years), to be monitored every 28 days for a continuous period of 140 days. Horses in group O had higher plasma protein and thiol concentrations and white blood cell and neutrophil counts than the other two groups. Serum α₂-globulin concentrations were positively correlated with total plasma cholesterol (r = 0.514; P < .001), alkaline phosphatase (r = 0.430; P < .001), aspartate aminotransferase (r = 0.339; P < .001), ceruloplasmin (r = 0.321; P < .001), glutamic pyruvate transaminase (r = 0.444; P < .001), reactive oxygen metabolites (r = 0.426; P < .001), and blood neutrophil counts (r = 0.344; P < .01), and negatively with plasma bilirubin (r = −0.522; P < .001) and creatinine (r = −0.400; P < .001). These results suggest differences in hematological and metabolic profile in Italian Saddle horses after 15 years of age, resulting mainly from changes in plasma proteins and inflammatory mediators. The α₂-globulins fraction seems a quick but reliable marker of an inflammatory situation that, successively, should be better investigated with specific metabolites or enzymes.     

Comparative genome sequencing identifies a prophage-associated genomic island linked to host adaptation of Lawsonia intracellularis infections

Lawsonia intracellularis is an obligate intracellular bacterium and the causative agent of proliferative enteropathy (PE). The disease is endemic in pigs, emerging in horses and has also been reported in a variety of other animal species, including nonhuman primates. Comparing the whole genome sequences of a homologous porcine L. intracellularis isolate cultivated for 10 and 60 passages in vitro, we identified a 18-kb prophage-associated genomic island in the passage 10 (pathogenic variant) that was lost in the passage 60 (non-pathogenic variant). This chromosomal island comprises 15 genes downstream from the prophage DLP12 integrase gene. The prevalence of this genetic element was evaluated in 12 other L. intracellularis isolates and in 53 infected animals and was found to be conserved in all porcine isolates cultivated for up to 20 passages and was lost in isolates cultivated for more than 40 passages. Furthermore, the prophage region was also present in 26 fecal samples derived from pigs clinically affected with both acute and chronic forms of the disease. Nevertheless, equine L. intracellularis isolates evaluated did not harbor this genomic island regardless of the passage in vitro. Additionally, fecal samples from 21 clinically affected horses and four wild rabbits trapped in horse farms experiencing PE outbreaks did not show this prophage-associated island. Although the presence of this prophage-associated island was not essential for a virulent L. intracellularis phenotype, this genetic element was porcine isolate-specific and potentially contributed to the ecological specialization of this organism for the swine host.     

Antibacterial efficacy of polypyrrole in textile applications

This paper describes application and evaluation of polypyrrole as an antibacterial polymer. Polypyrrole was produced embedding two doping agents: chloride and dicyclohexyl sulfosuccinate ions. Stability of the antibacterial efficacy of polypyrrole deposited on cotton fabrics was assessed before and after three different kinds of washing (namely, laundering with anionic and non-ionic detergents and dry-cleaning). Polypyrrole showed excellent antibacterial properties (100 % of bacterial reduction) against Escherichia coli for both doping agents. Treated fabrics were further characterised by scanning electron microscopy, energy dispersive X-ray analysis and infrared spectroscopy. The antibacterial efficacy diminished after launderings with anionic and non-ionic detergents because of two different mechanisms: the neutralisation of positive charges under alkali conditions (dedoping), and a partial removal of polypyrrole by abrasion and surfactant action. After dry-cleaning, polypyrrole embedding chloride and dicyclohexyl sulfosuccinate ions still showed excellent antibacterial efficacy. Moreover, scanning electron microscopy investigations were used to intuitively explain the bactericidal mechanism of polypyrrole on Escherichia coli bacteria.     

Effects of intertrochanteric varus osteotomy on Norberg angle and percent coverage of the femoral head in displastic dogs

This study was conducted to assess the effects of femoral varus osteotomy on joint congruency in dogs affected by early stage hip dysplasia. Preoperative planning to move the femoral head within the acetabulum was carried out. Varisation of the femoral inclination angle (fIA) was achieved by Intertrochanteric Osteotomy (ITO). Norberg angle (NA), percent coverage (PC) of the femoral head by the acetabulum and fIA was measured from preoperative, immediate postoperative and first and second recheck radiographs of seven dogs that underwent an ITO (joint n = 9). There was significant (p < 0.05) improvement of both NA and PC in all patients as indicated by a change in the mean ± standard deviation of 78.9° ± 7.5 and 36.9% ± 5.2 to 92.2° ± 6.7 and 50.6% ± 8.3, respectively. No significant difference (p < 0.05) was observed between the values of the planned femoral inclination angle (pfIA) of the femur and the effective femoral inclination angle (efIA) obtained after surgery (115.9° ± 2.5 and 111.3° ± 6.4, respectively). These findings could encourage the use of ITO in veterinary practice and indicate that intertrochanteric varus osteotomy should be re-considered for the treatment of early stage hip dysplasia in dogs with radiological signs of joint incongruency.     

Feasible application of a portable NIR-AOTF tool for on-field prediction of phenolic compounds during the ripening of olives for oil production

Olive fruits of three different cultivars (Moraiolo, Dolce di Andria, and Nocellara Etnea) were monitored during ripening up to harvest, and specific and total phenols were measured by HPLC (High Pressure Liquid Chromatography). On the same olive samples (n = 450), spectral detections were performed using a portable NIR (Near Infrared)-AOTF (Acousto Optically Tunable Filter) device in diffuse reflectance mode (1100-2300 nm). Prediction models were developed for the main phenolic compounds (e.g., oleuropein, verbascoside, and 3,4-DHPEA-EDA) and total phenols using Partial Least Squares (PLS). Internal cross-validation (leave-one-out method) was applied for calibration and prediction models developed on the data sets relative to each single cultivar. Validation of the models obtained as the sum of the three sample sets (total phenols, n = 162; verbascoside, n = 162; oleuropein, n = 148; 3,4-DHPEA-EDA, n = 162) were performed by external sets of data. Obtained results in term of R(2) (in calibration, prediction and cross-validation) ranged between 0.930 and 0.998, 0.874-0.942, and 0.837-0.992, respectively. Standard errors in calibration (RMSEC), cross-validation (RMSECV), and prediction (RMSEP) were calculated obtaining minimum error in prediction of 0.68 and maximum of 6.33 mg/g. RPD ratios (SD/SECV) were also calculated as references of the model effectiveness. This work shows how NIR-AOTF can be considered a feasible tool for the on-field and nondestructive measurement of specific and total phenols in olives for oil production.     

The Effect of the Organic Matter Composition on POP Accumulation in Soil

The effect of different humic fractions on polychlorinated biphenyl (PCB) contamination in soils was tested in the field by means of 53 soil samples from a high-altitude grassland plateau in the Italian Alps. Three humic fractions (humin, humic acids, and fulvic acids) were characterized in parallel by quantifying 12 PCB congeners to establish a direct relationship between PCB levels and humic fraction concentrations. Humin (the most hydrophobic fraction) appears to be the most closely correlated with the amount of PCBs in soil (R ²?=?0.83), while fulvic acid shows the lowest correlation (R ²?=?0.49). The idea of preferential sorption of hydrophobic compounds in the humin fraction is discussed, and the humin carbon content (f _huminC) is proposed as an improved parameter for evaluating the potential for POP accumulation in soils, replacing total organic carbon (f _oc). Congener studies revealed that penta- and hexa-substituted-CBs show the optimal combination of physicochemical properties for equilibrating with the humin content in soil. Moreover, f _huminC/f _oc is conceptually equivalent to the empirical coefficients used in predictive K _sa equations. In our samples, the f _huminC/f _oc was 0.55, a value in between the empirical coefficients proposed in the literature. In predictive equations, the use of f _huminC instead f _oc avoids the necessity of using an empirical parameter for a ??generic?? condition by introducing an experimental parameter (f _huminC) that takes into account local conditions (organic matter composition).     

Towards an ecologically-sustainable forestry in the Atlantic Forest

Since pristine Atlantic Forest remnants are vanishing, and biological reserves are in short, conservation of biodiversity will largely depend on proper management of the anthropogenic matrix. Here, we test (1) the effectiveness of ecologically-managed tree monocultures in maintaining Araucaria Forest biodiversity, (2) how this effectiveness change among taxa, and (3) we discuss management principles that can be used by the forestry industry in order to contribute positively to biodiversity conservation. The study was conducted in the São Francisco de Paula National Forest, southern Brazil, an environmental mosaic composed mostly of patches of Araucaria Forest and ecologically-managed monocultures of Araucaria, Pinus and Eucalyptus. Using standardized sampling methods in these four main habitats, we have recorded the richness and species composition of small mammals, birds, leaf-litter frogs, butterflies, galling insects, spiders, opiliones, flatworms, woody plants, epiphytic angiosperms, epiphytic ferns, lichens, and fruit-body producing fungi. Overall, we recorded 506 species in Araucaria Forest, 181 (36%) of which were exclusive of this habitat while 325 (64%) could be found in at least one monoculture. Distribution patterns of species richness and number of records across taxonomic groups showed that a large biodiversity can be found inside ecologically-managed plantations of Araucaria, Pinus, and Eucalyptus. For all studied taxa, except for epiphytic angiosperms and fruit-body producing fungi, more than half of the Araucaria Forest species could be found living on monocultures. We discuss how the actual management practices of the forest industry can be improved to contribute positively to the conservation of the Atlantic Forest biodiversity.     

PCV2-DNA in formalin-fixed and paraffin embedded lymph nodes of wild boar (Sus scrofa ssp. scrofa): one sampling approach for two laboratory techniques

Superficial inguinal lymph nodes from 72 wild boars examined in a previous immunohistochemical (IHC) study on porcine circovirus type 2 (PCV2) were selected for a PCV2 polymerase chain reaction (PCR) analysis. Four of these lymph nodes were PCV2-IHC strongly positive with PMWS histological lesions (outcome 1), 6 weak to mild PCV2-IHC positive without PMWS histological lesions (outcome 2) and 62 PCV2-IHC negative. Considering IHC the gold standard for diagnosis, the aims of the study were to evaluate the suitability of the PCV2-DNA extraction from formalin-fixed and paraffin-embedded (FFPE) tissue and the sensitivity and specificity of PCR under two IHC interpretations criteria: (A) the sample was considered positive if the result was outcome 1; (B) the sample was considered positive if the result was outcome 1 or 2. Under (A) criteria, sensitivity and specificity of PCR were 100% and 89.7%, respectively; the Cohen''s Kappa coefficient was 0.49. Under (B) criteria, sensitivity and specificity of PCR were 80.0% and 95.2%, respectively; the Cohen''s Kappa coefficient was 0.72. The high Cohen''s Kappa coefficient under the (B) interpretative criteria indicates good agreement between the two methods. In conclusion, 1) DNA extracted from FFPE specimens of wild boar is suitable for PCR and further represents a screening test for PCV2/PCVD (PCV2 Diseases) investigations in wild boar as well; 2) routine histological sampling can also be useful for PCV2 virological studies in wild boar.