Effect of high-pressure-moderate-temperature processing on the volatile profile of milk

The effects of high hydrostatic pressure on volatile generation in milk were investigated in this study. Raw milk samples were treated under different pressures (482, 586, and 620 MPa), temperatures (25 and 60 degrees C), and holding times (1, 3, and 5 min). Samples submitted to heat treatments alone (25, 60, and 80 degrees C for 1, 3, and 5 min) were used for comparison. Trace volatile sulfur compounds were analyzed using solid-phase microextraction (SPME) and gas chromatography (GC) with pulsed-flame photometric detection (PFPD), whereas the rest of the volatile compounds were analyzed using SPME-GC with flame ionization detection (FID). Multivariate analysis of variance (MANOVA) and principal component analysis (PCA) were used to study the effect of pressure, temperature, and time on volatile generation. Relative concentration increases of 27 selected volatile compounds were compared to an untreated sample. It was found that pressure, temperature, and time, as well as their interactions, all had significant effects (P < 0.001) on volatile generation in milk. Pressure and time effects were significant at 60 degrees C, whereas their effects were almost negligible at 25 degrees C. The PCA plot indicated that the volatile generation of pressure-heated samples at 60 degrees C was different from that of heated-alone samples. Heat treatment tended to promote the formation of methanethiol, hydrogen sulfide, methyl ketones, and aldehydes, whereas high-pressure treatment favored the formation of hydrogen sulfide and aldehydes.     

Biofilm-forming ability profiling of Staphylococcus aureus and Staphylococcus epidermidis mastitis isolates

Biofilm-forming ability has been increasingly recognized as an important virulence factor in Staphylococci, facilitating their persistence in the host, evading its defences and allowing bacterial survival at high antimicrobial concentrations. Staphylococcus aureus remains a major pathogen of chronic mastitis, but in the last years Staphylococcus epidermidis has emerged as a relevant mastitis pathogen. The present work aimed at the evaluation of the biofilm-forming ability of Staphylococci field isolates from bovine subclinical mastitis and at the development of a fluorescent in situ hybridisation (FISH) protocol that would allow the direct observation of biofilm formation in milk samples. The analysis of phenotypic expression in Congo Red Agar (CRA) and by FISH, showed that 37.5% of the S. aureus isolates produced biofilm, while by optical density measurement only 18.75% isolates revealed this phenotype. The results showed a fair agreement according to the kappa coefficient test (kappa = 0.259). Regarding S. epidermidis mastitis isolates, 37.5% revealed the ability to produce biofilm, but only four isolates were positive by all methods. This agreement was moderate (kappa = 0.467). The application of FISH to artificially contaminated milk samples allowed the direct observation of biofilm production by 37.5% isolates, showing total agreement with the CRA results. This method better mimics the in vivo conditions, especially in terms of the presence of calcium and iron, which in high concentrations, respectively, are known to inhibit or induce biofilm production.     

Evaluation of antiviral activity of phenolic compounds and derivatives against rabies virus

Human rabies is a viral disease with a great impact on public health, mainly on account of its fatal course in the majority of cases. Despite the well-established prophylaxis by immunization, rabies is believed to be responsible for 40,000-70,000 human deaths per year, mostly in endemic areas. Palliative support and experimental protocols to avoid death have been employed with no expressive results, with the exception of a recent human case of recovery from rabies. No antiviral drugs are currently available to fight against this infection. In combination with the prophylaxis, an antiviral drug would be useful for human rabies treatment, providing enhanced protection against the encephalitis caused by the virus. Phenolic compounds are derived from the secondary plant metabolism, although they can also be obtained by synthetic processes. Many studies have shown a great range of pharmacological effects for these substances, including vasodilatation, antiallergenic, antiinflammatory and antiviral properties, among others. In this study, the potential in-vitro anti-rabies activity of 24 synthetic phenolic compounds was evaluated using McCoy cells and PV rabies strain. The cytotoxicity (CC50) was assayed by the MTT method and the antiviral activity (IC50) was estimated by the inhibition of viral cytopathic effects. Isoprinosine and ketamine were used as positive controls. The tested compounds showed selectivity indices (SI=CC50/IC50) ranging from 1.0 to 3.9. Six phenolic compounds failed to inhibit the cytopathic effect to any degree, and four showed SI > or = 3.0. According to these results, some probable structure-activity relationships are suggested. It was observed that the presence of free hydroxyl and ether groups influenced the anti-rabies activity. However, additional studies are required to establish these relationships.     

Inhibition of hemoglobin- and iron-promoted oxidation in fish microsomes by natural phenolics

Natural phenolic antioxidants have been tested in hake (Merluccious merluccious) microsomes as inhibitors of lipid oxidation promoted by fish muscle prooxidants: hemoglobin (Hb), enzymatic NADH-iron and nonenzymatic ascorbate-iron. The phenolics selected were as follows: (a) a grape phenolic extract (OW), (b) a fraction (IV) with isolated grape procyanidins with a medium-low degree of polymerization and galloylation percentage, (c) hydroxytyrosol obtained from olive oil byproducts, and (d) a synthetic phenolic antioxidant, propyl gallate. All compounds delayed lipid oxidation activated by Hb, enzymatic NADH-iron, and nonenzymatic ascorbate-iron, excluding hydroxytyrosol that was not an effective antioxidant on oxidation promoted by nonenzymatic iron. The relative antioxidant efficiency was independent of the prooxidant system, IV > propyl gallate > OW > hydroxytyrosol, and showed a positive correlation with their incorporation into microsomes (p < 0.05). The reducing capacity or ability for donating electrons and the chelating properties may also contribute to the antioxidant activity of phenolics, although these factors were less decisive than their affinity for incorporating into the microsomes. Conversely, the inhibition of Hb oxidation by phenolics and their polarity did not seem to play an important role on antioxidant mechanism. These results stressed the importance of incorporating the exogenous antioxidants into the membranes where are located key substances for fish lipid oxidation (highly unsaturated phospholipids, iron-reducing enzymes, and endogenous alpha-tocopherol).     

Hemodynamic effects of methylprednisolone acetate administration in cats

OBJECTIVE: To investigate the mechanisms by which corticosteroid administration may predispose cats to congestive heart failure (CHF). ANIMALS: 12 cats receiving methylprednisolone acetate (MPA) for the treatment of dermatologic disorders. PROCEDURE: The study was conducted as a repeated-measures design. Various baseline variables were measured, after which MPA (5 mg/kg, IM) was administered. The same variables were then measured at 3 to 6 days and at 16 to 24 days after MPA administration. Evaluations included physical examination, systolic blood pressure measurement, hematologic analysis, serum biochemical analysis, thoracic radiography, echocardiography, and total body water and plasma volume determination. RESULTS: MPA resulted in a substantial increase in serum glucose concentration at 3 to 6 days after administration. Concurrently, RBC count, Hct, and hemoglobin concentration as well as serum concentrations of the major extracellular electrolytes, sodium and chloride, decreased. Plasma volume increased by 13.4% (> 40% in 3 cats), whereas total body water and body weight slightly decreased. All variables returned to baseline by 16 to 24 days after MPA administration. CONCLUSIONS AND CLINICAL RELEVANCE: These data suggest that MPA administration in cats causes plasma volume expansion as a result of an intra to extracellular fluid shift secondary to glucocorticoid-mediated extracellular hyperglycemia. This mechanism is analogous to the plasma volume expansion that accompanies uncontrolled diabetes mellitus in humans. Any cardiovascular disorders that impair the normal compensatory mechanisms for increased plasma volume may predispose cats to CHF following MPA administration.     

Differential induction of pathogenesis-related proteins in banana in response to Mycosphaerella fijiensis infection

Black leaf streak or “black Sigatoka” is one of the most important diseases affecting bananas and plantains worldwide. Very few studies have been published on the host-pathogen interaction of this pathosystem, particularly at the molecular level. The aim of this work was to analyze, under controlled conditions, the enzyme activity of peroxidase (POX), phenylalanine ammonia lyase (PAL), β-1, 3-glucanase (GLU) and chitinase (CHI) as well as the production of H₂O₂ in banana plants infected with Mycosphaerella fijiensis. Defence responses were examined and compared in a resistant (Calcutta 4) and a susceptible (Williams) cultivar. Plants were inoculated and tested for enzyme activity at 0, 6, 12, 18, 24, 48 and 72?h after infection (HAI) and 6, 9, 12, 15 and 18?days after inoculation (DAI). A rapid induction of PAL, POX and GLU was observed in the resistant cultivar at 6–18 HAI as well as H₂O₂ production at 72 HAI. In contrast, in the susceptible cultivar, induction of these enzymes was only observed from 6 DAI. These results suggest that the first 72 HAI are important in determining the response of the host to the disease. Further studies characterizing banana responses to M. fijiensis at the early stages of the infection are necessary in order to better understand this host-pathogen interaction.     

Biological control of postharvest diseases of fruit

Postharvest decay in harvested fruit causes considerable economical losses. Fungicides are the primary means to control these losses. Public concern in food safety and environmental issues and the increase of pathogen resistant populations have enhanced the interest in developing alternative methods to fungicides to control postharvest fruit decay. During the last two decades a huge information and advances concerning the selection of antagonists, mode of action, different approaches to enhance biocontrol activity, formulation and production have been achieved, and some biofungicides are already in the market. It is likely that several more products will enter the market in the near future, as the result of the biological control research programs worldwide. Nonetheless, it is necessary to continue finding new potential microorganisms, better understanding the mode of action, and pathogen, antagonist and host interactions, to increase the potential of biocontrol helping to become a real alternative to synthetic postharvest fungicides. This article presents an overview of postharvest biological control approaches and explores new research possibilities to improve biocontrol activity.     

Increasing energy and lysine in diets for growing-finishing pigs in hot environmental conditions: consequences on performance, digestibility, slurry composition, and gas emission

The influence of dietary nutrient concentration on growth performance, manure composition, and gas emission was studied in pigs in hot environmental conditions. A total of 64 intact males and 64 females [(Landrace × Large White) × Pietrain] weighing 63.1 ± 9.7 kg were divided into 2 dietary treatments: high (HD: 14.39 MJ of DE/kg and 1.11% Lys) and low (LD: 13.97 MJ of DE/kg and 1.01% Lys) in energy and Lys contents. Pigs were allocated to 32 split-sex pens with 4 pigs/pen and 16 pens/treatment. Average productive performance was recorded for 41 d (phase 1). After phase 1, 12 females of 103.3 ± 3.15 kg (6 per treatment) were selected and housed individually, and feces and slurry were collected during 3 and 4 consecutive days, respectively, to calculate nutrient digestibility and measure gas emissions (phase 2). For gas emission measurements, slurry was pooled by treatment and stored for 76 d. Initial composition of slurry and pH were analyzed. Maximum and minimum temperatures registered in the barn throughout the growing period were 35.1 and 18.1°C, respectively. Animals fed the HD diet grew more efficiently than pigs fed the LD diet (G:F, 0.43 vs. 0.40; SEM = 0.01; P < 0.05). Fat digestibility was greater in HD compared with LD pigs (88.0 vs. 84.9%; SEM = 0.9; P < 0.05). Slurry from pigs fed the LD diet showed greater DM, OM, total N, and VFA contents than slurry from pigs fed the HD diet. Cumulative NH(3), CO(2), and especially CH(4) emissions were greater in the HD slurry compared with the LD slurry (192.4 vs. 210.g of NH(3)/m(3); 2,712 vs. 3,210 g of CO(2)/m(3); 1,502 vs. 2,647 mL of CH(4)/kg of OM). Increasing feed density in the present study led to a more efficient growth, a decreased nutrient concentration in the slurry, and a greater gas emission.     

A data mining approach to improve multiple regression models of soil nitrate concentration predictions in Quercus rotundifolia montados (Portugal)

The changes in the soil nitrate concentration were studied during 2 years in a “montado” ecosystem, in the South of Portugal. Total rainfall, air and soil temperature and soil water content under and outside Quercus rotundifolia canopy were also evaluated. A cluster analysis was carried out using climatic and microclimatic parameters in order to maximize the intraclass similarity and minimize the interclass similarity. It was used the k-Means Clustering Method. Several cluster models were developed using k values ranging between 2 and 5. Thereafter, in each cluster, the data were divided according to their origin (soil under canopy and open areas, and from surface and deep layers). Multiple regression models were tested for each cluster, to assess the relationship between soil nitrate concentration and a set of climatic and microclimatic parameters and the results were compared with models assessed without clustering. The models achieved with data grouped in result of clustering analysis showed better performance than the models achieved without clustering, mostly for data from open areas soils. When temperature is low and/or water presents excess or scarcity levels, the data from soils in undercanopy areas, give rise to models with worst performance than models from open soil areas data. The results obtained for undercanopy area suggest that nitrification process in soil under Quercus rotundifolia trees influence is more complex than for open areas, and subject to other relevant factors beyond water and temperature.