Evaluation of alternative sanitizers to chlorine disinfection for reducing foodborne pathogens in fresh-cut apple

The risk of undesirable by-products from chlorine disinfection in fresh-cut industries, together with its limited efficacy, has led to a search for alternative agents. The aim of this study was to test several alternative putative antimicrobial substances to reduce Escherichia coli O157:H7, Salmonella spp. and Listeria spp. populations on fresh-cut apple. Carvacrol, vanillin, peroxyacetic acid, hydrogen peroxide, N-acetyl-l-cysteine and Citrox were selected for their results in in vitro assays against E. coli O157:H7 and Listeria spp., to be tested on fresh-cut apple plugs. Apple flesh was inoculated by dipping in a suspension of a mix of the studied pathogens at 10⁶ cfu mL^?1, and then treated with the antimicrobial substances. All treatments were compared to deionized water and a standard sodium hypochlorite treatment (SH, 100 mg L^?1, pH 6.5). Pathogen population on apple plugs was monitored for up to 6 days at 10 °C. Bacterial reductions obtained by peroxyacetic acid (80 and 120 mg L^?1), vanillin (12 g L^?1), hydrogen peroxide (5, 10, 20 mL L^?1) and N-acetyl-l-cysteine (5 and 10 g L^?1) were similar or higher than reduction obtained by SH. In addition, bacterial populations were maintained at low levels throughout storage. No cells of any of the pathogens were detected in the peroxyacetic acid, hydrogen peroxide, Citrox and SH washing solutions after apple treatment. Peroxyacetic acid, hydrogen peroxide and N-acetyl-l-cysteine could be potential disinfectants for the fresh-cut industry as an alternative to chlorine disinfection. However, their effect on sensory quality and effectiveness under commercial processing conditions should be evaluated.     

Renal handling of calcium and phosphorus in experimental renal hyperparathyroidism in dogs

Twenty-four hour urinary excretion, fractional excretion and the filtered load of calcium and phosphorus were monitored as hyperparathyroidism evolved in a model of progressive canine renal failure. Thirteen beagles of both sexes aged four and a half months were used. Nine of them were subjected to a renal damaging schedule (neomycine, 60 mg/kg/48 h, IM, 32 weeks) in order to induce chronic renal failure leading to secondary hyperparathyroidism (2HPT group). The remaining four were kept as the control group. The experiment was conducted over 32 weeks. Blood and 24 h urine were collected every four weeks. Calcium, phosphorus and creatinine were analyzed. Plasma parathormone and calcitonin were determined at weeks 0, 12, 24 and 32. The level of renal function in the 2HPT animals was reduced to 25% of that of the controls (endogenous creatinine clearance was 0.45 +/- 0.22 mL/min/kg as opposed to 1.81 +/- 0.54 mL/min/kg). Hyperparathyroidism was confirmed by a progressive increase in the levels of the parathyroid hormone. Calcitonin levels were not modified. A tendency to hypocalcaemia was observed, reaching statistically significant levels from the twenty-eighth week of the study, when hyperphosphataemia also became significant. Daily urinary excretion of calcium and phosphorus remained at values considered normal throughout the experiment with no alteration imputable to the impaired renal function. This is explained by the decrease in the filtered load of these elements (in both cases statistically significant from the 24th week on) being associated with an increase in their fractional excretion. Thus, calcium and phosphorus urinary excretion values could be maintained in a normal range up to the end of the experiment, showing that renal calcium handling in dogs with experimentally induced renal failure seems to differ from that observed in human patients.     

Thinning decreased soil respiration differently in two dryland Mediterranean forests with contrasted soil temperature and humidity regimes

The effects of a thinning treatment on soil respiration (Rs) were analysed in two dryland forest types with a Mediterranean climate in east Spain: a dry subhumid holm oak forest (Quercus ilex subsp. ballota) in La Hunde; a semiarid postfire regenerated Aleppo pine (Pinus halepensis) forest in Sierra Calderona. Two twin plots were established at each site: one was thinned and the other was the control. Rs, soil humidity and temperature were measured regularly in the field at nine points per plot distributed into three blocks along the slope for 3 years at HU and for 2 years at CA after forest treatment. Soil heterotrophic activity was measured in laboratory on soil samples obtained bimonthly from December 2012 to June 2013 at the HU site. Seasonal Rs distribution gave low values in winter, began to increase in spring before lowering as soil dried in summer. This scenario indicates that with a semiarid climate, soil respiration is controlled by both soil humidity and soil temperature. Throughout the study period, the mean Rs value in the HU C plot was 13% higher than at HU T, and was 26% higher at CA C than the corresponding CA T plot value, being the differences significantly higher in control plots during active growing periods. Soil microclimatic variables explain the biggest proportion of variability for Rs: soil temperature explained 24.1% of total variability for Rs in the dry subhumid forest; soil humidity accounted for 24.6% of total variability for Rs in the semiarid forest. As Mediterranean climates are characterised by wide interannual variability, Rs showed considerable variability over the years, which can mask the effect caused by thinning treatment.

     

Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test

An outbreak of human leishmaniosis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.     

Effects of Rapid Cooling Prior to Freezing on the Quality of Canine Cryopreserved Spermatozoa

The aim of this study was to evaluate the effects of rapid cooling prior to freezing on frozen-thawed canine sperm quality. In experiment 1, centrifuged ejaculates from 6 dogs were pooled, split into 4 aliquots and cryopreserved by the Uppsala procedure using different cooling rates (control, cooling speed 18 C/90 min and average cooling rate 0.2 C/min; rapid, cooling speed 18 C/8 min and average cooling rate 2.25 C/min) in combination with 2 glycerol addition protocols (fractionated or unfractionated). In experiment 2, centrifuged ejaculates from 4 dogs were processed individually using the same cooling rates described in experiment 1 in combination with an unfractionated glycerol addition protocol. Each of the experiments was replicated 5 times. Sperm quality was evaluated after 30 and 150 min of post-thawing incubation at 38 C. Total motility (TM), progressive motility (PM) and quality of movement parameters were assessed using a computerized system, and sperm viability (spermatozoa with intact plasma and acrosome membranes) was assessed using flow cytometry (H-42/PI/FITC-PNA). Values for TM, PM, viable spermatozoa and the quality of movement parameters after thawing were not significantly affected by the cooling rate. The interaction between the cooling rate and the added glycerol protocol was not significant. There were significant differences among the males (P<0.01) in the sperm quality parameters evaluated after thawing. The interaction between the males and the cooling rate was not significant. In conclusion, canine spermatozoa can be cryopreserved using the Uppsala method at an average cooling rate of 2.25 C/min prior to freezing together with addition of fractionated or unfractionated glycerol.     

Amino acid profile of the quinoa (Chenopodium quinoa Willd.) using near infrared spectroscopy and chemometric techniques

The high content of amino acids of the quinoa, especially essential amino acids (higher than other cereals) makes a food increasingly demanded by consumers. A total of twelve amino acids (arginine, cystine, isoleucine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine) were analyzed in quinoa samples from Chile by near infrared spectroscopy (NIR) with direct application to the samples of a remote fiber-optic reflectance probe. The calibration results using modified partial least squares (MPLS) regression satisfactorily allowed the determination of the concentrations of this amino acid group with high multiple correlation coefficients (RSQ = 0.97–0.71) and low standard prediction errors (SEPC = 0.07–0.20). The prediction capacity (RPD) for the arginine, the cystine, the isoleucine, the lysine, the serine, the threonine, the tryptophan, the tyrosine and the valine ranged between 2.6 and 5.2, for the rest of amino acids were higher to 1.8, indicating that the NIRS equations obtained were applicable to unknown samples. It has confirmed that NIRS technology is a method that may be useful to replace the traditional methods for routine analysis of some amino acids.     

Influence of Salinity in the Bioavailability of Zn in Sediments of the Gulf of Cádiz (Spain)

This study assesses the effect of salinity in bioavailability and toxicity of Zn by means of laboratory bioassays by observing contamination in both sediment and water, accumulation of Zn in biological tissues, and histopathological damage in the gills and guts tissues of Ruditapes philippinarum clams, which were exposed to different types of sediments from the Gulf of Cádiz (SW Spain) as well as two dilutions of toxic mud coming from an accidental mining spill. With this objective, the coefficients of distribution (K _D) for Zn between overlying water and sediments were calculated, the histopathological frequencies in the tissues of the gills and guts of clams were determined, and the biota-sediment bioaccumulation factors as well as the bioaccumulation factors were quantified in the different stations. Results showed that the greatest histopathological damages appeared when the salinity values decreased. Statistical results showed that salinity was inversely correlated with histopathological damage (p?<?0.01) for the lesion index for gills. The most outstanding results were observed in the two dilutions of toxic mud (0.3% and 7.9%) at a salinity value of 10. Salinity was inversely correlated with the concentration of Zn in biological tissues (p?<?0.05) and inversely correlated with the concentration of Zn in water and sediment. Zn mobilization to the overlying water is produced when salinity values decrease.     

Effect of a new thermal treatment in combination with saprobic fungal incubation on the phytotoxicity level of alperujo

Byproducts generated from food industries, such as olive oil mills, have been studied to decrease harmful pollution and their environmental consequences. In this work, a new thermal pretreatment and saprobic fungal incubation to detoxify alperujo (two-phase olive mill waste) have been evaluated in view of its use as fertilizer in agriculture. The sequential use of both methods simplifies the thermal conditions and incubation times of the fungal treatment. Optimization of the thermal treatment from 150 to 170 °C for 45 and 15 min, respectively, reduced the incubation time with Coriolpsis rigida from 20 to 10 weeks needed to reduce phytotoxic effects on tomato plants. Therefore, the combination of thermal and biological treatments will allow the development of the potential benefits of alperujo to improve nutrients in agricultural soil.     

Characterization of cider apples on the basis of their fatty acid profiles

In the current study, the fatty acids composition of 30 monovarietal apple juices from six cider apple varieties belonging to two categories was analyzed. The different apple juices were obtained from three consecutive harvests (1997, 1998, and 1999). The fatty acids concentration in apple juice together with chemometric techniques such as principal components analysis (PCA), soft independent modeling of class analogy (SIMCA), and linear discriminant analysis (LDA), allowed us to differentiate apple juices on the basis of the sweet or sharp category to which the cider apple variety belongs. Fatty acids such as the unsaturated oleic and linoleic acids, and saturated caprylic, capric, stearic, and palmitic acids were related to the sweet cider apple category, while pentadecanoic acid is related to the sharp class.     

Ocular lesions associated with Trypanosoma evansi in experimentally infected goats

Ocular lesions associated with Trypanosoma spp. infection have been described in man and many animal species. However, loss of vision has not been demonstrated in humans presenting Chagas disease or in animals affected by different trypanosome species. In order to assess the possible ocular disorders caused by Trypanosoma evansi infection, six goats were inoculated with 1 x 10(5) T. evansi and maintained for 12 months and four goats were used as control. The inoculated animals became positive at serological and parasitological tests at 1-month post-inoculation and showed a subclinical course of the disease. Unilateral superficial corneal ulceration and retinochoroiditis were observed in two inoculated animals. Data from ocular neurologic examination and electroretinography showed no significant differences between inoculated and non-inoculated goats. It could be concluded that Trypanosoma evansi can produce ocular lesion but without apparent loss of vision in goats.