Daily variations of plasma serotonin levels in 2-year-old horses

Serotonin (5-HT) is a neurotransmitter implicated in the regulation of circadian rhythms. Synthesis and release of 5-HT are subjected to daily fluctuations, but there are no data on plasma serotonin levels in 2-year-old horses. Aim of this study was to investigate the daily variations in 5-HT levels in platelet-poor plasma from 2-year-old horses, so as to understand whether plasma 5-HT could be considered as an additional variable to assess the natural adaptability of horses to training. The research was carried out on 6 clinically healthy crossbred mares. Data showed higher levels of platelet-poor plasma 5-HT at 5 p.m. (P < 0.001) than at 1 p.m. and lower values at 5 a.m. (P < 0.001) than at 1 a.m. One-way repeated-measures analysis of variance showed a significant effect of sampling time (P < 0.0001) on plasma 5-HT changes. Results suggest that the increase of 5-HT levels could be related to an enhanced 5-HT output and release, and that their daily variations could be considered as an additional variable to assess the natural adaptability of horses for being trained. Tryptophan levels, the essential amino acid precursor of 5-HT, seemed to hint at slight fluctuations at regular intervals, with the highest amounts at 9 a.m. and at 9 p.m. and lowest at 1 a.m. and at 1 p.m., in agreement with data reported in adult horses. Cortisol levels showed wide fluctuations, with the highest amounts at 1 a.m. and the lowest amounts at 9.00 p.m.     

Autophagy-dependent anticancer immune responses induced by chemotherapeutic agents in mice

Antineoplastic chemotherapies are particularly efficient when they elicit immunogenic cell death, thus provoking an anticancer immune response. Here we demonstrate that autophagy, which is often disabled in cancer, is dispensable for chemotherapy-induced cell death but required for its immunogenicity. In response to chemotherapy, autophagy-competent, but not autophagy-deficient, cancers attracted dendritic cells and T lymphocytes into the tumor bed. Suppression of autophagy inhibited the release of adenosine triphosphate (ATP) from dying tumor cells. Conversely, inhibition of extracellular ATP-degrading enzymes increased pericellular ATP in autophagy-deficient tumors, reestablished the recruitment of immune cells, and restored chemotherapeutic responses but only in immunocompetent hosts. Thus, autophagy is essential for the immunogenic release of ATP from dying cells, and increased extracellular ATP concentrations improve the efficacy of antineoplastic chemotherapies when autophagy is disabled.     

Arbuscular mycorrhizal fungi of a Mediterranean island (Pianosa), within a UNESCO Biosphere Reserve

In this work we have determined the community composition of spore-forming arbuscular mycorrhizal fungi (AMF) in a maquis site on Pianosa island, a protected area within the Tuscan Islands UNESCO Biosphere Reserve, Italy. We have analysed rhizosphere soil of the dominant plant species Pistacia lentiscus, Smilax aspera, Rosmarinus officinalis and of the endemic plant Helichrysum litoreum. The AMF species recovered were: Scutellospora dipurpurescens, Glomus coronatum, Glomus mosseae, Glomus etunicatum, Glomus geosporum, Glomus viscosum, Entrophospora sp., Pacispora sp. and Glomus rubiforme. The identification of native S. dipurpurescens and G. coronatum was carried out on spores isolated from rhizosphere soil of H. litoreum, by combining morphological traits and 18S (SSU) and ITS rDNA sequences. Therefore, AMF species of Pianosa rhizosphere soils represent an important repository for the conservation and maintenance in their natural habitat of such beneficial symbionts, key microorganisms of soil fertility.     

Pulmonary rhodococcosis in a cat

Feline Rhodococcus equi infection is rare, despite the bacteria is widespread in the environment. R equi infection is typically observed in equine species but the infection has also been reported in dogs, cats and other domestic animals. There are a few reports regarding pulmonary R equi infection in cats and the disease appears to be limited to the skin and the subcutaneous tissue. This report describes the pathological, microbiological and the virulence features associated with an acute necrosuppurative pneumonia in a cat. To the best of our knowledge, this is the first report of feline pulmonary R equi infection in Italy.     

Distribution,abundance, and movements of the bottlenose dolphin (Tursiops truncatus) in the Pelagos Sanctuary MPA (north‐west Mediterranean Sea)

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Molecular characterization of Prototheca in 11 symptomatic dogs

Protothecosis is an uncommon disease caused by algae of the genus Prototheca. In dogs, the infection is usually first localized to the colon but has the propensity to later disseminate hematogenously to many other organs, with marked tropism for the eyes and central nervous system. Diagnosis is established by culture and/or evidence of Prototheca organisms in cytologic or histologic preparations. Species characterization, however, requires molecular investigations. Our laboratory set up a real-time PCR targeting portion D1/D2 of the 28S rRNA for identification of Prototheca species from both positive cultures (of rectal swabs and urine) and formalin-fixed, paraffin-embedded tissue. Prototheca bovis, P. ciferrii, and P. wickerhamii were characterized in 11 dogs with systemic or cutaneous protothecosis. Prototheca identifications were phylogenetically consistent with the new taxonomy proposed for this genus based on the mitochondrial cytochrome b gene. As a pilot study, we screened feces and rectal scrapes from 200 asymptomatic dogs, using 2 cohorts of stray and owned animals, to determine the prevalence of intestinal carriage of Prototheca spp. The Prototheca-negative results from both cohorts of healthy dogs suggest that predisposing factors related to the host probably contribute more to the acquisition of clinical disease than exposure to contaminated environments.     

Effects of municipal solid waste compost amendments on soil enzyme activities and bacterial genetic diversity

Municipal solid waste (MSW) composts have been used to maintain the long-term productivity of agroecosystems and to protect the soil environment from overcropping, changes in climatic conditions and inadequate management; they also have the additional benefit of reducing waste disposal costs. Since MSW may contain heavy metals and other toxic compounds, amendments cannot only influence soil fertility, but may also affect the composition and activity of soil microorganisms. The effects of MSW compost and mineral N amendments in a 6-year field trial on some physical-chemical properties, enzyme activities and bacterial genetic diversity of cropped plots (Beta vulgaris-Triticum turgidum rotation) and uncropped plots were investigated. The compost was added at the recommended and twice the recommended dosage (12, 24 t ha⁻¹). Amendments of cropped plots with MSW compost increased the contents of organic C from 13.3 to 15.0 g kg⁻¹ soil and total N from 1.55 to 1.65 g kg⁻¹ soil. There were significant increases in dehydrogenase (9.6%), β-glucosidase (13.5%), urease (15.4%), nitrate reductase (21.4%) and phosphatase (9.7%) activities. A significant reduction in protease activity (from 3.6 to 2.8 U g⁻¹ soil) was measured when a double dose of compost was added to the cropped plots. No dosage effect was detected for the other enzymes. Changes in the microbial community, as a consequence of MSW amendment, were minimal as determined using denaturing gradient gel electrophoresis, rDNA internal spacer analysis and amplified ribosomal DNA restriction analysis of bacteria, archaea, actinomycetes, and ammonia oxidizers. This indicates that there was no significant variation in the overall bacterial communities nor in selected taxonomic groups deemed to be essential for soil fertility.     

Development and validation of an ultrasensitive chemiluminescent enzyme immunoassay for aflatoxin M1 in milk

A fast and ultrasensitive chemiluminescent enzyme immunoassay for aflatoxin M(1) in milk samples has been developed and validated. The method is an indirect competitive type format involving the immobilization of an aflatoxin M(1)-bovine serum albumin conjugate on 384 well black polystyrene microtiter plates and the use of a secondary antibody labeled with horseradish peroxidase detected with a luminol-based substrate. Aflatoxin M(1) standard solutions were prepared in milk-based buffer, and milk samples were analyzed without any cleanup procedure. The limit of quantification was 1 ppt, the coefficient of variation was below 9% for both intra- and interassay precision, and the recovery ranged from 96 to 122%. The method is specific, and other aflatoxins do not significantly cross-react with the antibody. Twenty-four milk samples were analyzed, and a good correlation was observed (y = 0.98x + 1.71, r(2) = 0.98, n = 24) when the data were compared with a reference high-performance liquid chromatography method with a fluorescent detector. The developed method is suitable for an accurate, sensitive, and high-throughput screening of aflatoxin M(1) in milk samples with a reduction of costs and increased detectability, as compared with previously developed immunoassays.     

Molecular approaches to investigate herbicide-induced bacterial community changes in soil microcosms

Since biochemical and microbiological methods used to study microbial community changes induced by anthropogenic activities can be biased, the impact of two herbicides on soil microorganisms was investigated by culture-independent molecular techniques. The effect of three different amounts (the recommended field dose, tenfold, and 100-fold the dose) of propanil or prometryne on the bacterial community of a clay soil, two modalities of incubation (soil moisture at 70% of the field capacity and a soil-herbicide suspension, 1:10, w:v), and time of incubation were investigated by denaturing gradient gel electrophoresis (DGGE) and amplified rDNA restriction analysis (ARDRA). Two sets of primers for 16S rDNA were used to amplify total soil DNA. Sterile and non-sterile samples were used to determine, by HPLC, the amounts of herbicides adsorbed on soil and transformed by soil microorganisms. Prometryne persisted in soil longer than propanil. Propanil was removed significantly more by non-sterile than by sterile samples, while for prometryne, slight differences were observed. 3,4-Dichloroaniline, a product of propanil hydrolysis, was detected in non-sterile samples and increased with incubation time. Propanil did not affect soil bacteria significantly as indicated by DGGE and ARDRA, with the only exception being the soil-herbicide suspension. Despite a lower utilization of prometryne by soil microorganisms, DGGE analysis showed a more diverse banding than with propanil. Some bands were also detected in the DNA sample extracted from the soil-prometryne suspension, and could be representative of bacterial species utilizing the herbicide as a carbon source, in two very different soil microcosms.     

Kinetics and mechanism of cymoxanil degradation in buffer solutions

The kinetics and mechanism(s) of the hydrolytic degradation of a compound are needed to evaluate a compound's abiotic degradation in the environment. In this paper, the hydrolysis of cymoxanil [2-cyano-N-[(ethylamino)carbonyl]-2-(methoxyimino) acetamide] was investigated in dark sterile aqueous solutions under a variety of pH conditions (pH 2.8-9.2) and temperatures (15-50 degrees C). Hydrolysis of cymoxanil was described by first-order kinetics, which was dependent on pH and temperature. Cymoxanil degraded rapidly at pH 9 (half-life = 31 min) and relatively slowly at pH 2.8 (half-life = 722 days). The effect of temperature on the rate of cymoxanil degradation was characterized using the Arrhenius equation with an estimated energy of activation of 117.1 kJ mol(-)(1). An increase in temperature of 10 degrees C resulted in a decrease in half-life by a factor of approximately 5. Three competing degradation pathways are proposed for the hydrolysis of cymoxanil, with two of the pathways accounting for approximately 90% of cymoxanil degradation. These two pathways involved either initial cyclization to 1-ethyldihydro-6-imino-2,3,5(3H)-pyrimidinetrione-5-(O-methyloxime) (1, Figure 1) or direct cleavage of the C-1 amide bond to form cyano(methoxyimino) acetic acid (7). The third pathway of degradation involved initial cyclization to 3-ethyl-4-(methoxyimino)-2,5-dioxo-4-imidazolidinecarbonitrile (8), which rapidly degrades into 1-ethyl-5-(methoxyimino)-2,4-imidazoline-2,4-dione (9). All three pathways eventually lead to the formation of the polar metabolite oxalic acid.