Uracil as an index of lactic acid bacteria contamination of tomato products

The aim of this research was to evaluate the suitability of uracil as an hygienic quality index of tomato products. Whereas uridine was naturally present throughout tomato fruits' ripening, uracil appeared only after microbial contamination. In tomato pulp inoculated with nine different microbial strains, all five lactic acid bacteria (LAB) studied released relevant quantities of uracil (150-1040 mg/kg of dm), with a correlated partial or total decrease of uridine. Uracil production by yeasts and molds was very low or nonexistent; the starting uridine concentration (approximately 960 mg/kg of dm) remained constant or increased. Uracil thermostability was also verified. Twenty-six samples of tomato paste (30 degrees Brix) were collected from bag-in-drums produced in an industrial processing plant, some with evident swelling symptoms. All of the samples with high microbial count presented uracil. Uracil was also present in samples with microbial contamination under the detection limit and Howard mold count below legislation limits, implying the reprocessing, at least partial, of altered tomato product. The results indicate that uracil presence in tomato products is an index of LAB contamination that has occurred before heat treatment.     

Responses of Atlantic cod <Emphasis Type="Italic">Gadus morhua</Emphasis> head kidney leukocytes to phytase produced by gastrointestinal-derived bacteria

This study identified phytase-producing bacteria that were previously isolated from the gastrointestinal tract of Atlantic cod, Gadus morhua and determined its effect on head kidney leukocytes. Out of the 216 bacterial strains tested, the two phytase producers were identified as Pseudomonas sp. and Psychrobacter sp. based on their 16S rDNA sequence. Crude phytase from these two bacterial strains was produced employing the shake flask method. Even though the total protein of the crude phytase was not significantly different for the two bacteria, the phytase activity of the crude enzyme produced by Pseudomonas sp. (97.1 ± 16.7 U) was significantly higher than that of the enzyme from Psychrobacter sp. (75.9 ± 2.4 U). When cod head kidney leukocytes were incubated with the crude phytase (50 μg ml⁻¹), it resulted in enhanced cell proliferation, higher myeloperoxidase, and acid phosphatase activities. Extracellular responses—respiratory burst activity and hydrogen peroxide production were not enhanced by the crude enzyme. As a consequence, the growth of two pathogenic bacteria Aeromonas salmonicida and Vibrio anguillarum was not suppressed by the supernatants obtained from head kidney leukocytes incubated with the crude bacterial phytase. Thus, the enzyme from phytase-producing intestinal bacteria of Atlantic cod can stimulate intracellular head kidney leukocyte activities but not the production of extracellular substances that are involved in antibacterial response. These have implications on the potential use of bacterial phytase as feed supplement to boost cellular immune response of the fish and could be employed as a health management strategy in culture systems.     

Volatile Organic Compounds,Indole, and Biogenic Amines Assessment in Two Mediterranean Irciniidae (Porifera,Demospongiae)

Solid phase microextraction (SPME) coupled to gas chromatography-mass spectrometry (GC-MS) was employed for the headspace determination of the volatile organic fraction emitted by two of the most common Mediterranean demosponges, Ircinia variabilis and Sarcotragus spinosulus, and of indole and some biogenic amines released by sponges in an aqueous medium. A total of 50/30 µm divinylbenzene/carboxen/polydimethylsiloxane and 75 µm carboxen/polydimethylsiloxane fibers were used for the headspace extraction of low molecular weight sulfur compounds from a hermetically sealed vial containing sponge fragments, while the direct immersion determination of indole and biogenic amines was performed. The biogenic amines were extracted after in-solution derivatization with isobutyl chloroformate. All analytical parameters (linearity, limits of detection, and quantification, precision, and recovery) were evaluated for indole and biogenic amines. SPME-GC-MS proved to be a reliable means of highlighting the differences between molecules released by different sponges, principally responsible for their smell. The combined approaches allowed the identification of several volatile compounds in the headspace and other molecules released by the sponges in an aqueous medium, including indole and the BAs cadaverine, histamine, isobutylamine, isopentylamine, propylamine, 2-phenylethylamine, putrescine and tryptamine. The results obtained represent a further contribution to the picture of odoriferous molecules secreted by sponges.     

Pig slurry treatment modifies slurry composition,N2O,and CO2 emissions after soil incorporation

The treatment of manures may improve their agricultural value and environmental quality, for instance with regards to greenhouse gases mitigation and enhancement of carbon (C) sequestration. The present study verified whether different pig slurry treatments (i.e. solid/liquid separation and anaerobic digestion) changed slurry composition. The effect of the slurry composition on N₂O and CO₂ emissions, denitrification and soil mineral nitrogen (N), after soil incorporation, was also examined during a 58-day mesocosm study. The treatments included a non-treated pig slurry (NT), the solid fraction (SF), and the liquid fraction (LF) of a pig slurry and the anaerobically digested liquid fraction (DG). Finally, a non-fertilized (N0) and a treatment with urea (UR) were also present.The N₂O emissions measured represented 4.8%, 2.6%, 1.8%, 1.0% and 0.9% of N supplied with slurry/fertilizer for NT, LF, DG, SF and UR, respectively. Cumulative CO₂ emissions ranged from 0.40 g CO₂-C kg^?1 soil (0.38 Mg CO₂-C ha^?1) to 0.80 g CO₂-C kg^?1 soil (0.75 Mg CO₂-C ha^?1). They were highest for SF (56% of C applied), followed by NT (189% of C applied), LF (337% of C applied) and DG (321% of C applied). Ammonium was detected in the soil for all treatments only at day one, while nitrate concentration increased linearly from day 15 to day 58, at a rate independent of the type of slurry/fertilizer applied. The nitrate recovery at day 58 was 39% of the N applied for NT, 19% for SF, 52% for LF, 67% for DG, and 41% for UR. The solid fraction generally produced higher potential denitrification fluxes (75.3 for SF, 56.7 for NT, 53.6 for LF, 47.7 for DG and 39.7 mg N₂O + N₂-N kg^?1 soil for UR). The high variability of actual denitrification results obfuscated any treatment effect.We conclude that treatment strongly affects slurry composition (mainly its C, fibre and NH₄⁺ content), and hence N₂O and CO₂ emission patterns as well as denitrification processes and nitrate availability. In particular, the solid fraction obtained after mechanical separation produced the most pronounced difference, while the liquid fraction and the anaerobically digested liquid fraction did not show significant difference with respect to the original slurry for any of the measured parameters. Combining data from the different fractions we showed that separation of slurry leads to reduced N₂O emissions, irrespective of whether the liquid fraction is digested or not. Furthermore, our results suggested that the default emission factor for N₂O emissions inventory is too low for both the non-treated pig slurry and its liquid fraction (digested or not), and too high for the separated solid fraction and urea.     

Experimental infection of cattle, sheep and pigs with 'Hobi'-like pestivirus

To date, limited information is available on the ability of 'Hobi'-like pestiviruses (putative bovine viral diarrhoea 3) to infect and cause disease in animal species traditionally affected by pestiviruses. In order to obtain new insights into host range and pathogenic potential of this atypical pestivirus, BVDV-seronegative calves (n=5), lambs (n=5) and piglets (n=5) were experimentally infected with the European 'Hobi'-like strain Italy-1/10-1, whereas two animals per species served as uninfected controls. Appearance of clinical signs, leukopenia, viremia, viral shedding and seroconversion were monitored for 28 days post-infection. Calves and lambs were successfully infected, displaying respiratory signs (nasal discharge), moderate hyperthermia and leukopenia, viremia and viral shedding through the nasal and faecal routes. Antibody responses were observed in both animal species by ELISA and virus neutralisation assays. In contrast, inoculated piglets did not display any clinical signs nor leukopenia and viral RNA was not detected in any biological samples. Nevertheless, the presence of detectable antibodies by virus neutralisation accounted for a successful, albeit limited infection of these animals.     

Tenovaginoscopic approach to the common digital flexor tendon sheath of adult cattle: technique, normal findings and preliminary results in four clinical cases

The aim of this study was to describe the tenovaginoscopic approach to the bovine common digital flexor tendon sheath (CDFTS). A comparative anatomical, ultrasonographic and endoscopic study was undertaken using 26 healthy cadaver feet from adult dairy cows. Tenovaginoscopy was performed using a rigid, 30° arthroscope (length 18 cm; outer diameter 4mm) enabling a direct view of the synovial cavity and the following structures: digital flexor tendons, digital annular ligaments, lateral and medial pouches, three mesotendons, the vinculum of the superficial digital flexor tendon, and a slot-shaped opening in the manicaflexoria of the hind feet. Additionally, four clinical cases of septic tenosynovitis treated with lavage under tenovaginoscopic control were examined. Tenovaginoscopy represents a feasible, minimally invasive method for the diagnosis and treatment of septic tenosynovitis of the CDFTS, which allows the degree of alterations of the normal structures to be evaluated.     

Pathogen translocation and histopathological lesions in an experimental model of Salmonella Dublin infection in calves receiving lactic acid bacteria and lactose supplements

The purpose of this study was to evaluate the capacity of a lactic acid bacteria (LAB) inoculum to protect calves with or without lactose supplements against Salmonella Dublin infection by evaluating histopathological lesions and pathogen translocation. Fifteen calves were divided into three groups [control group (C-G), a group inoculated with LAB (LAB-G), and a group inoculated with LAB and given lactose supplements (L-LAB-G)] with five, six, and four animals, respectively. The inoculum, composed of Lactobacillus (L.) casei DSPV 318T, L. salivarius DSPV 315T, and Pediococcus acidilactici DSPV 006T, was administered with milk replacer. The LAB-G and L-LAB-G received a daily dose of 10⁹ CFU/kg body weight of each strain throughout the experiment. Lactose was provided to the L-LAB-G in doses of 100 g/day. Salmonella Dublin (2 × 10¹⁰ CFU) was orally administered to all animals on day 11 of the experiment. The microscopic lesion index values in target organs were 83%, 70%, and 64.3% (p < 0.05) for the C-G, LAB-G, and L-LAB-G, respectively. Administration of the probiotic inoculum was not fully effective against infection caused by Salmonella. Although probiotic treatment was unable to delay the arrival of pathogen to target organs, it was evident that the inoculum altered the response of animals against pathogen infection.     

Effects of exposure to overcrowding on rodlet cells of the teleost fish <Emphasis Type="Italic">Dicentrarchus labrax</Emphasis> (L.)

Farmed fish are usually exposed to routine procedures which have strong effects on stress responses. Rodlet cells may represent an useful biomarker for studies on the presence of stressors in aquaculture. This work focused on the localization of rodlet cells by light and electron microscopy in gills of sea bass (Dicentrarchus labrax) subjected to different conditions of overcrowding. In general, a significant increase in number of rodlet cells has been observed in all animals subjected to overcrowding stress. In gills of control group rare rodlet cells were detected at the level of both primary and secondary lamellae, whereas in stressed group clusters of rodlet cells have been found in the epithelium of primary and secondary lamellae indicating that these cells are influenced by stocking density.