Uncommonly thorough hydrolysis of peptides during ripening of Ragusano cheese revealed by tandem mass spectrometry     

Gagnaire V  Carpino S  Pediliggieri C  Jardin J  Lortal S  Licitra G 《Journal of agricultural and food chemistry》2011,59(23):12443-12452

Ragusano is a pasta filata cheese produced from raw milk in Sicily. The proteolysis was extensively analyzed after stretching (day 0), at 4 and 7 months of ripening through soluble nitrogen, urea-PAGE, and peptide identification by tandem mass spectrometry. After stretching, 123 peptides were identified: 72 arising from β-casein, 34 from α(s1)-casein, and 17 from α(s2)-casein. The main protein splitting corresponded to the action of plasmin, chymosin, cathepsin D, cell envelope proteinase, and peptidase activities of lactic acid bacteria. Unlike other types of cheeses, <10% residual β- and α(s)-caseins remained intact at 7 months, indicating original network organization based on large casein fragments. The number of identified soluble peptides also dramatically decreased after 4 and 7 months of ripening, to 47 and 25, respectively. Among them, bioactive peptides were found, that is, mineral carrier, antihypertensive, and immunomodulating peptides and phosphopeptides.  相似文献   

Influence of shearing on oxidative stress and some physiological parameters in ewes     

Giuseppe PICCIONE  Francesco FAZIO  Stefania CASELLA  Pietro PENNISI  Giovanni CAOLA 《Animal Science Journal》2011,82(3):481-485

The aim of the present study was to evaluate the effect of shearing on physiological and oxidative stress parameters in ewes. Twenty Comisana ewes were used and divided into two groups. Ten ewes were left unshorn as a control group (Group A) and 10 ewes were shorn (Group B). All measurements were taken before and after shearing, and repeated 8 h after shearing and 1, 2, 3, 4, 5, 10 and 15 days after shearing. Reactive oxygen species (dROMs), antioxidant barrier (oxy‐adsorbent), thiol antioxidant barrier (SHp) and packed cell volume (PCV) were assessed in blood samples collected by means of jugular venipuncture. Rectal temperature (RT), respiratory rate (RR) and heart rate (HR) were also measured. Two‐way repeated measures analysis of variance (ANOVA), followed by Bonferroni's test, was used for the assessment of significant effects due to shearing and time. The statistical analysis showed significant increases (P < 0.01) of dROMs, oxy‐adsorbent, SHp, and a significant decrease (P < 0.01) of RT and RR associated with time and shearing. Our results indicate that shearing causes a change in the ewe's homeostatic balance that leads to oxidative stress.  相似文献   

Improving virgin olive oil quality by means of innovative extracting biotechnologies     

Ranalli A  Gomes T  Delcuratolo D  Contento S  Lucera L 《Journal of agricultural and food chemistry》2003,51(9):2597-2602

Three major virgin olive oil varieties (Dritta, Leccino, and Coratina) extracted by a modern centrifugation system aided with a new plant enzyme preparation (having prevalently pectolytic activity) were characterized. These oils showed a clearly enhanced quality standard, owing to higher levels of some important minor components (phenolics, volatiles, tocopherols, carotenes, and chlorophylls) and to frequently lower concentrations of oxidized triglycerides and diglycerides. The oils were therefore characterized by lower susceptibility to oxidation and longer shelf life, and their flavor, aroma, and color features appeared to be significantly improved. The saponifiable fraction was practically not affected as the enzymatic effects involved only the membranes of the oil droplets, where the nonglyceridic compounds are essentially located.  相似文献   

Pseudomonas syringae pv. coryli, the Causal Agent of Bacterial Twig Dieback of Corylus avellana     

Scortichini M  Rossi MP  Loreti S  Bosco A  Fiori M  Jackson RW  Stead DE  Aspin A  Marchesi U  Zini M  Janse JD 《Phytopathology》2005,95(11):1316-1324

ABSTRACT Thirty-eight bacterial strains isolated from hazelnut (Corylus avellana) cv. Tonda Gentile delle Langhe showing a twig dieback in Piedmont and Sardinia, Italy, were studied by a polyphasic approach. All strains were assessed by fatty acids analysis and repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using BOX and ERIC primer sets. Representative strains also were assessed by sequencing the 16S rDNA and hrpL genes, determining the presence of the syrB gene, testing their biochemical and nutritional characteristics, and determining their pathogenicity to hazelnut and other plants species or plant organs. Moreover, they were compared with reference strains of other phytopathogenic pseudomonads. The strains from hazelnut belong to Pseudomonas syringae (sensu latu), LOPAT group Ia. Both fatty acids and repetitive-sequence-based PCR clearly discriminate such strains from other Pseudomonas spp., including P. avellanae and other P. syringae pathovars as well as P. syringae pv. syringae strains from hazelnut. Also, the sequencing of 16S rDNA and hrpL genes differentiated them from P. avellanae and from P. syringae pv. syringae. They did not possess the syrB gene. Some nutritional tests also differentiated them from related P. syringae pathovars. Upon artificial inoculation, these strains incited severe twig diebacks only on hazelnut. Our results justify the creation of a new pathovar because the strains from hazelnut constitute a homogeneous group and a discrete phenon. The name of P. syringae pv. coryli is proposed and criteria for routine identification are presented.  相似文献   

Three cases of prolapse of the nictitans gland in cats     

Chahory S  Crasta M  Trio S  Clerc B 《Veterinary ophthalmology》2004,7(6):417-419

We report three cases of adult cats showing a prolapse of the gland of the third eyelid. Three different breeds were affected: Burmese, Persian and Domestic Short-haired. In all cases, the disorder occurred spontaneously, without any other ocular sign. Surgical replacement of the gland was performed using the Morgan pocket technique. Good esthetic results were obtained, and no recurrence occurred.  相似文献   

Evaluation of phenolic compounds in virgin olive oil by direct injection in high-performance liquid chromatography with fluorometric detection     

Selvaggini R  Servili M  Urbani S  Esposto S  Taticchi A  Montedoro G 《Journal of agricultural and food chemistry》2006,54(8):2832-2838

Hydrophilic phenols are the most abundant natural antioxidants of virgin olive oil (VOO), in which tocopherols and carotenes are also present. The prevalent classes of hydrophilic phenols found in VOO are phenyl alcohols, phenolic acids, secoiridoids such as the dialdehydic form of decarboxymethyl elenolic acid linked to (3,4-dihydroxyphenyl)ethanol or (p-hydroxypheny1)ethanol (3,4-DHPEA-EDA or p-HPEA-EDA) and an isomer of the oleuropein aglycon (3,4-DHPEA-EA), lignans such as (+)-1-acetoxypinoresinol and (+)-pinoresinol, and flavonoids. A new method for the analysis of VOO hydrophilic phenols by direct injection in high-performance liquid chromatography (HPLC) with the use of a fluorescence detector (FLD) has been proposed and compared with the traditional liquid-liquid extraction technique followed by the HPLC analysis utilizing a diode array detector (DAD) and a FLD. Results show that the most important classes of phenolic compounds occurring in VOO can be evaluated using HPLC direct injection. The efficiency of the new method, as compared to the liquid-liquid extraction, was higher to quantify phenyl alcohols, lignans, and 3,4-DHPEA-EA and lower for the evaluation of 3,4-DHPEA-EDA and p-HPEA-EDA.  相似文献   

Treatment of cereal products with a tailored preparation of trichoderma enzymes increases the amount of soluble dietary fiber     

Napolitano A  Lanzuise S  Ruocco M  Arlotti G  Ranieri R  Knutsen SH  Lorito M  Fogliano V 《Journal of agricultural and food chemistry》2006,54(20):7863-7869

Nutritionists recommend increasing the intake of soluble dietary fiber (SDF), which is very low in most cereal-based products. Conversion of insoluble DF (IDF) into SDF can be achieved by chemical treatments, but this affects the sensorial properties of the products. In this study, the possibility of getting a substantial increase of SDF from cereal products using a tailored preparation of Trichoderma enzymes is reported. Enzymes were produced cultivating Trichoderma using durum wheat fiber (DWF) and barley spent grain (BSG) as unique carbon sources. Many Trichoderma strains were screened, and the hydrolysis conditions able to increase by enzymatic treatment the amount of SDF in DWF and BSG were determined. Results demonstrate in both products that it is possible to triple the amount of SDF without a marked decrease of total DF. The enzymatic treatment also causes the release of hydroxycinnamic acids, mainly ferulic acid, that are linked to the polysaccharides chains. This increases the free phenolic concentration, the water-soluble antioxidant activity, and, in turn, the phenol compounds bioavailability.  相似文献   

Genetic analysis and molecular characterisation of laboratory and field mutants of Botryotinia fuckeliana (Botrytis cinerea) resistant to QoI fungicides     

De Miccolis Angelini RM  Rotolo C  Masiello M  Pollastro S  Ishii H  Faretra F 《Pest management science》2012,68(9):1231-1240

BACKGROUND: QoI fungicides, inhibitors of mitochondrial respiration, are considered to be at high risk of resistance development. In several phytopathogenic fungi, resistance is caused by mutations (most frequently G143A) in the mitochondrial cytochrome b (cytb) gene. The genetic and molecular basis of QoI resistance were investigated in laboratory and field mutants of Botryotinia fuckeliana (de Bary) Whetz. exhibiting in vitro reduced sensitivity to trifloxystrobin. RESULTS: B. fuckeliana mutants highly resistant to trifloxystrobin were obtained in the laboratory by spontaneous mutations in wild‐type strains, or from naturally infected plants on a medium amended with 1–3 mg L^?1 trifloxystrobin and 2 mM salicylhydroxamic acid, an inhibitor of alternative oxidase. No point mutations were detected, either in the complete nucleotide sequences of the cytb gene or in those of the aox and Rieske protein genes of laboratory mutants, whereas all field mutants carried the G143A mutation in the mitochondrial cytb gene. QoI resistance was always maternally inherited in ascospore progeny of sexual crosses of field mutants with sensitive reference strains. CONCLUSIONS: The G143A mutation in cytb gene is confirmed to be responsible for field resistance to QoIs in B. fuckeliana. Maternal inheritance of resistance to QoIs in progeny of sexual crosses confirmed that it is caused by extranuclear genetic determinants. In laboratory mutants the heteroplasmic state of mutated mitochondria could likely hamper the G143A detection, otherwise other gene(s) underlying different mechanisms of resistance could be involved. Copyright © 2012 Society of Chemical Industry  相似文献   

Towards a rapid molecular identification of the common phlebotomine sand flies in the Mediterranean region     

Latrofa MS  Annoscia G  Dantas-Torres F  Traversa D  Otranto D 《Veterinary parasitology》2012,184(2-4):267-270

The present study reports two simple molecular approaches allowing a rapid identification of the most prevalent species of phlebotomine sand flies in the Mediterranean region. A PCR protocol for the amplification of ITS2 ribosomal region and a PCR-RFLP on a mitochondrial DNA fragment (cytb-nd1) were settled in order to identify and discriminate among Phlebotomus perniciosus, Phlebotomus neglectus, Phlebotomus perfiliewi, Phlebotomus papatasi and Sergentomyia minuta. The ITS2 regions showed a certain degree of interspecific variability, which led to PCR amplicons of different sizes, i.e., 450, 490, 460, 480 and 530 bp for P. perniciosus, P. neglectus, P. perfiliewi, P. papatasi, and S. minuta, respectively. Analogously, the digestion of a mitochondrial DNA amplicon with Ase I enzyme showed five different restriction profiles, which allowed the unequivocal differentiation of the sand fly species examined. These methods might represent useful tools for a molecular large scale screening of phlebotomine sand fly species caught in areas where leishmaniasis is endemic, in order to plan appropriate epidemiological surveillance programs for both Leishmania spp. and their vectors.  相似文献   

A duplex real-time polymerase chain reaction assay for the detection of and differentiation between Dirofilaria immitis and Dirofilaria repens in dogs and mosquitoes     

Latrofa MS  Dantas-Torres F  Annoscia G  Genchi M  Traversa D  Otranto D 《Veterinary parasitology》2012,185(2-4):181-185

This study describes a duplex real-time polymerase chain reaction (PCR) assay for the detection and differentiation between Dirofilaria immitis and Dirofilaria repens in dog blood and mosquitoes. Regions of a cytochrome oxidase 1 (cox1) mitochondrial DNA fragment and the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA were amplified from microfilariae and adult worm samples, using a sensitive SsoFast? EvaGreen(?) based real-time PCR method coupled with melting-curve analysis. The limit of the real-time PCR in detecting microfilaria and adult worm DNA was also tested both in dog blood and in artificially infected microfilarial. Two peaks at different melting temperatures (T(m)) for D. immitis (mean ± SD=75.7 ± 0.3°C) and D. repens (mean ± SD=70 ± 0.7°C), respectively, were obtained for microfilarial and adult positive controls of both species when examined separately and together. The real-time PCR protocol was also efficient in detecting microfilarial and adult DNA of both species when tested in samples spiked with DNA from Aedes albopictus, in Aedes aegypti experimentally infected by D. repens and in Culex pipiens naturally infected by D. repens and D. immitis. The high sensitivity of real-time PCR confirmed its reliability in detecting small amounts of genomic DNA either in dog blood or mosquitoes (2.5 pg/μl and 3 × 10(-1)pg/μl for D. immitis and D. repens, respectively). This assay is proposed as a tool for the epidemiological surveillance of the two most important Dirofilaria species in areas where they are endemic and sympatric.  相似文献