Study of the Oxidative State,as a Marker of Welfare,on Gilthead Sea Bream,Sparus aurata,Subjected to Handling Stress

In this study, we investigated the effect of repeated handling on the oxidative state of gilthead sea bream, Sparus aurata. We determined the activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPX), glutathione transferase (GST), and DT‐diaphorase (DTD) in liver, digestive tract, brain, gills, and white muscle. We also measured the lipid‐peroxidation level (malondialdehyde – MDA – concentration) and the glucose 6‐phosphate dehydrogenase activity (G6PDH). The results show that repeated low‐intensity handling on gilthead sea bream alter the oxidative status of tissues such as liver, digestive tract, and muscle in the last term can affect its welfare. The brain in a situation of handling stress is protected from oxidative damage by the activity of antioxidant enzymes. Finally, the concentration of MDA in tissues can be a good candidate as a biomarker of stress and welfare in fish.     

Effect of pyriproxyfen on developing stages and embryogenesis of California red scale (CRS), Aonidiella aurantii

During the past decade, the insect growth regulator pyriproxyfen has been the most used insecticide for controlling the California red scale (CRS), Aonidiella aurantii (Maskell), an important pest in citrus groves worldwide. The aim of the present research was to develop bioassays for determining the effect of pyriproxyfen on developing stages and embryogenesis of CRS that can serve as baselines for monitoring resistance. Pyriproxyfen was very effective against first- and second-instar nymphs of a susceptible strain of CRS; 0.1 and 0.2 mg AI L(-1), resulting in 80 and 100% mortality of first-instar nymphs, and in 62.5 and 91.7% mortality of second-instar nymphs respectively. According to probit analysis, the LC(50) and LC(90) values of pyriproxyfen for first-instar nymphs were 0.03 and 0.15 mg AI L(-1), and for second-instar nymphs they were 0.05 and 0.25 mg AI L(-1) respectively. Pyriproxyfen was quite effective on first-instar nymphs of a CRS field strain originating from a citrus grove where two successive applications of the compound had been applied. Mortality rates were 86 and 100% at concentrations of 0.2 and 0.4 mg AI L(-1) respectively. Pyriproxyfen suppressed embryogenesis when CRS females were treated with pyriproxyfen. Young CRS females treated with 1.0 and 5.0 mg AI L(-1) pyriproxyfen produced 6.6 N(1)/F and 2.4 N(1)/F as compared with 10.9 N(1)/F in untreated control.     

Effect of positive end-expiratory pressure on oxygen delivery during 1-lung ventilation for thoracoscopy in normal dogs

OBJECTIVE: To evaluate the effect of positive end-expiratory pressure (PEEP) on oxygen delivery (DO(2)) with 1-lung ventilation during thoracoscopy in normal anesthetized dogs. STUDY DESIGN: Prospective, controlled experimental study. ANIMALS: Eight, adult, intact Walker Hound dogs weighing 25.6-29.2 kg. METHODS: Anesthetized dogs had 1-lung ventilation during an open-chest condition. A Swan-Ganz catheter was used to measure pulmonary hemodynamic variables and to obtain mixed venous blood samples for blood gas analysis. A dorsal pedal catheter was used for measurement of systemic arterial pressure and to obtain arterial blood samples for blood gas analysis. Oxygen delivery was calculated and used to assess the effect of 0, 2.5, and 5 cm H(2)O PEEP during 1-lung ventilation on cardiopulmonary function. Each dog was its own control at 0 cm H(2)O PEEP. A randomized block ANOVA for repeated measures was used to evaluate the effect of the treatment on hemodynamic and pulmonary variables. RESULTS: Use of 5 cm H(2)O PEEP induced a significant augmentation in the arterial partial pressure of oxygen (PaO(2)). Shunt fraction (Q(s)/Q(t)), physiologic dead space (V(D)/V(T)), and the alveolar-arterial oxygen difference (P(A-a)O(2)) decreased significantly after 5 cm H(2)O PEEP, compared with 1-lung ventilation without PEEP. Use of 2.5 cm H(2)O PEEP had no significant effect on cardiopulmonary variables. Use of PEEP had no significant effect on arterial oxygen saturation (SaO(2)), DO(2), and hemodynamic variables in normal dogs. CONCLUSIONS: PEEP had no effect on DO(2) in normal dogs undergoing open-chest 1-lung ventilation because it had no adverse effect on hemodynamic variables. CLINICAL RELEVANCE: PEEP in normal dogs during open-chest 1-lung ventilation for thoracoscopy is not detrimental to cardiac output and can be recommended in clinical patients.     

Experimental infection of South American camelids with bluetongue virus serotype 8

Bluetongue (BT) is an infectious, non-contagious disease of wild and domestic ruminants. It is caused by bluetongue virus (BTV) and transmitted by Culicoides biting midges. Since 1998, BT has been emerging throughout Europe, threatening not only the na?ve ruminant population. Historically, South American camelids (SAC) were considered to be resistant to BT disease. However, recent fatalities related to BTV in captive SAC have raised questions about their role in BTV epidemiology. Data on the susceptibility of SAC to experimental infection with BTV serotype 8 (BTV-8) were collected in an animal experiment. Three alpacas (Vicugna pacos) and three llamas (Lama glama) were experimentally infected with BTV-8. They displayed very mild clinical signs. Seroconversion was first measured 6-8 days after infection (dpi) by ELISA, and neutralising antibodies appeared 10-13 dpi. BTV-8 RNA levels in blood were very low, and quickly cleared after seroconversion. However, spleens collected post-mortem were still positive for BTV RNA, over 71 days after the last detection in blood samples. Virus isolation was only possible from blood samples of two alpacas by inoculation of highly sensitive interferon alpha/beta receptor-deficient (IFNAR(-/-)) mice. An in vitro experiment demonstrated that significantly lower amounts of BTV-8 adsorb to SAC blood cells than to bovine blood cells. Although this experiment showed that SAC are generally susceptible to a BTV-8 infection, it indicates that these species play a negligible role in BTV epidemiology.     

Chilling and cooking rate effects on some myofibrillar determinants of tenderness of beef

Our objectives were to examine the effects of prerigor excision and rapid chilling vs. conventional carcass chilling of two muscles on proteolysis and tenderness during the postmortem storage, as well as the effects of fast and slow rates of cooking on myofibrillar characteristics and tenderness. The longissimus thoracis (LT) and triceps brachii (TB), long head muscles were removed 45 min after exsanguination from the left side of 12 carcasses and chilled in an ice bath to induce cold shortening (excised, rapidly chilled). At 24 h postmortem, the corresponding muscles were removed from the right side (conventionally chilled). All muscles were cut into 2.54-cm-thick steaks and assigned to one of two postmortem times (1 or 14 d), and to raw and cooking treatments. Steaks were cooked at 260 degrees C (FAST) or 93 degrees C (SLOW) in a forced-air convection oven to an internal temperature of 70 degrees C. Cooking loss, cooking time, and Warner-Bratzler shear force (WBSF) were measured on cooked steaks. Sarcomere length (SL) and the extent of proteolysis of desmin were measured on raw and cooked steaks. As expected, the excised, rapidly chilled muscles had a much more rapid (P < 0.05) temperature decline than those that were conventionally chilled. The excised, rapidly chilled treatment resulted in shorter (P < 0.05) SL, and SL was shorter (P < 0.05) in LT than in TB steaks. Raw steaks had longer (P < 0.05) SL than cooked steaks, regardless of chilling treatment. The FAST cooking resulted in shorter (P < 0.05) SL than SLOW cooking in conventionally chilled steaks, but cooking rate had no effect (P > 0.05) on SL of rapidly chilled steaks. Generally, TB steaks required longer (P < 0.05) cooking times and had higher (P < 0.05) cooking losses than LT steaks, and FAST-cooked steaks had greater (P < 0.05) cooking losses than SLOW-cooked steaks. Rapidly chilled steaks had less (P < 0.05) degradation of desmin than conventionally chilled steaks (31 vs. 41%). Aging for 14 d increased (P < 0.05) desmin degradation. Rapid chilling of muscles resulted in much higher (P < 0.05) WBSF values, whereas aging resulted in lower (P < 0.05) WBSF values. The SLOW-cooked TB steaks were more tender (P < 0.05) than FAST-cooked TB steaks and LT steaks cooked at either rate. Excised, rapidly chilled muscles underwent proteolysis, but it occurred at a slower rate during the first 24 h postmortem than it did in conventionally chilled muscles. Cooking rate did not affect tenderness of LT steaks, but SLOW cooking resulted in more tender TB steaks.     

Expression of cyclooxygenase-2 in canine epithelial nasal tumors

Cyclooxygenase-2 (COX-2) is an enzyme upregulated in some human and animal tumors. Enzymatic products are associated with tumorigenic activities. Given the poor response of canine nasal tumors to radiation, we considered the possibility that some of this resistance may be associated with COX-2 expression. To test this, 21 formalin-fixed, paraffin-embedded, and archived biopsy samples from canine epithelial nasal tumors were analyzed for COX-2 expression using immunohistochemistry. The biopsies were collected from dogs prior to radiation therapy. COX-2 expression was present in 17 of 21 (81%) tumors. The expression was observed in several different tumor types, including nasal carcinomas, adenocarcinomas, and squamous cell carcinomas. Samples from five control dogs without nasal neoplasia were also analyzed for COX-2 staining. These specimens were characterized by varying degrees of lymphoplasmacytic rhinitis with scattered regions of COX-2 positive respiratory epithelial and stromal cells. Whether the intensity and distribution of COX-2 expression in nasal tumors can be used as a prognostic marker requires further investigation. A combination therapy of irradiation and a selective COX-2 inhibitor appears worthy of clinical investigation in the treatment of canine epithelial nasal tumors.     

Spatial variability in herbicide degradation in the subsurface environment of a groundwater protection zone

The aim of this study was to investigate the spatial variability in degradation and mineralization of atrazine and isoproturon in subsurface samples taken from sandy loam soils overlying gravel terraces which form part of a groundwater protection zone. Percussion drilling was used to obtain samples from 11 boreholes (maximum depth 3 m). Unlabelled atrazine or isoproturon, and ring-14C-labelled atrazine or isoproturon were added to samples, incubated at 25 degrees C for up to 16 weeks, and analyzed for the residual herbicide or [14C]carbon dioxide. All samples showed the potential to degrade these herbicides, although the percentage degradation decreased by a factor of 2-3 from the surface soil to a depth of 3 m. This was associated with a decrease in organic matter content, but there was no change in the potential to mineralize acetate, indicating that specific changes in the catabolic ability of the microbial population occurred with depth. The capacity of samples to mineralize atrazine and isoproturon to carbon dioxide decreased markedly with depth, with no mineralization potential observed at a depth of 80 cm.     

A comparative study of the effects of high pressure on proteolytic degradation of sardine and blue whiting muscle

ABSTRACT:   High-pressure technology is used as an alternative to heat processing because of its inactivating effect on microorganisms and enzymes. However, it can also alter the structure of other muscle proteins. The present study compares the effects of high pressure (300 MPa, 7°C, 20 min) on the proteolytic degradation and alterations in the myofibrillar proteins of sardine and blue whiting muscle. Also, muscle homogenates and enzyme extracts were pressurized in order to evaluate the high-pressure effects on unprotected proteolytic enzymes outside the whole muscle structure. Peak proteolytic activity was found to occur at 55°C in both species. The peak activity pH was pH 3 for the sardine and pH 8 for the blue whiting; the main enzyme families being aspartic proteases in the former and alkaline serine proteases in the latter. Pressurization lowered activity levels at the peak activity pH and temperature in the fish muscle (by 30.8% in the sardine and by 9.5% in the blue whiting) and also slightly in the enzyme extracts (by 16.8% in the sardine and by 19.4% in the blue whiting). The electrophoretic profiles disclosed higher protein degradation in the pressurized muscle. Overall, the observed changes in proteolytic activity can be attributed not only to the effect of high pressure on the enzymes, but also and mainly, to the effect on other muscle proteins.     

Crystal and molecular structure of piceatannol; scavenging features of resveratrol and piceatannol on hydroxyl and peroxyl radicals and docking with transthyretin

The mechanism by which the naturally occurring polyphenolic compounds resveratrol (RES), C(14)H(12)O(3), and its metabolite piceatannol (PIC), C(14)H(12)O(4), scavenge free radicals is studied using experimental and density functional theory (DFT) methods. PIC's crystal structure shows a strong intermolecular hydrogen bond network, which, through a concerted motion of the hydroxyl hydrogen atoms, can produce a second hydrogen bond chain. This reorganization offers a low-energy pathway for the transfer of hydrogen atoms and is a contributing factor to PIC's biological activity. Additionally, DFT calculations describing the entire reaction mechanism of RES, PIC, and 3,3',4',5,5'-pentahydroxystilbene with hydroxyl and peroxyl radicals agree with experimental results, showing that increased hydroxylation aids in scavenging activity. PIC is more efficient than RES because (i) by sharing its 3'-OH hydrogen atom with its adjacent neighbor, O-4', the abstraction and transfer of the 4'-H atom to the free radical becomes easier and (ii) the resulting PIC semiquinone radical is more stable. As a result of the reaction with OH(*), both RES and PIC form water; with the peroxyl radical, both RES and PIC form hydrogen peroxide. Also, docking of PIC onto the protein transthyretin suggests better performance than RES and confirms its possible application in neurodegenerative conditions such as Alzheimer's disease.