Fasting combined with long catch duration modifies the physio‐metabolic response and flesh quality of rainbow trout

Pre‐slaughter handling involves fasting fish and catching them, which can affect fish welfare and flesh quality, but few studies have considered their combined effects. In this study, adult rainbow trout (320 ± 10 g average weight) were fasted for 7 days (135.6 degree days) and subjected to a long catch duration (20 min), compared with controls (no fasting or short catch duration). Condition factor, organ weight indexes and carcass yield decreased with fasting but not catch duration. Plasma concentrations of cortisol, glucose and lactate increased after a long catch, while plasma triglycerides decreased with fasting. Liver glycogen concentration was lower in fasted fish, and liver luminosity and chroma were higher after fasting with a long catch. Regarding flesh quality, rigor mortis resolved more slowly and final muscle pH at 48 hr post‐mortem was higher for fasted fish with a long catch time. Muscle glycogen concentration was higher in fasted fish, where chroma was also lower. Fasted fish had lower lipid oxidation, but there were no differences in fat content in muscle. Fasted fish with a long catch duration also had less monounsaturated and more saturated fatty acids. In conclusion, a long catch triggered a stress response that had negative effects on flesh quality, independently of fasting.     

Prevalence and serotyping of Listeria monocytogenes in Portuguese live bivalve molluscs sampled in various steps along the sanitary control process

The prevalence and contamination levels of Listeria monocytogenes were investigated in live bivalve molluscs for human consumption, collected in various steps of the commercial and sanitary circuits in the North of Portugal. Samples of different species were collected per lot before and after depuration treatment in two depuration units and further, when placed in retail markets. Listeria monocytogenes was isolated from 4% of the samples although with very low contamination levels (less than 100 MPN g^?1). None of the environmental (non‐depurated) samples were contaminated with the food‐borne pathogen. The positive samples involved depurated and commercialized samples from two lots, one in each circuit. Two serotypes (1/2a and 3b) were detected among the isolated strains. This study demonstrated the possibility of occurrence of L. monocytogenes contamination of live bivalve molluscs along the sanitary control circuit, including at the retail market level. As L. monocytogenes grows well at refrigerated temperatures and in high salt matrixes such as seafood its presence in these products should not be overlooked.     

Use of probiotic bacteria in the rearing of Senegalese sole (Solea senegalensis) larvae

Three candidate probiotics, which had shown antimicrobial activity in vitro against two fish pathogens, were used in the rearing of Senegalese sole larvae and postlarvae. These probiotics improved the survival of starved sole yolk‐sac larvae. A feeding experiment of sole larvae and postlarvae comprised three treatments: A, B and C. Cultures of a Vibrio sp. (2J18) were added to treatment A, whereas a gram‐positive (J84) and a Shewanella sp. strain (2J27), were added to treatment B, while in a control treatment C no bacteria were added. Addition of bacteria in treatment B increased survival of larvae in the first phase of the experiment [0–20 days after hatching (DAH) (P<0.05)] and decreased the numbers of colony‐forming units (CFU) in larval gut 5 DAH compared with the control treatment (P<0.05). No differences were observed in survival (25–47%) during the second phase of the experiment (20–60 DAH). Nevertheless, the total numbers of CFU in fish gut 40 DAH were significantly lower in treatment B (P<0.05). The colonization rates of the added bacteria were the highest 5 and 11 DAH, and the highest average values reached were 65%, 82% and 17% of the total CFU count for the strains 2J18, 2J27 and J84 respectively.     

Does the presence of microalgae influence fish larvae prey capture?

The green water technique has been widely shown to improve fish larvae growth, survival and feed ingestion. Therefore, fish larvae (Sparus aurata L. and Solea senegalensis Kaup) feeding behaviour was studied through gut content analysis, when using different species of microalgae, as the ‘green water’ technique. Six treatments were used: Stain – food green stain; Tetra – microalgae Tetraselmis chuii; Iso – microalgae Isochrysis galbana; Tetra Sup –T. chuii supernatant (obtained from centrifugation); Phyto – a microalgae paste, Nannochloropsis oculata, (Phytobloom^®); and C water – clear water, as control. At 9, 16 and 23 days after hatching (DAH) for S. aurata, and 4, 9 and 14 DAH for S. senegalensis, 40 unfed fish larvae were transferred to 3 L experimental tanks, filled with the different ‘green water’ technique. Fish larvae were sampled 2 h after being fed with live prey, anaesthetized and fixed in buffered formaldehyde for posterior gut content determination. Feeding was evaluated by the feeding rate, percentage of larvae with prey items in the digestive tract and feeding intensity, number of prey in each larva digestive tract. Fish larvae feeding ability was influenced by the interaction between light conditions and substances provided by the presence of microalgae during fish larvae development. Sparus aurata was more dependent on microalgae addition than S. senegalensis larvae, which may be related to the type of prey, larval behaviour, ontogeny and physiology. The presence of microalgae influenced the selection of larger prey (Artemia over rotifers) by S. aurata aged 23 DAH.     

Relationship between assays of inflammation and coagulation: A novel interpretation of the canine activated clotting time

The processes of inflammation and coagulation are known to be interconnected through several mechanisms; however, the influence of inflammation on the interpretation of coagulation assays remains unknown. Blood was collected from 87 dogs admitted to a tertiary referral intensive care unit (ICU) and 15 control dogs. The association between 2 markers of inflammation [mature neutrophil count and C-reactive protein (CRP)] and 5 coagulation parameters [activated clotting time (ACT), prothrombin time (PT), activated partial thromboplastin time (aPTT), antithrombin (AT), and platelet count (plt)] were evaluated through correlation analysis. The study population was then divided into 4 groups based on severity of ACT prolongation with comparisons to all other variables assessed through an analysis of variance (ANOVA) test. A strong correlation for a biological system was demonstrated between ACT and CRP (r = 0.66; P < 0.0001). Statistically significant results were also found between aPTT and AT with the markers of inflammation, but the correlations were weaker. Within ACT groups of increasing severity, higher CRP concentrations (P < 0.0001) and lower AT activities (P < 0.0001) were identified. This study provides evidence for an association between assays of inflammation and coagulation and suggests that modification of our traditional interpretations of coagulation assays may be required. As a point-of-care test, ACT is a simple and inexpensive tool that can be used to assess an underlying inflammatory or hemostatic process.     

Evaluation of a technique using the carbon dioxide laser for the treatment of aural hematomas

A new technique using the carbon dioxide (CO2) laser for the treatment of aural hematomas is described. The laser is used to make an incision into the hematoma to allow for evacuation of the blood, and then multiple, small incisions are made over the surface of the hematoma to stimulate adhesions between the tissue layers. The CO2 laser was used in this fashion to treat 10 aural hematomas in eight dogs. Follow-up ranged from 1 to 23 months. Owners evaluated the cosmetic results following CO2 laser surgery as excellent in three ears, good in five ears, and fair in two ears. Hematomas were resolved in all 10 cases, although two cases developed serosanguineous fluid accumulation that required percutaneous drainage in one case and a second laser procedure in the other case.     

Does the Intensity of Anisakis Infection Affect the Quality of Hake Muscle?

The distribution of Anisakis larvae in fresh caught hake was studied, and the influence of three infection levels of the viscera (low, medium, high) on quality changes of deep frozen muscle was followed over 220 days. Differences in quality were attributed more to the size of the individuals than to the infection degree of the three groups. The removal of belly flaps diminished the risk of consumers’ infection and sensitization as prevalence and intensity was reduced to 75.6 and 5.9%, respectively. Allergens were detected after 220 days frozen storage, which represents a risk for consumers already allergic to Anisakis.     

Evaluation of micronized lupin seed meal as an alternative protein source in diets for gilthead sea bream Sparus aurata L. juveniles

A 12‐week growth trial was performed to evaluate the effect of lupin seed meal as a protein source in diets for gilthead sea bream (Sparus aurata) juveniles. Six experimental diets were formulated to be isonitrogenous and isoenergetic and to contain 10%, 20% and 30% of raw lupin (Lupinus angustifolius) seed meal protein or 20% and 30% lupin (L. angustifolius) seed meal processed by infrared radiation (micronized) in place of fish meal protein, the only protein source of the control diet. Fish accepted all diets well and no significant differences in feed utilization among groups were noticed during the trial. Final weight of fish fed the experimental diets was identical or higher than the control group. Final weight of fish fed diets including 20% micronized lupin protein was even significantly higher than that of fish fed the fish meal‐based control diet. Moreover, at the same dietary lupin seed meal protein inclusion levels, final weight of fish fed diets including micronized lupin was significantly higher than with raw lupin. A trend was also noticed for a decrease of final weight with the increase in lupin seed meal in the diets. At the end of the trial no significant differences in proximate whole‐body composition, hepatosomatic and visceral indices were observed among groups. It is concluded that lupin seed meal can replace up to 30% fish meal protein in diets for gilthead sea bream juveniles with no negative effects on growth performance. Furthermore, micronization of lupin seeds improves its dietary value for gilthead sea bream juveniles. At the same dietary lupin inclusion levels, diets including micronized lupin seeds promote significantly higher growth rates than raw lupin seeds.     

Soil microbiome biomass,activity, composition and CO2 emissions in a long-term organic and conventional farming systems

The implementation of environmentally friendly agricultural policies has increased the need to compare agricultural aspects of conventional (CON) and organic farming (ORG) systems. The objective of the present work was to compare the effects of an organic and conventional long-term experiment on bacterial and fungal biomass and activity, as well as soil CO₂ emission and readily available nitrogen forms in a soil cultivated with Helianthus annuus L. The microbial biomass was more active and abundant in ORG as well as soil CO₂ emission. Despite being less abundant, fungi were more active than bacteria in both ORG and CON experiments. 16S rRNA gene sequencing showed that the ORG treatment had a significantly greater bacterial richness than CON. Cyanobacteria, Actinobacteria and Proteobacteria were the most abundant phyla contributing more than others to the differences between the two systems. Moreover, the soil ${\mathit{NH}}_4^{+}$ and ${\mathit{NO}}_2^{-}$ content was not significantly different between ORG and CON, while ${\mathit{NO}}_3^{-}$ was less in ORG. ORG sunflower yield was significantly less compared with CON. While much remains to be discovered about the effects of these agricultural practices on soil chemical properties and microbial diversity, our findings may contribute to this type of investigation.     

Standardization of canine meningioma grading: Validation of new guidelines for reproducible histopathologic criteria

Canine meningiomas are currently graded using the human grading system. Recently published guidelines have adapted the human grading system for use in dogs. The goal of this study was to validate the new guidelines for canine meningiomas. To evaluate the inter-observer agreement, 5 veterinary surgical pathologists graded 158 canine meningiomas following the human grading system alone or with the new guidelines. The inter-observer agreement for histologic grade and each of the grading criteria (mitotic grade, invasion, spontaneous necrosis, macronucleoli, small cells, hypercellularity, pattern loss and anaplasia) was evaluated using the Fleiss kappa index. The diagnostic accuracy (sensitivity and specificity) was assessed by comparing the diagnoses obtained with the 2 grading systems with a consensus grade (considered the reference classification). The consensus histologic grade was obtained by agreement between 4 experienced veterinary neuropathologists following the guidelines. Compared with the human grading alone, the canine-specific guidelines increased the inter-observer agreement for: histologic grade (κ = 0.52); invasion (κ = 0.67); necrosis (κ = 0.62); small cells (κ = 0.36); pattern loss (κ = 0.49) and anaplasia (κ = 0.55). Mitotic grade agreement remained substantial (κ = 0.63). The guidelines improved the sensitivity in identifying grade 1 (95.6%) and the specificity in identifying grade 2 (96.2%) meningiomas. In conclusion, the new grading guidelines for canine meningiomas are associated with an overall improvement in the inter-observer agreement and higher diagnostic accuracy in diagnosing grade 1 and grade 2 meningiomas.