Piglets performance, nutrient digestibility and gut health in response to feeding Ulva lactuca seaweed supplemented with a recombinant ulvan lyase or a commercial carbohydrase mixture.

Ulva lactuca, a green seaweed, may be an alternative source of nutrients and bioactive compounds for weaned piglets. However, it has a recalcitrant cell wall rich in a sulphated polysaccharide - ulvan - that is indigestible to monogastrics. The objective of this study was to evaluate the effect of dietary incorporation of 7% U. lactuca, combined with carbohydrases supplementation (commercial carbohydrase mixture or recombinant ulvan lyase), on growth performance, nutrient digestibility and gut health parameters (morphology and microbiota) of weaned piglets. The experiment was conducted over 14 days using 40 weaned piglets randomly allocated to one of four experimental diets: a control diet based on wheat-maize-soybean meal, a diet with 7% U. lactuca replacing the control diet (UL), a diet with UL supplemented with 0.005% Rovabio® Excel AP, and a diet with UL supplemented with 0.01% of a recombinant ulvan lyase. The dietary treatments had no major effects on growth performance, nitrogen balance and gut content variables, as well as histological measurements. Contrarily, dry matter and organic matter digestibility decreased with dietary seaweed inclusion, while hemicellulose digestibility increased, suggesting a high fermentability of this cell wall fraction independently of carbohydrases supplementation. Some beneficial microbial populations increased as a consequence of enzymatic supplementation (e.g., Prevotella), while seaweed diets as a whole led to an increased abundance of Shuttleworthia, Anaeroplasma and Lachnospiraceae_NK3A20_group, all related with a healthier gut. It also decreased Lactobacillus when compared to controls, which is possibly related to increased bioavailability of seaweed zinc. This study indicates that, under these experimental conditions, up to 7% dietary U. lactuca has no detrimental effect on piglet growth, despite decreasing acid detergent fibre digestibility. Carbohydrases supplementation of Ulva diets is not required at this incorporation level.

Trends in abundance indices derived from commercial fisheries for priority marine stocks in the Azores.

Large-scale assessments of fish and shellfish populations may be valuable for identifying the impacts of fishing pressure on stocks, but they are rarely possible due to a lack of long-term datasets. In this study, a two-part mixed-effect model for lognormal data was used to obtain historical abundance indices for 16 priority fish and shellfish stocks in the Azores region derived from catch per unit effort (kg per day at sea per vessel) and landing per unit effort (kg per landing per vessel). This data was obtained over the past 30 years under the EC Data Collection Framework and, in addition to information on effort and catch, included details about the fishing operation such as quarter, vessel length and fishing gear. This information was analysed to investigate how abundance indices changed over the years, verify if trends from different data sources were different and, if possible, relate these results to the population and fishery dynamics. The abundance indices derived from both datasets were generally in agreement with each other. Declining trends have been observed for some commercially important species, such as forkbeard Phycis phycis, European conger Conger conger, parrotfish Sparisoma cretense, red scorpionfish Scorpaena scrofa, offshore rockfish Pontinus kuhlii, common spiny lobster Palinurus elephas, splendid alfonsino Beryx splendens and alfonsino B. decadactylus, which is concerning because they are particularly slow-growing, long-lived and have low natural mortality, making them susceptible to overfishing. The results of this study are very important and should help future stock assessment and management initiatives.

Life History and Fishing Aspects of the Deep-Sea Silver Scabbardfish Lepidopus caudatus in the Azores.

Deep-sea fisheries are of important economic value. Therefore, it is necessary to generate biologically and ecologically based fishing plans to make this fishery sustainable over time. The silver scabbardfish, Lepidopus caudatus, is a worldwide-distributed demersal fish, commercially exploited in the Azores. Despite L. caudatus high landings and discharges in the region, information about its ecology, population structure, biology and fisheries remains little-known. This work analyzed scientific survey and fishery-dependent data from the past 30 years to understand the changes in abundance, spatial distribution and life history of this species. Lepidopus caudatus spatial distribution was associated with depths between 200 and 400 m, close to seamounts and islands, and on sandy bottoms. The size structure varied yearly, and the population was dominated by females (sex ratio, M:F = 0.46:1). Growth rates were between the estimated values in previous studies in the Azores and indicated a slow-grower species (L∞ = 171.62 cm, k = 0.12 year-1, Φ' = 3.52). The abundance indices highlighted a declining tendency, and this result was backed by the high exploitation rate for the fish in the region (E = 0.53). Lack of management measurements and the species' vulnerability could lead to the depletion of this species.

Effect of dietary Spirulina (Arthrospira platensis) on the intestinal function of post-weaned piglet: An approach combining proteomics, metabolomics and histological studies.

The effect of dietary Spirulina (Arthrospira platensis) and CAZyme supplementation was assessed on the gut of weaned piglets, using an integrated NMR-metabolomics approach combined with Tandem Mass Tag labelled proteomics. Thirty weaned male piglets were assigned to one of the three following diets (n = 10): cereal and soybean meal basal diet (Control), basal diet with 10% Spirulina inclusion (SP) and SP diet supplemented with 0.01% lysozyme (SP + L). The experiment lasted 4 weeks and, upon slaughter, small intestine samples were collected for histological, metabolomic and proteomic analysis. No significant differences were found for the histology and metabolomics analysis between the three experimental groups. Lactate, glutamate, glycine and myo-inositol were the most abundant metabolites. Proteomics results showed 1502 proteins identified in the intestine tissue. A total of 23, 78, 27 differentially abundant proteins were detected respectively for the SP vs. Control, SP + L vs. Control and SP + L vs. SP comparisons. The incorporation of Spirulina and supplementation of lysozyme in the piglet's diets is associated to intestinal proteomic changes. These include increased protein synthesis and abundance of contractile apparatus proteins, related with increased nutrient availability, which has beneficial (increased glucose uptake) and detrimental (increased digesta viscosity) metabolic effects. SIGNIFICANCE: The use of conventional feedstuffs becomes increasingly prohibitive due to its environmental toll. To increase the sustainability of the livestock sector, novel feedstuffs such as microalgae need to be considered. However, its recalcitrant cell wall has antinutritional effects that can inhibit high dietary inclusion levels. The supplementation with CAZymes is a possible solution to this issue. The small intestine is a central piece in monogastric digestion and of particular importance for the weaned piglet. Studying the effect of dietary Spirulina and CAZyme supplementation on its histomorphology, metabolome and proteome allows studying relevant physiological adaptations to these diets.

Growth Parameters and Mortality Rates Estimated for Seven Data-Deficient Fishes from the Azores Based on Length-Frequency Data.

Given the scarcity of information suitable for stock assessments, the growth and mortality of seven exploited marine fishes in Azorean waters were estimated based on length-frequency data. The studied species were Trachurus picturatus, Sparisoma cretense, Scomber colias, Scorpaena scrofa, Serranus atricauda, Seriola spp. and Aphanopus carbo. The growth parameters L∞ (cm), k (year-1) and ϕ' estimated through the ELEFAN_GA_boot routine were set at 55.87, 0.08 and 2.39 for T. picturatus; 55.03, 0.11 and 2.53 for S. cretense; 55.93, 0.18 and 2.76 for S. colias; 61.11, 0.11 and 2.61 for S. scrofa; 52.10, 0.12 and 2.51 for S. atricauda; 107.33, 0.12 and 3.18 for Seriola spp.; and 133.16, 0.09 and 3.19 for A. carbo; respectively. The total mortality rate estimated using the length-converted catch curve method was 0.22, 0.35, 0.58, 0.32, 0.31, 0.39 and 0.22 year-1; the natural mortality included 0.15, 0.20, 0.30, 0.20, 0.21, 0.21 and 0.16 year-1; and fishing mortality rate 0.07, 0.15, 0.28, 0.12, 0.10, 0.18 and 0.06 year-1, respectively, for the species mentioned. The relatively large sizes and slow growth with a low natural mortality rate indicate a high vulnerability to overfishing. Therefore, assessment and management initiatives are highly encouraged to ensure the sustainability of the resources.

Influence of Chlorella vulgaris on growth, digestibility and gut morphology and microbiota of weaned piglet.

The purpose of this study was to evaluate the impact of Chlorella vulgaris (5% in the diet), supplemented or not with two exogenous carbohydrase mixtures on piglets' performance, nutrient digestibility and gut morphology, fermentation products and microbiota. Forty-four male piglets weaned at 28 days of age, with 11.2 ± 0.46 kg of live weight, were used and assigned to 1 of 4 dietary treatments: cereal and soybean meal based-diet (control, n = 11), control diet with 5% of C. vulgaris (CH, n = 10), CH diet supplemented with 0.005% of Rovabio® Excel AP (CH + R, n = 10) and CH diet supplemented with 0.01% of a recombinant 4-carbohydrase mixture (CH + M, n = 11). Growth performance was not changed by the of C. vulgaris inclusion during 21 days of trial. However, total tract apparent digestibility of nutritional fractions was negatively impacted by the inclusion. In addition, the viscosity of duodenum plus jejunum contents slightly increased in all groups fed with the microalga. In contrast, dietary microalga increased duodenum villus height and promoted a healthier gut microbiota, with higher abundance of some specific bacterial taxa (Colidextribacter, Oscillospira and Lactobacillus). This study indicates that the dietary inclusion of 5% C. vulgaris improves piglets' gut health without impairing performance. Data also indicate that C. vulgaris reduces nutrient digestibility but promotes compensatory developments of gut mucosa and prebiotic effects. Dietary supplementation with exogenous carbohydrases does not seem to be necessary for this inclusion level. Therefore, the incorporation of CH as a sustainable feed ingredient in piglets' nutrition is a viable alternative approach.

Demographics and Yield-Per-Recruit Assessment of the Vulnerable Spiny Lobster Palinurus elephas in the Azores-Implications for Conservation and Fisheries Management.

The spiny lobster Palinurus elephas has been intensively harvested across its range and is generally considered overfished, with global landings declining sharply from an average of 820 t in 1960-79 to 385 t in 2000-19. Despite its economic and ecological importance, limited information is available to perform stock assessments and make robust management decisions. In this study, demographics and exploitation levels of P. elephas were determined from coastal areas of the Azores, and the relevance of these data for conservation planning was discussed. Carapace length varied between 39.6 and 174.3 mm, with mean sizes decreasing by depth. Males reached larger sizes and grew faster than females but were less dominant. Lifespan was 43 years for females and 60 years for males. The estimated mean length at first capture was 101.65 mm-around 58% of its asymptotic length. Fishing mortality and exploitation levels were close to the optimal values, indicating an uncertain future for Azorean populations if stock assessment and management initiatives are not focused on this species. Conservation and management strategies may benefit from these results under the 'precautionary approach' principle. However, up-to-date and accurate catch and fishing effort data need to be urgently collected.

Freeze-dried Nannochloropsis oceanica biomass protects eicosapentaenoic acid (EPA) from metabolization in the rumen of lambs.

Eicosapentaenoic acid (EPA) from freeze-dried biomass of Nannochloropsis oceanica microalgae resists ruminal biohydrogenation in vitro, but in vivo demonstration is needed. Therefore, the present study was designed to test the rumen protective effects of N. oceanica in lambs. Twenty-eight lambs were assigned to one of four diets: Control (C); and C diets supplemented with: 1.2% Nannochloropsis sp. oil (O); 12.3% spray-dried N. oceanica (SD); or 9.2% N. oceanica (FD), to achieve 3 g EPA /kg dry matter. Lambs were slaughtered after 3 weeks and digestive contents and ruminal wall samples were collected. EPA concentration in the rumen of lambs fed FD was about 50% higher than lambs fed SD or O diets. Nevertheless, the high levels of EPA in cecum and faeces of animals fed N. oceanica biomass, independently of the drying method, suggests that EPA was not completely released and absorbed in the small intestine. Furthermore, supplementation with EPA sources also affected the ruminal biohydrogenation of C18 fatty acids, mitigating the shift from the t10 biohydrogenation pathways to the t11 pathways compared to the Control diet. Overall, our results demonstrate that FD N. oceanica biomass is a natural rumen-protected source of EPA to ruminants.

Biological Knowledge of Thornback Ray (Raja clavata) from the Azores: Improving Scientific Information for the Effectiveness of Species-Specific Management Measures.

Elasmobranchs are globally recognized as vulnerable due to their life-history characteristics, fishing pressure, and habitat degradation. Among the skates and rays caught by commercial fisheries, the thornback ray Raja clavata is one of the most economically important in Northwest European seas. However, the scarcity of biological knowledge about this species in Azorean waters has limited the stock assessment types that can be conducted. To improve information on its habitat preferences, spatial distribution and movement pattern, growth, sex ratio, mortality, and reproduction, as well as to investigate long-term changes in abundance and size, this study analyzed approximately 25 years of fishery-dependent and independent data from the Azores. Raja clavata was mainly caught at depths up to 250 m. Most of the tagged fish were recaptured near the release point. A larger-deeper trend was found, and females were larger and more abundant than males. Life-history parameters showed that R. clavata has a long lifespan, large size, slow growth, and low natural mortality. The sustainability of its population is of concern to fisheries management and, while our findings suggested a relatively healthy stock in the Azores, a thorough increase in data quality is required to better understand the stock condition and prevent overexploitation.

Contributions to Management Strategies in the NE Atlantic Regarding the Life History and Population Structure of a Key Deep-Sea Fish (Mora Moro).

With the commercial fishery expansion to deeper waters, some vulnerable deep-sea species have been increasingly captured. To reduce the fishing impacts on these species, exploitation and management must be based on detailed and precise information about their biology. The common mora Mora moro has become the main deep-sea species caught by longliners in the Northeast Atlantic at depths between 600 and 1200 m. In the Azores, landings have more than doubled from the early 2000s to recent years. Despite its growing importance, its life history and population structure are poorly understood, and the current stock status has not been assessed. To better determine its distribution, biology, and long-term changes in abundance and size composition, this study analyzed a fishery-dependent and survey time series from the Azores. M. moro was found on mud and rock bottoms at depths below 300 m. A larger-deeper trend was observed, and females were larger and more abundant than males. The reproductive season took place from August to February. Abundance indices and mean sizes in the catch were marked by changes in fishing fleet operational behavior. M. moro is considered vulnerable to overfishing because it exhibits a long life span, a large size, slow growth, and a low natural mortality.

Influence of Dietary Supplementation with an Amino Acid Mixture on Inflammatory Markers, Immune Status and Serum Proteome in LPS-Challenged Weaned Piglets.

In order to investigate the effect of a dietary amino acid mixture supplementation in lipopolysaccharide (LPS)-challenged weaned piglets, twenty-seven 28-day-old (8.2 ± 1.0 kg) newly weaned piglets were randomly allocated to one of three experimental treatments for five weeks. Diet 1: a CTRL treatment. Diet 2: an LPS treatment, where piglets were intraperitoneally administered LPS (25 µg/kg) on day 7. Diet 3: an LPS+MIX treatment, where piglets were intraperitoneally administered LPS on day 7 and fed a diet supplemented with a mixture of 0.3% of arginine, branched-chain amino acids (leucine, valine, and isoleucine), and cystine (MIX). Blood samples were drawn on day 10 and day 35, and serum was analysed for selected chemical parameters and proteomics. The LPS and LPS+MIX groups exhibited an increase in haptoglobin concentrations on day 10. The LPS group showed an increased cortisol concentration, while this concentration was reduced in the LPS+MIX group compared to the control group. Similarly, the LPS+MIX group showed a decreased haptoglobin concentration on day 35 compared to the two other groups. Immunoglobulin concentrations were affected by treatments. Indeed, on day 10, the concentrations of IgG and IgM were decreased by the LPS challenge, as illustrated by the lower concentrations of these two immunoglobulins in the LPS group compared to the control group. In addition, the supplementation with the amino acid mixture in the LPS+MIX further decreased IgG and increased IgM concentrations compared to the LPS group. Although a proteomics approach did not reveal important alterations in the protein profile in response to treatments, LPS-challenged piglets had an increase in proteins linked to the immune response, when compared to piglets supplemented with the amino acid mixture. Overall, data indicate that LPS-challenged piglets supplemented with this amino acid mixture are more protected against the detrimental effects of LPS.

Can we consider the stocks of alfonsinos Beryx splendens and Beryx decadactylus from the Azores a discrete fishery management unit?

This paper summarises the detailed information on catch, fork length, weight, sex and maturity of alfonsinos Beryx splendens and Beryx decadactylus collected from targeted surveys in Azores waters, complemented by fishery data, over the past c. 20 years. To date, it was not possible to define if the Azorean component of the population can be considered as a discrete local management unit. The reason for this is the lack of available information for these resources covering the entire spatial distribution of the species in the North Atlantic Ocean. Additionally, there are some conflicts between the different genetic results available, poor understanding about the species movements and data from the Azores show some discrepancies in aspects of reproduction. Consequently, there is no analytical assessment for this resource and the stock of Beryx spp. is currently managed based on the precautionary approach. Outputs of recent analyses are presented in this study and possible assumptions and strategies for the assessment are discussed.

Preclinical randomized controlled trial of bilateral discectomy versus bilateral discopexy in Black Merino sheep temporomandibular joint: TEMPOJIMS - Phase 1- histologic, imaging and body weight results.

INTRODUCTION: The role of temporomandibular joint (TMJ) surgery is not well defined due to a lack of quality randomized controlled clinical trials, comparing different TMJ surgical treatments with medical and placebo interventions. The temporomandibular joint interposal study (TEMPOJIMS) is a rigorous preclinical trial divided in 2 phases. In phase 1 the authors investigated the role of the TMJ disc and in phase 2 the authors evaluated 3 different interposal materials. The present work of TEMPOJIMS - phase 1, aims to evaluate histopathologic and imaging changes of bilateral discectomy and discopexy in Black Merino sheep TMJ, using a high-quality trial following the ARRIVE guidelines. MATERIAL AND METHODS: This randomized, blinded and controlled preclinical trial was conducted in 9 Black Merino sheep to investigate histopathologic (primary outcome), imaging and body weight (secondary outcomes) changes after bilateral discectomy, discopexy and sham surgery. RESULTS: Significant changes were noticed in discectomy group, both in imaging and histopathologic analyses. Body weight changes were most pronounced in the discectomy group in the first 4 months after surgery with recovery to baseline weight 6 months after surgery. Discopexy induced nonsignificant changes in histopathologic, imaging and body weight analyses. CONCLUSIONS: This study reinforces the importance of developing an effective interposal material to substitute the TMJ disc and the need to explore the molecular mechanisms that underlie TMJ cartilage degeneration. The study design proposed in TEMPOJIMS represents an important progress towards future rigorous TMJ investigations.

Bioengineered Temporomandibular Joint Disk Implants: Study Protocol for a Two-Phase Exploratory Randomized Preclinical Pilot Trial in 18 Black Merino Sheep (TEMPOJIMS).

BACKGROUND: Preclinical trials are essential to test efficacious options to substitute the temporomandibular joint (TMJ) disk. The contemporary absence of an ideal treatment for patients with severe TMJ disorders can be related to difficulties concerning the appropriate study design to conduct preclinical trials in the TMJ field. These difficulties can be associated with the use of heterogeneous animal models, the use of the contralateral TMJ as control, the absence of rigorous randomized controlled preclinical trials with blinded outcomes assessors, and difficulties involving multidisciplinary teams. OBJECTIVE: This study aims to develop a new, reproducible, and effective study design for preclinical research in the TMJ domain, obtaining rigorous data related to (1) identify the impact of bilateral discectomy in black Merino sheep, (2) identify the impact of bilateral discopexy in black Merino sheep, and (3) identify the impact of three different bioengineering TMJ discs in black Merino sheep. METHODS: A two-phase exploratory randomized controlled preclinical trial with blinded outcomes is proposed. In the first phase, nine sheep are randomized into three different surgical bilateral procedures: bilateral discectomy, bilateral discopexy, and sham surgery. In the second phase, nine sheep are randomized to bilaterally test three different TMJ bioengineering disk implants. The primary outcome is the histological gradation of TMJ. Secondary outcomes are imaging changes, absolute masticatory time, ruminant time per cycle, ruminant kinetics, ruminant area, and sheep weight. RESULTS: Previous preclinical studies in this field have used the contralateral unoperated side as a control, different animal models ranging from mice to a canine model, with nonrandomized, nonblinded and uncontrolled study designs and limited outcomes measures. The main goal of this exploratory preclinical protocol is to set a new standard for future preclinical trials in oromaxillofacial surgery, particularly in the TMJ field, by proposing a rigorous design in black Merino sheep. The authors also intend to test the feasibility of pilot outcomes. The authors expect to increase the quality of further studies in this field and to progress in future treatment options for patients undergoing surgery for TMJ disk replacement. CONCLUSIONS: The study has commenced, but it is too early to provide results or conclusions.

Incomplete Dll4/Notch signaling inhibition promotes functional angiogenesis supporting the growth of skin papillomas.

BACKGROUND: In invasive malignancies, Dll4/Notch signaling inhibition enhances non-functional vessel proliferation and limits tumor growth by reducing its blood perfusion. METHODS: To assess the effects of targeted Dll4 allelic deletion in the incipient stages of tumor pathogenesis, we chemically induced skin papillomas in wild-type and Dll4 (+/-) littermates, and compared tumor growth, their histological features, vascularization and the expression of angiogenesis-related molecules. RESULTS: We observed that Dll4 down-regulation promotes productive angiogenesis, although with less mature vessels, in chemically-induced pre-cancerous skin papillomas stimulating their growth. The increase in endothelial activation was associated with an increase in the VEGFR2 to VEGFR1 ratio, which neutralized the tumor-suppressive effect of VEGFR-targeting sorafenib. Thus, in early papillomas, lower levels of Dll4 increase vascularization through raised VEGFR2 levels, enhancing sensitivity to endogenous levels of VEGF, promoting functional angiogenesis and tumor growth. CONCLUSION: Tumor promoting effect of low-dosage inhibition needs to be considered when implementing Dll4 targeting therapies.

Cell phenotypic change due to Cryptosporidium parvum infection in immunocompetent mice.

Cryptosporidium parvum is an intracellular parasite causing enteritis which can become life-threatening in immunocompromised host. Immunoregulatory T cells play a central role in the regulatory network of the host. Here, we proposed to characterize the populations of immune cells during infection and reinfection with C. parvum. Four-week-old BALB/C mice were inoculated with oocysts of C. parvum at days 0 and 22. Fecal and blood samples, spleens, and small intestines were collected for analysis. Peripheral blood and spleen cell populations were characterized by flow cytometry. After infection (days 0 to 21), mice presented higher values of neutrophils, eosinophils, NK cells and CD4(+)CD25(high) T cells in peripheral blood. After reinfection, this upward trend continued in the following days for all four populations in infected mice. At day 35, infected mice presented similar values to the control group, except for CD4(+)CD25(high) T cells, which remained higher in infected mice. A possible correlation between alterations in blood and spleen cell populations was also studied, but no consistent association could be established. Small intestine sections were screened for intracellular stages of the parasite but no evidence of pathology was observed. Here, we report information which may be important for the understanding of the specific cell-mediated response in immunocompetent mice to C. parvum infection. Although some questions remain unanswered and complementary studies are needed, our results are expected to contribute to a better understanding of innate and Treg cells role in the clearance process of this parasite.

Contrasting cellularity and fatty acid composition in fat depots from Alentejana and Barrosã bovine breeds fed high and low forage diets.

During the finishing phase of bovines, large amounts of subcutaneous and visceral fats are deposited leading to production inefficiencies with major impact on meat quality. A better understanding of the cellularity features of the main fat depots could provide strategies for adipose tissue manipulation. This study assessed the effect of feeding diets with distinct forage to concentrate ratios on the cellularity of two fat depots of beef cattle and their implications on the fatty acid profile. Thus, two phylogenetically distant Portuguese bovine breeds, Alentejana and Barrosã, were selected. The results did not show differences in subcutaneous fat deposition nor in visceral fat depots partitioning. Plasma adipokines concentration failed to show a consistent relationship with fatness, as leptin remained constant in all experimental groups, whereas interleukin-6 was influenced by breed. Fat depot seems to determine the area and number of adipocytes, with larger adipocytes and a lower number of cells in subcutaneous fat than in mesenteric fat. Neither breed nor diet influenced adipocytes area and number. The contents of total fatty acids, partial sums of fatty acids and conjugated linoleic acid isomeric profile were affected by breed and fat depot. The incorporation of saturated fatty acids (SFA), trans fatty acids, polyunsaturated fatty acids (PUFA) and branched chain fatty acids (BCFA) was higher in mesenteric fat depot, whereas subcutaneous fat depot had greater percentages of monounsaturated fatty acids (MUFA). In addition, SFA and MUFA proportions seem to be breed-related. In spite of the less relevant role of diet, the percentages of PUFA and BCFA were influenced by this factor. Under these experimental conditions, the effect of fat depot on cellularity and fatty acid composition prevails over breed or diet, as reinforced by the principal component analysis.

Contrasting apoptotic responses of conjugated linoleic acid in the liver of obese Zucker rats fed palm oil or ovine fat.

We hypothesized that reducing weight properties of conjugated linoleic acid (CLA) are due to adipocyte apoptosis and that CLA differentially modulates the apoptotic responses in hepatic lipotoxicity from rats fed saturated fat diets. Obese Zucker rats were fed atherogenic diets (2%w/w of cholesterol) formulated with high (15%w/w) saturated fat, from vegetable or animal origin, supplemented or not with 1% of a mixture (1:1) of cis-9, trans-11 and trans-10, cis-12 CLA isomers for 14 weeks. CLA induced no changes on retroperitoneal fat depot weight, which was in line with similar levels of apoptosis. Interestingly, CLA had a contrasting effect on cell death in the liver according to the dietary fat. CLA increased hepatocyte apoptosis, associated with upregulation of Fas protein in rats fed palm oil, compared to rats receiving palm oil alone. However, rats fed ovine fat alone displayed the highest levels of hepatic cell death, which were decreased in rats fed ovine fat plus CLA. This reducing effect of CLA was related to positively restoring endoplasmic reticulum (ER) ATF-6α, BiP and CHOP protein levels and increasing phosphorylated c-Jun NH(2)-terminal kinase (JNK) and c-Jun, thus suggesting an adaptive response of cell survival. These findings reinforce the role of CLA as regulator of apoptosis in the liver. Moreover, the dietary fat composition is a key factor in activation of apoptosis.

Diet supplementation with the cis-9,trans-11 conjugated linoleic acid isomer affects the size of adipocytes in Wistar rats.

Previous reports have demonstrated that conjugated linoleic acid (CLA) acts on body fat accumulation in a variety of animal models. The aim of the present study was to investigate the effect of cis (c)-9,trans (t)-11 and t10,c12 CLA isomers on the number and size of adipocytes from the inguinal and retroperitoneal fats in Wistar male rats. A 5.1% palm oil-based diet was supplemented with CLA isomers as follows: 0.6% of c9,t11, 0.6% of t10,c12, 1.3% of c9,t11 and t10,c12 isomers in mixture, and a control nonsupplemented group for comparative purposes. Fat tissues were prepared on microscope slides for histologic examination using an image-analysis software to count the number of adipocytes and measure cell sizes. The results showed that CLA isomers did not affect (P > .05) either final body and fat depot weights or serum lipids (with the exception of triacylglycerols) and adipocytokines (leptin and adiponectin). Animals fed the c9,t11 CLA isomer diet showed larger adipocytes when compared to other groups. Independently of the CLA dietary treatment, retroperitoneal fat showed larger adipocytes (3319 microm(2)) and therefore a smaller number of adipocytes per unit of area, compared to inguinal fat (3055 microm(2)). Taken together, the data suggest that a palm oil-based diet supplemented with the c9,t11 CLA isomer in Wistar rats, in contrast to the t10,c12 isomer and the mixture of both isomers, increases adipocyte dimensions in inguinal and retroperitoneal fat depots, while having a minor effect in serum lipids and adipocytokines.

Intra- and inter-specific variability in total and methylmercury bioaccumulation by eight marine fish species from the Azores.

We relate fish biological and ecological characteristics to total and organic mercury concentrations to determine whether accumulation is influenced by trophic level, Hg concentration in the diet, and vertical distribution. Levels of total mercury and methylmercury were determined in the muscle tissue of eight species of fish: Pagellus acarne, Trachurus picturatus, Phycis phycis, P. blennoides, Polyprion americanus, Conger conger, Lepidopus caudatus and Mora moro, caught in the Azores. All such fishes are commercially valuable and were selected to include species from a wide range of vertical distributions from epipelagic (<200 m) to mesopelagic (>300 m) environments. Methylmercury was the major form accumulated in all species, comprising an average of 88.1% of total mercury. Concentrations of mercury (total and methylmercury) increased with age, length and weight. Based on data from other studies, mercury concentrations in fish diet were estimated. Mercury levels in food ranged from 0.08 to 0.32 ppm, dry weight. Hg concentrations in the food and in muscle tissue from different species were positively correlated. Total Hg levels in the muscles were approximately nine times those estimated in food. Total mercury concentrations in muscle were positively correlated with both trophic level and median depth. Such enhanced mercury bioaccumulation in relation to depth appears to be determined primarily by concentrations in food and ultimately by water chemistry, which controls mercury speciation and uptake at the base of the food chain.