Efficacy of Doxycycline as Pre-exposure and/or Post-exposure Prophylaxis to Prevent Sexually Transmitted Diseases: A Systematic Review and Meta-analysis.

INTRODUCTION: Bacterial sexually transmitted infections (STIs), specifically infection by Chlamydia trachomatis, Neisseria gonorrhoeae, and Treponema pallidum, have an important burden worldwide. The use of doxycycline as Pre-Exposure Prophylaxis (PREP or Doxy-PREP) or Post-Exposure Prophylaxis (PEP or Doxy-PEP) might be effective as prophylaxis since it is effective against C. trachomatis and T. pallidum. AIMS: Our objective was to evaluate the efficacy of doxycycline as PREP or PEP against bacterial STIs (C. trachomatis, N. gonorrhoeae, and Treponema pallidum). METHODS: A systematic review and meta-analysis of randomized clinical trials of a high-risk group of individuals was conducted to evaluate whether doxycycline is as effective as PREP or PEP in preventing bacterial STIs. The PubMed-MEDLINE, Cohrane, Scielo, and LILACS databases were searched for randomized clinical trials published up to March 2024. Data were extracted from published reports. Hazard Ratios (HRs) and Risk Ratios (RR) with 95% CI were pooled across trials. MAIN OUTCOME MEASURE: The primary endpoints were any incidence of bacterial STIs and individual STI infections. RESULTS: A total of four studies were included in the analysis, three of which evaluated Doxy-PEP and one evaluated Doxy-PREP. In the Doxy-PEP group, a total of 1182 participants were evaluated. In the pooled analysis of Doxy-PEP studies, the incidence of the first STI was lower in the Doxy-PEP group (HR = 0.538 [95% CI = 0.337-0.859]) (I2 = 77%; P-value <0.05). Regarding individual infections, only two studies were included. In the Doxy-PEP group, the incidence of individual infection of C. trachomatis was lower compared to controls (RR = 0.291 [95% CI = 0.093-0.911]) (I2 = 89%; P-value <0.05). Since only one study evaluated Doxy-PREP, it was not possible to calculate a meta-analysis index; however, the use of doxycycline as pre-exposure prophylaxis was associated with a decrease in the rate of any STI. CONCLUSIONS: The use of Doxy-PEP might reduce the first STI, mainly C. trachomatis, if used within 72 h after condomless sex. The use of Doxy-PREP might also decrease the chance of any STI, however, only one study was evaluated.

The Use of Information Entropy and Expert Opinion in Maximizing the Discriminating Power of Composite Indicators.

This research offers a solution to a highly recognized and controversial problem within the composite indicator literature: sub-indicators weighting. The research proposes a novel hybrid weighting method that maximizes the discriminating power of the composite indicator with objectively defined weights. It considers the experts' uncertainty concerning the conceptual importance of sub-indicators in the multidimensional phenomenon, setting maximum and minimum weights (constraints) in the optimization function. The hybrid weighting scheme, known as the SAW-Max-Entropy method, avoids attributing weights that are incompatible with the multidimensional phenomenon's theoretical framework. At the same time, it reduces the influence of assessment errors and judgment biases on composite indicator scores. The research results show that the SAW-Max-Entropy weighting scheme achieves greater discriminating power than weighting schemes based on the Entropy Index, Expert Opinion, and Equal Weights. The SAW-Max-Entropy method has high application potential due to the increasing use of composite indicators across diverse areas of knowledge. Additionally, the method represents a robust response to the challenge of constructing composite indicators with superior discriminating power.

Effect of Dietary Incorporation of Yellow Mealworm as a Partial Fishmeal Replacer on Growth, Metabolism, and Intestinal Histomorphology in Juvenile Meagre (Argyrosomus regius).

Efforts have been made to find alternatives to fish meal (FM), as the sustainability of aquaculture depends on it. Insect meal (IM) is a potential candidate to partially replace FM, being more sustainable and economically viable. In this experimental trial, three diets were tested with different yellow mealworm incorporation: a control diet with no IM, a diet with an inclusion of 10% IM (Ins10), and a diet with an incorporation of 20% IM (Ins20). The diets were tested on 10.5 g meagre for 47 days. The results showed that an IM inclusion higher than 10% affected both growth (2.6 vs. 2.2) and FCR (1.5 vs. 1.9) of meagre juveniles. However, this reduction in growth did not result from lower protein retention or changes in muscle fibre area or density. Little differences were observed in the activity of pancreatic and intestinal enzymes except for aminopeptidase total activity which was higher in the control and Ins10 compared to Ins20 (3847 vs. 3540 mU/mg protein), suggesting no limitations in protein synthesis. Also, the alkaline phosphatase intestinal maturation index was higher in the control group compared to the IM groups (437 vs. 296). On the contrary, several differences were also found in the proteolytic activity in the hepatic and muscle tissues of meagre juveniles fed the Ins10 diet. The inclusion of IM had no impact on intestine histomorphology but changes were detected in the enterocytes of fish from control and Ins10 which showed hypervacuolization and nucleus misplacement compared to the Ins20 treatment. Nevertheless, a higher percentage of Vibrionaceae was recorded for meagre fed on the Ins20 diet. Since no signs of inflammation were observed in the distal intestine, this suggests IM incorporation could have had an important impact on intestinal health due to its antimicrobial properties. This is supported by an increase in the haematocrit in the treatments where IM was added (20 to 25%). In conclusion, incorporations of IM at percentages up to 10% do not seem to have a negative impact on meagre performance at this age but can enhance the fish immune system and protection against intestinal inflammation.

Obesity markers in patients with drug allergy and body fat as a predictor.

BACKGROUND: Obesity is a chronic low-grade inflammation state associated with several diseases. OBJECTIVE: To investigate a potential link between drug allergy and obesity, exploring whether the association depends on the type (immediate vs nonimmediate) or the severity of the reaction. METHODS: Anthropometric measurements, bioimpedance, and biochemical analysis, including serum adipokines, were performed in 90 consecutive adult patients studied for suspected drug allergy. Logistic regression models were developed to identify predictors of drug allergy. RESULTS: A total of 84 patients completed the diagnostic workup (78.6% women; mean age 39.58 ± 13.3 years). Drug allergy was confirmed in 39 patients and excluded in 45 (controls). Regarding body mass index, 42.2% had normal weight and 55.3% were overweight/obese. A total of 58% of women and 41% of men fulfilled the criteria for central obesity. Patients with drug allergy exhibited considerably higher body mass index, waist and hip circumferences, waist-hip ratio, fat mass, body fat percentage (BFP), trunk fat mass, leptin levels, and leptin-adiponectin ratio than controls. Similar results were obtained in the subgroup with immediate reactions, compared with the nonimmediate or unknown reactions. The higher the BFP and the number of reactions, the greater the odds of drug allergy (odds ratio [OR], 1.07; 95% confidence interval [CI], 1.01-1.14 and OR, 2.82; 95% CI, 1.31-6.10, respectively). An immediate reaction was also a predictor of drug allergy (OR, 3.81; 95% CI, 1.30-11.14, P = .02), compared with nonimmediate or unknown reactions. In patients with drug allergy, BFP was a predictor of having an immediate reaction (OR, 1.12; 95% CI, 1.02-1.24, P = .02). CONCLUSION: Our study illustrates, for the first time, evidence of a link between obesity and drug allergy, particularly immediate reactions. The BFP emerged as a potential predictor of drug allergy.

Adipocytes and macrophages secretomes coregulate catecholamine-synthesizing enzymes.

Obesity associates with macrophage accumulation in adipose tissue where these infiltrating cells interact with adipocytes and contribute to the systemic chronic metabolic inflammation present in immunometabolic diseases. Tyrosine hydroxylase (TH) and phenylethanolamine N-methyltransferase (PNMT) are two of the main enzymes of catecholamines (CA) synthesis. Adipocytes and macrophages produce, secrete and respond to CA, but the regulation of their synthesis in the interplay between immune and metabolic systems remains unknown. A model of indirect cell coculture with conditioned medium (CM) from RAW 264.7 macrophages with or without LPS-activation and 3T3-L1 adipocytes and preadipocytes was established to study the effect of cellular secretomes on the expression of the above enzymes. During the adipocyte differentiation process, we found a decrease of TH and PNMT expression. The secretome from LPS-activated macrophages downregulated TH and PNMT expression in preadipocytes, but not in mature adipocytes. Mature adipocytes CM induced a decrease of PNMT levels in RAW 264.7 macrophages. Pre and mature adipocytes showed a similar pattern of TH, PNMT and peroxisome proliferator-activated receptor gamma expression after exposure to pro and anti-inflammatory cytokines. We evidenced macrophages and adipocytes coregulate the expression of CA synthesis enzymes through secretome, with non-inflammatory signaling networks possibly being involved. Mediators released by macrophages seem to equally affect CA production by adipocytes, while adipocytes secretome preferentially affect AD production by macrophages. CA synthesis seems to be more determinant in early stages of adipogenic differentiation. Our results suggest that CA are key signaling molecules in the regulation of immune-metabolic crosstalk within the adipose tissue.

Communication Interface Manager for Improving Performance of Heterogeneous UAV Networks.

Exchanging messages with stable connections in missions composed of multiple unmanned aerial vehicles (UAV) remains a challenge. The variations in UAV distances from each other, considering their individual trajectories, and the medium dynamic factors are important points to be addressed.In this context, to increase the stability of UAV-to-UAV (U2U) communication with link quality, this paper presents an interface manager (IM) that is capable of improving communication in multi-UAV networks.Given a predefined set of available individual wireless interfaces, the proposed IM dynamically defines the best interface for sending messages based on on-flight conditions sensed and calculated dynamically from the wireless medium. Different simulation scenarios are generated using a complex and realistic experimental setup composed of traditional simulators such as NS-3, Gazebo, and GzUAV. IEEE 802.11n 2.4 GHz and 802.11p 5 GHz interfaces are used for the IM selection. The IM performance is evaluated in terms of metrics from the medium-access-control (MAC) and physical layers, which aim to improve and maintain the connectivity between the UAVs during the mission, and from the application layer, which targets the reliability in the delivery of messages. The obtained results show that compared with the cases where a single interface is used, the proposed IM is able to increase the network throughput and presents the best proportion of transmitted and received packets, reception power (-60 dBm to -75 dBm), and loss (-80 dB to -85 dB), resulting in a more efficient and stable network connections.

Potential biomarkers of ductal carcinoma in situ progression.

BACKGROUND: Ductal carcinoma in situ is a non-obligate precursor of invasive breast carcinoma and presents a potential risk of over or undertreatment. Finding molecular biomarkers of disease progression could allow for more adequate patient treatment. We aimed to identify potential biomarkers that can predict invasiveness risk. METHODS: In this epithelial cell-based study archival formalin-fixed paraffin-embedded blocks from six patients diagnosed with invasive lesions (pure invasive ductal carcinoma), six with in-situ lesions (pure ductal carcinoma in situ), six with synchronous lesions (invasive ductal carcinoma with an in-situ component) and three non-neoplastic breast epithelium tissues were analyzed by gene expression profiling of 770 genes, using the nCounter® PanCancer Pathways panel of NanoString Technologies. RESULTS: The results showed that in comparison with non-neoplastic tissue the pure ductal carcinoma in situ was one with the most altered gene expression profile. Comparing pure ductal carcinoma in situ and in-situ component six differentially expressed genes were found, three of them (FGF2, GAS1, and SFRP1), play a role in cell invasiveness. Importantly, these genes were also differentially expressed between invasive and noninvasive groups and were negatively regulated in later stages of carcinogenesis. CONCLUSIONS: We propose these three genes (FGF2, GAS1, and SFRP1) as potential biomarkers of ductal carcinoma in situ progression, suggesting that their downregulation may be involved in the transition of stationary to migrating invasive epithelial cells.

High-fat diet promotes adrenaline production by visceral adipocytes.

PURPOSE: Catecholamines (CA) play a major role in metabolism and immune response. Recent reports showing adipose tissue can synthetize CA enlighten new roles for these amines in obesity. This study aimed to evaluate the expression of both tyrosine hydroxylase (TH) and phenylethanolamine N-methyltransferase (PNMT) and CA content along preadipocytes differentiation, under normal and obesigenic conditions. METHODS: 8-9 week-old male C57BL/6 mice were divided in two groups: one fed with a high-fat diet (HFD) and other with a standard diet (SD) for 20 weeks. Afterwards, both TH and PNMT expression, localization, and CA content in adipocytes, were evaluated. RESULTS: qPCR results showed no changes for TH and PNMT expression during the differentiation process for visceral and subcutaneous preadipocytes from mice fed with SD. Comparing to SD, HFD increased TH gene expression of subcutaneous preadipocytes and PNMT gene expression of both visceral preadipocytes and adipocytes. HPLC-ED analyses revealed HFD increased visceral adipocytes noradrenaline intracellular content comparing with preadipocytes (p = 0.037). When compared with SD, HFD raised and decreased noradrenaline content, respectively, in visceral adipocytes (p = 0.004) and subcutaneous preadipocytes (p = 0.001). Along the differentiation process, HFD increased visceral adrenaline intracellular content comparing with SD (p < 0.001). HFD increased visceral comparing to subcutaneous adrenaline content for both preadipocytes (p = 0.004) and adipocytes (p = 0.001). CONCLUSIONS: TH and PNMT expression in adipose tissue is differently modulated in visceral and subcutaneous adipose depots, and seems to depend on diet. Differences observed in visceral adipose CA handling in HFD-fed mice might uncover novel pharmacological/nutritional strategies against obesity and cardiovascular risk.

Catechol-O-Methyltransferase (COMT): An Update on Its Role in Cancer, Neurological and Cardiovascular Diseases.

Catechol-O-methyltransferase (COMT) is an enzyme that catalyses the methylation of catechol substrates, classically in catecholamine metabolism, but also acting upon other substrates such as oestrogen and polyphenols. Although its classical function has been established for more than five decades, an ever expanding COMT role in other pathways and diseases has become a subject of active study in recent years. The most highlighted domains are related with COMT involvement in neuropsychiatric disorders and its role in the neurobiology of cognition, behaviour, emotions, pain processing and perception, sleep regulation, addictive behaviour and neurodegeneration. Nonetheless, great attention is also being devoted to a possible COMT contribution to the development of cardiovascular disorders and hormonally influenced diseases, including cancer. This review aims to update the role of COMT function and its involvement in cardiovascular and neurological disorders.

Effect of copper exposure and recovery period in reared Diplodus sargus.

The aquaculture growth can be followed by the occurrence of more and new pathogenic agents, since the production leads to higher fish densities in confined areas more appropriate to the appearance and propagation of pathologies. Copper sulfate has been widely used in preventing and controlling fish parasites. The objective of this study is to investigate the effects of copper treatments in the fish tissues (bioaccumulation and histological changes in different organs), mortality and evaluate what happens during the recovery period. White sea bream (Diplodus sargus) were exposed to copper sulfate (0.25 and 0.5 mg L-1) during 60 days followed with a 75-day recovery period. The results showed that the concentration of copper in fish liver was significantly higher in the 0.5 mg L-1 treatment than in the 0.25 mg L-1 treatment. Conversely, copper load in the muscle did not differ significantly between treatments and control. Copper levels in muscle, and especially in liver, increased during copper exposure (up to 60 days). In summary, at higher concentrations copper sulfate treatment (0.5 mg L-1) might be toxic to fish, which showed histological alterations and copper accumulation in their tissues, mainly in the liver. Nevertheless, individuals returned to their original state after a 75-day recovery period and the tested copper concentrations does not represents risk for food safety.

Preliminary studies on haematological and plasmatic parameters in gilthead sea bream (Sparus aurata) held under day/night temperature variations.

This study aimed to evaluate to what extent diel water temperature oscillations over a short period of time (41 h) affected haematologic and plasmatic parameters of gilthead sea bream. Sea bream juveniles (160 fish; 177.2 ± 20.9 g) were divided in two homogenous groups, experimental and control. The experimental group was exposed to water at 22.3 ± 0.6 °C between 5 a.m. and 5 p.m. and to colder water from 5 p.m. to 5 a.m. (temperature variation of 8.6 ± 0.3 °C) during 41h period, whereas for the control group, water temperature was maintained constant (22.4 ± 0.7 °C) for the same period. Fish were fed three times per day (10:30 a.m., 2 p.m., 4 p.m.). Blood was collected from fish caudal veins (n = 6) at different time points (10 a.m., 1 p.m., 5 p.m., 6:30 p.m., 8 p.m. and 10 a.m. of the following day) from each group, for haematologic (haematocrit and haemoglobin) and plasmatic (glucose, cortisol, triglycerides and total protein) determinations. All parameters studied tended to vary throughout the study period, both for experimental and control groups. Significant differences were observed between the two groups for all the studied parameters at different sampling points. These differences were observed near the influence of the cold-water period or during the cooling-water period. Results suggested that colder water temperature cycles depressed fish metabolic activity and affected the levels of the analysed blood-circulating metabolites. Overall, results suggested that stress response was similar for experimental and control groups and that gilthead sea bream were able to cope with diel water temperature variations.

ß2 Adrenoceptors are underexpressed in peripheral blood mononuclear cells and associated with a better metabolic profile in central obesity.

Background: Central obesity (CO) is an inflammatory disease. Because immune cells and adipocytes are catecholamines(CA)-producing cells, we studied the expression of adrenoceptors (AR) in peripheral blood mononuclear cells (PBMCs) hypothesizing a distinct adrenergic pattern in inflammatory obesity. Methods: AR expression was assessed in blood donors categorized by waist circumference (WC) (CO: WC≥0.80 m in women and ≥0.94 m in men). Following a pilot study for all AR subtypes, we measured ß2AR expression in fifty-seven individuals and correlated this result with anthropometric, metabolic and inflammatory parameters. A ratio (R) between AR mRNA of CO and non-CO<0.5 was considered under and >2.0 over expression. Results: The pilot study revealed no differences between groups, except for ß2AR mRNA. CO individuals showed underexpression of ß2AR relatively to those without CO (R=0.08; p=0.009). ß2AR expression inversely correlated with triacylglycerol (r=-0.271; p=0.041), very low-density lipoprotein-cholesterol (r=-0.313; p=0.018) and leptin (r=-0.392; p=0.012) and positively with high-density lipoprotein-cholesterol (r=0.310: p=0.045) plasma levels. Multiple logistic regression analysis showed a protective effect of ß2AR expression (≥2x10-6) [odds ratio (OR) 0.177 with respective confidence interval of 95% (95% CI) (0.040- 0.796)] for the occurrence of CO. A higher association was found for women as compared to men (Ξ9:1) [OR 8.972 (95% CI) (1.679-47.949)]. Conclusion: PBMCs ß2AR, underexpressed in centrally obese, are associated with a better metabolic profile and showed a protective role for the development of CO. The discovery of ß2AR as a new molecular marker of obesity subphenotypes in PBMCs might contribute to clarify the adrenergic immunomodulation of inflammatory obesity.

Performance of a core of transversal skills: self-perceptions of undergraduate medical students.

BACKGROUND: There is an increasingly growing trend towards integrating scientific research training into undergraduate medical education. Communication, research and organisational/learning skills are core competences acquired by scientific research activity. The aim of this study was to assess the perceived performance of a core of transversal skills, related with scientific research, by Portuguese medical students. METHODS: A cross-sectional study was conducted in 611 Portuguese students attending the first, fourth and sixth years of the medical course, during the same academic year. A validated questionnaire was applied for this purpose. RESULTS: Medical students felt confident regarding the majority of the analyzed transversal skills, particularly regarding team work capacity (72.7% perceived their own capacity as good). On the other hand, the perceived ability to manage information technology, time and to search literature was classified only as sufficient by many of them. The progression over the medical course and participation in research activities were associated with an increasing odds of a good perceived performance of skills such as writing skills (research activity: OR = 2.00; 95% CI: 1.34-2.97) and English proficiency (research activity: OR = 1.59; 95% CI: 1.06-2.38/final year medical students: OR = 3.63; 95% CI: 2.42-5.45). CONCLUSIONS: In this line, the early exposure to research activities along undergraduate medical education is an added value for students and the implementation of an integrated research program on medical curriculum should be considered.

Lipoxin Inhibits Fungal Uptake by Macrophages and Reduces the Severity of Acute Pulmonary Infection Caused by Paracoccidioides brasiliensis.

Cysteinyl leukotrienes (CysLTs) and lipoxins (LXs) are lipid mediators that control inflammation, with the former inducing and the latter inhibiting this process. Because the role played by these mediators in paracoccidioidomycosis was not investigated, we aimed to characterize the role of CysLT in the pulmonary infection developed by resistant (A/J) and susceptible (B10.A) mice. 48 h after infection, elevated levels of pulmonary LTC4 and LXA4 were produced by both mouse strains, but higher levels were found in the lungs of susceptible mice. Blocking the CysLTs receptor by MTL reduced fungal loads in B10.A, but not in A/J mice. In susceptible mice, MLT treatment led to reduced influx of PMN leukocytes, increased recruitment of monocytes, predominant synthesis of anti-inflammatory cytokines, and augmented expression of 5- and 15-lipoxygenase mRNA, suggesting a prevalent LXA4 activity. In agreement, MTL-treated macrophages showed reduced fungal burdens associated with decreased ingestion of fungal cells. Furthermore, the addition of exogenous LX reduced, and the specific blockade of the LX receptor increased the fungal loads of B10.A macrophages. This study showed for the first time that inhibition of CysLTs signaling results in less severe pulmonary paracoccidioidomycosis that occurs in parallel with elevated LX activity and reduced infection of macrophages.

Comparative analysis on natural spawning of F1 meagre, Argyrosomus regius, with wild broodstock spawns in Portugal.

The purpose of this study was to describe and compare the reproductive success and egg and larvae quality between wild and first-generation (F1) breeders of Argyrosomus regius. Wild broodstock were adapted to captivity, and good-quality spawns were obtained in 2009-2010, after GnRH treatment. In 2012, the F1 meagre (3 years old) spawned naturally at IPMA's Aquaculture Research Station facilities. From each spawning event, the following parameters were determined: number of floating and non-floating eggs, egg size, hatching success and larval total length. Eggs size and percentage of hatching obtained from F1 breeders (1.04 ± 0.10 mm and 90.5 ± 6.4%) were significantly higher when compared with wild breeders (0.97 ± 0.13 mm and 17.0 ± 12.7%). Although wild breeder spawns exhibited 2.7 ± 0.2 mm for larval total length, F1 breeder spawns presented 2.6 ± 0.2 mm. The wild and F1 breeder spawns exhibit a good egg and larval quality, indicating a promising starting point for a successful meagre hatchery production.

Problem-based learning in pharmaceutical education: a systematic review and meta-analysis.

OBJECTIVE: To assess the effects of problem-based learning (PBL) on the learning achievements of pharmacy students. METHODS: We searched for controlled studies that compared PBL to traditional learning in pharmacy courses (graduate and undergraduate) in the major literature databases up to January 2014. Two independent researchers selected the studies, extracted the data, and assessed the quality of the studies. Meta-analyses of the outcomes were performed using a random effects model. RESULTS: From 1,988 retrieved records, five were included in present review. The studies assessed students' impressions about the PBL method and compared student grades on the midterm and final examinations. PBL students performed better on midterm examinations (odds ratio [OR] = 1.46; confidence interval [IC] 95%: 1.16, 1.89) and final examinations (OR = 1.60; IC 95%: 1.06, 2.43) compared with students in the traditional learning groups. No difference was found between the groups in the subjective evaluations. CONCLUSION: pharmacy students' knowledge was improved by the PBL method. Pharmaceutical education courses should consider implementing PBL.

Critical thinking pedagogical practices in medical education: a systematic review.

Introduction: The development of critical thinking (CT) has been a universal goal in higher education. A systematic review of the literature was conducted to evaluate the effectiveness of currently used pedagogical practices to foster CT/ clinical reasoning (CR)/ clinical judgment (CJ) skills and/or dispositions in undergraduate medical students. Methods: PubMed, Web of Science and Scopus databases were searched from January 2010 to April 2021 with a predefined Boolean expression. Results: Of the 3221 articles originally identified, 33 articles were included by using PICOS methodology. From these, 21 (64%) reported CR pedagogical practices and 12 (36%) CT pedagogical practices. Discussion: Overall, pedagogical practices such as cognitive/visual representation, simulation, literature exposure, test-enhancing and team-based learning, clinical case discussion, error-based learning, game-based learning seem to enhance CT/CR skills and/or dispositions. Further research is required to identify the optimal timing, duration and modality of pedagogical interventions for effectively foster CT/CR in medical education.

Will the white blood cells tell? A potential novel tool to assess broiler chicken welfare.

This study assessed qualitative and quantitative leukocyte evaluation as potential broiler chicken welfare indicators, contributing to the limited literature on white blood cell (WBC) morphology as a diagnostic tool for welfare. Broiler chicken welfare within four poultry houses (PH) 1 to 4, each on a different farm, was assessed using on-field indicators of affective states and health, and WBC morphology was examined. Affective states were evaluated using the Qualitative Behavior Assessment (QBA), with 25 behavioral expressions scored on a visual analogue scale (VAS) and grouped into two categories. Health indicators included assessments of lameness, footpad dermatitis, dermatitis on the breast and abdominal areas, hock burn, and feather cleaning. Blood samples were collected, differential leukocyte counts were performed, and a cell score was created for the recognition, classification, and interpretation of morphologic diversity of heterophils and lymphocytes. The heterophil to lymphocyte ratio (H/L) was also determined. Descriptive statistics and generalized linear models for binomial responses were used to analyze the results. PH4 differed from the other farms, showing a higher frequency of birds within QBA group 1 ('Attentive'to 'Desperate'), while birds in PH1, PH2, and PH3 were more frequent in QBA group 2 ('Relaxed' to 'Positively occupied'). Elevated proportions of heterophils in birds from PH4 (0.61, CI95%: 0.58; 0.64) and PH3 (0.60, CI95%: 0.57; 0.63) suggested higher stress levels and inflammatory responses. Birds in PH2 and PH4 exhibited higher frequencies of health issues such as dermatitis and lameness, and higher proportions of abnormalities in WBC number and morphology. PH3 and PH4 exhibited higher H/L ratios of 3.03 and 2.58, respectively, consistent with the on-field health and behavioral indicators. Blood samples from birds in PH2 and PH4 showed a proportion of 90% toxic change in heterophils, while in PH1 and PH3 it was 70%, indicating high levels of abnormal WBC morphology across all PHs. The findings emphasize the multifactorial nature of welfare impairments, including environmental conditions, health, and affective states. This highlights the need for indicators that reflect multiple welfare impacts, such as WBC counts and morphological alterations, which can serve as powerful tools in the complex task of assessing animal welfare.

Early life stage mechanisms of an active fish species to cope with ocean warming and hypoxia as interacting stressors.

Ocean's characteristics are rapidly changing, modifying environmental suitability for early life stages of fish. We assessed whether the chronic effects of warming (24 °C) and hypoxia (<2-2.5 mg L-1) will be amplified by the combination of these stressors on mortality, growth, behaviour, metabolism and oxidative stress of early stages of the white seabream Diplodus sargus. Combined warming and hypoxia synergistically increased larval mortality by >51%. Warming induced faster growth in length and slower gains in weight when compared to other treatments. Boldness and exploration were not directly affected, but swimming activity increased under all test treatments. Under the combination of warming and hypoxia, routine metabolic rate (RMR) significantly decreases when compared to other treatments and shows a negative thermal dependence. Superoxide dismutase and catalase activities increased under warming and were maintained similar to control levels under hypoxia or under combined stressors. Under hypoxia, the enzymatic activities were not enough to prevent oxidative damages as lipid peroxidation and DNA damage increased above control levels. Hypoxia reduced electron transport system activity (cellular respiration) and isocitrate dehydrogenase activity (aerobic metabolism) below control levels. However, lactate dehydrogenase activity (anaerobic metabolism) did not differ among treatments. A Redundancy Analysis showed that ∼99% of the variability in mortality, growth, behaviour and RMR among treatments can be explained by molecular responses. Mortality and growth are highly influenced by oxidative stress and energy metabolism, exhibiting a positive relationship with reactive oxygen species and a negative relationship with aerobic metabolism, regardless of treatment. Under hypoxic condition, RMR, boldness and swimming activity have a positive relationship with anaerobic metabolism regardless of temperature. Thus, seabreams may use anaerobic reliance to counterbalance the effects of the stressors on RMR, activity and growth. The outcomes suggests that early life stages of white seabream overcame the single and combined effects of hypoxia and warming.

Effect of dietary l-glutamine supplementation on the intestinal physiology and growth during Solea senegalensis larval development.

The maturation of the intestinal digestive and absorptive functions might limit the amount of absorbed nutrients to fulfil the high requirements of the fast-growing marine fish larva. Glutamine (Gln) has been described to improve intestinal epithelium functions, due to its involvement in energy metabolism and protein synthesis. The purpose of this study was to evaluate dietary 0.2% Gln supplementation on aspects of intestinal physiology, protein metabolism and growth-related genes expression in Senegalese sole larvae. Experiment was carried out between 12 and 33 days post hatching (DPH) and fish were divided into two experimental groups, one fed Artemia spp. (CTRL) and the other fed Artemia spp. supplemented with Gln (GLN). GLN diet had two times more Gln than the CTRL diet. Samples were collected at 15, 19, 26 and 33 DPH for biometry, histology, and digestive enzymes activity, and at 33 DPH for gene expression, protein metabolism and AA content determination. Growth was significantly higher for Senegalese sole fed GLN diet, supported by differences on protein metabolism and growth-related gene expression. Slight differences were observed between treatments regarding the intestinal physiology. Overall, GLN diet seems to be directed to enhance protein metabolism leading to higher larval growth.