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A quantitative risk assessment model was developed to estimate the annual probability of introducing bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BoHV-1) at the farm level through animal movements. Data from 2017 official animal movements, biosecurity questionnaires, scientific literature, and expert opinion from field veterinarians were taken into consideration for model input parameters. Purchasing or introducing cattle, rearing replacement heifers offsite, showing cattle at competitions, sharing transport vehicles with other herds, and transporting cattle in vehicles that have not been cleaned and disinfected were considered in the model. The annual probability of introducing BVDV or BoHV-1 through infected animals was very heterogeneous between farms. The median likelihoods of BVDV and BoHV-1introduction were 12 and 9%, respectively. Farms that purchased cattle from within their region (i.e., local movements) and shared transport with other farms had a higher probability for BVDV and BoHV-1 introduction. This model can be a useful tool to support decision-making on biosecurity measures that should be prioritized to reduce the probability of introduction of these 2 diseases in dairy herds This work was supported by a Grant from the Ministerio de Ciencia e Innovación of Spain (AGL2016-77269-C2-1-R and AGL2016-77269-C2-2-R). The Universidad de Nariño (Pasto, Colombia) provided funding to the first author (BBB) SI

Spotted wolffishAnarhichas minorreproduction in captivity is dependent on in vitro fertilization. However, it is often challenging to acquire sufficient fresh sperm to fertilize the eggs that are obtained. In this study, we evaluate the possibility to store spotted wolffish sperm by refrigeration. Spotted wolffish sperm has the particularity that is already motile on stripping, and currently it is not possible to immobilize and reactivate. Thus, sperm refrigeration protocols should focus in extending this motility period that usually lasts up to 2 days. In a first experiment, we evaluated the possibility that the motility period of the sperm was limited by contamination with urine. The urea concentration in the sperm obtained both by stripping (17.10 ± 1.98 mg/dL) and directly from the testis (12.59 ± 2.37 mg/dL) was similar (p > 0.05), which indicate that the sperm collection method used avoid contamination with urine. Afterwards, we tested the possibility that the sperm motility period was limited by energy stores. The ATP concentration (initial value 5.65 ± 0.86 nmol/109cells) remained stable (p = 0.099) during 30 h after sperm collection, and similar values (p = 0.329) were recorded at end of sperm storage in both diluted (3.88 ± 1.35 nmol/109cells) and undiluted samples (4.76 ± 1.08 nmol/109). This indicates that the low intracellular ATP consumption, derived from the slow sperm motility, can probably be compensated rapidly enough by mitochondrial synthesis of ATP in the spotted wolffish sperm. In both experiments, diluted sperm kept higher percentage of motile cells during the storage time.

Life cycle impact assessment (LCIA) is a lively field of research, and data and models are continuously improved in terms of impact pathways covered, reliability, and spatial detail. However, many of these advancements are scattered throughout the scientific literature, making it difficult for practitioners to apply the new models. Here, we present the LC‐IMPACT method that provides characterization factors at the damage level for 11 impact categories related to three areas of protection (human health, ecosystem quality, natural resources). Human health damage is quantified as disability adjusted life years, damage to ecosystem quality as global species extinction equivalents (based on potentially disappeared fraction of species), and damage to mineral resources as kilogram of extra ore extracted. Seven of the impact categories include spatial differentiation at various levels of spatial scale. The influence of value choices related to the time horizon and the level of scientific evidence of the impacts considered is quantified with four distinct sets of characterization factors. We demonstrate the applicability of the proposed method with an illustrative life cycle assessment example of different fuel options in Europe (petrol or biofuel). Differences between generic and regionalized impacts vary up to two orders of magnitude for some of the selected impact categories, highlighting the importance of spatial detail in LCIA. This article met the requirements for a gold – gold JIE data openness badge described at http://jie.click/badges., Journal of Industrial Ecology, 24 (6), ISSN:1088-1980, ISSN:1530-9290

Background The automated collection of non-specific data from livestock, combined with techniques for data mining and time series analyses, facilitates the development of animal health syndromic surveillance (AHSyS). An example of AHSyS approach relates to the monitoring of bovine fallen stock. In order to enhance part of the machinery of a complete syndromic surveillance system, the present work developed a novel approach for modelling in near real time multiple mortality patterns at different hierarchical administrative levels. To illustrate its functionality, this system was applied to mortality data in dairy cattle collected across two Spanish regions with distinct demographical, husbandry, and climate conditions. Results The process analyzed the patterns of weekly counts of fallen dairy cattle at different hierarchical administrative levels across two regions between Jan-2006 and Dec-2013 and predicted their respective expected counts between Jan-2014 and Jun- 2015. By comparing predicted to observed data, those counts of fallen dairy cattle that exceeded the upper limits of a conventional 95% predicted interval were identified as mortality peaks. This work proposes a dynamic system that combines hierarchical time series and autoregressive integrated moving average models (ARIMA). These ARIMA models also include trend and seasonality for describing profiles of weekly mortality and detecting aberrations at the region, province, and county levels (spatial aggregations). Software that fitted the model parameters was built using the R statistical packages. Conclusions The work builds a novel tool to monitor fallen stock data for different geographical aggregations and can serve as a means of generating early warning signals of a health problem. This approach can be adapted to other types of animal health data that share similar hierarchical structures.

Mammary serum amyloid A3 (M-SAA3) has shown potential in stimulating innate immunity during intramammary infections, at calving and at dryoff. In this study, we produced recombinant caprine M-SAA3 to test its ability to reduce intramammary infections with Staphylococcus aureus, Streptococcus uberis, Streptococcus dysgalactiae, and Escherichia coli, which are all common mastitis-producing pathogens. Recombinant production of M-SAA3 (followed by lipopolysaccharide removal to avoid lipopolysaccharide-nonspecific stimulation of the immune system) was successfully achieved. Mammary serum amyloid A3 stimulated the expression of IL-8 in a dose-dependent manner in primary mammary cultures. Although a direct killing effect on Staph. aureus by M-SAA3 was not detected, this acute phase protein was able to reduce Staph. aureus, Strep. uberis, and Strep. dysgalactiae infections by up to 50% and induced a reduction in E. coli counts of 67%. In general, the best concentration of caprine M-SAA3 for inhibiting infections was the lowest concentration tested (10 μg/mL), although higher concentrations (up to 160 μg/mL) increased its antimicrobial potential against some pathogens. info:eu-repo/semantics/acceptedVersion

The effect of spray‐dried porcine plasma (SDPP) on the intestinal histological organization and autochthonous microbiota composition was evaluated in Sparus aurata. Fish were fed a basal diet (51 g/kg protein, 17 g/kg fat, 20.6 MJ/kg gross energy) and a diet containing 3 g/kg SDPP for 95 days (initial body weight, BW = 9.5 ± 0.2g, mean ± SD). The inclusion of SDPP promoted growth (p < .05), being fish fed the SDPP diet 6.2% (BW = 88.2 ± 1.6 g) heavier than the control (BW = 82.7 ± 3.2 g). SDPP increased the density of intestinal goblet cells (p < .05), whereas no differences in villi height were found (p > .05) between both groups. Intestinal microbiota was dominated by Proteobacteria (>85%) and Firmicutes (5%–12%), whereas Bacteroidetes never represented more than 1.5%. γ‐Proteobacteria, and Bacilli and Clostridia were the predominant classes. The short administration of SDPP (20 days) resulted in changes in microbiota diversity and richness associated with an increase in the sequences of the genus Lactobacillus and to a decrease in the genus Vibrio, whereas these changes were reverted at 95 days. Intestinal goblet cell density was not correlated to microbiota diversity and richness changes rather than to the immunostimulatory effect of the SDPP. info:eu-repo/semantics/acceptedVersion

The objective of this study was to evaluate the potential benefits of supplementing glutamic acid in milk replacers (MR) with respect to calf performance, intestinal permeability, and metabolism. Sixty Holstein male calves (3 ± 1.3 d old and 45 ± 5.9 kg body weight) were individually housed and fed a control MR without AA supplementation (24.8% crude protein and 19.1% fat, dry matter basis), or MR supplemented with 0.3% glutamic acid (25.1% crude protein and 20.3% fat, dry matter basis). Animals followed the same MR feeding program and were weaned at 56 d of the study. The amount of starter concentrate offered was restricted to limit the effect of concentrate intake on calf metabolism. Individual daily consumption and weekly body weight were measured, and 4 h after the morning feeding, blood samples were obtained at 14 and 35 d to determine general biochemical parameters and plasma AA concentrations. On d 10 of the study, we conducted an intestinal permeability test by including 21 g of lactulose and 4.2 g of d-mannitol as markers in the MR. We found no differences in calf performance or in intestinal permeability (measured as lactulose:mannitol ratio). Serum glucose concentration was greater in unsupplemented calves than in Glu-supplemented calves. At 14 d, the proportion of plasma Leu was greater in Glu-supplemented calves; the proportion of Ile tended to be greater in Glu-supplemented calves; and the proportion of Met tended to be greater in unsupplemented calves. We observed no other differences. Small changes occurred in AA metabolism when supplementing calf MR with 0.3% glutamic acid, without leading to improvements in calf performance or changes in intestinal permeability. info:eu-repo/semantics/publishedVersion

Background Virus neutralization test (VNT) is widely used for serological survey of classical swine fever (CSF) and efficacy evaluation of CSF vaccines. However, VNT is a time consuming procedure that requires cell culture and live virus manipulation. C-strain CSF vaccine is the most frequently used vaccine for CSF control and prevention. In this study, we presented a neutralizing monoclonal antibody (mAb) based competitive enzyme-linked immunosorbent assay (cELISA) with the emphasis on the replacement of VNT for C-strain post–vaccination monitoring. Results One monoclonal antibody (6B211) which has potent neutralizing activity against C-strain was generated. A novel cELISA was established and optimized based on the strategy that 6B211 can compete with C-strain induced neutralizing antibodies in pig serum to bind capture antigen C-strain E2. By testing C-strain VNT negative pig sera (n = 445) and C-strain VNT positive pig sera (n = 70), the 6B211 based cELISA showed 100% sensitivity (95% confidence interval: 94.87 to 100%) and 100% specificity (95% confidence interval: 100 to 100%). The C-strain antibody can be tested in pigs as early as 7 days post vaccination with the cELISA. By testing pig sera (n = 139) in parallel, the cELISA showed excellent agreement (Kappa = 0.957) with VNT. The inhibition rate of serum samples in the cELISA is highly correlated with their titers in VNT (r2 = 0.903, p Conclusions This novel cELISA demonstrated excellent agreement and high level correlation with VNT. It is a reliable tool for sero-monitoring of C-strain vaccination campaign because it is a rapid, simple, safe and cost effective assay that can be used to monitor vaccination-induced immune response at the population level.

The effectiveness of using essential oils against fish parasites and pathogenic bacteria as environmentally friendly phytotherapeutic agents in the aquaculture industry is demonstrated. A commercial additive composed of garlic essential oil, thymol and carvacrol (AROTEC-G®) was added to a diet for gilthead seabream (Sparus aurata) as a protection strategy against Sparicotyle chrysophrii. The intense aromatic properties of these essential oils might lead to the need of a suppression time to reverse possible changes in the organoleptic quality of the fish. Two experimental groups were set up: an experimental group fed a commercial feed supplemented with AROTEC-G® (diet A) during the first 4 weeks of the nutritional assay, and after which the treatment was suspended for 14, 21 and 28 days after a first sampling performed (day 0); and a control group, fed the same basal diet without AROTEC-G® supplementation (diet C). A physicochemical evaluation measured the pH, colour and texture, and a descriptive quantitative sensory analysis (QDA) was carried out using a trained panel for gilthead seabream fillets. Overall, the findings showed that the use of dietary essential oils did not result in significant sensory differences after day 0 of the study, although slight differences were observed in some of the physicochemical parameters analysed. info:eu-repo/semantics/acceptedVersion

The pandemic caused by the new coronavirus SARS-CoV-2 has made evident the need for broad-spectrum, efficient antiviral treatments to combat emerging and re-emerging viruses. Plitidepsin is an antitumor agent of marine origin that has also shown a potent pre-clinical efficacy against SARS-CoV-2. Plitidepsin targets the host protein eEF1A (eukaryotic translation elongation factor 1 alpha) and affects viral infection at an early, post-entry step. Because electron microscopy is a valuable tool to study virus-cell interactions and the mechanism of action of antiviral drugs, in this work we have used transmission electron microscopy (TEM) to evaluate the effects of plitidepsin in SARS-CoV-2 infection in cultured Vero E6 cells 24 and 48h post-infection. In the absence of plitidepsin, TEM morphological analysis showed double-membrane vesicles (DMVs), organelles that support coronavirus genome replication, single-membrane vesicles with viral particles, large vacuoles with groups of viruses and numerous extracellular virions attached to the plasma membrane. When treated with plitidepsin, no viral structures were found in SARS-CoV-2-infected Vero E6 cells. Immunogold detection of SARS-CoV-2 nucleocapsid (N) protein and double-stranded RNA (dsRNA) provided clear signals in cells infected in the absence of plitidepsin, but complete absence in cells infected and treated with plitidepsin. The present study shows that plitidepsin blocks the biogenesis of viral replication organelles and the morphogenesis of virus progeny. Electron microscopy morphological analysis coupled to immunogold labeling of SARS-CoV-2 products offers a unique approach to understand how antivirals such as plitidepsin work., This research was funded by Pharma Mar, which commercializes Aplidin/Plitidepsin. The authors also acknowledge the crowdfunding initiative #Yomecorono (https://www.yomecorono.com) and the CBIG consortium supported by Grifols. N.I–U. is supported by grant PID2020-117145RB-I00 from the Spanish Ministry of Science and Innovation. C.R. is supported by grant RTI2018-094445-B-I00 (MCI/AEI/FEDER, UE) from the Spanish Ministry of Science and Innovation.

Enterococcus faecalis (E. faecalis) was identified among the most relevant antimicrobial-resistant (AMR) bacteria in the EU for poultry in a previous scientific opinion. Thus, it has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of Article 7 on disease profile and impacts, Article 5 on its eligibility to be listed, Annex IV for its categorisation according to disease prevention and control rules as in Article 9 and Article 8 for listing animal species related to the bacterium. The assessment has been performed following a methodology previously published. The outcome is the median of the probability ranges provided by the experts, which indicates whether each criterion is fulfilled (lower bound ≥ 66%) or not (upper bound ≤ 33%), or whether there is uncertainty about fulfilment. Reasoning points are reported for criteria with uncertain outcome. According to the assessment here performed, it is uncertain whether AMR E. faecalis can be considered eligible to be listed for Union intervention according to Article 5 of the AHL (33–66% probability). According to the criteria in Annex IV, for the purpose of categorisation related to the level of prevention and control as in Article 9 of the AHL, the AHAW Panel concluded that the bacterium does not meet the criteria in Sections 1, 2 and 4 (Categories A, B and D; 0–5%, 5–10% and 1–10% probability of meeting the criteria, respectively) and the AHAW Panel is uncertain whether it meets the criteria in Sections 3 and 5 (Categories C and E, 33–66% and 33–66% probability of meeting the criteria, respectively). The animal species to be listed for AMR E. faecalis according to Article 8 criteria are mostly birds of the orders Galliformes and Anseriformes, but also mammals and reptiles can serve as reservoirs., The AHAW Panel wishes to thank Rikke Heidemann Olsen and Peter Panduro Damborg from the University of Copenhagen for conducting the extensive literature review under the contract OC/EFSA/ALPHA/2020/02 – LOT 1. The AHAW Panel also wishes to thank Friederike Hilbert from the University of Veterinary Medicine, Vienna, Ana R. Freitas from the University Institute of Health Sciences, Gandra, Paul McMullin from Poultry Health International, and Verena Oswaldi from EFSA for the support provided for this scientific output.

The spray-dried porcine plasma (SDPP) is an abattoir by-product used in animal nutrition with beneficial effects reported in livestock and commercial aquatic species. Previous results have found that the dietary inclusion of SDPP in gilthead sea bream (Sparus aurata) increased the density of intestinal goblet cells, and it did not result in significant changes in the autochthonous microbiota. However, there is no comprehensive data on the mechanisms that could take place on the intestine of gilthead sea bream fed with an SDPP-supplemented diet. For this reason, this study aimed to unveil the biological mechanisms modulated in response to the dietary administration of SDPP in the gilthead sea bream gut. To achieve this goal, we made a microarrays-based transcriptomic approach in gut samples from gilthead sea bream fed with an SDPP-supplemented diet for 95 days. As control diet, we used a protein-rich commercial feed (51% crude protein, 17% crude fat, and 20.6 MJ/kg gross energy) which was supplemented with 3% SDPP at the expense of LT70 fishmeal. The microarray analyses showed a total of 803 (468 up- and 335 down-regulated) differential expressed genes (DEGs). The functional network analysis revealed that dietary inclusion of SDPP induced sustained changes in 120 biological processes, grouped in 12-clusters. Among them, the metabolic-related process (cellular catabolic process, organic substance catabolic process, protein metabolism process), protein transport, and leukocyte mediated immunity interacted in the leading interactome network. This evidence confirms the previous evidence of the enhancement of the mucosal health status in response to the dietary administration of SDPP and provides further understanding of the mode of action of this ingredient in aquafeeds.

The transcriptomic response of the head kidney, the main lymphohematopoietic tissue of the body, was evaluated in Atlantic salmon (Salmo salar) smolts fed a functional feed containing a phytogenic rich in verbascoside and triterpenic compounds like ursolic acid. Fish (initial body weight = 55.0 ± 0.1 g) were fed two experimental diets (40% crude protein, 22% crude fat; 21.6 MJ/kg gross energy) that only differed in the phytogenic content (0.1% inclusion). Each diet has six replicates and was tested over a period of 133 days. The tested zootechnical feed additive a medicinal plant leaf extract (MPLE) obtained from sage (Salvia officinalis) and lemon verbena (Lippia citriodora). At the end of the trial, smolts fed the MPLE diet were heavier than their congeners from the control group (271.5 ± 7.9 g vs. 240.2 ± 19.3 g, respectively; P < 0.05). Feed conversion ratio (FCR) values in fish fed the control diet were higher than those in fish fed the MPLE diet (FCRcontrol diet = 1.27 ± 0.08 vs. FCR0.1% MPLE diet = 1.08 ± 0.05; P < 0.05). The immunomodulatory properties of the functional diet were evaluated by means of an in vivo challenge with Aeromonas salmonicida subsp. salmonicida (1 × 107 CFU mL–1). The microarray analysis of head kidney samples from both dietary groups revealed 1,178 differentially expressed genes (802 upregulated and 376 downregulated). Among them, several biological processes related to immunity were identified in fish fed the MPLE diet (i.e., interferon-gamma-mediated signaling pathway, antigen processing and presentation of peptide antigen via MHC class II, autophagy, regulation of i-kappaB kinase/NF-kappaB signaling, and leukocyte activation). Results from the bacterial challenge showed that survival rates were higher in smolts from the MPLE group (90.6 ± 6.4%) in comparison to the control group (60.7 ± 13.5%), confirming the functional benefits of the phytogenic in terms of host’s immunity and disease resistance. Biological processes such as cytoskeleton organization and regulation of cellular protein metabolic process detected in fish fed the MPLE diet supported the metabolic changes related to increased somatic growth promoted. The present findings showed that the inclusion at 0.1% of the tested MPLE obtained from sage and lemon verbena in diets for Atlantic salmon smolts promoted somatic growth, and enhanced their systemic immune response and reduced mortality when fish were challenged with A. salmonicida cumulative, the causative agent of furunculosis in salmonids.

Dinoflagellates of the genera Prorocentrum and Dinophysis are known producers of toxic compounds belonging to the okadaic acid (OA) group. The ingestion of shellfish contaminated with these toxins causes a human disease named diarrheic shellfish poisoning (DSP). In this study, the first record of Prorocentrum panamense, a potential toxin-producer species, was reported in the Canary Islands together with its toxicological characterization. Prorocentrum panamense cells were collected during April 2017 from natural pools located in the Northeastern part of Gran Canaria. This new record represents an expansion of P. panamense distribution area, previously restricted to the Pacific Ocean, Indian Ocean, Arabian Gulf and the Caribbean, and its introduction mechanisms are discussed. Laboratory cultures of P. panamense were settled and toxin production was assessed in both cell pellets and culture media at four different growth phases (latency, exponential, early stationary and late stationary) implementing LC-MS/MS and neuro-2a cell-based assay (CBA). LC-MS/MS allowed the identification of OA in the fraction corresponding to the late stationary phase, and tests performed on neuro-2a cells showed, for most of the fractions, OA-like activity observable by both cell morphology changes and cell mortality. This information is fundamental for a better understanding of the genus Prorocentrum global distribution, its ecology, and risks associated to toxic producing species. info:eu-repo/semantics/acceptedVersion

There is a need to develop cost-effective and non-invasive approaches to sample large populations to evaluate the disease status of breeding herds. In this study we assessed the detection of the M. hyopneumoniae genetic material in environmental surfaces and air of farrowing rooms, and skin (udder, snout and vagina) of lactating sows at weaning, in farms having different M. hyopneumoniae infection status (negative, positive sub-clinically infected and positive clinically affected). Mycoplasma hyopneumoniae was detected in air, air deposition particles, dam and stall surfaces of the positive clinically affected herd. Mycoplasma hyopneumoniae could only be detected in dam and stall surfaces in sub-clinically infected herds. Mycoplasma hyopneumoniae was not detected in all samples collected in the negative herd. The cycle threshold of the positive PCR samples were not statistically different between sample types or farms. However, a significant difference (p < 0.05) was observed in the percentage of positive samples between the positive clinically affected farm and the rest. Likewise, M. hyopneumoniae was detected in the environment and surfaces at weaning in positive breeding herds. Further testing and validation is recommended for environmental and surface samples before they can be employed as part of the M. hyopneumoniae diagnostic process. In addition, results from this study highlight potential sources of M. hyopneumoniae infection for piglets in breeding herds, especially during an outbreak. info:eu-repo/semantics/acceptedVersion

This article also appears in: Animal Health Law: Assessment of the control measures of the category A diseases of terrestrial animals., EFSA received a mandate from the European Commission to assess the effectiveness of some of the control measures against diseases included in the Category A list according to Regulation (EU) 2016/429 on transmissible animal diseases (‘Animal Health Law’). This opinion belongs to a series of opinions where these control measures will be assessed, with this opinion covering the assessment of control measures for Contagious Caprine Pleuropneumonia (CCPP). In this opinion, EFSA and the AHAW Panel of experts review the effectiveness of: (i) clinical and laboratory sampling procedures, (ii) monitoring period, (iii) the minimum radius of the protection and surveillance zones and iv) the minimum length of time the measures should be applied in these zones. The general methodology used for this series of opinions has been published elsewhere. Several scenarios for which these control measures had to be assessed were designed and agreed prior to the start of the assessment. Different clinical and laboratory sampling procedures are proposed depending on the scenarios considered. The monitoring period of 45 days was assessed as effective in affected areas where high awareness is expected, and when the index case occurs in an area where the awareness is low the monitoring period should be at least 180 days (6 months). Since transmission kernels do not exist and data to estimate transmission kernels are not available, a surveillance zone of 3 km was considered effective based on expert knowledge, while a protection zone should also be developed to include establishments adjacent to affected ones. Recommendations, provided for each of the scenarios assessed, aim to support the European Commission in the drafting of further pieces of legislation, as well as for plausible ad hoc requests in relation to CCPP., European Commission: EFSA-Q-2020-0080

Background Every year, more than 700,000 people die from vector-borne diseases, mainly transmitted by mosquitoes. Vector surveillance plays a major role in the control of these diseases and requires accurate and rapid taxonomical identification. New approaches to mosquito surveillance include the use of acoustic and optical sensors in combination with machine learning techniques to provide an automatic classification of mosquitoes based on their flight characteristics, including wingbeat frequency. The development and application of these methods could enable the remote monitoring of mosquito populations in the field, which could lead to significant improvements in vector surveillance. Methods A novel optical sensor prototype coupled to a commercial mosquito trap was tested in laboratory conditions for the automatic classification of mosquitoes by genus and sex. Recordings of > 4300 laboratory-reared mosquitoes of Aedes and Culex genera were made using the sensor. The chosen genera include mosquito species that have a major impact on public health in many parts of the world. Five features were extracted from each recording to form balanced datasets and used for the training and evaluation of five different machine learning algorithms to achieve the best model for mosquito classification. Results The best accuracy results achieved using machine learning were: 94.2% for genus classification, 99.4% for sex classification of Aedes, and 100% for sex classification of Culex. The best algorithms and features were deep neural network with spectrogram for genus classification and gradient boosting with Mel Frequency Cepstrum Coefficients among others for sex classification of either genus. Conclusions To our knowledge, this is the first time that a sensor coupled to a standard mosquito suction trap has provided automatic classification of mosquito genus and sex with high accuracy using a large number of unique samples with class balance. This system represents an improvement of the state of the art in mosquito surveillance and encourages future use of the sensor for remote, real-time characterization of mosquito populations. Graphical abstract

To understand the determinants of long-term immune responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the concurrent impact of vaccination and emerging variants, we follow a prospective cohort of 332 patients with coronavirus disease 2019 (COVID-19) over more than a year after symptom onset. We evaluate plasma-neutralizing activity using HIV-based pseudoviruses expressing the spike of different SARS-CoV-2 variants and analyze them longitudinally using mixed-effects models. Long-term neutralizing activity is stable beyond 1 year after infection in mild/asymptomatic and hospitalized participants. However, longitudinal models suggest that hospitalized individuals generate both short- and long-lived memory B cells, while the responses of non-hospitalized individuals are dominated by long-lived B cells. In both groups, vaccination boosts responses to natural infection. Long-term (>300 days from infection) responses in unvaccinated participants show a reduced efficacy against beta, but not alpha nor delta, variants. Multivariate analysis identifies the severity of primary infection as an independent determinant of higher magnitude and lower relative cross-neutralization activity of long-term neutralizing responses., Graphical abstract, Pradenas and Trinité et al. analyze the kinetics of neutralizing antibodies in response to natural SARS-CoV-2 infection and evaluate the long-term (beyond 1 year) stability and breadth of the neutralizing response before subsequent vaccination.

African swine fever (ASF) is currently causing a major pandemic affecting the swine industry and protein availability from Central Europe to East and South Asia. No commercial vaccines are available, making disease control dependent on the elimination of affected animals. Here, we show that the deletion of the African swine fever virus (ASFV) E184L gene from the highly virulent ASFV Georgia 2010 (ASFV-G) isolate produces a reduction in virus virulence during the infection in swine. Of domestic pigs intramuscularly inoculated with a recombinant virus lacking the E184L gene (ASFV-G-ΔE184L), 40% experienced a significantly (5 days) delayed presentation of clinical disease and, overall, had a 60% rate of survival compared to animals inoculated with the virulent parental ASFV-G. Importantly, all animals surviving ASFV-G-ΔE184L infection developed a strong antibody response and were protected when challenged with ASFV-G. As expected, a pool of sera from ASFV-G-ΔE184L-inoculated animals lacked any detectable antibody response to peptides partially representing the E184L protein, while sera from animals inoculated with an efficacious vaccine candidate, ASFV-G-ΔMGF, strongly recognize the same set of peptides. These results support the potential use of the E184L deletion for the development of vaccines able to differentiate infected from vaccinated animals (DIVA). Therefore, it is shown here that the E184L gene is a novel ASFV determinant of virulence that can potentially be used to increase safety in preexisting vaccine candidates, as well as to provide them with DIVA capabilities. To our knowledge, E184L is the first ASFV gene product experimentally shown to be a functional DIVA antigenic marker. IMPORTANCE No commercial vaccines are available to prevent African swine fever (ASF). The ASF pandemic caused by the ASF virus Georgia 2010 (ASFV-G) strain is seriously affecting pork production in a contiguous geographical area from Central Europe to East Asia. The only effective experimental vaccines are viruses attenuated by deleting ASFV genes associated with virus virulence. Therefore, identification of such genes is of critical importance for vaccine development. Here, we report the discovery of a novel determinant of ASFV virulence, the E184L gene. Deletion of the E184L gene from the ASFV-G genome (ASFV-G-ΔE184L) produced a reduction in virus virulence, and importantly, animals surviving infection with ASFV-G-ΔE184L were protected from developing ASF after challenge with the virulent parental virus ASFV-G. Importantly, the virus protein encoded by E184L is highly immunogenic, making a virus lacking this gene a vaccine candidate that allows the differentiation of infected from vaccinated animals (DIVA). Here, we show that unlike what is observed in animals inoculated with the vaccine candidate ASFV-G-ΔMGF, ASFV-G-ΔE184L-inoculated animals do not mount a E184L-specific antibody response, indicating the feasibility of using the E184L deletion as the antigenic marker for the development of a DIVA vaccine in ASFV. info:eu-repo/semantics/acceptedVersion

Equine Herpesvirus-1 infection has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of: Article 7 on disease profile and impacts, Article 5 on the eligibility of the disease to be listed, Article 9 for the categorisation of the disease according to disease prevention and control measures as in Annex IV and Article 8 on the list of animal species related to Equine Herpesvirus-1 infection. The assessment has been performed following a methodology composed of information collection and compilation, and expert judgement on each criterion at individual and collective level. The outcome is the median of the probability ranges provided by the experts, which indicates whether the criterion is fulfilled (66–100%) or not (0–33%), or whether there is uncertainty about fulfilment (33–66%). For the questions where no consensus was reached, the different supporting views are reported. According to the assessment performed, Equine Herpesvirus-1 infection can be considered eligible to be listed for Union intervention according to Article 5 of the Animal Health Law with 33–90% certainty. According to the criteria as in Annex IV of the AHL related to Article 9 of the AHL for the categorisation of diseases according to the level of prevention and control, it was assessed with less than 1% certainty that EHV-1 fulfils the criteria as in Section 1 (category A), 1–5% for the criteria as in Section 2 (category B), 10–66% for the criteria as in Section 3 (category C), 66–90% for the criteria as in Section 4 (category D) and 33–90% for the criteria as in Section 5 (category E). The animal species to be listed for EHV-1 infection according to Article 8(3) criteria are the species belonging to the families of Equidae, Bovidae, Camelidae, Caviidae, Cervidae, Cricetidae, Felidae, Giraffidae, Leporidae, Muridae, Rhinocerontidae, Tapiridae and Ursidae., European Commission. Question number: EFSA-Q-2021-00183

This handbook aims to support higher education institutions with the integration of FAIR-related content in their curricula and teaching. It was written and edited by a group of about 40 collaborators in a series of six book sprint events that took place between 1 and 10 June 2021. The document provides practical material, such as competence profiles, learning outcomes and lesson plans, and supporting information. It incorporates community feedback received during the public consultation which ran from 27 July to 12 September 2021. A print version as well as an Open Access PDF version (most up-to-date version) were published by Göttingen University Press in May 2022. Both are available via: https://doi.org/10.17875/gup2022-1915. A GitBook version is available at: https://fairsfair.gitbook.io/fair-teaching-handbook/ (associated GitHub repository: https://github.com/FAIRsFAIR/FAIRteachinghandbook).

The Canary Islands are a ciguatoxin (CTX) hotspot with an established official monitoring for the detection of CTX in fish flesh from the authorised points of first sale. Fish caught by recreational fishermen are not officially tested and the consumption of toxic viscera or flesh could lead to ciguatera poisoning (CP). The objectives of this study were to determine the presence of CTX-like toxicity in relevant species from this archipelago, compare CTX levels in liver and flesh and examine possible factors involved in their toxicity. Sixty amberjack (Seriola spp.), 27 dusky grouper (Epinephelus marginatus), 11 black moray eels (Muraena helena) and 11 common two-banded seabream (Diplodus vulgaris) were analysed by cell-based assay (CBA) and Caribbean ciguatoxin-1 (C-CTX1) was detected by liquid chromatography mass spectrometry (LC-MS/MS) in all these species. Most of the liver displayed higher CTX levels than flesh and even individuals without detectable CTX in flesh exhibited hepatic toxicity. Black moray eels stand out for the large difference between CTX concentration in both tissues. None of the specimens with non-toxic liver showed toxicity in flesh. This is the first evidence of the presence of C-CTX1 in the common two-banded seabream and the first report of toxicity comparison between liver and muscle from relevant fish species captured in the Canary Islands. European Food Safety Authority | Ref. GP/EFSA/AFSCO/2015/03 Ministerio de Educación | Ref. FPU17/04022 Ministerio de Ciencia e Innovación | Ref. PID2019-108781RR-C21 Ministerio de Ciencia e Innovación | Ref. PID2019 108781RR C22 Xunta de Galicia | Ref. ED481A-2018/207

To provide practical feeding management guidelines preceding a stressful episode during farming practices, European sea bass juveniles (initial weight: 72.3 g) were fed for 60-days different fish meal (FM) and fish oil (FO) dietary levels [high (30% FM, 15% FO, FM30/FO15), intermediate (20% FM, 7% FO, FM20/FO7), and low (10% FM, 3% FO, FM10/FO3)] in triplicate conditions. Fish were then fasted for 36 h and exposed to a 2-h acute crowding (80 kg m–3 biomass). Plasma biochemistry, skin mucus parameters and gene expression of stress and immune-related genes were performed before, at 2 and 24 h after crowding. At the end of the trial, the FM10/FO3 group showed lower final body weight, weight gain, and specific growth rate compared to the other treatments. Most of the plasma parameters were mainly affected by crowding condition rather than diet; however, after stress, lactate was higher in the FM30/FO15 group compared to the other treatments. Similarly, protease, antiprotease, peroxidase and lysozyme in skin mucus were mostly affected by crowding conditions, while fish fed FM10/FO3 displayed higher skin mucosal IgM and bactericidal activity against Vibrio anguillarum and V. harveyi. Most of the stress-related genes considered (hsp70 and gr-1 in the brain; hsp70, gr-1 and gr-2 in the head kidney), showed an overall expression pattern that increased over time after stress, in addition, hsp70 in the head kidney was also up-regulated in fish fed FM30/FO15 after stress. Higher plasmatic lactate together with the up-regulation of some stress-related transcripts suggest a higher reactivity to acute crowding of the stress-response mechanism in fish fed high FM and FO dietary levels. Otherwise, the higher skin mucosal IgM and bactericidal activity observed in fish fed FM10/FO3 dietary levels seems to indicate that acute crowding was able to activate a higher pro-inflammatory response in this treatment. Overall, the results of the present study seem to indicate that 10% FM and 3% FO dietary levels might affect stress and immune responses.

Porcine circoviruses (PCVs) are distributed in swine herds worldwide and represent a threat to the health of domestic pigs and the profits of the swine industry. Currently, four PCV species, including PCV-1, PCV-2, PCV-3 and PCV-4, have been identified in China. Considering the ubiquitous characteristic of PCVs, the new emerged PCV-4 and the large scale of swine breeding in China, an overall analysis on codon usage bias for Chinese PCV sequences was performed by using the major proteins coding sequences (ORF1 and ORF2) to better understand the relationship of these viruses with their host. The data from genome nucleotide frequency composition and relative synonymous codon usage (RSCU) analysis revealed an overrepresentation of AT pair and the existence of a certain codon usage bias in all PCVs. However, the values of an effective number of codons (ENC) revealed that the bias was of low magnitude. Principal component analysis, ENC-plot, parity rule two analysis and correlation analysis suggested that natural selection and mutation pressure were both involved in the shaping of the codon usage patterns of PCVs. However, a neutrality plot revealed a stronger effect of natural selection than mutation pressure on codon usage patterns. Good host adaptation was also shown by the codon adaptation index analysis for all these viruses. Interestingly, obtained data suggest that PCV-4 might be more adapted to its host compared to other PCVs. The present study obtained insights into the codon usage pattern of PCVs based on ORF1 and ORF2, which further helps the understanding the molecular evolution of these swine viruses. info:eu-repo/semantics/publishedVersion

The growing emergence of microorganisms resistant to antibiotics has prompted the development of alternative antimicrobial therapies. Among them, the antimicrobial peptides produced by innate immunity, which are also known as host defense peptides (HDPs), hold great potential. They have been shown to exert activity against both Gram-positive and Gram-negative bacteria, including those resistant to antibiotics. These HDPs are classified into three categories: defensins, cathelicidins, and histatins. Traditionally, HDPs have been chemically synthesized, but this strategy often limits their application due to the high associated production costs. Alternatively, some HDPs have been recombinantly produced, but little is known about the impact of the bacterial strain in the recombinant product. This work aimed to assess the influence of the Escherichia coli strain used as cell factory to determine the activity and stability of recombinant defensins, which have 3 disulfide bonds. For that, an α-defensin [human α-defensin 5 (HD5)] and a β-defensin [bovine lingual antimicrobial peptide (LAP)] were produced in two recombinant backgrounds. The first one was an E. coli BL21 strain, which has a reducing cytoplasm, whereas the second was an E. coli Origami B, that is a strain with a more oxidizing cytoplasm. The results showed that both HD5 and LAP, fused to Green Fluorescent Protein (GFP), were successfully produced in both BL21 and Origami B strains. However, differences were observed in the HDP production yield and bactericidal activity, especially for the HD5-based protein. The HD5 protein fused to GFP was not only produced at higher yields in the E. coli BL21 strain, but it also showed a higher quality and stability than that produced in the Origami B strain. Hence, this data showed that the strain had a clear impact on both HDPs quantity and quality., This work was funded by Ministerio de Ciencia, Innovación y Universidades Grants (PID2019-107298RB-C21) to AA and EG-F, PID2019-105622RB-I00 to IR and through the ‘‘Severo Ochoa’’ Programme for Centers of Excellence in R&D (FUNFUTURE CEX2019-000917-S and SEV-2017-0706). The authors are also grateful to Marató de TV3 foundation for the Grant 201812-30-31-32-33 to EG-F and IR, and the Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN). AL-C received a pre-doctoral fellowship from Generalitat de Catalunya (FI-AGAUR), EG-F a post-doctoral fellowship from INIA and JG a Ramón y Cajal fellowship from MINN (RyC-2017-22614)., With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000917-S).

This study aimed to assess the efficacy of a heat-inactivated Mycobacterium caprae (HIMC) vaccine in goats experimentally challenged with the same strain of M. caprae. Twenty-one goats were divided into three groups of seven: vaccinated with heat-inactivated Mycobacterium bovis (HIMB), with HIMC and unvaccinated. At 7 weeks post-vaccination all animals were endobronchially challenged with M. caprae. Blood samples were collected for immunological assays and clinical signs were recorded throughout the experiment. All goats were euthanized at 9 weeks post-challenge. Gross pathological examination, analysis of lung pathology using computed tomography, and bacterial load quantification in pulmonary lymph nodes (LN) by qPCR were carried out. Only HIMC vaccinated goats showed a significant reduction of lung lesions volume and mycobacterial DNA load in LN compared to unvaccinated controls. Both vaccinated groups showed also a significant reduction of the other pathological parameters, an improved clinical outcome and a higher proportion of IFN-γ-producing central memory T cells after vaccination. The results indicated that homologous vaccination of goats with HIMC induced enhanced protection against M. caprae challenge by reducing lung pathology and bacterial load compared to the heterologous vaccine (HIMB). Further large-scale trials are necessary to assess the efficacy of autovaccines under field conditions.

Feed restriction and fasting experienced during commercial production negatively affect unweaned calves' behavior and health status. Transportation and stays at assembly centers are the main factors generating these disorders. For this study, 20 unweaned Angus-Holstein bull calves [44.1 ± 2.04 kg of body weight (BW) and 14.7 ± 0.63 d of age (± standard error)] were used to evaluate the effects of feed restriction and fasting on performance, energy status [serum concentration of glucose, β-hydroxybutyrate (BHB), and nonesterified fatty acids], and gastrointestinal permeability [serum concentration of citrulline, chromium (Cr)-EDTA, lactulose, and d-mannitol]. Calves were randomly assigned to 1 of 4 treatments that simulated the feed restrictions of an assembly center situation on one hand, and the fasting hours during transportation on the other. Treatments were as follows. Control (CT): from d −4 to −1, calves were fed 2.5 L of milk replacer (MR) twice daily; concentrate and straw were offered ad libitum. Mild (MD): calves were fed only MR (d −4 to −1) as described for CT, and on d −1 calves were subjected to a 9-h feed withdrawal. Moderate (MO): calves were fed only MR (d −4 to −1) as described for CT and on d −1 subjected to a 19-h feed withdrawal. Severe (SV): calves were fed only 2.5 L of a rehydrating solution twice daily (d −4 to −1) and on d −1 subjected to a 19-h feed withdrawal. From d 0 to d 42 (weaning) all calves were fed the same feeding program (MR, concentrate, and straw ad libitum). Results showed that BW was greater for the CT treatment compared with the others from d 0 to d 7, whereas BW of SV was lesser compared with the others from d −1 to d 7. No differences among treatments were observed at weaning. At d 2 concentrate intakes of MD, MO, and SV were lesser compared with CT. By d 4, concentrate intake of SV was similar to that for CT and greater than MD and MO. Similarly to BW, no differences in concentrate intake among treatments were observed at weaning on d 42 of the study. At d −1 for SV and d 0 in all restricted calves, serum glucose concentration was lesser compared with CT. At d −1 and 0, nonesterified fatty acids and BHB serum concentrations were greater in the SV calves compared with the other treatments. By d 2, serum concentrations of nonesterified fatty acids, BHB, and glucose were restored to CT levels. At d −1 serum citrulline concentration was lesser in SV and greater in MD calves. The CT calves had lower serum concentrations of Cr-EDTA (d −1 and d 0), lactulose (d 0), and d-mannitol (d 0) compared with the other restricted calves. Results showed that degree of dietary restriction, type of liquid diet (MR or rehydrating solution), and fasting hours (9 vs. 19 h) affected calves' BW, concentrate intake, and serum concentration of markers indicative of energy status and gastrointestinal permeability. info:eu-repo/semantics/publishedVersion

Pseudoplatystoma punctifer is a highly appreciated fish species native to the Amazon basin, whose commercial farming has been hampered by low survival during early life stages due to the high incidence of cannibalism and the low acceptability of compound diets at weaning. Dietary DHA is known to promote digestive system development and maturation and growth, whereas its deficiency induces stress. The aim of this study was to evaluate the effect of dietary DHA supplementation on the incidence of cannibalism, digestive physiology, and growth performance during the early life stages of P. punctifer to improve current feeding protocols. Four dietary treatments were generated using a commercial enrichment product with high DHA content. Fish larvae were fed non-enriched or enriched Artemia from 4 to 15 days post fertilization (dpf) and fed a non-enriched or enriched compound diet from 15 to 26 dpf, coinciding with the start of the juvenile stage. Growth, survival, incidence of cannibalism, proximate and fatty acid composition, histology of the intestine and liver, and quantitative gene expression of the main digestive enzymes (amy, try, ctr, pga, pla2, and lpl) were analyzed in the different dietary groups at the end of each feeding period. Results showed that dietary DHA supplementation influenced P. punctifer in a developmental stage-dependent manner. In particular, DHA-enriched Artemia provided during the larval stage contributed to reduce the incidence of cannibalism and improved survival at the early juvenile stage, while enriching the compound diet improved growth. The expression level of genes involved in protein and carbohydrate digestion (ctr, amy) was higher in groups fed enriched Artemia despite that both enriched and non-enriched Artemia displayed similar proximate composition, suggesting that different dietary fatty acid profiles may modulate the expression of these digestive enzyme precursors. The group transitioning from enriched Artemia to non-enriched compound diet showed a higher expression of most genes at the early juvenile stage. At the histological level, the group fed non-enriched Artemia and compound diet showed a significant accumulation of lipids in the intestine relative to the liver, contrary to the rest of the groups that showed similar amounts of lipids in both tissues, indicating a more balanced lipid metabolism. The group fed both DHA-enriched Artemia and compound diet provided the best results in terms of growth, survival, incidence of cannibalism, and digestive physiology. In conclusion, this study showed that the nutritional history during the larval period affected fish nutrition and behavior during the early juvenile stage. info:eu-repo/semantics/acceptedVersion

In this study, we assessed the potential synergistic effect of the Erns RNase activity and the poly-U insertion in the 3′ untranslated region (UTR) of the low-virulence classical swine fever virus (CSFV) isolate Pinar de Rio (PdR) in innate and adaptive immunity regulation and its relationship with classical swine fever (CSF) pathogenesis in pigs. We knocked out the Erns RNase activity of PdR and replaced the long polyuridine sequence of the 3′ UTR with 5 uridines found typically at this position, resulting in a double mutant, vPdR-H30K-5U. This mutant induced severe CSF in 5-day-old piglets and 3-week-old pigs, with higher lethality in the newborn (89.5%) than in the older (33.3%) pigs. However, the viremia and viral excretion were surprisingly low, while the virus load was high in the tonsils. Only alpha interferon (IFN-α) and interleukin 12 (IL-12) were highly and consistently elevated in the two groups. Additionally, high IL-8 levels were found in the newborn but not in the older pigs. This points toward a role of these cytokines in the CSF outcome, with age-related differences. The disproportional activation of innate immunity might limit systemic viral spread from the tonsils and increase virus clearance, inducing strong cytokine-mediated symptoms. Infection with vPdR-H30K-5U resulted in poor neutralizing antibody responses compared with results obtained previously with the parent and RNase knockout PdR. This study shows for the first time the synergistic effect of the 3′ UTR and the Erns RNase function in regulating innate immunity against CSFV, favoring virus replication in target tissue and thus contributing to disease severity. IMPORTANCE CSF is one of the most relevant viral epizootic diseases of swine, with high economic and sanitary impact. Systematic stamping out of infected herds with and without vaccination has permitted regional virus eradication. However, the causative agent, CSFV, persists in certain areas of the world, leading to disease reemergence. Nowadays, low- and moderate-virulence strains that could induce unapparent CSF forms are prevalent, posing a challenge for disease eradication. Here, we show for the first time the synergistic role of lacking the Erns RNase activity and the 3′ UTR polyuridine insertion from a low-virulence CSFV isolate in innate immunity disproportional activation. This might limit systemic viral spread to the tonsils and increase virus clearance, inducing strong cytokine-mediated symptoms, thus contributing to disease severity. These results highlight the role played by the Erns RNase activity and the 3′ UTR in CSFV pathogenesis, providing new perspectives for novel diagnostic tools and vaccine strategies. info:eu-repo/semantics/publishedVersion

The potential beneficial and harmful social impacts generated by the introduction of novel technologies, in general, and those concerning nutrient recovery and the improvement of nutrient efficiency in agriculture, in particular, have received little attention, as shown in the literature. This study investigated the current social impacts of agricultural practices in Belgium, Germany and Spain, and the potential social impacts of novel technologies introduced in agriculture to reduce nutrient losses. Based on 65 indicators used in the PSILCA database, the greatest impacts in the baselines are related to fair salaries, biomass consumption, industrial water depletion and public sector corruption. The potential social impacts of the technologies were assessed using 17 midpoint indicators that have a potential to affect social endpoints. The potential benefits of novel agricultural technologies were the creation of more attractive jobs in agriculture, and a better and healthier environment for local communities, workers and society. However, their harmful effects mainly related to workers and local community health, due to the substances used in the technologies and the potential gases emitted. Given the current lack of Social Life Cycle Assessment (S-LCA) studies on novel technologies in agriculture, this study is the first to use the PSILCA database to assess different technologies for nutrient recovery in agriculture in an initial and prospective assessment of their potential social impacts. Further work is required for a site-specific assessment of the technologies when a higher level of social adaptation is achieved. info:eu-repo/semantics/publishedVersion

A total of 2,880 one-day-old male and female broiler chicks from two breeds, Ross308 and Cobb500 were randomly assigned to 72 pens. Broilers were offered three diets: a wheat-soybean diet without (CO), or with either a probiotic (probiotic; 2.4 x 109 CFU/kg diet of Bacillus subtilis DSM32324 and DSM32325 and B. amyloliquefaciens DSM25840) or a phytobiotic (phytobiotic; grape extract with 165 ppm procyanidin and 585 ppm polyphenol) product. The trial was conducted with a 3 × 2 × 2 factorial arrangement of diet, breed and sex in a completely randomized design and consisted of 6 replicate-pens per treatment (40 birds per pen). At day 7, 21, and 35, one chicken per pen was slaughtered for caecal sampling to quantify bacterial metabolites (digesta) as well as evaluate mRNA abundance and histomorphology (tissue). Data were subjected to ANOVA using GLM procedure to evaluate age, diet, breed and sex and their interactions. Spearman’s correlation (r) was analyzed between metabolite concentration and mRNA abundance. Overall, the concentration of short chain fatty acids increased with age, while lactate decreased from day 7 to 21 (p < 0.05). The mRNA abundance of IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17α, IL-18, IFN-γ and TGF-β2 increased with age but IL-1β and TNF-α increased in abundance from day 7 to 21 and then decreased (p < 0.05). Abundance of MUC2 and CLDN5 increased after day 21 (p < 0.05). Caecal crypt depth increased with age (p < 0.05). Acidic goblet cell (GC) number peaked at day 21 (p < 0.05), while mixed GC number was not affected by age. A few impacts of breed, diet and interactions on the investigated variables showed no meaningful biological pattern. Propionate positively correlated with all cytokines investigated (r = 0.150–0.548), except TNF-α. Lactate negatively correlated with pro-inflammatory cytokines like IL-1β (r = −0.324). Aging affected caecal histomorphology, bacterial activity and genes responsible for barrier integrity and inflammatory response. This effect could be attributed to the interaction between gut microbiota and immune system as well as the direct effect of metabolites on gut histomorphology and cytokine mRNA abundance.

Plitidepsin, a marine-derived cyclic-peptide, inhibits SARS-CoV-2 replication at nanomolar concentrations by targeting the host protein eukaryotic translation elongation factor 1A. Here, we show that plitidepsin distributes preferentially to lung over plasma, with similar potency against across several SARS-CoV-2 variants in preclinical studies. Simultaneously, in this randomized, parallel, open-label, proof-of-concept study (NCT04382066) conducted in 10 Spanish hospitals between May and November 2020, 46 adult hospitalized patients with confirmed SARS-CoV-2 infection received either 1.5 mg (n = 15), 2.0 mg (n = 16), or 2.5 mg (n = 15) plitidepsin once daily for 3 d. The primary objective was safety; viral load kinetics, mortality, need for increased respiratory support, and dose selection were secondary end points. One patient withdrew consent before starting procedures; 45 initiated treatment; one withdrew because of hypersensitivity. Two Grade 3 treatment-related adverse events were observed (hypersensitivity and diarrhea). Treatment-related adverse events affecting more than 5% of patients were nausea (42.2%), vomiting (15.6%), and diarrhea (6.7%). Mean viral load reductions from baseline were 1.35, 2.35, 3.25, and 3.85 log10 at days 4, 7, 15, and 31. Nonmechanical invasive ventilation was required in 8 of 44 evaluable patients (16.0%); six patients required intensive care support (13.6%), and three patients (6.7%) died (COVID-19-related). Plitidepsin has a favorable safety profile in patients with COVID-19., This work was supported by grants from the Government of Spain (PIE_INTRAMURAL_ LINEA 1 - 202020E079; PIE_INTRAMURAL_CSIC-202020E043). The research of CBIG consortium (constituted by IRTA-CReSA, BSC, & IrsiCaixa) is supported by Grifols pharmaceutical. We also acknowledge the crowdfunding initiative #Yomecorono (https://www.yomecorono.com). N Izquierdo-Useros has nonrestrictive funding from PharmaMar to study the antiviral effect of Plitidepsin. NJ Krogan was funded by grants from the National Institutes of Health (P50AI150476, U19AI135990, U19AI135972, R01AI143292, R01AI120694, and P01AI063302); by the Excellence in Research Award (ERA) from the Laboratory for Genomics Research (LGR), a collaboration between the University of California, San Francisco (UCSF), University of California, Berkley (UCB), and GlaxoSmithKline (GSK) (#133122P); by the Roddenberry Foundation, and gifts from QCRG philanthropic donors. This work was supported by the Defense Advanced Research Projects Agency (DARPA) under Cooperative Agreement #HR0011-19-2-0020. The views, opinions, and/or findings contained in this material are those of the authors and should not be interpreted as representing the official views or policies of the Department of Defense or the U.S. Government. This research was partly funded by Center for Research for Influenza Pathogenesis and Transmission (CRIPT), a National Institute of Allergy and Infectious Diseases (NIAID) supported Center of Excellence for Influenza Research and Response (CEIRS, contract # 75N93021C00014), by DARPA grant HR0011-19-2-0020, by supplements to NIAID grants U19AI142733, U19AI135972, and DoD grant W81XWH-20-1-0270, and by the generous support of the JPB Foundation, the Open Philanthropy Project (research grant 2020-215611 (5384)), and anonymous donors to A García-Sastre. S Yildiz received funding from a Swiss National Foundation Early Postdoc Mobility fellowship (P2GEP3_184202).

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of animal welfare legislation. The present Opinion deals with the protection of small ruminants (sheep and goats) during transport. The main focus is on welfare of sheep during transport by road but other means of transport and concerns for welfare of goats during transport are also covered. Current practices related to transport of sheep during the different stages (preparation, loading and unloading, transit and journey breaks) are described. Overall, 11 welfare consequences were identified as being highly relevant for the welfare of sheep during transport based on severity, duration and frequency of occurrence: group stress, handling stress, heat stress, injuries, motion stress, predation stress, prolonged hunger, prolonged thirst, restriction of movement, resting problems and sensory overstimulation. These welfare consequences and their animal-based measures are described. A wide variety of hazards, mainly relating to inappropriate or aggressive handling of animals, structural deficiencies of vehicles and facilities, unfavourable microclimatic and environmental conditions and poor husbandry practices, leading to these welfare consequences were identified. The Opinion contains general and specific conclusions in relation to the different stages of transport. Recommendations to prevent hazards and to correct or mitigate welfare consequences have been developed. Recommendations were also developed to define quantitative thresholds for microclimatic conditions within the means of transport and spatial thresholds (minimum space allowance). The development of welfare consequences over time were assessed in relation to maximum journey time. The Opinion covers specific animal transport scenarios identified by the European Commission relating to the export of sheep by livestock vessels, export of sheep by road, roll-on-roll-off vessels and ‘special health status animals’, and lists welfare concerns associated with these. info:eu-repo/semantics/publishedVersion

Respiratory diseases in weaned pigs are a common problem, with a complex etiology involving both viruses and bacteria. In the present study, we investigated the presence of eleven viruses in nasal swabs, collected from nurseries (55 cases) under the suspicion of swine influenza A virus (swIAV) and submitted by swine veterinarians for diagnosis. The other ten viruses included in the study were influenza B (IBV) and D (IDV), Porcine reproductive and respiratory syndrome virus (PRRSV), Porcine respiratory coronavirus (PRCV), Porcine cytomegalovirus (PCMV), Porcine circovirus 2 (PCV2), 3 (PCV3) and 4 (PCV), Porcine parainfluenza 1 (PPIV1) and Swine orthopneumovirus (SOV). Twenty-six swIAV-positive cases and twenty-nine cases of swIAV-negative respiratory disease were primarily established. While IBV, IDV, PCV4 and PPIV1 were not found in any of the cases, PRCV, SOV, and PCMV were more likely to be found in swIAV-positive nurseries with respiratory disease (p < 0.05). Overall, PCV3, PRRSV, and PCMV were the most frequently detected agents at herd level. Taken individually, virus prevalence was: swIAV, 48.6%; PRCV, 48.0%; PRRSV, 31.6%; SOV, 33.8%; PCMV, 48.3%, PCV2, 36.0%; and PCV3, 33.0%. Moreover, low Ct values (p < 0.05). Also, PRCV and SOV were positively correlated between them and negatively with PCMV. Besides, the analysis of suckling pig samples, collected in subclinically infected farrowing units under an influenza monitoring program, showed that circulation of PRCV, PCMV, SOV, and PCV3 started during the early weeks of life. Interestingly, in those subclinically infected units, none of the pathogens was found to be correlated to any other. Overall, our data may contribute to a better understanding of the complex etiology and epidemiology of respiratory diseases in weaners. This is the first report of SOV in Spain and shows, for the first time, the dynamics of this pathogen in swine farms.

Este artículo se encuentra disponible en la siguiente URL: https://www.mdpi.com/2306-7381/9/6/264 En este artículo de investigación también participan: Alicia Manzanares, Teresa Ayats, Aida Mencía-Gutiérrez, Fernando González, Carlos Rojo-Solís, Carlos Barros y Daniel García-Párraga. Este artículo pertenece al número especial "Epidemiology of Wildlife Infectious Diseases". Salmonella is mostly noted as a food-borne pathogen, but contact with chelonians has also been reported as a source of infection. Moreover, high levels of antimicrobial resistance (AMR) have been reported in Salmonella isolated from wild and captive reptiles. The aim of this study was to assess the occurrence of Salmonella AMR carriage by chelonians admitted to two zoological institutions in Spain, characterizing the isolates to assess the Salmonella AMR epidemiology in wildlife. To this end, 152 chelonians from nine species were sampled upon their arrival at the zoological nuclei. Salmonella identification was based on ISO 6579-1:2017 (Annex D), isolates were serotyped and their AMR analysed according to the EU Decision 2013/652. Moreover, the genetic relationship of the isolates was assessed by pulsed-field gel electrophoresis (PFGE). Results showed 19% (29/152) of the chelonians positive to Salmonella, all of them tortoises. For all isolates, 69% (20/29) were resistant and 34% (10/29) multidrug-resistant (MDR) strains. PFGE clustered isolates according to the serovar, confirming a low genetic diversity. In conclusion, this study shows a high presence of MDR Salmonella strains in tortoises at their entry into zoological nuclei. This condition highlights the need to establish Salmonella detection protocols for the entry of animals into these centres.

This article also appears in: Assessment of diseases according to Animal Health Law criteria., Staphylococcus pseudintermedius (S. pseudintermedius) was identified among the most relevant antimicrobial-resistant (AMR) bacteria in the EU for dogs and cats in a previous scientific opinion. Thus, it has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of Article 7 on disease profile and impacts, Article 5 on its eligibility to be listed, Annex IV for its categorisation according to disease prevention and control rules as in Article 9, and Article 8 for listing animal species related to the bacterium. The assessment has been performed following a methodology previously published. The outcome is the median of the probability ranges provided by the experts, which indicates whether each criterion is fulfilled (lower bound ≥ 66%) or not (upper bound ≤ 33%), or whether there is uncertainty about fulfilment. Reasoning points are reported for criteria with uncertain outcome. According to the assessment here performed, it is uncertain whether AMR S. pseudintermedius can be considered eligible to be listed for Union intervention according to Article 5 of the AHL (30–90% probability). According to the criteria in Annex IV, for the purpose of categorisation related to the level of prevention and control as in Article 9 of the AHL, the AHAW Panel concluded that the bacterium does not meet the criteria in Sections 1, 2 and 4 (Categories A, B and D; 0–1%, 1–10% and 10–33% probability of meeting the criteria, respectively) and the AHAW Panel is uncertain whether it meets the criteria in Sections 3 and 5 (Categories C and E, 5–66% and 30–90% probability of meeting the criteria, respectively). The animal species to be listed for AMR S. pseudintermedius according to Article 8 criteria are mostly species belonging to the families of Canidae and Felidae, such as dogs and cats., The AHAW Panel wishes to thank Peter Panduro Damborg from the University of Copenhagen for conducting the extensive literature review under the contract OC/EFSA/ALPHA/2020/02 – LOT 1. The AHAW Panel also wishes to thank Luca Guardabassi from the University of Copenhagen and Verena Oswaldi from EFSA for the support provided for this scientific output.

In this Opinion, the antimicrobial-resistant bacteria responsible for transmissible diseases that constitute a threat to the health of certain kept fish species have been assessed. Atlantic salmon (Salmo salar), carp (Cyprinus spp.), rainbow trout (Oncorhynchus mykiss), sea bream (Sparus aurata) and tilapia (Oreochromis spp.), selected as representative of the most important fish species and production systems that are commercially reared in fresh and saltwater farms, were the focus of this assessment. The assessment was performed following a methodology based on information collected by an extensive literature review and expert judgement. Details of the methodology used for this assessment are explained in a separate Opinion. The global state of play of antimicrobial resistance in Aeromonas hydrophila, Aeromonas salmonicida, Flavobacterium psychrophilum and Flavobacterium columnare is provided. Among these bacteria, none was identified as being among the most relevant antimicrobial-resistant bacteria in the assessed kept fish species in the EU due to the very limited scientific evidence available. info:eu-repo/semantics/publishedVersion

Sentinel-2 offers great potential for monitoring water quality in inland and coastal waters. However, atmospheric correction in these waters is challenging, and there is no standardized approach yet, but different methods coexist under constant development. The atmospheric correction Case 2 Regional Coast Colour (C2RCC) processor has been recently updated with the C2X-COMPLEX (C2XC). This study is one of the first attempts at exploring its performance, in comparison with C2RCC and C2X, in inland and coastal waters in the east of the Iberian Peninsula, in retrieving water surface reflectance and estimating chlorophyll-a ([Chl-a]), total suspended matter ([TSM]), and Secchi disk depth (ZSD). The relationship between in situ ZSD and Kd_z90max product (i.e., the depth of the water column from which 90% of the water-leaving irradiance is derived) of the C2RCC processors demonstrated the potential of this product for estimating water clarity (r > 0.75). However, [TSM] and [Chl-a] derived from the different processors with default calibration factors were not suitable within the targeted scenarios, requiring recalibration based on optical water types or a shift to dynamic algorithm blending approaches. This would benefit from switching between C2RCC and C2XC, which extends the potential for improving surface reflectance estimates to a wide range of scenarios and suggests a promising future for C2-Nets in operational monitoring of water quality. Jesús Soriano held a pre-doctoral grant funded by Agència de Gestió d’Ajuts Universitaris i de Recerca (2020FI_B2 00148). Esther Patricia Urrego holds a pre-doctoral grant funded by Ministerio de Ciencia, Innovación y Universidades (PRE2019-088232). This research was partially funded by the European Union—ERDF and the Ministry of Science and Innovation and the State Research Agency of Spain under project RTI2018-098651-B-C51 (FLEXL3L4—Advanced Products L3 and L4 for the FLEX-S3 mission).

Porcine circovirus 3 (PCV-3) has been associated with several pig diseases. Despite the pathogenicity of this virus has not been completely clarified, reproductive disorders are consistently associated with its infection. The aim of the present work was to analyze the presence of PCV-3 DNA in tissues from pig fetuses from different gestational timepoints. The fetuses were obtained either from farms with no reproductive problems (NRP, n = 249; all of them from the last third of gestation) or from a slaughterhouse (S, n = 51; 49 of the second-third of gestation and 2 from the third one). Tissues collected included brain, heart, lung, kidney, and/or spleen. Overall, the frequency of detection of PCV-3 was significantly higher in fetuses from the last third of the gestation (69/251, 27.5%) when compared to those from the second-third (5/49, 10.2%), although the viral loads were not significantly different. Moreover, the frequency of detection in NRP fetuses (69/249, 27.7%) was significantly higher than in S ones (5/51, 9.8%). Furthermore, PCV-3 DNA was detected in all tissue types analyzed. In conclusion, the present study demonstrates a higher frequency of PCV-3 DNA detection in fetuses from late periods of the gestation and highlights wide organ distributions of the virus in pig fetuses. info:eu-repo/semantics/publishedVersion

16 Pàg., This work was supported by grants S2013/ABI-2906 (PLATESA), P2018/BAA-4370 (PLATESA2) from Comunidad de Madrid/FEDER and AGL2017-83326-R from Ministerio de Economía y Competitividad and PdC2021-121717-I00 funded by MCIN/AEI /10.13039/501100011033 and by Unión Europea Next Generation EU/ PRTR. AL-V has a pre-doctoral fellowship FPI 2017, Ministerio de Ciencia, Innovación y Universidades from the Spanish government.

[Background] The effect of the cecal microbiome on growth of rabbits that were fed under different regimes has been studied previously. However, the term “effect” carries a causal meaning that can be confounded because of potential genetic associations between the microbiome and production traits. Structural equation models (SEM) can help disentangle such a complex interplay by decomposing the effect on a production trait into direct host genetics effects and indirect host genetic effects that are exerted through microbiota effects. These indirect effects can be estimated via structural coefficients that measure the effect of the microbiota on growth while the effects of the host genetics are kept constant. In this study, we applied the SEM approach to infer causal relationships between the cecal microbiota and growth of rabbits fed under ad libitum (ADG) or restricted feeding (ADG)., [Results] We identified structural coefficients that are statistically different from 0 for 138 of the 946 operational taxonomic units (OTU) analyzed. However, only 15 and 38 of these 138 OTU had an effect greater than 0.2 phenotypic standard deviations (SD) on ADG and ADG, respectively. Many of these OTU had a negative effect on both traits. The largest effects on ADG were exerted by an OTU that is taxonomically assigned to the Desulfovibrio genus (− 1.929 g/d, CSS-normalized OTU units) and by an OTU that belongs to the Ruminococcaceae family (1.859 g/d, CSS-normalized OTU units). For ADG, the largest effect was from OTU that belong to the S24-7 family (− 1.907 g/d, CSS-normalized OTU units). In general, OTU that had a substantial effect had low to moderate estimates of heritability., [Conclusions] Disentangling how direct and indirect effects act on production traits is relevant to fully describe the processes of mediation but also to understand how these traits change before considering the application of an external intervention aimed at changing a given microbial composition by blocking/promoting the presence of a particular microorganism., This study was part of the Feed-a-Gene project that received funding from the European Union’s H2020 program under Grant agreement no. 633531, the Spanish project RTI2018-097610R-I00 and the project PID2021-128173OR-C21 (GENEF3). MMF is a recipient of a “Formación de Personal Investigador (FPI)” associated with the research project RTI2018-097610R-I00. YRC is recipient of a Ramon y Cajal post-doctoral fellowship (RYC2019-027244-I) from the Spanish Ministry of Science and Innovation.

The excessive use of antibiotics has led to the emergence of resistant bacteria, mainly from the Enterobacterales group, with high pathogenic/zoonotic potentials that can lead to problems in public health. The increasing presence in freshwater ecosystems highlights the need to evaluate potential sentinel species as risk indicators for both ecosystem and human health. The freshwater mussels provide several ecosystem services, may represent potential sentinel species due to their ability to flter water and retain both organic and inorganic particles. We tested the capability of U. mancus to retain Escherichia coli as a model bacterial organism. Under experimental conditions, the mussels could clear suspended E. coli, facilitating its rapid elimination from water within the frst 24 h after exposure. The species also presented a maximum retention time of 4 days. We also provide allometric equations correlating the fltering capacity with the length and the weight of mussel body parts often used in biometric studies. We provide a frst assessment of the potential of the bivalve Unio mancus to act as a sentinel species for the detection of Enterobacterales and demonstrate the ability to act as a water cleaner., Tis study has been fnanced by the Girona Provincial Council and the Obra Social de la Caixa within the framework of the collaboration project between the Girona Provincial Council and la Caixa regarding natural spaces 2017 (approval by the Governing Board of April 17, 2018).

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes respiratory disease, but it can also affect other organs including the central nervous system. Several animal models have been developed to address different key questions related to Coronavirus Disease 2019 (COVID-19). Wild-type mice are minimally susceptible to certain SARS-CoV-2 lineages (beta and gamma variants), whereas hACE2-transgenic mice succumb to SARS-CoV-2 and develop a fatal neurological disease. In this article, we aimed to chronologically characterize SARS-CoV-2 neuroinvasion and neuropathology. Necropsies were performed at different time points, and the brain and olfactory mucosa were processed for histopathological analysis. SARS-CoV-2 virological assays including immunohistochemistry were performed along with a panel of antibodies to assess neuroinflammation. At 6 to 7 days post inoculation (dpi), brain lesions were characterized by nonsuppurative meningoencephalitis and diffuse astrogliosis and microgliosis. Vasculitis and thrombosis were also present and associated with occasional microhemorrhages and spongiosis. Moreover, there was vacuolar degeneration of virus-infected neurons. At 2 dpi, SARS-CoV-2 immunolabeling was only found in the olfactory mucosa, but at 4 dpi intraneuronal virus immunolabeling had already reached most of the brain areas. Maximal distribution of the virus was observed throughout the brain at 6 to 7 dpi except for the cerebellum, which was mostly spared. Our results suggest an early entry of the virus through the olfactory mucosa and a rapid interneuronal spread of the virus leading to acute encephalitis and neuronal damage in this mouse model. The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: The CBIG Consortium (constituted by IRTA-CReSA, BSC, and IrsiCaixa) is supported by Grifols pharmaceutical. The authors also acknowledge the crowdfunding initiative of https://www.yomecorono.com. Peer Reviewed "Article signat per 12 autors/es: Enric Vidal*, Carlos López-Figueroa*, Jordi Rodon, Mónica Pérez, Marco Brustolin, Guillermo Cantero, Víctor Guallar, Nuria Izquierdo-Useros, Jorge Carrillo, Julià Blanco, Bonaventura Clotet, Júlia Vergara-Alert, Joaquim Segalés"

This article also appears in: Assessment of diseases according to Animal Health Law criteria., Pseudomonas aeruginosa (P. aeruginosa) was identified among the most relevant antimicrobial-resistant (AMR) bacteria in the EU for dogs and cats in a previous scientific opinion. Thus, it has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of Article 7 on disease profile and impacts, Article 5 on its eligibility to be listed, Annex IV for its categorisation according to disease prevention and control rules as in Article 9, and Article 8 for listing animal species related to the bacterium. The assessment has been performed following a methodology previously published. The outcome is the median of the probability ranges provided by the experts, which indicates whether each criterion is fulfilled (lower bound ≥ 66%) or not (upper bound ≤ 33%), or whether there is uncertainty about fulfilment. Reasoning points are reported for criteria with uncertain outcome. According to the assessment here performed, it is uncertain whether AMR P. aeruginosa can be considered eligible to be listed for Union intervention according to Article 5 of the AHL (33–90% probability). According to the criteria in Annex IV, for the purpose of categorisation related to the level of prevention and control as in Article 9 of the AHL, the AHAW Panel concluded that the bacterium does not meet the criteria in Sections 1, 2, 3 and 4 (Categories A, B, C and D; 0–5%, 1–5%, 5–33% and 5–33% probability of meeting the criteria, respectively) and the AHAW Panel was uncertain whether it meets the criteria in Section 5 (Category E, 33–90% probability of meeting the criteria). The animal species to be listed for AMR P. aeruginosa according to Article 8 criteria are mainly dogs and cats., The AHAW Panel wishes to thank Joanne Leri Fothergill from the University of Liverpool for conducting the extensive literature review under the contract EOI/EFSA/SCIENCE/2020/01. The AHAW Panel also wishes to Verena Oswaldi from EFSA for the support provided for this scientific output.

Background Tuberculosis (TB) due to Mycobacterium caprae is endemic in goat herds and free-ranging wild boars in Spain, causing infections in other livestock or wild animals to a lesser extent. TB infection in foxes is infrequently reported and they are usually considered spillover hosts of TB. Case presentation A blind, depressed and severely emaciated red fox (Vulpes vulpes) was admitted to a rehabilitation center. After clinical examination it was humanely sacrificed. At necropsy, generalized TB lesions were observed that were subsequently confirmed by histopathology along with a co-infection with canine distemper virus. M. caprae was isolated from mycobacterial culture and spoligotype SB0415 was identified. Whole genome sequencing (WGS) of the isolated M. caprae was carried out and single nucleotide polymorphisms (SNP) were compared with other sequences of M. caprae isolated from livestock and wildlife of the same area throughout the last decade. Conclusions This is the first reported case of TB due to M. caprae in a fox in the Iberian Peninsula. WGS and SNP analysis, together with spatial-temporal investigations, associated this case with recent M. caprae outbreaks in cattle and goat herds of the area. The results indicated transmission of M. caprae between livestock and the fox, suggesting that this species may occasionally play a role in the epidemiology of animal TB.

EFSA received a mandate from the European Commission to assess the effectiveness of some of the control measures against diseases included in the Category A list according to Regulation (EU) 2016/429 on transmissible animal diseases ('Animal Health Law’). This opinion belongs to a series of opinions where these control measures will be assessed, with this opinion covering the assessment of control measures for glanders. In this opinion, EFSA and the AHAW Panel of experts review the effectiveness of: (i) clinical and laboratory sampling procedures, (ii) monitoring period and (iii) the minimum radius of the protection and surveillance zone, and the minimum length of time the measures should be applied in these zones. The general methodology used for this series of opinions has been published elsewhere. Considering the epidemiology and distribution of glanders, it was foreseen that three different situations could lead to a suspicion of the disease. Sampling procedures were defined for each of the three different suspicion types, which can also be applied in most of the other scenarios assessed. The monitoring period (6 months) was assessed as effective in all scenarios. The AHAW Panel of experts considered the minimum radius and duration of the existing protection and surveillance zone, set at the establishment level, effective. Recommendations provided for each of the scenarios assessed aim to support the European Commission in the drafting of further pieces of legislation, as well as for plausible ad hoc requests in relation to glanders. info:eu-repo/semantics/publishedVersion

The presence of Dolomedes plantarius (Clerck, 1757) (Araneae: Pisauridae) in the Ebro Delta extends its distribution in the Iberian Peninsula southwards. The rare Dolomedes plantarius has been observed in a permanent pond (“ullal”) in the Ebro Delta (Catalonia, north-eastern Spain). This is the most meridional record registered in the Iberian Peninsula, and perhaps in western Europe. Thus, the presence of this semiaquatic species may vary our assumptions on its rarity. Such finding evidences the relevance of aquatic environments in general and “ullals” in particular for biodiversity conservation, as well as the importance of considering both the semiaquatic and the terrestrial species inhabiting them. info:eu-repo/semantics/publishedVersion

Altres ajuts: acords transformatius de la UAB Altres ajuts: BBVA Foundation; Grifols; National Agency for Research and Development of Chile, Grant/Award Number: 72180406; La Marató TV3, Grant/Award Number: 342/C/2021 Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the coronavirus infectious disease 2019 (COVID-19) pandemic in humans, is able to infect several domestic, captive and wildlife animal species. Since reverse zoonotic transmission to pets has been demonstrated, it is crucial to determine their role in the epidemiology of the disease to prevent further spillover events and major spreads of SARS-CoV-2. In the present study, we determined the presence of virus and the seroprevalence to SARS-CoV-2, as well as the levels of neutralizing antibodies (nAbs) against several variants of concern (VOCs) in pets (cats, dogs and ferrets) and stray cats from North-Eastern of Spain. We confirmed that cats and dogs can be infected by different VOCs of SARS-CoV-2 and, together with ferrets, are able to develop nAbs against the ancestral (B.1), Alpha (B.1.1.7), Beta (B.1.315), Delta (B.1.617.2) and Omicron (BA.1) variants, with lower titres against the latest in dogs and cats, but not in ferrets. Although the prevalence of active SARS-CoV-2 infection measured as direct viral RNA detection was low (0.3%), presence of nAbs in pets living in COVID-19 positive households was relatively high (close to 25% in cats, 10% in dogs and 40% in ferrets). It is essential to continue monitoring SARS-CoV-2 infections in these animals due to their frequent contact with human populations, and we cannot discard the probability of a higher animal susceptibility to new potential SARS-CoV-2 VOCs.

Salinity is changing in aquatic systems due to anthropogenic activities (like irrigation or dam management) and climate change. Although there are studies on the effects of salinity variations on individual species, little is known about the effects on overall ecosystems, these impacts being more uncertain in transitional waters such as estuaries or fiords. The few works that do address this topic have considered these impacts using ecotoxicity models. However, these models state that an increase in the concentration of a pollutant generates an increase in the impacts, disregarding the effects of water freshening. The present research work introduces a general framework to address the impacts of salinity variations, including emission-related positive effects. We validated this framework by applying it to an estuarine area in Galicia (northwestern Spain), where sharp drops in the salt concentration have caused mass mortalities of shellfish in recent decades. This research work addresses for the first time the potential effects on the environment derived from a decrease in the concentration of essential substances, where the effects of an emission can also generate positive impacts. Moreover, it is expected that the framework can also be applied to model the environmental impacts of other essential substances in life cycle assessment (LCA), such as metals and macronutrients. info:eu-repo/semantics/acceptedVersion

Antimicrobial susceptibility testing is necessary to carry out antimicrobial stewardship but a limited number of drugs belonging to each antimicrobial family has to be tested for technical limitations and economic resources. In this study, we have determined the minimal inhibitory concentration, using microdilution following international standards (CLSI), for 490 Actinobacillus pleuropneumoniae, 285 Pasteurella multocida, 73 Bordetella bronchiseptica, 398 Streptococcus suis and 1571 Escherichia coli strains from clinical cases collected in Spain between 2018 and 2020. The antimicrobial susceptibility pattern was deciphered using a principal component analysis for each bacterium and a matrix correlation (high > 0.8, medium 0.5-0.8 and low < 0.5) was obtained for each pair of antimicrobials. No significant associations were observed between MIC patterns for different antimicrobial families, suggesting that co-selection mechanisms are not generally present in these porcine pathogens. However, a high correlation was observed between the fluroquinolones (marbofloxacin and enrofloxacin) for all mentioned pathogens and for ceftiofur and cefquinome for E. coli and S. suis. Moreover, a significant association was also observed for tetracyclines (doxycycline and oxytetracycline) and B. bronchiseptica and tildipirosin/tulathromycin for P. multocida. These results suggest that generally, a representative drug per antimicrobial class cannot be selected, however, for some drug-bug combinations, MIC values from one representative drug could be extrapolated to the whole antimicrobial family. info:eu-repo/semantics/publishedVersion