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A human-ACE2 knock-in mouse model for SARS-CoV-2 infection recapitulates respiratory disorders but avoids neurological disease associated with the transgenic K18-hACE2 model
No full textAnimal models have been instrumental in elucidating the pathogenesis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and in testing coronavirus disease 2019 (COVID-19) vaccines and therapeutics. Wild-type (WT) mice are not susceptible to many SARS-CoV-2 variants, and therefore, transgenic K18-hACE2 mice have emerged as a standard model system. However, this model is characterized by a severe disease, particularly associated with neuroinfection, which leads to early humane endpoint euthanasia. Here, we established a novel knock-in (KI) mouse model by inserting the original K18-hACE2 transgene into the collagen type I alpha chain (COL1A1) locus using a recombinase-mediated cassette exchange (RMCE) system. Once the Col1a1-K18-hACE2 mouse colony was established, animals were challenged with a B.1 SARS-CoV-2 (D614G) isolate and were monitored for up to 14 days. Col1a1-K18-hACE2 mice exhibited an initial weight loss similar to the K18-hACE2 transgenic model but did not develop evident neurologic clinical signs. The majority of Col1a1-K18-hACE2 mice did not reach the pre-established humane endpoint, showing a progressive weight gain 9 days postinfection (dpi). Importantly, despite this apparent milder pathogenicity of the virus in this mouse model compared to the K18-hACE2 transgenic model, high levels of viral RNA were detected in the lungs, oropharyngeal swab, and nasal turbinates. Moreover, the remaining lesions and inflammation in the lungs were still observed 14 dpi. In contrast, although low-level viral RNA could be detected in a minority of Col1a1-K18-hACE2 animals, no brain lesions were observed at any timepoint. Overall, Col1a1-K18-hACE2 mice constitute a new model for investigating SARS-CoV-2 pathogenesis and treatments, with potential implications for studying long-term COVID-19 sequelae.The authors would like to acknowledge Jorge Díaz, Yaiza Rosales-Salgado, Rosa María Ampudia-Carrasco, Sergi Sunyé-Casas, and Mireia Martínez from the CMCiB for their essential help in the BSL3 facility and the K18-hACE2 mouse colony. We also thank Marisa Larramona from Parc Científic de Barcelona for her invaluable help with the knock-in mouse colony. We thank the Dormeur Fondation for their financial support for the acquisition of the QuantStudio-5 real-time PCR system. A.P.-G. was supported by a predoctoral grant from Generalitat de Catalunya and Fons Social Europeu (2022 FI_B 00698). This study was funded by Grifols, the Departament de Salut of the Generalitat de Catalunya (grant nos. SLD016 to J.B. and SLD015 to J.C.), the Spanish Health Institute Carlos III, CERCA Programme/Generalitat de Catalunya 2021 SGR 00452, and the crowdfunding initiatives #joemcorono, BonPreu/Esclat, and Correos. N.I.-U. is supported by the Spanish Ministry of Science and Innovation (grants PID2023-147498OB-I00, PID2020-117145RB-I00, and 10.13039/501100011033, Spain), the EU HORIZON-HLTH-2021CORONA-01 (grant 101046118, European Union), and the institutional funding of Pharma Mar, Grifols, HIPRA, and Amassence. The funders had no role in the study design, data collection and interpretation, or the decision to submit the work for publication. A.P.-G., F.T.-F., B. Trinité, and J.B. conceived and designed the experiments. A.P.-G., F.T.-F., and B. Trinité performed the animal procedures. A.P.-G., F.T.-F., M.P., E.R.-M., D.R.-R., D.P.-Z., J.M.-B., E.P., J.S., and B. Trinité performed the analytical experiments. A.P.-G., B. Trinité, J.S., J.V.-A., V.U., and J.B. analyzed and interpreted the data. S.F. and B. Tondelli established and provided the knock-in mouse colony. S.C. established communication between animal facilities and contributed to the veterinary report verification to allow the knock-in animal shipment to the CMCiB facility. D.P.-Z., J.M.-B., D.R.-R., and N.I.-U. provided key reagents. A.P.-G., B. Trinité, and J.B. wrote the paper. All authors contributed to the article and approved the submitted version.info:eu-repo/semantics/publishedVersio
Inactivated Rothia nasimurium promotes a persistent antiviral immune status in porcine alveolar macrophages
Get PDFGlobalization has increased the incidence of infectious diseases in livestock, further aggravated by the reduction of antibiotic usage. To minimize the resulting economic consequences to the meat production industry, as well as the risk of zoonotic events, the use of immunostimulants has emerged as a potential strategy to enhance animal resilience to diseases. In particular, the capability of bacterial-based immunostimulants to modulate innate immune cells functionality makes them cost-effective candidates as vaccine adjuvants, antimicrobials, or preventive immunostimulators inducing long-term innate immune memory in livestock. However, further research is required to identify novel bacterial strains with immunostimulatory properties. Here we characterized in vitro the immunostimulatory properties of Rothia nasimurium isolated from warthog fecal microbiota. Stimulation with heat-inactivated Rothia induced cytokine production by porcine immune cells, and a robust innate immune transcriptomic signature in porcine alveolar macrophages. Interestingly, the bacteria induced inflammasome activation and IL-1β production, thus confirming its pro-inflammatory properties, and suggesting its potential as vaccine adjuvant. Importantly, this immunostimulatory status functionally resulted in an antimicrobial state, enhancing the phagocytic capability of alveolar macrophages, and hampering the replication levels of two major porcine viral pathogens: the porcine reproductive and respiratory syndrome virus (PRRSV) and the African swine fever virus (ASFV). Moreover, macrophages showed an enhanced cytokine response upon ASFV infection several days after heat-inactivated Rothia stimulation, suggesting the induction of an innate immune memory phenotype. This nonspecific response resulted in a significant reduction of ASFV replication kinetics, demonstrating the capacity of the bacteria to induce a more resistant state in macrophages against a virus infection. Altogether, these results demonstrate the immunostimulatory capability of heat-inactivated R. nasimurium in porcine macrophages, showing potential to enhance animal resilience to diseases through the modulation of innate immune cells responsiveness to infections.The author(s) declare that financial support was received for the research and/or publication of this article. This work is part of the projects PDC2021-120987-I00 (FR) and PID2022-136312OB-I00 (FR&JA), both funded by the MCIN/AEI/10.13039/501100011033/FEDER, UE. YZ is supported by a fellowship from the China Scholarship Council Visiting Scholar Program (202305960008). Institutional support to CNAG was provided by the Spanish Ministry of Science and Innovation through the Instituto de Salud Carlos III, and by the Generalitat de Catalunya through the Departament de Salut and the Departament de Recerca i Universitats. KK is supported by funding from the Spanish Ministry of Research and Innovation (RYC2021-033035-I/AEI/10.13039/501100011033).info:eu-repo/semantics/publishedVersio
Impact of zinc oxide on gut health, immunity, and growth in weaned piglets: exploring potential modes of action
Get PDFZinc oxide (ZnO) has been used at pharmacological levels to promote gut health
and growth performance in the critical postweaning (PW) phase of piglets. The
pharmacological use of ZnO in piglet diets has been banned in Europe and other
countries due to antimicrobial resistance and environmental concerns. Therefore,
understanding its mode of action, including its molecular mechanisms, is crucial for
developing effective and sustainable alternatives. We investigated the mechanisms
by which dietary supplementation with 3,000 mg/kg ZnO supports gut health and
improves growth performance of piglets during the first 14 days PW. During the
2 weeks of trial (0–14 d PW), ZnO fed piglets had higher average daily gain (165
vs. 123 g/d; p < 0.01), and tended to have increased average daily feed intake (204
vs. 181 g/d; p < 0.1) and improved gain-to-feed ratio (0.669 vs. 0.774; p < 0.05)
compared to control piglets. Feces from piglets in the ZnO group were also more
consistent during the 2 weeks of trial (p < 0.01). At day 14 PW, ZnO piglets had
lower calprotectin concentrations in serum (p < 0.01). Dietary ZnO downregulated
several genes, involved in immune, oxidative and inflammatory responses, in
jejunal (GPX2, REG3G, IL-8, IL-6, IL-22, and TGFβ1) and ileal (REG3G, IL-17A, IL-1β,
and TLR2) mucosa (p < 0.05). It also downregulated the expression of the zinc
transporter SLC39A4, that is associated with zinc homeostasis, in both tissues.
Notably, PPARGC1A, which promotes energy production and lipid metabolism
through fatty acid oxidation, was upregulated by ZnO in ileum. In conclusion,
the current results suggest that high dietary levels of ZnO reduce the expression
of inflammatory cytokines, the oxidative enzyme GPX2, pathogen recognition
proteins, and zinc transporters while promoting the expression of PPARGC1A gene
related with energy metabolism in the intestine. Therefore, ZnO can facilitate a
smoother weaning transition to reduce weaning related gut health disturbances,
ultimately contributing to gut homeostasis and improved performanceThe author(s) declare that financial support was received for the research and/or publication of this article. ZN received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 945413 and from the Universitat Rovira i Virgili (URV) and the Institute of Agrifood Research and Technology (IRTA).info:eu-repo/semantics/publishedVersio
Floració: La promesa de la propera collita d’olis, a punt de fer-se realitat
Get PDFSi passegeu pel camp o visiteu algun jardí on hi hagi oliveres, veureu de lluny que han canviat: es veuen
les branques plenes de flors a punt d’obrir-seinfo:eu-repo/semantics/publishedVersio
Adventively established Leptopilina japonica: a new opportunity for augmentative biocontrol of Drosophila suzukii
Get PDFThe invasive spotted-wing drosophila, Drosophila suzukii, has emerged as a significant global pest over the past decade, threatening fruit production worldwide. The parasitoid Leptopilina japonica, presumed native to East Asia, has established adventive populations in Europe and North America and is increasingly recognized for its ability to parasitize substantial proportions of D. suzukii larvae across diverse habitats. Here, we provide a broad review of the biology, establishment, distribution, and potential impacts of L. japonica. Using field data from international monitoring programs, we document the seasonal dynamics of plant–host–parasitoid associations and assess evidence for L. japonica’s impact on D. suzukii and non-target organisms. Findings indicate that L. japonica has successfully established in several areas where D. suzukii is present in Europe and North America, showing promise as a biological control agent to support sustainable pest management. Current data suggest it provides some suppression of D. suzukii populations with minimal non-target effects. However, long-term studies are necessary to clarify its food web interactions and efficacy as a biological control agent. In areas where L. japonica has been established, we propose its use in augmentative biological control programs to enhance its impacts in specific agricultural settings. Case-specific evaluations of its ecological effects and role in integrated pest management, supported by continued monitoring, are essential. The case of L. japonica illustrates the need for clear, research-informed policies to guide the use of adventively established non-indigenous natural enemies in pest management.Open access funding provided by Fondazione Edmund Mach - Istituto Agrario di San Michele all'Adige within the CRUI-CARE Agreement. This research was supported by multiple funding sources. In the USA, funding were provided by USDA Animal and Plant Health Inspection Service (APHIS) Farm Bill Award No. 60–8010-4–001, USDA base funds: CRIS 2072–22000-044-00D, USDA Agricultural Research Service (ARS) Areawide Program (National Program Leader: S. Young), USDA Specialty Crop Research Initiative (SCRI) Award No. 2020–51181-32140, USDA Crop Protection and Pest Management (CPPM) Award No. 2021–70006-35312, USDA National Institute of Food and Agriculture (NIFA) Awards No. 2021–70006-35312 and 2020–51181-32140, USDA Specialty Crop Block Grant Program (SCBGP), USDA Northwest Center for Small Fruit Research, Oregon Blueberry Commission, Oregon Blackberry Commission, New Jersey Blueberry and Cranberry Research Council, Michigan Blueberry Commission and the “GREEEN—Generating Research and Extension to meet Economic and Environmental Needs” project. In Canada, funding was provided by the Dekaban Scholars program, the Natural Sciences and Engineering Research Council of Canada (NSERC) and the Agriculture and Agri-Food Canada: Projects J-003401 and J-002839. In France, field surveys by INRAE ISA were part of the Ecophyto SUZoCARPO project, funded by the French Ministry of Agriculture and Food. In Switzerland, field surveys were funded by the Swiss Federal Office for Agriculture. In Italy, funding were provided by the Samurai Wasp Action Team (SWAT) project, funded by the Autonomous Province of Trento, the project "URCOFI—Unità Regionale Coordinamento Fitosanitario," funded by the Campania Region government (Grant No. CUP B29I22001290009), the project “SUccess of Specialist versus generalist parasitoid in Hampering the spread of an Invasive pest (SUSHI),” funded by the Italian Ministry of University and Research (MUR) (Grant No. 20227ZYCH3, CUP Master B53D2301184 0006).info:eu-repo/semantics/publishedVersio
A lexicon obtained and validated by a data-driven approach for organic residues valorization in emerging and developing countries
Get PDFThe author(s) declare that financial support was received for the research and/or publication of this article. This work was supported by European Union Feder funding (EU ERDF: GURTDI 20151501-0000735).info:eu-repo/semantics/publishedVersio
Production of biogenic amines and changes in the microbiome composition of dry-cured fish during dynamic storage up to the end of the shelf life
Get PDFThe aim of this study was to quantify and identify biogenic amines in vacuum packed dry-cured salmon, swordfish and tuna, stored under dynamic abuse temperature conditions during their shelf life. The same samples were also submitted to shotgun metagenomic sequencing to characterize their microbiome in terms of taxonomic groups and functional genes linked to biogenic amines production. In tuna and swordfish, the biogenic amines were always below the quantification limit. In salmon, cadaverine was 6.98 mg/kg at T0 and increased up to 535.69 mg/kg. Histamine, tyramine and putrescine were detected from day 32–61. The salmon microbiome was mostly represented by Photobacterium up to 19 days and then replaced by Latilactobacillus. Both genera included species, such as Photobacterium phosphoreum and Latilactobacillus curvatus, producing biogenic amines. The tuna microbiome was stable over time, while in swordfish Staphylococcus increased from day 32, becoming the most represented genus at the end of the shelf life. The functional genes associated with biogenic amines metabolism were more abundant in salmon than in tuna and swordfish, thus reflecting their biogenic profile. The results of this study highlight the importance of preventing the presence of biogenic amines in raw fish before to the dry-curing process, in order to avoid their accumulation during shelf life. Moreover, they may inform risk assessments and support the development of comprehensive regulatory standards for biogenic amines in dry-cured fish products.info:eu-repo/semantics/publishedVersio
Predation by Native Mediterranean Species on the Invasive Blue Crab: Experimental Evidence from the Common Octopus and the Loggerhead Sea Turtle
Get PDFThe rapid spread of the invasive blue crab (Callinectes sapidus) in Mediterranean coastal ecosystems poses ecological and socio-economic challenges, highlighting the need to explore natural control mechanisms. We conducted controlled aquarium experiments to assess the functional size preference and feeding rates of two native Mediterranean predators: the common octopus (Octopus vulgaris) and the loggerhead sea turtle (Caretta caretta). Two sets of trials were performed: (1) size preference experiments, in which individual predators were simultaneously offered crabs of varying sizes; and (2) ad libitum feeding experiments to estimate daily consumption rates. Octopuses consumed only the soft tissues biomass (STB, ~54% of total crab biomass), reaching maximum intake rates of 454 ± 71 gWW·d−1 in adult individuals. Sea turtles ingested whole crabs, including the exoskeleton, with total consumption rates up to 815 ± 592 gWW·d−1 of total biomass (TB) in adults. Subadult octopuses exhibited shorter handling times for small crabs, although neither predator showed a statistically significant size preference. Our findings indicate that both predators can consume substantial amounts of blue crab biomass, but O. vulgaris—due to its higher feeding efficiency and ecological abundance—might play a more prominent role as a natural biocontrol agent. These results provide valuable insights for integrated management strategies aimed at mitigating the ecological impact of C. sapidus in the Mediterranean.This research was funded by the Ministry of Science and Innovation (Grant PID 2020-118476RR-C21 by MCIN/AEI/10.13039/501100011033). P. Prado was contracted under the INIA-CCAA cooperative research program for postdoctoral incorporation from the Spanish National Institute for Agricultural and Food Research and Technology (INIA).info:eu-repo/semantics/publishedVersio
Principales problemas sensoriales de las carnes y de los preparados de carne (II)
No full textEn este artículo continuamos con la serie sobre los problemas sensoriales que pueden presentar las carnes y los preparados de carne, en este caso los relacionados con el color y la estabilidad del color de la carne fresca.info:eu-repo/semantics/publishedVersio
El futur de la detecció de les plagues d'insectes en productes emmagatzemats
No full textDesprés de la collita, els productes agrícoles com els cereals i les llegums fan un llarg viatge fins arribar al consumidor. Aquests es guarden en sitges i magatzems durant períodes de fins a diversos mesos abans d’arribar al molí o altres instal·lacions de processament i envasat per, finalment, comercialitzar-los.
Durant l’emmagatzematge, és comú que aquests productes siguin atacats per plagues, cosa que pot causar pèrdues econòmiques importants i deteriorar-ne la qualitat nutricional i sanitària.info:eu-repo/semantics/publishedVersio