A rapaza das cabras

El artículo cuenta con la colaboración de Mikel Oleaga (Agranda la Olla), Edu Balsells Martínez (Asociación La Gaiata), Daniel Morante e Isara Belinfanta (Cel i Sol), Aitor Azkarate (Jauregia Esnekiak), Daniel Mercader (Camp de la sort) y Lidia Fanjul y Sebastián Burch.Engarzados por lo que nos transmiten, presentamos en este artículo los sentimientos y motivos que a mucha gente joven mueven hacia el campo como medio de vida. Porque estaban, o porque quieren disfrutarlo. Se puede apreciar en cada palabra

Biosafety steps in the manufacturing process of spray-dried plasma : a review with emphasis on the use of ultraviolet irradiation as a redundant biosafety procedure

Ajuts: Secretaria de Universitats i Recerca del Departament d'Economia i Coneixement de la Generalitat de Catalunya 014 DI 066Spray dried plasma (SDP) is a functional protein source obtained from blood of healthy animals, approved by the veterinary authorities from animals declared to be fit for slaughter for human consumption. Blood of these animals is collected at the slaughterhouse, treated with an anticoagulant, chilled and transported to industrial facilities in which blood is centrifuged to separate the red blood cells from the plasma fraction. Plasma is then concentrated, and spray dried at high temperatures (80 °C throughout its substance) to convert it in a powder. Such method preserves the biological activity of its proteins, mainly albumins and globulins. SDP is mainly used in pig feed diets to significantly improve daily gain, feed intake, production efficiency, and to reduce post-weaning lag caused by the appearance of post-weaning diarrhea. Although SDP is considered a safe product and its manufacturing process consists of several biosafety steps, the security of the SDP is often questioned due to its nature as raw blood by-product, especially when emergent or re-emergent pathogens appear. This review provides an evaluation and validation of the different safety steps present in the manufacturing process of SDP, with special focus on a new redundant pathogen inactivation step, the UV-C irradiation, that may be implemented in the manufacturing process of the SDP. Overall results showed that the manufacturing process of SDP is safe and the UV-C radiation was effective in inactivating a wide range of bacteria and viruses spiked and naturally present in commercially collected liquid animal plasma and it can be implemented as a redundant biosafety step in the manufacturing process of the SDP

Estimated quantity of swine virus genomes based on quantitative PCR analysis in spray-dried porcine plasma samples collected from multiple manufacturing plants

This survey was conducted to estimate the incidence and level of potential viral contamination in commercially collected porcine plasma. Samples of spray dried porcine plasma (SDPP) were collected over a 12- month period from eight spray drying facilities in Spain, England, Northern Ireland, Brazil, Canada, and the United States. In this survey, viral load for several porcine pathogens including SVA, TGEV, PRRSV (EU and US strains), PEDV, PCV-2, SIV, SDCoV and PPV were determined by qPCR. Regression of Ct on TCID of serial diluted stock solution of each virus allowed the estimate of potential viral level in SDPP and unprocessed liquid plasma (using typical solids content of commercially collected porcine plasma). In this survey SVA, TGEV or SDCoV were not detected in any of the SDPP samples. Brazil SDPP samples were free of PRRSV and PEDV. Samples of SDPP from North America primarily contained the PRRSV-US strain while the European samples contained the PRRSV-EU strain (except for one sample from each region containing a relatively low estimated level of the alternative PRRSV strain). Estimated viral level tended to be in the range from <1.0 log TCID to <2.5 log TCID. Estimated level of SIV was the exception with a very low incidence rate but higher estimated viral load <3.9 log TCID. In summary, the incidence of potential viral contamination in commercially collected porcine plasma was variable and estimated virus level in samples containing viral DNA/RNA was relatively low compared with that occurring at the peak viremia during an infection for all viruses or when considering the minimal infectious dose for each of them

Caso inusual de fuga en un tubo endotraqueal por defecto de fabricación.

One of the fields where more progress has been made in recent decades is security. In the surgical setting, standardized protocols are particularly needed to increase detection capabilities. The surgical checklist includes material review. A case is presented in which high leaks were observed during mechanical ventilation after anesthetic induction. After reviewing the endotracheal tube, a perforation was observed. It is recommended to check the walls of the tube and the correct passage of air through it, checking only the pneumo-plug entails not detecting latent errors that can trigger a critical event.Uno de los campos donde más avances se han producido en las últimas décadas es la seguridad.  En el entorno quirúrgico son particularmente necesarios protocolos estandarizados para aumentar la capacidad de detección. El listado de verificación quirúrgica incluye la revisión del material. Se presenta un caso en el que se objetivaron fugas elevadas durante la ventilación mecánica tras inducción anestésica. Tras revisar el tubo endotraqueal, se objetivó una perforación. Se recomienda revisar las paredes del tubo y el correcto paso de aire a su través, revisar únicamente el neumotaponamiento conlleva no detectar errores latentes que pueden desencadenar un evento crítico

Evaluation of the effectiveness of the surepure turbulator ultraviolet-C irradiation equipment on inactivation of different enveloped and non-enveloped viruses inoculated in commercially collected liquid animal plasma

This study was partly supported by Secretaria de Universitats i Recerca del Departament d'Economia i Coneixement of the Generalitat de Catalunya. Author who received the award:EB. Grant number: 2014 DI 066. Full name of funder: Secretaria de Universitats i Recerca del Departament d'Economia i Coneixement of the Generalitat de Catalunya. URL funder website: universitatsirecerca.gencat.cat/ca/inici. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. APC EUROPE provided support in the form of salaries for authors EB, CR, JR and JPolo retrospectively, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. We want to thank the Secretaria de Universitats i Recerca del Departament d'Economia i Coneixement de la Generalitat de Catalunya (2014 DI 066) for its colaboration.The objective of this study was to evaluate the effectiveness of the SurePure Turbulator ultraviolet-C (UV-C, 254 nm wavelength) irradiation equipment on inactivation of different enveloped and non-enveloped viruses in commercially collected liquid animal plasma. Specifically, Pseudorabies virus (PRV), Porcine reproductive and respiratory syndrome virus (PRRSV), Porcine epidemic diarrhea virus (PEDV), Bovine viral diarrhea virus (BVDV), Classical swine fever virus (CSFV), Swine influenza virus (SIV) as enveloped viruses and Porcine parvovirus (PPV), Swine vesicular disease virus (SVDV), Porcine circovirus type 2 (PCV-2) and Senecavirus A (SVA) as non-enveloped viruses, were inoculated in bovine or porcine plasma and subjected to different UV-C irradiation doses (0, 750, 1500, 3000, 6000 and 9000 J/L) using an UV-C device developed for opaque liquid working under turbulent flow. The enveloped viruses tested were inactivated at < 3000 J/L of UV-C, being the dose needed to inactivate 4 log TCID (4D) of 1612 J/L for PRV,1004 J/L for PRRSV, 1953 J/L for PEDV, 1639 J/L for SIV, 1641 J/L for CSFV and 1943 J/L for BVDV. The non-enveloped viruses tended to have higher 4D values: 2161 J/L for PPV, 3223 J/L for SVA and 3708 J/L for SVDV. Because the initial viral concentration was <4.0 Log for PCV-2, it was not possible to calculate the 4D value for this virus. In conclusion, these results demonstrated that the SurePure Turbulator UV-C treatment system is capable of inactivating significant levels of swine viruses inoculated in commercially collected porcine or bovine plasma. It was concluded that irradiation with UV-C can provide an additional redundant biosafety feature in the manufacturing process of spray-dried animal plasma

Combined effects of spray-drying conditions and postdrying storage time and temperature on Salmonella choleraesuis and Salmonella typhimurium survival when inoculated in liquid porcine plasma

Altres ajuts: GC/2014DI066The objective of this study was to determine the effectiveness of the spray-drying process on the inactivation of Salmonella choleraesuis and Salmonella typhimurium spiked in liquid porcine plasma and to test the additive effect of immediate postdrying storage. Commercial spray-dried porcine plasma was sterilized by irradiation and then reconstituted (1:9) with sterile water. Aliquots of reconstituted plasma were inoculated with either S. choleraesuis or S. typhimurium, subjected to spray-drying at an inlet temperature of 200°C and an outlet temperature of either 71 or 80°C, and each spray-drying temperature combinations were subjected to either 0, 30 or 60 s of residence time () as a simulation of residence time typical of commercial dryers. Spray-dried samples were stored at either 4·0 ± 3·0°C or 23·0 ± 0·3°C for 15 days. Bacterial counts of each Salmonella spp., were completed for all samples. For both Salmonella spp., spray-drying at both outlet temperatures reduced bacterial counts about 3 logs at 0 s, while there was about a 5·5 log reduction at 60 s. Storage of all dried samples at either 4·0 ± 3·0°C or 23·0 ± 0·3°C for 15 days eliminate all detectable bacterial counts of both Salmonella spp. Safety of raw materials from animal origin like spray-dried porcine plasma (SDPP) may be a concern for the swine industry. Spray-drying process and postdrying storage are good inactivation steps to reduce the bacterial load of Salmonella choleraesuis and Salmonella typhimurium. For both Salmonella spp., spray-drying at 71°C or 80°C outlet temperatures reduced bacterial counts about 3 log at residence time (RT) 0 s, while there was about a 5.5 log reduction at RT 60 s. Storage of all dried samples at either 4.0 ± 3.0°C or 23.0 ± 0.3°C for 15 days was effective for eliminating detectable bacterial counts of both Salmonella spp. Significance and Impact of the Study: Safety of raw materials from animal origin like spray-dried porcine plasma (SDPP) may be a concern for the swine industry. Spray-drying process and postdrying storage are good inactivation steps to reduce the bacterial load of Salmonella choleraesuis and Salmonella typhimurium. For both Salmonella spp., spray-drying at 71°C or 80°C outlet temperatures reduced bacterial counts about 3 log at residence time (RT) 0 s, while there was about a 5.5 log reduction at RT 60 s. Storage of all dried samples at either 4.0 ± 3.0°C or 23.0 ± 0.3°C for 15 days was effective for eliminating detectable bacterial counts of both Salmonella spp

Effectiveness of an intervention for improving drug prescription in primary care patients with multimorbidity and polypharmacy:Study protocol of a cluster randomized clinical trial (Multi-PAP project)

This study was funded by the Fondo de Investigaciones Sanitarias ISCIII (Grant Numbers PI15/00276, PI15/00572, PI15/00996), REDISSEC (Project Numbers RD12/0001/0012, RD16/0001/0005), and the European Regional Development Fund ("A way to build Europe").Background: Multimorbidity is associated with negative effects both on people's health and on healthcare systems. A key problem linked to multimorbidity is polypharmacy, which in turn is associated with increased risk of partly preventable adverse effects, including mortality. The Ariadne principles describe a model of care based on a thorough assessment of diseases, treatments (and potential interactions), clinical status, context and preferences of patients with multimorbidity, with the aim of prioritizing and sharing realistic treatment goals that guide an individualized management. The aim of this study is to evaluate the effectiveness of a complex intervention that implements the Ariadne principles in a population of young-old patients with multimorbidity and polypharmacy. The intervention seeks to improve the appropriateness of prescribing in primary care (PC), as measured by the medication appropriateness index (MAI) score at 6 and 12months, as compared with usual care. Methods/Design: Design:pragmatic cluster randomized clinical trial. Unit of randomization: family physician (FP). Unit of analysis: patient. Scope: PC health centres in three autonomous communities: Aragon, Madrid, and Andalusia (Spain). Population: patients aged 65-74years with multimorbidity (≥3 chronic diseases) and polypharmacy (≥5 drugs prescribed in ≥3months). Sample size: n=400 (200 per study arm). Intervention: complex intervention based on the implementation of the Ariadne principles with two components: (1) FP training and (2) FP-patient interview. Outcomes: MAI score, health services use, quality of life (Euroqol 5D-5L), pharmacotherapy and adherence to treatment (Morisky-Green, Haynes-Sackett), and clinical and socio-demographic variables. Statistical analysis: primary outcome is the difference in MAI score between T0 and T1 and corresponding 95% confidence interval. Adjustment for confounding factors will be performed by multilevel analysis. All analyses will be carried out in accordance with the intention-to-treat principle. Discussion: It is essential to provide evidence concerning interventions on PC patients with polypharmacy and multimorbidity, conducted in the context of routine clinical practice, and involving young-old patients with significant potential for preventing negative health outcomes. Trial registration: Clinicaltrials.gov, NCT02866799Publisher PDFPeer reviewe

Role of age and comorbidities in mortality of patients with infective endocarditis

[Purpose]: The aim of this study was to analyse the characteristics of patients with IE in three groups of age and to assess the ability of age and the Charlson Comorbidity Index (CCI) to predict mortality. [Methods]: Prospective cohort study of all patients with IE included in the GAMES Spanish database between 2008 and 2015.Patients were stratified into three age groups:<65 years,65 to 80 years,and ≥ 80 years.The area under the receiver-operating characteristic (AUROC) curve was calculated to quantify the diagnostic accuracy of the CCI to predict mortality risk. [Results]: A total of 3120 patients with IE (1327 < 65 years;1291 65-80 years;502 ≥ 80 years) were enrolled.Fever and heart failure were the most common presentations of IE, with no differences among age groups.Patients ≥80 years who underwent surgery were significantly lower compared with other age groups (14.3%,65 years; 20.5%,65-79 years; 31.3%,≥80 years). In-hospital mortality was lower in the <65-year group (20.3%,<65 years;30.1%,65-79 years;34.7%,≥80 years;p < 0.001) as well as 1-year mortality (3.2%, <65 years; 5.5%, 65-80 years;7.6%,≥80 years; p = 0.003).Independent predictors of mortality were age ≥ 80 years (hazard ratio [HR]:2.78;95% confidence interval [CI]:2.32–3.34), CCI ≥ 3 (HR:1.62; 95% CI:1.39–1.88),and non-performed surgery (HR:1.64;95% CI:11.16–1.58).When the three age groups were compared,the AUROC curve for CCI was significantly larger for patients aged <65 years(p < 0.001) for both in-hospital and 1-year mortality. [Conclusion]: There were no differences in the clinical presentation of IE between the groups. Age ≥ 80 years, high comorbidity (measured by CCI),and non-performance of surgery were independent predictors of mortality in patients with IE.CCI could help to identify those patients with IE and surgical indication who present a lower risk of in-hospital and 1-year mortality after surgery, especially in the <65-year group

Ultraviolet light (UV-C) as a redundant biosafety step for pathogen inactivation in the manufacturing process of spray dried plasma from animal origin

El plasma dessecat per atomització (SDP) és una font de proteïnes funcionals obtinguda a partir de sang d’animals sans, aprovats per les autoritats veterinàries per al consum humà. La sang es recull a l'escorxador, es tracta amb anticoagulant, es refreda i es transporta a instal·lacions industrials on es centrifuga per separar els glòbuls vermells de la fracció plasmàtica. Després, el plasma es concentra i s'asseca a altes temperatures (80ºC) per convertir-lo en pols. Aquest mètode conserva l’activitat biològica de les proteïnes. El SDP s’utilitza principalment en dietes de porcí per millorar significativament el guany diari, la ingesta, l’eficiència productiva i per reduir el retard causat per l’aparició de diarrea després del deslletament. Tot i que el SDP es considera un producte segur i el seu procés de fabricació consta diverses etapes de bioseguretat, la seva seguretat sovint es qüestiona donada la seva naturalesa de subproducte hemoderivat, especialment quan apareixen patògens emergents o reemergents. Aquesta tesi doctoral es va centrar en l'avaluació i la validació d'una nova etapa redundant d'inactivació de patògens per a implementar en el procés de producció de SDP, la irradiació UV-C. El treball ha consistit en avaluar l’efectivitat del tractament per irradiació UV-C mitjançant un dispositiu de flux turbulent, SurePure TurbulatorTM, a l’irradiar un plasma natiu inoculat artificialment amb diferents patògens d’interès per a la indústria porcina. Als estudis 1 i 2 es va avaluar l’efecte UV-C sobre la supervivència de Salmonella typhimurium, S. choleraesuis, Enterococcus faecium i Escherichia coli K88 i K99 sotmesos a diferents dosis UV-C. Es van trobar cinètiques d'inactivació no lineal amb un valor de reducció de 4 log10 (4D), proper a 3000 J/L en tots els casos. A l’estudi 3 es va analitzar l’efecte de l’UV-C sobre diferents virus d’interès en la indústria del porcí. La selecció de virus amb envolta va incloure el Virus de la pseudoràbia (PRV), el Virus de la síndrome respiratòria i reproductiva porcina (PRRSV), el Virus de la diarrea epidèmica porcina (PEDV), el Virus de la diarrea viral bovina, el Virus de la grip porcina (SIV) i el Virus de la pesta porcina clàssica (CSFV); i, com a virus sense envolta, el Parvovirus porcí (PPV), Virus de la malaltia vesicular de porc (SVDV), Circovirus porcí 2 (PCV-2) i Senecavirus A (SVA). Tots aquests virus van ser sotmesos a diferents dosis UV-C i, mitjançant la titulació de les mostres, es va construir la seva corba d'inactivació. En termes generals, els resultats van mostrar que els virus amb envolta tenen una sensibilitat superior a l’UV-C, amb valors 4D inferiors a 2000 J/L, que els que no presenten envolta, presentant valors propers a 3000 J/L o més. A l’estudi 4 es va dur a terme un bioassaig utilitzant diferents grups de garrins, inoculats intraperitonealment amb plasma irradiat per UV-C a 0 J/L (plasma no tractat), 3000 i 9000 J/L. Els resultats van mostrar que cap dels porcs dels grups que rebien el plasma irradiat per UV-C es va infectar ni va seroconvertir en front als virus detectats al plasma inicial (PCV-2, PRRSV (soques europees), SIV, PPV, Virus de l’hepatitis E i Rotavirus A), confirmant així l’eficàcia de l’UV-C. Alhora, es varen dissenyar mètodes convencionals de PCR per amplificar amplicons llargs dels genomes de PCV-2 i PEDV. Mitjançant la comparació dels resultats amb els de PCRs quantitatives, es va demostrar que l’UV-C va ser capaç de danyar el genoma viral. Els resultats globals de la present tesi doctoral mostren que l’aparell d’UV-C SurePure TurbulatorTM és eficaç inactivant un ampli ventall de bacteris i virus presents de forma natural en plasma animal líquid recollit d’escorxadors comercials.El plasma secado por atomización (SDP) es una fuente de proteína funcional obtenida de la sangre de animales sanos, aprobada por las autoridades veterinarias para el consumo humano. La sangre se recoge en el matadero, se trata con anticoagulante, se enfría y se transporta a instalaciones industriales en las que se centrifuga para separar los glóbulos rojos de la fracción plasmática. Luego, el plasma se concentra y se seca por atomización a altas temperaturas (80°C) para convertirlo en polvo. Dicho método conserva la actividad biológica de sus proteínas. El SDP se usa principalmente en dietas porcinas para mejorar significativamente la ganancia diaria, la ingesta, la eficiencia de producción y para reducir el retraso causado por la aparición de diarrea posterior al destete. Aunque el SDP se considera un producto seguro y su proceso de fabricación consta de varias etapas de bioseguridad, su seguridad a menudo se cuestiona debido a su naturaleza de subproducto hemoderivado, especialmente cuando aparecen patógenos emergentes o reemergentes en las poblaciones animales. Esta tesis doctoral se centró en la evaluación y validación de una nueva etapa redundante de inactivación de patógenos a implementar en el proceso de fabricación de SDP, la radiación UV-C. El trabajo ha consistido en evaluar la efectividad del tratamiento de irradiación con UV-C utilizando un dispositivo de flujo turbulento, SurePure TurbulatorTM, al irradiar plasma inoculado artificialmente con diferentes patógenos de interés para la industria porcina. En los estudios 1 y 2, el efecto UV-C sobre la supervivencia bacteriana se evaluó sobre Salmonella typhimurium, S. choleraesuis, Enterococcus faecium y Escherichia coli K88 y K99, sometidas a diferentes dosis de UV-C. Todas las bacterias analizadas mostraron una cinética de inactivación no lineal con un valor de reducción de 4 log10 (4D) cercano a 3000 J/L en todos los casos. En el estudio 3, se analizó el efecto de UV-C en diferentes virus de interés. La selección de virus con envuelta incluyó el Virus de la pseudorabia (PRV), Virus del síndrome respiratorio y reproductivo porcino (PRRSV), Virus de la diarrea epidémica porcina (PEDV), Virus de la diarrea viral bovina (BVDV), Virus de la influenza porcina A (SIV) y el virus de la peste porcina clásica (CSFV). El Parvovirus porcino (PPV), Virus de la enfermedad vesicular porcina, Circovirus porcino 2 (PCV-2) y Senecavirus A (SVA) fueron elegidos como virus sin envuelta. Los virus se sometieron a diferentes dosis de UV-C y, mediante la titulación de las muestras, se construyó su curva de inactivación. Los resultados mostraron que los virus envueltos tienen una mayor sensibilidad al l UV-C, con valores 4D inferiores a 2000 J/L, que los no envueltos, con valores 4D alrededor de 3000 J/L o superiores. En el estudio 4 se realizó un bioensayo utilizando diferentes grupos de lechones inoculados por vía intraperitoneal con plasma irradiado por UV-C a 0 (plasma no tratado), 3000 y 9000J/L. Ninguno de los cerdos en los grupos que recibieron el plasma irradiado por UV-C presentó infección o seroconversión frente a los virus cuyo genoma se detectó en el plasma inicial (PCV-2, PRRSV (cepas europeas), SIV, PPV, Virus de la hepatitis E y Rotavirus A), lo que confirma la eficacia de UV-C. Además, se diseñaron PCR convencionales para amplificar amplicones largos de los genomas de PCV-2 y PEDV. Al comparar los resultados con los obtenidos por PCR cuantitativa en tiempo real, se demostró que la UV-C podía dañar el genoma viral. Los resultados generales de la presente tesis doctoral demuestran que el diseño del turbulador SurePure UV-C fue eficaz para inactivar una amplia gama de bacterias y virus presentes de forma natural en el plasma animal líquido comercial.Spray dried plasma (SDP) is a functional protein source obtained from blood of healthy animals, approved by the veterinary authorities to be fit for slaughter for human consumption. Blood is collected at the slaughterhouse, treated with an anticoagulant, chilled and transported to industrial facilities in which blood is centrifuged to separate the red blood cells from the plasma fraction. Plasma is then concentrated and spray dried at high temperatures (80ºC throughout its substance) to convert it in a powder. Such method preserves the biological activity of its proteins. SDP is mainly used in pig feed diets to significantly improve daily gain, feed intake, production efficiency, and to reduce post-weaning lag caused by the appearance of post-weaning diarrhea. Although SDP is considered a safe product and its manufacturing process consists of several biosafety steps, the security of the SDP is often questioned due to its nature as raw blood by-product, especially when emergent or re-emergent pathogens appear in animal populations. This PhD Thesis focused on the evaluation and validation of a new redundant pathogen inactivation step that may be implemented in the manufacturing process of SDP, the UV-C irradiation. The work has consisted in evaluating the effectiveness of the UV-C irradiation treatment using a turbulent flow device, SurePure TurbulatorTM, when irradiating raw plasma artificially inoculated with different pathogens of interest for the swine industry. In studies 1 and 2 the UV-C effect on bacterial survival was assessed on Salmonella typhimurium, S. choleraesuis, Enterococcus faecium, and Escherichia coli K88 and K99 strains subjected to different UV-C doses. All tested bacteria showed non-linear inactivation kinetics with 4 log10 (4D) reduction value in all cases close to 3000 J/L. In study 3, the effect of UV-C on different viruses of interest in the swine industry was analyzed. The selection of enveloped viruses included Pseudorabies virus, Porcine reproductive and respiratory syndrome virus (PRRSV), Porcine epidemic diarrhea virus (PEDV), Bovine viral diarrhea virus, Swine inflenza A virus (SIV) and Classical swine fever virus. On the other hand, Porcine parvovirus (PPV), Swine vesicular disease virus, Porcine circovirus 2 (PCV-2) and Senecavirus A (SVA) were chosen as non-enveloped viruses. All these viruses were subjected to different UV-C doses and, by titration of the samples at each UV-C dose, the inactivation curve for each virus was constructed. In general terms, results showed that enveloped viruses have a higher sensitivity to UV-C than non-enveloped ones, being the 4D values less than 2000 J/L for enveloped viruses and around 3000 J/L or higher for non-enveloped ones. To validate the effectiveness of the plasma UV-C irradiation measured in previous studies, a bioassay was carried out in the study 4 using different groups of piglets inoculated intraperitoneally with UV-C irradiated plasma at 0 (untreated plasma), 3000, and 9000J/L. The results showed that none of the pigs in the groups that received the plasma irradiated by UV-C were infected or seroconverted against the viruses which genome was detected in the initial plasma (PCV-2, PRRSV (European strains), SIV, PPV, Hepatitis E virus and Rotavirus A), thus confirming the efficacy of UV-C. Furthermore, in the study 4, conventional PCR methods able to generate long amplicons were designed to amplify fragments of approximately 1.7 kb of PCV-2 and PEDV genomes. By comparison of the results with those of real time quantitative PCRs to detect the same viruses (using short amplicons), it was demonstrated that UV-C was able to damage the viral genome. Overall results of the present PhD Thesis showed that the SurePure UV-C Turbulator design was effective in inactivating a wide range of bacteria and viruses spiked and naturally present in commercially collected liquid animal plasma

Ultraviolet light (UV-C) as a redundant biosafety step for pathogen inactivation in the manufacturing process of spray dried plasma from animal origin

El plasma dessecat per atomització (SDP) és una font de proteïnes funcionals obtinguda a partir de sang d’animals sans, aprovats per les autoritats veterinàries per al consum humà. La sang es recull a l'escorxador, es tracta amb anticoagulant, es refreda i es transporta a instal·lacions industrials on es centrifuga per separar els glòbuls vermells de la fracció plasmàtica. Després, el plasma es concentra i s'asseca a altes temperatures (80ºC) per convertir-lo en pols. Aquest mètode conserva l’activitat biològica de les proteïnes. El SDP s’utilitza principalment en dietes de porcí per millorar significativament el guany diari, la ingesta, l’eficiència productiva i per reduir el retard causat per l’aparició de diarrea després del deslletament. Tot i que el SDP es considera un producte segur i el seu procés de fabricació consta diverses etapes de bioseguretat, la seva seguretat sovint es qüestiona donada la seva naturalesa de subproducte hemoderivat, especialment quan apareixen patògens emergents o reemergents. Aquesta tesi doctoral es va centrar en l'avaluació i la validació d'una nova etapa redundant d'inactivació de patògens per a implementar en el procés de producció de SDP, la irradiació UV-C. El treball ha consistit en avaluar l’efectivitat del tractament per irradiació UV-C mitjançant un dispositiu de flux turbulent, SurePure TurbulatorTM, a l’irradiar un plasma natiu inoculat artificialment amb diferents patògens d’interès per a la indústria porcina. Als estudis 1 i 2 es va avaluar l’efecte UV-C sobre la supervivència de Salmonella typhimurium, S. choleraesuis, Enterococcus faecium i Escherichia coli K88 i K99 sotmesos a diferents dosis UV-C. Es van trobar cinètiques d'inactivació no lineal amb un valor de reducció de 4 log10 (4D), proper a 3000 J/L en tots els casos. A l’estudi 3 es va analitzar l’efecte de l’UV-C sobre diferents virus d’interès en la indústria del porcí. La selecció de virus amb envolta va incloure el Virus de la pseudoràbia (PRV), el Virus de la síndrome respiratòria i reproductiva porcina (PRRSV), el Virus de la diarrea epidèmica porcina (PEDV), el Virus de la diarrea viral bovina, el Virus de la grip porcina (SIV) i el Virus de la pesta porcina clàssica (CSFV); i, com a virus sense envolta, el Parvovirus porcí (PPV), Virus de la malaltia vesicular de porc (SVDV), Circovirus porcí 2 (PCV-2) i Senecavirus A (SVA). Tots aquests virus van ser sotmesos a diferents dosis UV-C i, mitjançant la titulació de les mostres, es va construir la seva corba d'inactivació. En termes generals, els resultats van mostrar que els virus amb envolta tenen una sensibilitat superior a l’UV-C, amb valors 4D inferiors a 2000 J/L, que els que no presenten envolta, presentant valors propers a 3000 J/L o més. A l’estudi 4 es va dur a terme un bioassaig utilitzant diferents grups de garrins, inoculats intraperitonealment amb plasma irradiat per UV-C a 0 J/L (plasma no tractat), 3000 i 9000 J/L. Els resultats van mostrar que cap dels porcs dels grups que rebien el plasma irradiat per UV-C es va infectar ni va seroconvertir en front als virus detectats al plasma inicial (PCV-2, PRRSV (soques europees), SIV, PPV, Virus de l’hepatitis E i Rotavirus A), confirmant així l’eficàcia de l’UV-C. Alhora, es varen dissenyar mètodes convencionals de PCR per amplificar amplicons llargs dels genomes de PCV-2 i PEDV. Mitjançant la comparació dels resultats amb els de PCRs quantitatives, es va demostrar que l’UV-C va ser capaç de danyar el genoma viral. Els resultats globals de la present tesi doctoral mostren que l’aparell d’UV-C SurePure TurbulatorTM és eficaç inactivant un ampli ventall de bacteris i virus presents de forma natural en plasma animal líquid recollit d’escorxadors comercials.El plasma secado por atomización (SDP) es una fuente de proteína funcional obtenida de la sangre de animales sanos, aprobada por las autoridades veterinarias para el consumo humano. La sangre se recoge en el matadero, se trata con anticoagulante, se enfría y se transporta a instalaciones industriales en las que se centrifuga para separar los glóbulos rojos de la fracción plasmática. Luego, el plasma se concentra y se seca por atomización a altas temperaturas (80°C) para convertirlo en polvo. Dicho método conserva la actividad biológica de sus proteínas. El SDP se usa principalmente en dietas porcinas para mejorar significativamente la ganancia diaria, la ingesta, la eficiencia de producción y para reducir el retraso causado por la aparición de diarrea posterior al destete. Aunque el SDP se considera un producto seguro y su proceso de fabricación consta de varias etapas de bioseguridad, su seguridad a menudo se cuestiona debido a su naturaleza de subproducto hemoderivado, especialmente cuando aparecen patógenos emergentes o reemergentes en las poblaciones animales. Esta tesis doctoral se centró en la evaluación y validación de una nueva etapa redundante de inactivación de patógenos a implementar en el proceso de fabricación de SDP, la radiación UV-C. El trabajo ha consistido en evaluar la efectividad del tratamiento de irradiación con UV-C utilizando un dispositivo de flujo turbulento, SurePure TurbulatorTM, al irradiar plasma inoculado artificialmente con diferentes patógenos de interés para la industria porcina. En los estudios 1 y 2, el efecto UV-C sobre la supervivencia bacteriana se evaluó sobre Salmonella typhimurium, S. choleraesuis, Enterococcus faecium y Escherichia coli K88 y K99, sometidas a diferentes dosis de UV-C. Todas las bacterias analizadas mostraron una cinética de inactivación no lineal con un valor de reducción de 4 log10 (4D) cercano a 3000 J/L en todos los casos. En el estudio 3, se analizó el efecto de UV-C en diferentes virus de interés. La selección de virus con envuelta incluyó el Virus de la pseudorabia (PRV), Virus del síndrome respiratorio y reproductivo porcino (PRRSV), Virus de la diarrea epidémica porcina (PEDV), Virus de la diarrea viral bovina (BVDV), Virus de la influenza porcina A (SIV) y el virus de la peste porcina clásica (CSFV). El Parvovirus porcino (PPV), Virus de la enfermedad vesicular porcina, Circovirus porcino 2 (PCV-2) y Senecavirus A (SVA) fueron elegidos como virus sin envuelta. Los virus se sometieron a diferentes dosis de UV-C y, mediante la titulación de las muestras, se construyó su curva de inactivación. Los resultados mostraron que los virus envueltos tienen una mayor sensibilidad al l UV-C, con valores 4D inferiores a 2000 J/L, que los no envueltos, con valores 4D alrededor de 3000 J/L o superiores. En el estudio 4 se realizó un bioensayo utilizando diferentes grupos de lechones inoculados por vía intraperitoneal con plasma irradiado por UV-C a 0 (plasma no tratado), 3000 y 9000J/L. Ninguno de los cerdos en los grupos que recibieron el plasma irradiado por UV-C presentó infección o seroconversión frente a los virus cuyo genoma se detectó en el plasma inicial (PCV-2, PRRSV (cepas europeas), SIV, PPV, Virus de la hepatitis E y Rotavirus A), lo que confirma la eficacia de UV-C. Además, se diseñaron PCR convencionales para amplificar amplicones largos de los genomas de PCV-2 y PEDV. Al comparar los resultados con los obtenidos por PCR cuantitativa en tiempo real, se demostró que la UV-C podía dañar el genoma viral. Los resultados generales de la presente tesis doctoral demuestran que el diseño del turbulador SurePure UV-C fue eficaz para inactivar una amplia gama de bacterias y virus presentes de forma natural en el plasma animal líquido comercial.Spray dried plasma (SDP) is a functional protein source obtained from blood of healthy animals, approved by the veterinary authorities to be fit for slaughter for human consumption. Blood is collected at the slaughterhouse, treated with an anticoagulant, chilled and transported to industrial facilities in which blood is centrifuged to separate the red blood cells from the plasma fraction. Plasma is then concentrated and spray dried at high temperatures (80ºC throughout its substance) to convert it in a powder. Such method preserves the biological activity of its proteins. SDP is mainly used in pig feed diets to significantly improve daily gain, feed intake, production efficiency, and to reduce post-weaning lag caused by the appearance of post-weaning diarrhea. Although SDP is considered a safe product and its manufacturing process consists of several biosafety steps, the security of the SDP is often questioned due to its nature as raw blood by-product, especially when emergent or re-emergent pathogens appear in animal populations. This PhD Thesis focused on the evaluation and validation of a new redundant pathogen inactivation step that may be implemented in the manufacturing process of SDP, the UV-C irradiation. The work has consisted in evaluating the effectiveness of the UV-C irradiation treatment using a turbulent flow device, SurePure TurbulatorTM, when irradiating raw plasma artificially inoculated with different pathogens of interest for the swine industry. In studies 1 and 2 the UV-C effect on bacterial survival was assessed on Salmonella typhimurium, S. choleraesuis, Enterococcus faecium, and Escherichia coli K88 and K99 strains subjected to different UV-C doses. All tested bacteria showed non-linear inactivation kinetics with 4 log10 (4D) reduction value in all cases close to 3000 J/L. In study 3, the effect of UV-C on different viruses of interest in the swine industry was analyzed. The selection of enveloped viruses included Pseudorabies virus, Porcine reproductive and respiratory syndrome virus (PRRSV), Porcine epidemic diarrhea virus (PEDV), Bovine viral diarrhea virus, Swine inflenza A virus (SIV) and Classical swine fever virus. On the other hand, Porcine parvovirus (PPV), Swine vesicular disease virus, Porcine circovirus 2 (PCV-2) and Senecavirus A (SVA) were chosen as non-enveloped viruses. All these viruses were subjected to different UV-C doses and, by titration of the samples at each UV-C dose, the inactivation curve for each virus was constructed. In general terms, results showed that enveloped viruses have a higher sensitivity to UV-C than non-enveloped ones, being the 4D values less than 2000 J/L for enveloped viruses and around 3000 J/L or higher for non-enveloped ones. To validate the effectiveness of the plasma UV-C irradiation measured in previous studies, a bioassay was carried out in the study 4 using different groups of piglets inoculated intraperitoneally with UV-C irradiated plasma at 0 (untreated plasma), 3000, and 9000J/L. The results showed that none of the pigs in the groups that received the plasma irradiated by UV-C were infected or seroconverted against the viruses which genome was detected in the initial plasma (PCV-2, PRRSV (European strains), SIV, PPV, Hepatitis E virus and Rotavirus A), thus confirming the efficacy of UV-C. Furthermore, in the study 4, conventional PCR methods able to generate long amplicons were designed to amplify fragments of approximately 1.7 kb of PCV-2 and PEDV genomes. By comparison of the results with those of real time quantitative PCRs to detect the same viruses (using short amplicons), it was demonstrated that UV-C was able to damage the viral genome. Overall results of the present PhD Thesis showed that the SurePure UV-C Turbulator design was effective in inactivating a wide range of bacteria and viruses spiked and naturally present in commercially collected liquid animal plasma