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

Spray 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.info:eu-repo/semantics/publishedVersio

Evaluation of potential biomarkers to determine adequate colostrum provision in male dairy-beef calves upon arrival at the rearing facility beyond 14 days of age

Colostrum consumption is crucial for passive immunization and development of the newborn calf. However, the incidence on failed transfer of passive immunity in male calves destined to dairy-beef production remains high to date. In addition, the lack of an automated procedure to validate the immunization status upon arrival at rearing facilities in calves beyond 14 d of age impedes the identification of failed transfer of passive immunity, and therefore, of those calves at high risk of suffering diseases. For this study, 82 newborn male Holstein calves (43.3 ± 0.86 kg of body weight; mean ± standard error) from a commercial dairy farm were used to investigate potential serum biomarkers of colostrum provision. The potential biomarkers selected were IgG, IgG1, cholesterol, alkaline phosphatase, gamma-glutamyl transferase (GGT), and total protein (TP). Treatments were as follows: high-colostrum (HC; n = 49), in which calves received 4 L of colostrum within the first 2 h after birth and 2 L of colostrum in the next 3 feedings within the first 24 h after birth, for a total of 10 L of colostrum; and low-colostrum (LC; n = 33), in which calves received only 2 L of colostrum within the first 2 h after birth. After colostrum consumption, calves were allocated to individual hutches and fed 2 L of milk replacer twice daily at a concentration of 125 g/L as fed. Starter feed and water were offered ad libitum. At approximately 14 d of age (14.2 ± 0.81 d of age; mean ± standard error) calves were transported 2.5 h to a research unit at IRTA (Torre Marimon, Spain) simulating the arrival to a rearing facility. Blood samples were collected before feeding at birth, 48 h after birth, and at arrival to the rearing facility. Results on the serum concentrations of the potential biomarkers at arrival to the rearing facility showed that IgG, IgG1, GGT, and TP were greater for the HC calves compared with the LC calves. Serum concentrations of cholesterol and alkaline phosphatase did not show differences between treatment groups. Additionally, body weight losses from birth until arrival to the rearing facility were greater for the LC treatment compared with the HC. Because of their low cost, quickness, and ease of measurement, GGT and TP were good indicators of colostrum intake in calves arriving at rearing facilities beyond 14 d of age.info:eu-repo/semantics/publishedVersio

Feeding Spray-Dried Porcine Plasma to Pigs Reduces African Swine Fever Virus Load in Infected Pigs and Delays Virus Transmission—Study 1

The objective of this study was to evaluate the potential benefits of feeding spray-dried porcine plasma (SDPP) to pigs infected with African swine fever virus (ASFV). Two groups of twelve weaned pigs each were fed with CONVENTIONAL or 8% SDPP enriched diets. Two pigs (trojans)/group) were injected intramuscularly with the pandemic ASFV (Georgia 2007/01) and comingled with the rest of the pigs (1:5 trojan:naïve ratio) to simulate a natural route of transmission. Trojans developed ASF and died within the first week after inoculation, but contact pigs did not develop ASF, viremia, or seroconversion. Therefore, three more trojans per group were introduced to optimize the ASFV transmission (1:2 trojan:naïve ratio). Blood, nasal, and rectal swabs were weekly harvested, and at end of the study ASFV-target organs collected. After the second exposure, rectal temperature of conventionally fed contact pigs increased >40.5 °C while fever was delayed in the SDPP contact pigs. Additionally, PCR Ct values in blood, secretions, and tissue samples were significantly lower (p < 0.05) for CONVENTIONAL compared to SDPP contact pigs. Under these study conditions, contact exposed pigs fed SDPP had delayed ASFV transmission and reduced virus load, likely by enhanced specific T-cell priming after the first ASFV-exposure.Funding for this study was provided by APC Europe, S.L.U., Granollers, Spain, and APC LLC, Ankeny, US. Both companies manufacture animal blood products for animal consumption. The companies provided support in the form of salaries for authors EB, JC, and J. Polo retrospectively, but none of these companies did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. We also acknowledge support of the Spanish Ministry of Science and Innovation (grant reference PID2019-107616RB-I00).info:eu-repo/semantics/publishedVersio

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

Commercial feed containing porcine plasma spiked with African swine fever virus is not infective in pigs when administered for 14 consecutive days

The objective of this study was to determine if commercially collected liquid porcine plasma contaminated with African swine fever virus (ASFV) and fed for 14 consecutive days would infect pigs. Commercially collected liquid porcine plasma was mixed with the serum from an ASFV experimentally infected pig. To simulate the potential of pigs slaughtered being ASFV viremic but asymptomatic and passing antemortem inspection, the ratio of liquid plasma from healthy animals to serum from an ASFV infected pig used in this study represented 0.4% or 2.0% of the pigs slaughtered being viremic (Studies 1 or 2, respectively). The contaminated liquid plasma was mixed on commercial feed and pigs were fed for 14 consecutive days providing to each pig 104.3 or 105.0 TCID50 ASFV daily (Studies 1 or 2, respectively). Pigs were observed for an additional 5 or 9 days (Studies 1 or 2, respectively). In both experiments, the pigs did not become infected with ASFV during the 14d feeding period or during the subsequent observation period. In these experiments, unprocessed liquid plasma contaminated with ASFV mixed on commercial feed and fed for 14 consecutive days did not infect pigs. From our results we can conclude that the infectious dose of ASFV on feed is much higher than that previously reported, at least with ASFV-spiked raw plasma.info:eu-repo/semantics/publishedVersio

Neutralizing antibodies against porcine circovirus type 2 in liquid pooled plasma contribute to the biosafety of commercially manufactured spray-dried porcine plasma

Neutralizing antibodies (NA) inherently present in pooled plasma collected at commercial abattoirs may provide some protection against potential porcine circovirus type 2 (PCV2) infectivity of plasma. Moreover, these NA may also contribute to the biosafety of spray-dried porcine plasma (SDPP). The objective of the study was to characterize and quantify the PCV2 antibody neutralizing capacity in pooled liquid porcine plasma and SDPP samples collected from industrial spray-drying facilities located in the Southeast and Midwest regions of the United States and the Northeast region of Spain. In the United States, PCV2 NA was determined in 1 sample of pooled liquid plasma from commercial spray-drying plants in the Southeast and 1 from the Midwest region. Obtained results were compared with those of a plasma sample from a PCV2 vaccinated sow and 1 from a PCV2 antibody negative sow. In Spain, 15 pooled liquid porcine plasma samples and 10 SDPP samples were collected at a commercial spray-drying plant total and NA against PCV2 were determined. Results with pooled liquid porcine plasma from commercial spray-drying facilities in the United States indicated that NA titers against PCV2 in these samples (log2 8.33 ± 0.41 and 9.0 ± 0.0) were similar or greater than the plasma from the PCV2-vaccinated sow (log2 6.33 ± 0.41). The analysis of U.S. samples indicated that liquid plasma diluted to 1:256 (10–2.41) was able to neutralize between 100 to 200 PCV2 virus particles or about 4 logs10 median tissue culture infective dose (TCID50) per milliliter. Similarly, samples from the Spanish pooled liquid plasma and the SDPP samples indicated an increased amount of NA activity against PCV2. Specifically, a dilution of 10–2.47 ± 0.33 of plasma was able to inactivate 100 PCV2 virus particles; therefore, the inactivation capacity of commercial liquid plasma was greater than 104 TCID50/mL. The calculated 90% reduction in infected cells because of NA in pooled plasma samples (log2 8.2 ± 0.38) was less (P < 0.05) than in its concentrate form of SDPP (mean, log2 10.2 ± 0.85). In conclusion, PCV2 NA contained in liquid pooled plasma from market pigs was detected at greater concentrations than that from a vaccinated sow and that after spray-drying biological neutralizing activity was conserved, which implies that the inherent NA in liquid plasma may have an important role in the biosafety of commercially produced SDPP.info:eu-repo/semantics/publishedVersio

Epidemiological surveillance of Schmallenberg virus in small ruminants in southern Spain

Schmallenberg virus (SBV) is an emerging Culicoides‐borne Orthobunyavirus that affects ruminant species. Between 2011 and 2013, it was responsible for a large‐scale epidemic in Europe. In the present study, we aimed to determine the seroprevalence, spatial distribution and risk factors associated with SBV exposure in sheep and goats in the region where the first Schmallenberg disease outbreak in Spain was reported. Blood samples from 1,796 small ruminants from 120 farms were collected in Andalusia (southern Spain) between 2015 and 2017. Antibodies against SBV were detected in 536 of 1,796 animals (29.8%; 95%CI: 27.7–32.0) using a commercial blocking ELISA. The individual seroprevalence according to species was 31.1% (280/900; 95%CI: 28.1–34.1) in sheep and 28.6% (256/896; 95%CI: 25.6–31.5) in goats. The farm prevalence was 76.7% (95%CI: 69.1–84.2). Seropositivity to SBV was confirmed in both sheep and goats in all provinces by virus neutralization test. Two significant (p 14ºC) and altitude (<400 metres above sea level) were risk factors associated with SBV exposure in small ruminants. Our results highlight widespread but not homogeneous circulation of SBV in small ruminant populations in Spain.info:eu-repo/semantics/acceptedVersio

On the activity of the γ -Ursae Minorids meteoroid stream in 2010 and 2011

Accurate orbital data obtained for the recently discovered γ -Ursae Minorids meteoroid stream during the 2010 and 2011 Spanish Meteor Network and Finnish Fireball Network observing ampaigns are presented. In particular, we focus on an outburst detected in 2010 and on the analysis of the first emission spectrum recorded for a member of this meteoroid stream. An array of high-sensitivity CCD video devices operating from different locations in Spain and Finland was used to perform this study. We have obtained precise trajectory, radiant and orbital information for seven members of this stream. Considerations about its likely parent body based on orbital dissimilarity criteria are made. We also present an estimation of the tensile strength for these meteoroids and a unique emission spectrum of a γ -Ursae inorid fireball that reveals that the main rocky components have chondritic abundances.Ministerio de Ciencia e Innovación AYA2009-13227, AYA2009-14000-C03-01, AYA2011-26522, AYA2011-30106-C02-01CSIC #201050I043Junta de Andalucía P09-FQM-455

Long-Term Dynamics of Bluetongue Virus in Wild Ruminants : relationship with Outbreaks in Livestock in Spain, 2006-2011

Wild and domestic ruminants are susceptible to Bluetongue virus (BTV) infection. Three BTV serotypes (BTV-4, BTV-1 and BTV-8) have been detected in Spain in the last decade. Even though control strategies have been applied to livestock, BTV circulation has been frequently detected in wild ruminant populations in Spain. The aim of the present study is to assess the role for wild ruminants in maintaining BTV after the vaccination programs in livestock in mainland Spain. A total of 931 out 1,914 (48.6%) serum samples, collected from eight different wild ruminant species between 2006 and 2011, were BTV positive by ELISA. In order to detect specific antibodies against BTV-1, BTV-4 and BTV-8, positive sera were also tested by serumneutralisation test (SNT). From the ELISA positive samples that could be tested by SNT (687 out of 931), 292 (42.5%) showed neutralising antibodies against one or two BTV serotypes. For each BTV seroptype, the number of outbreaks in livestock (11,857 outbreaks in total) was modelled with pure autoregressive models and the resulting smoothed values, representing the predicted number of BTV outbreaks in livestock at municipality level, were positively correlated with BTV persistence in wild species. The strength of this relationship significantly decreased as red deer (Cervus elaphus) population abundance increased. In addition, BTV RNA was detected by real time RT-PCR in 32 out of 311 (10.3%) spleen samples from seropositive animals. Although BT outbreaks in livestock have decreased substantially after vaccination campaigns, our results indicated that wild ruminants have been exposed to BTV in territories where outbreaks in domestic animals occurred. The detection of BTV RNA and spatial association between BT outbreaks in livestock and BTV rates in red deer are consistent with the hypothesis of virus circulation and BTV maintenance within Iberian wild ruminant populations