Characterization of contemporary 2010.1 H3N2 swine influenza A viruses circulating in United States pigs
Not Available
Powell, Joshua D. (ORCID:0000000308693481)↗
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Quantitative cardiac phosphoproteomics profiling during ischemia-reperfusion in an immature swine model
Ischemia-reperfusion (I/R) results in altered metabolic and molecular responses, and phosphorylation is one of the most noted regulatory mechanisms mediating signaling mechanisms during physiological stresses. To expand our knowledge of the potential phosphoproteomic changes in the myocardium during I/R, we used Isobaric Tags for Relative and Absolute Quantitation-based analyses in left ventricular samples obtained from porcine hearts under control or I/R conditions. The data are available via ProteomeXchange with identifier PXD006066. We identified 1,896 phosphopeptides within left ventricular control and I/R porcine samples. Significant differential phosphorylation between control and I/R groups was discovered in 111 phosphopeptides from 86 proteins. Analysis of the phosphopeptides using Motif-x identified five motifs: (..R..S..), (..SP..), (..S.S..), (..S…S..), and (..S.T..). Semiquantitative immunoblots confirmed site location and directional changes in phosphorylation for phospholamban and pyruvate dehydrogenase E1, two proteins known to be altered by I/R and identified by this study. Novel phosphorylation sites associated with I/R were also identified. Functional characterization of the phosphopeptides identified by our methodology could expand our understanding of the signaling mechanisms involved during I/R damage in the heart as well as identify new areas to target therapeutic strategies.
60 APPLIED LIFE SCIENCES↗
ISU FLUture: a veterinary diagnostic laboratory web-based platform to monitor the temporal genetic patterns of Influenza A virus in swine
Not Available
60 APPLIED LIFE SCIENCES↗
Integrated transcriptomic and metabolomic investigation of the genes and metabolites involved in swine follicular cyst formation
Follicular cysts are a common reproductive disorder in mammals that is usually caused by stress. However, the pathogenesis of follicular cysts in sows remains unclear. To provide new insights into the mechanisms of follicular cyst formation in pigs, we conducted a combined transcriptomic and metabolomic analysis on theca interna and mural granulosa cells of follicular cysts and mature follicles. We identified 2,533 up-regulated and 1,355 down-regulated genes in follicular cysts, compared with mature follicles. These differentially expressed genes were mainly found in signaling pathways related to tumor formation and cortisol synthesis and secretion as shown by Ingenuity Pathway Analysis, which predicted 4,362 upstream regulatory factors. The combined gene expression and pathway analysis identified the following genes as potential biomarkers for porcine follicular cysts: cytochrome P450 family 2 subfamily C polypeptide 18, L-lactate dehydrogenase, carbamoyl-phosphate synthase, fibroblast growth factor 7, integrin binding sialoprotein, interleukin 23 receptor, prolactin receptor, epiregulin, interleukin 1 receptor type II, arginine vasopressin receptor 1A, fibroblast growth factor 10, claudin 7, G Protein Subunit Gamma 3, cholecystokinin B receptor and cytosolic phospholipase A2 . Metabolomics analysis found significant differences in 87 metabolites, which were enriched in unsaturated fatty acid biosynthesis, and sphingolipid signaling pathways. These results provide valuable information on the molecular mechanisms of follicular cyst formation, which may facilitate the development of new therapeutics to prevent and treat follicular cysts.
Dai, Jiage↗
The Impact of Surficial Biochar Treatment on Acute H2S Emissions during Swine Manure Agitation before Pump-Out: Proof-of-the-Concept
Acute releases of hydrogen sulfide (H2S) are of serious concern in agriculture, especially when farmers agitate manure to empty storage pits before land application. Agitation can cause the release of dangerously high H2S concentrations, resulting in human and animal fatalities. To date, there is no proven technology to mitigate these short-term releases of toxic gas from manure. In our previous research, we have shown that biochar, a highly porous carbonaceous material, can float on manure and mitigate gaseous emissions over extended periods (days–weeks). In this research, we aim to test the hypothesis that biochar can mitigate H2S emissions over short periods (minutes–hours) during and shortly after manure agitation. The objective was to conduct proof-of-the-concept experiments simulating the treatment of agitated manure. Two biochars, highly alkaline and porous (HAP, pH 9.2) made from corn stover and red oak (RO, pH 7.5), were tested. Three scenarios (setups): Control (no biochar), 6 mm, and 12 mm thick layers of biochar were surficially-applied to the manure. Each setup experienced 3 min of manure agitation. Real-time concentrations of H2S were measured immediately before, during, and after agitation until the concentration returned to the initial state. The results were compared with those of the Control using the following three metrics: (1) the maximum (peak) flux, (2) total emission from the start of agitation until the concentration stabilized, and (3) the total emission during the 3 min of agitation. The Gompertz’s model for determination of the cumulative H2S emission kinetics was developed. Here, 12 mm HAP biochar treatment reduced the peak (1) by 42.5% (p = 0.125), reduced overall total emission (2) by 17.9% (p = 0.290), and significantly reduced the total emission during 3 min agitation (3) by 70.4%. Further, 6 mm HAP treatment reduced the peak (1) by 60.6%, and significantly reduced overall (2) and 3 min agitation’s (3) total emission by 64.4% and 66.6%, respectively. Moreover, 12 mm RO biochar treatment reduced the peak (1) by 23.6%, and significantly reduced overall (2) and 3 min total (3) emission by 39.3% and 62.4%, respectively. Finally, 6 mm RO treatment significantly reduced the peak (1) by 63%, overall total emission (2) by 84.7%, and total emission during 3 min agitation (3) by 67.4%. Biochar treatments have the potential to reduce the risk of inhalation exposure to H2S. Both 6 and 12 mm biochar treatments reduced the peak H2S concentrations below the General Industrial Peak Limit (OSHA PEL, 50 ppm). The 6 mm biochar treatments reduced the H2S concentrations below the General Industry Ceiling Limit (OSHA PEL, 20 ppm). Research scaling up to larger manure volumes and longer agitation is warranted.
Chen, Baitong (ORCID:0000000266071253)↗
Evaluation of Cross-Protection of African Swine Fever Vaccine ASFV-G-ΔI177L Between ASFV Biotypes
Background/Objectives: Vaccine development for the prevention of ASF has been very challenging due to the extensive genetic and largely unknown antigenic diversity. Inactivated vaccines, using different inactivation methods and a variety of adjuvants, have been consistently inefficacious. Historically, animals recovering from an infection with an attenuated virus became protected from the development of a clinical disease caused by an antigenically related strain. Therefore, immunization of susceptible animals with attenuathe ted virus strains has become a common method of vaccination with the first two commercially available vaccines based on recombinant live-attenuated viruses (LAVs). An important limitation is that the efficacy of the LAV is restricted to those strains that are antigenically related and, in most cases, only provide protection against homologous strains. Due to the unknown antigenic heterogeneity among all ASFV field isolates, the development of broad-spectrum vaccines is a challenge. Besides the anecdotal data, there is not a large amount of information describing patterns of cross-protection between different ASFV strains. Methods: We evaluated the cross-protection induced by the ASFV live-attenuated vaccine ASFV-G-ΔI177L against different biotypes of ASFV and compared their genomic sequences to determine potential genetic mutations that could cause the lack of cross-protection. Results: Results presented here demonstrate different patterns of protection when ASFV-G-ΔI177L vaccinated pigs were challenged with six different ASFV field isolates belonging to different biotypes. Conclusions: The presence of cross-protection cannot be predicted solely by the classical methodology for genotyping-based B646L ORF only. Biotyping, considering the entire virus proteome, appears to be a more promising prediction tool, although additional gathering of experimental data will be necessary to fully validate it; until then, the presence of cross-protection needs to be confirmed in efficacy trials challenging vaccinated animals.
Immunology↗
Life cycle analysis of polylactic acids from different wet waste feedstocks
Producing a valuable chemical product through diversion of wet wastes can simultaneously resolve the problems associated with increasing wastes and greenhouse gas emissions from conventional chemical production processes. In this work, we investigated the life-cycle greenhouse gas emissions, water, and fossil-fuel consumption for waste-derived polylactic acids (PLA) from three different waste feedstocks, namely wastewater sludge, food waste, and swine manure, using the Greenhouse Gases, Regulated Emissions, and Energy Use in Technologies (GREET) model. The decarbonization potential of replacing fossil-based resins with the waste-derived polymer was also investigated. The results show that swine manure-to-PLA pathway was the least carbon intensive (—1.4 kgCO 2 e/kg) among the three waste-to-PLA pathways on a cradle-to-grave basis, followed by the food waste case (—1.3 kgCO 2 e/kg) and then by the wastewater sludge case (0.6 kgCO 2 e/kg). In the baseline scenario, all three waste-to-PLA pathways were less carbon intensive than both fossil-based PET and HDPE on a cradle-to-grave basis: 66% (vs. PET) and 56% (vs. HDPE), 171 and 192%, 181 and 205% reduction in GHG emissions for wastewater sludge-, food waste-, and swine manure-to-PLA pathway, respectively. For all sensitivity cases investigated, the food waste- and swine manure-to-PLA pathways were significantly less carbon intensive than their fossil-counterparts. In terms of the annual decarbonization potential of replacing fossil-based PET or HDPE, the wastewater sludge- and food waste-pathway showed higher mitigation potential than the swine manure-pathway: i) 18–28 kilotons CO 2 e-reduction per year for wastewater sludge pathway; ii) 23–26 kTCO 2 e-reduction/yr for food waste pathway; and iii) about 5 kTCO 2 e-reduction/yr for swine manure pathway depending on the type of conventional resin replaced. However, given the abundant availability of the swine manure feedstocks across the United States, the decarbonization potential of swine manure-based pathway can also increase as the plant capacity or the number of plants grow.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
Techno-economic Analysis and Life-Cycle Analysis of Renewable Diesel Fuels Produced with Waste Feedstocks
Wet waste feedstocks represent an important category of resources that could be utilized to produce biofuels. Diversion of the wet waste resources from going through conventional waste management practices to utilization as feedstocks for energy production also benefits from avoided cost and pollutant emissions of waste management and disposal. This study investigates the economic and environmental implications of producing bio-blendstocks for mixing controlled compression ignition (MCCI) engines from two waste-to-fuel pathways: hydroprocessed esters and fatty acids (HEFA) from yellow grease, and swine manure hydrothermal liquefaction (HTL) followed by biocrude upgrading. Detailed process models were developed for both pathways, which informed the techno-economic analysis (TEA) and life-cycle analysis (LCA). Conventional swine manure management practice was also modeled in detail as the business-as-usual (BAU) scenario for the swine manure HTL pathway. The estimated minimum fuel selling prices (MFSP) were $4.47/gasoline gallon equivalent (GGE) and $3.57/GGE for the yellow grease to HEFA and swine manure HTL pathways, respectively. The life-cycle greenhouse gas (GHG) emissions of the two pathways were 11.2 and -33.3 g CO2e/MJ, respectively, for the yellow grease to HEFA and swine manure HTL pathways. The credits of avoided emissions from conventional swine manure management were the main reason for the negative GHG emissions of the swine manure HTL pathway. The marginal GHG emissions abatement costs were estimated to be $100 – $253/tonne CO2e and $5 – $103/tonne CO2e for the yellow grease HEFA and swine manure HTL pathways, respectively, for a diesel price ranging between $2.0/GGE and $3.5/GGE. Since the yellow grease HEFA pathway is already commercialized, it can benefit from the $200/tonne carbon credit in the California Low Carbon Fuel Standard (LCFS) market, which could help the yellow grease HEFA pathway to achieve near-zero marginal GHG emissions abatement cost.
renewable diesel, techno economic analysis, life c↗
Bivalent hemagglutinin and neuraminidase influenza replicon particle vaccines protect pigs against influenza a virus without causing vaccine associated enhanced respiratory disease
Alphavirus-derived RNA replicon particle (RP) vaccines represent the next generation of swine influenza A virus (IAV) vaccines, as they were shown to be safe, effective, and offer advantages over traditional vaccine platforms. IAV is a significant respiratory pathogen of swine and there is a critical need to improve current commercial swine IAV vaccine platforms. Adjuvanted whole inactivated virus (WIV) IAV swine vaccines provide limited heterologous protection and may lead to vaccine-associated enhanced respiratory disease (VAERD). This study investigated the ability of RP IAV hemagglutinin (HA) vaccines to avoid VAERD and evaluated experimental multivalent HA and neuraminidase (NA) RP vaccines. RP vaccines were formulated with HA or NA heterologous or homologous to the challenge virus in monovalent HA or HA and NA bivalent combinations (HA/NA bivalent). Pigs were vaccinated with an HA RP, HA/NA bivalent RP, or heterologous HA WIV, followed by IAV challenge and necropsy 5 days post infection. RP vaccines provided homologous protection from challenge and induced robust peripheral and local antibody responses. The RP vaccine did not induce VAERD after challenge with a virus containing the heterologous HA, in contrast to the traditional WIV vaccine. The HA monovalent and HA/NA bivalent RP vaccines showed superior protection compared to traditional WIV. Additionally, the RP platform allows greater flexibility to adjust HA and NA content to reflect circulating IAV in swine antigenic diversity.
60 APPLIED LIFE SCIENCES↗
Conserved HA-peptide NG34 formulated in pCMV-CTLA4-Ig reduces viral shedding in pigs after a heterosubtypic influenza virus SwH3N2 challenge
Swine influenza viruses (SIVs), the causal agents of swine influenza, are not only important to control due to the economic losses in the swine industry, but also can be pandemic pathogens. Vaccination is one of the most relevant strategies to control and prevent influenza infection. Current human vaccines against influenza induce strain-specific immunity and annual update is required due to the virus antigenic shift phenomena. Previously, our group has reported the use of conserved hemagglutinin peptides (HA-peptides) derived from H1-influenza virus as a potential multivalent vaccine candidate. Immunization of swine with these HA-peptides elicited antibodies that recognized and neutralized heterologous influenza viruses in vitro and demonstrated strong hemagglutination-inhibiting activity. In the present work, we cloned one HA-peptide (named NG34) into a plasmid fused with cytotoxic T lymphocyte-associated antigen (CTLA4) which is a molecule that modifies T cell activation and with an adjuvant activity interfering with the adaptive immune response. The resulting plasmid, named pCMV-CTLA4-Ig-NG34, was administered twice to animals employing a needle-free delivery approach. Two studies were carried out to test the efficacy of pCMV-CTLA4-Ig-NG34 as a potential swine influenza vaccine, one in seronegative and another in seropositive pigs against SIV. The second one was aimed to evaluate whether pCMV-CTLA4-Ig-NG34 vaccination would overcome maternally derived antibodies (MDA). After immunization, all animals were intranasally challenged with an H3N2 influenza strain. A complete elimination or significant reduction in the viral shedding was observed within the first week after the challenge in the vaccinated animals from both studies. In addition, no challenged heterologous virus load was detected in the airways of vaccinated pigs. Overall, it is suggested that the pCMV-CTLA4-Ig-NG34 vaccine formulation could potentially be used as a multivalent vaccine against influenza viruses. © 2019 Sisteré-Oró et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
60 APPLIED LIFE SCIENCES↗
Transmission of Human Influenza A Virus in Pigs Selects for Adaptive Mutations on the HA Gene
Influenza A viruses (FLUAV) cause respiratory diseases in many host species, including humans and pigs. The spillover of FLUAV between swine and humans has been a concern for both public health and the swine industry. With the emergence of the triple reassortant internal gene (TRIG) constellation, establishment of human-origin FLUAVs in pigs has become more common, leading to increased viral diversity. However, little is known about the adaptation processes that are needed for a human-origin FLUAV to transmit and become established in pigs. Here, we generated a reassortant FLUAV (VIC11pTRIG) containing surface gene segments from a human FLUAV strain and internal gene segments from the 2009 pandemic and TRIG FLUAV lineages and demonstrated that it can replicate and transmit in pigs. Sequencing and variant analysis identified three mutants that emerged during replication in pigs, which were mapped near the receptor binding site of the hemagglutinin (HA). The variants replicated more efficiently in differentiated swine tracheal cells compared to the virus containing the wildtype human-origin HA, and one of them was present in all contact pigs. These results show that variants are selected quickly after replication of human-origin HA in pigs, leading to improved fitness in the swine host, likely contributing to transmission.
60 APPLIED LIFE SCIENCES↗
Comparative Virulence and Genomic Analysis of Streptococcus suis Isolates
Streptococcus suis is a zoonotic bacterial swine pathogen causing substantial economic and health burdens to the pork industry. Mechanisms used by S. suis to colonize and cause disease remain unknown and vaccines and/or intervention strategies currently do not exist. Studies addressing virulence mechanisms used by S. suis have been complicated because different isolates can cause a spectrum of disease outcomes ranging from lethal systemic disease to asymptomatic carriage. The objectives of this study were to evaluate the virulence capacity of nine United States S. suis isolates following intranasal challenge in swine and then perform comparative genomic analyses to identify genomic attributes associated with swine-virulent phenotypes. No correlation was found between the capacity to cause disease in swine and the functional characteristics of genome size, serotype, sequence type (ST), or in vitro virulence-associated phenotypes. A search for orthologs found in highly virulent isolates and not found in non-virulent isolates revealed numerous predicted protein coding sequences specific to each category. While none of these predicted protein coding sequences have been previously characterized as potential virulence factors, this analysis does provide a reliable one-to-one assignment of specific genes of interest that could prove useful in future allelic replacement and/or functional genomic studies. Collectively, this report provides a framework for future allelic replacement and/or functional genomic studies investigating genetic characteristics underlying the spectrum of disease outcomes caused by S. suis isolates.
59 BASIC BIOLOGICAL SCIENCES↗
Characterization of Senecavirus A Isolates Collected From the Environment of U.S. Sow Slaughter Plants
Vesicular disease caused by Senecavirus A (SVA) is clinically indistinguishable from foot-and-mouth disease (FMD) and other vesicular diseases of swine. When a vesicle is observed in FMD-free countries, a costly and time-consuming foreign animal disease investigation (FADI) is performed to rule out FMD. Recently, there has been an increase in the number of FADIs and SVA positive samples at slaughter plants in the U.S. The objectives of this investigation were to: (1) describe the environmental burden of SVA in sow slaughter plants; (2) determine whether there was a correlation between PCR diagnostics, virus isolation (VI), and swine bioassay results; and (3) phylogenetically characterize the genetic diversity of contemporary SVA isolates. Environmental swabs were collected from three sow slaughter plants (Plants 1-3) and one market-weight slaughter plant (Plant 4) between June to December 2020. Of the 426 samples taken from Plants 1-3, 304 samples were PCR positive and 107 were VI positive. There was no detection of SVA by PCR or VI at Plant 4. SVA positive samples were most frequently found in the summer (78.3% June-September, vs. 59.4% October-December), with a peak at 85% in August. Eighteen PCR positive environmental samples with a range of C t values were selected for a swine bioassay: a single sample infected piglets ( n = 2). A random subset of the PCR positive samples was sequenced; and phylogenetic analysis demonstrated co-circulation and divergence of two genetically distinct groups of SVA. These data demonstrate that SVA was frequently found in the environment of sow slaughter plants, but environmental persistence and diagnostic detection was not indicative of whether a sampled was infectious to swine. Consequently, a more detailed understanding of the epidemiology of SVA and its environmental persistence in the marketing chain is necessary to reduce the number of FADIs and aide in the development of control measures to reduce the spread of SVA.
59 BASIC BIOLOGICAL SCIENCES↗
Absence of clinical disease and contact transmission of HPAI H5NX clade 2.3.4.4 from North America in experimentally infected pigs
Background In the fall of 2014, highly pathogenic avian influenza (HPAI) subtype H5N8 clade 2.3.4.4 was introduced into North America by migrating waterfowl from Asia where, through reassortment, novel HPAI H5N2 and H5N1 viruses emerged. Objectives Assess the susceptibility of pigs to HPAI H5N1, H5N2, and H5N8 clade 2.3.3.3 from North America. Methods Pigs and trachea explants were inoculated with a representative panel of H5NX clade 2.3.4.4 HPAI viruses from North America. Nasal swabs, BALF, and sera were collected to assess replication and transmission in challenged and direct contact pigs by RRT‐PCR, virus isolation, hemagglutination inhibition, and ELISA. Results Limited virus replication was restricted to the lower respiratory tract of challenged pigs, though absent in the nasal passages and trachea cultures, as determined by RRT‐PCR in all samples. Seroconversion of inoculated pigs was detected by NP ELISA but was not reliably detected by antigen‐specific hemagglutination inhibition. Boost with adjuvanted virus was required for the production of neutralizing antibodies to assess cross‐reactivity between wild‐type avian strains. All RRT‐PCR and serology tests were negative for contact animals indicating a failure of transmission from primary inoculated pigs. Conclusions H5NX clade 2.3.4.4 strains can replicate in the lower respiratory tract of swine upon high titer inoculation, though appear to be incapable of replication in swine nasal epithelium in vivo or ex vivo in trachea explants in culture. Infected pigs did not produce high levels of serum antibodies following infection. Collectively, our data show HPAI H5NX clade 2.3.4.4 viruses to be poorly adapted for replication and transmission in swine.
H5↗
The Presence of Virus Neutralizing Antibodies Is Highly Associated with Protection against Virulent Challenge in Domestic Pigs Immunized with ASFV live Attenuated Vaccine Candidates
African swine fever virus (ASFV) is currently producing a pandemic affecting a large area of Eurasia, and more recently, the Dominican Republic in the Western Hemisphere. ASFV is a large and structurally complex virus with a large dsDNA genome encoding for more than 150 genes. Live attenuated virus strains can induce protection in domestic swine against disease produced by homologous virulent parental viruses. The roles of the different immune mechanisms induced by the attenuated strains in protection still need to be understood. In particular, the role of ASFV neutralizing antibody in protection still is an important controversial issue to be elucidated. Here we present the development of a novel methodology to detect virus neutralizing antibodies based on the reduction of virus infectivity in a Vero cell adapted ASFV strain. The described method was used to assess levels of virus neutralizing antibodies in domestic swine inoculated with live attenuated ASFV. Results demonstrated a high association between the presence of virus neutralizing antibodies and protection in 84 animals immunized with the recombinant vaccine candidates ASFV-G-Δ9GL/ΔUK or ASFV-G-ΔI177L. To our knowledge, this is the first report demonstrating an association between virus neutralizing antibodies and protection against virulent challenge in such a large number of experimental individuals.
59 BASIC BIOLOGICAL SCIENCES↗