Free Pigs Research Articles

1 Microbiomes from Theory to Application

Abstract Microbiome research has gained significant momentum in the last decade. Although many studies have investigated livestock microbiome to improve animal health and productivity, only a few microbiome manipulation approaches are currently used in the industry. This is partly due to the limited understanding of livestock microbiome and how such knowledge can be applied to improve animal health and performance. Additionally, the potential role of the microbiome is poorly characterized, limiting the opportunities available to improve animal health and productivity through microbiome manipulation. Here we describe and demonstrate how current microbiome theory such as “colonization history” can be utilized to establish beneficial microbiomes in livestock species. Our data suggest that the sow microbiome greatly impacts the early microbiome establishment in piglets and represents as much as 67% of the community even after 21 days following birth. This suggests early colonization provides unique opportunities to establish “beneficial” microbiomes. Additionally, we describe how microbiome-wide association studies (MWAS) can be used to quantify the microbial role in animal performance using a cattle population consisting of more than 750 animals. Finally, we describe how livestock animal models can be used to understand key concepts of microbial establishment and how such models can be used to investigate the human microbiome using germ free pig models. USDA is an equal opportunity provider and employer.

Analysis of PERV-C superinfection resistance using HA-tagged viruses

BackgroundUsing pigs as organ donors has advanced xenotransplantation to the point that it is almost ready for clinical use. However, there is still a zoonotic risk associated with xenotransplantation, and the potential transmission of porcine endogenous retroviruses needs to be surveyed. Despite significant attempts to eliminate this risk, by the selection of PERV-C free pigs with low expression of PERV-A, -B, and by the genome-wide inactivation of PERV using CRISPR/Cas9, the impact of superinfection resistance (SIR) was not investigated. SIR is a viral trait that prevents reinfection (superinfection). For PERV, the underlying mechanism is unclear, whether and how cells, that harbor functional PERV, are protected. Using PERV-C(5683) as a reference virus, we investigated SIR in a newly developed in vitro model to pursue the mechanism and confirm its protective effect.ResultsWe developed three PERV-C constructs on the basis of PERV-C(5683), each of which carries a hemagglutinin tag (HA-tag) at a different position of the envelope gene (SP-HA, HA-VRA, and RPep-HA), to distinguish between primary infection and superinfection. The newly generated PERV-C(5683)-HA viruses were characterized while quantifying the viral RNA, reverse transcriptase activity, protein expression analysis, and infection studies. It was demonstrated that SP-HA and RPep-HA were comparable to PERV-C(5683), whereas HA-VRA was not replication competent. SP-HA and RPep-HA were chosen to challenge PERV-C(5683)-positive ST-IOWA cells demonstrating that PERV-C-HA viruses are not able to superinfect those cells. They do not integrate into the genome and are not expressed.ConclusionsThe mechanism of SIR applies to PERV-C. The production of PERV-C particles serves as a defense mechanism from superinfection with exogenous PERV-C. It was demonstrated by newly generated PERV-C(5683)-HA clones that might be used as a cutting-edge tool. The HA-tagging of PERV-C is novel, providing a blueprint for the tagging of other human tropic PERV viruses. The tagged viruses are suitable for additional in vitro and in vivo infection studies and will contribute, to basic research on viral invasion and pathogenesis. It will maintain the virus safety of XTx.

Characterization analysis of Rongchang pig population based on the Zhongxin-1 Porcine Breeding Array PLUS.

To carry out a comprehensive production planning of the existing Rongchang pig population from both environmental and genetic aspects, and to establish a closed population with stable genetic diversity and strict pathogen control, it is necessary to fully understand the genetic background of the population. We genotyped 54 specific pathogen free (SPF) Rongchang pigs using the Zhongxin-1 Porcine Breeding Array PLUS, calculated their genetic diversity parameters and constructed their families. In addition, we also counted the runs of homozygosity (ROH) of each individual and calculated the value of inbreeding coefficient based on ROH for each individual. Firstly, the results of genetic diversity analysis showed that the effective population size (Ne) of this population was 3.2, proportion of polymorphic markers (PN) was 0.515, desired heterozygosity (He) and observed heterozygosity (Ho) were 0.315 and 0.335. Ho was higher than He, indicating that the heterozygosity of all the selected loci was high. Secondly, combining the results of genomic relatedness analysis and cluster analysis, it was found that the existing Rongchang pig population could be divided into four families. Finally, we also counted the ROH of each individual and calculated the inbreeding coefficient value accordingly, whose mean value was 0.09. Due to the limitation of population size and other factors, the genetic diversity of this Rongchang pig population is low. The results of this study can provide basic data to support the development of Rongchang pig breeding program, the establishment of SPF Rongchang pig closed herd and its experimental utilization.

A split influenza vaccine formulated with a combination adjuvant composed of alpha-d-glucan nanoparticles and a STING agonist elicits cross-protective immunity in pigs

BackgroundSwine influenza A viruses (SwIAVs) pose an economic and pandemic threat, and development of novel effective vaccines is of critical significance. We evaluated the performance of split swine influenza A virus (SwIAV) H1N2 antigens with a plant-derived nanoparticle adjuvant alone (Nano-11) [Nano11-SwIAV] or in combination with the synthetic stimulator of interferon genes (STING) agonist ADU-S100 (NanoS100-SwIAV). Specific pathogen free (SPF) pigs were vaccinated twice via intramuscular (IM) or intradermal (ID) routes and challenged with a virulent heterologous SwIAV H1N1-OH7 virus.ResultsAnimals vaccinated IM or ID with NanoS100-SwIAV had significantly increased cross-reactive IgG and IgA titers in serum, nasal secretion and bronchoalveolar lavage fluid at day post challenge 6 (DPC6). Furthermore, NanoS100-SwIAV ID vaccinates, even at half the vaccine dose compared to their IM vaccinated counterparts, had significantly increased frequencies of CXCL10+ myeloid cells in the tracheobronchial lymph nodes (TBLN), and IFNγ+ effector memory T-helper/memory cells, IL-17A+ total T-helper/memory cells, central and effector memory T-helper/memory cells, IL-17A+ total cytotoxic T-lymphocytes (CTLs), and early effector CTLs in blood compared with the Nano11-SwIAV group demonstrating a potential dose-sparing effect and induction of a strong IL-17A+ T-helper/memory (Th17) response in the periphery. However, the frequencies of IFNγ+ late effector CTLs and effector memory T-helper/memory cells, IL-17A+ total CTLs, late effector CTLs, and CXCL10+ myeloid cells in blood, as well as lung CXCL10+ plasmacytoid dendritic cells were increased in NanoS100-SwIAV IM vaccinated pigs. Increased expression of IL-4 and IL-6 mRNA was observed in TBLN of Nano-11 based IM vaccinates following challenge. Furthermore, the challenge virus load in the lungs and nasal passage was undetectable in NanoS100-SwIAV IM vaccinates by DPC6 along with reduced macroscopic lung lesions and significantly higher virus neutralization titers in lungs at DPC6. However, NanoS100-SwIAV ID vaccinates exhibited significant reduction of challenge virus titers in nasal passages and a remarkable reduction of challenge virus in lungs.ConclusionsDespite vast genetic difference (77% HA gene identity) between the H1N2 and H1N1 SwIAV, the NanoS100 adjuvanted vaccine elicited cross protective cell mediated immune responses, suggesting the potential role of this combination adjuvant in inducing cross-protective immunity in pigs.Graphical

Plant derived Nano-11 particle adsorbed with stimulator of interferon genes adjuvant and split influenza virus antigens elicits the cross-protective immunity in pigs

Abstract We evaluated the performance of split swine influenza A virus (SwIAV) H1N2 antigens with a plant-derived nanoparticle adjuvant (Nano-11) or its combination with a synthetic STING agonist ADU-S100 (NanoS100). Specific pathogen free pigs were vaccinated twice via intramuscular (IM) and intradermal (ID) routes and challenged with a virulent heterologous H1N1. Animals vaccinated IM or ID with H1N2-ADU-S100-Nano-11 had significantly increased cross-reactive IgG and IgA titers in serum, nasal secretion and bronchoalveolar lavage fluid. There was a significant increase of IFNγ+ effector memory T-helper/memory cells and late effector cytotoxic T cells (CTLs), IL-17A+ central and effector memory T-helper/memory cells, IL-17A+ early effector CTLs, and CXCL10+ plasmacytoid dendritic cells in the PBMCs of the ID injected H1N2-ADU-S100-Nano-11 compared to H1N2-Nano-11 group. The frequency of IFNγ+ late effector CTLs and effector memory T-helper/memory cells and IL-17A+ late effectorCTLs, as well as in lungs CXCL10+ plasmacytoid dendritic cells was increased in PBMCs of IM injected H1N2-ADUS100-Nano-11 pigs. Increased expression of IL-4 and IL-6 mRNA was observed in tracheobronchial lymph nodes of IM Nano-11 based vaccinates following challenge. Furthermore, the challenge virus load in the lungs and nasal passage was undetectable in IM H1N2+ADU-S100-Nano-11 vaccinates by day 6 post infection along with reduced macroscopic lung lesions. In conclusion, despite vast genetic difference (77% HA gene identity) between the H1N2 and H1N1 SwIAV, the NanoS100 adjuvanted vaccine elicited cross protective immune responses, suggesting the potential role of this combination adjuvant in inducing cross-protective immunity in pigs. Supported by USDA National Institute of Food and Agriculture, AFRI project 2019-67015-29814

Current Topics of Relevance to the Xenotransplantation of Free Pig Islets.

Pig islet xenotransplantation is a potential treatment for patients with type 1 diabetes. Current efforts are focused on identifying the optimal pig islet source and overcoming the immunological barrier. The optimal age of the pig donors remains controversial since both adult and neonatal pig islets have advantages. Isolation of adult islets using GMP grade collagenase has significantly improved the quantity and quality of adult islets, but neonatal islets can be isolated at a much lower cost. Certain culture media and coculture with mesenchymal stromal cells facilitate neonatal islet maturation and function. Genetic modification in pigs affords a promising strategy to prevent rejection. Deletion of expression of the three known carbohydrate xenoantigens (Gal, Neu5Gc, Sda) will certainly be beneficial in pig organ transplantation in humans, but this is not yet proven in islet transplantation, though the challenge of the ‘4th xenoantigen’ may prove problematic in nonhuman primate models. Blockade of the CD40/CD154 costimulation pathway leads to long-term islet graft survival (of up to 965 days). Anti-CD40mAbs have already been applied in phase II clinical trials of islet allotransplantation. Fc region-modified anti-CD154mAbs successfully prevent the thrombotic complications reported previously. In this review, we discuss (I) the optimal age of the islet-source pig, (ii) progress in genetic modification of pigs, (iii) the immunosuppressive regimen for pig islet xenotransplantation, and (iv) the reduction in the instant blood-mediated inflammatory reaction.

Efficacy of the Synergy Between Live-Attenuated and Inactivated PRRSV Vaccines Against a NADC30-Like Strain of Porcine Reproductive and Respiratory Syndrome Virus in 4-Week Piglets.

The NADC30-like strain of porcine reproductive and respiratory syndrome virus (PRRSV) is a novel strain responsible for substantial economic losses to swine production in China. This study evaluated the cross-protective efficacy of the synergy between live-attenuated and inactivated PRRSV vaccines compared with a single vaccination with PRRS modified-live virus (MLV) vaccine against challenge with NADC30-like strain, v2016/ZJ/09-03. A total of 45 PRRSV free pigs were randomly divided into five groups: (1) strict control (SC); (2) positive control (PC); (3) single MLV dose (M1); (4) primed intramuscularly with MLV and boosted with killed vaccine 3 weeks later (MK1); and (5) intramuscular prime MLV boosted subcutaneously with killed vaccine B 3 weeks later (MK2). Serological tests in MK groups revealed no differences in both anti-N and anti-GP protein antibodies compared with M1 group, and failed to provide further protection against clinical signs, virus shedding, and gross lesions. However, the viremic titer, gross lung lesions, and average daily weight gain were significantly improved in the MLV vaccinated groups, suggesting that MLV provides substantial cross-protection against the NADC30-like virus. Thus, as a booster, the killed vaccine confers minimal additional protection in NADC30-like infected piglets.

Freedom to Grow: Improving Sow Welfare also Benefits Piglets.

Simple SummaryThis study identified positive effects on piglet welfare and growth rates when housed in free lactation pens compared with conventional farrowing crates. Free lactation pens allow piglets unrestricted interaction with the sow and increased space allowance; this improved behavioural development of piglets thus resulted in fewer instances of damaging behaviour toward littermates in this study. Piglets also benefit from better access to the udder for feeding in free lactation pens, with those piglets reared in such pens having better growth rates than those in conventional farrowing crates.Piglet mortality, especially due to crushing, is a concern in pig production. While current systems use farrowing crates to reduce mortality, they present major animal welfare problems for the sow. This study investigated the effects of free lactation farrowing accommodation on the welfare of piglets born and reared in such accommodation when compared with conventional farrowing crates. Piglets were born to sows in one of two farrowing accommodation treatments and followed from birth to slaughter. Treatments were conventional farrowing crates (control, n = 24 litters) and free lactation pens, which were larger and allowed the sow freedom of movement both pre and post farrowing, (free, n = 22 litters) (675 total piglets). Individual weights were recorded from birth to slaughter. Hoof score was recorded at weaning. Behaviour was recorded during lactation and during the weaner stage. Pre-weaning percentage mortality was equal in treatments (free = 15.95 ± 2.31, control = 14.42 ± 2.15, p = 0.61). Final weight was influenced by treatment (p < 0.05) with pigs from free lactation pens significantly heavier at 114.73 kg compared to 110.82 kg for control pigs. Free pigs took fewer days than control pigs to reach the target weight of 105 kg (147.6 vs. 149.23 days). Throughout lactation, fewer instances of damaging behaviour (ear and tail biting) were observed in free pigs (p = 0.07). Improved growth rates and a decrease in damaging behaviours provide evidence to suggest that pigs from free lactation pens experience improved welfare when compared with control pigs.

How electrically evoked compound action potentials in chronically implanted guinea pigs relate to auditory nerve health and electrode impedance.

This study examined how multiple measures based on the electrically evoked compound action potential (ECAP) amplitude-growth functions (AGFs) were related to estimates of neural [spiral ganglion neuron (SGN) density and cell size] and electrode impedance measures in 34 specific pathogen free pigmented guinea pigs that were chronically implanted (4.9-15.4 months) with a cochlear implant electrode array. Two interphase gaps (IPGs) were used for the biphasic pulses and the effect of the IPG on each ECAP measure was measured ("IPG effect"). When using a stimulus with a constant IPG, SGN density was related to the across-subject variance in ECAP AGF linear slope, peak amplitude, and N1 latency. The SGN density values also help to explain a significant proportion of variance in the IPG effect for AGF linear slope and peak amplitude measures. Regression modeling revealed that SGN density was the primary dependent variable contributing to across-subject variance for ECAP measures; SGN cell size did not significantly improve the fitting of the model. Results showed that simple impedance measures were weakly related to most ECAP measures but did not typically improve the fit of the regression model.

One-Tube Nested Real-Time PCR Assay for Rapid Screening of Porcine Cytomegalovirus in Clinical Samples.

Porcine cytomegalovirus (PCMV) is a pathogen that must be removed from pigs for use as organ donors in xenotransplantation. Recently, it has been found that when donor pigs are infected with PCMV, a pig-to-non-human-primate xenotransplantation lower transplant survival by 2–3 times. Therefore, highly sensitive methods are needed to maintain designated pathogen free (DPF) pig and screen for xenografts. The purpose of this study was to evaluate the performance of commercially available method with one-tube nested real-time PCR assay to quickly detect PCMV infection in clinical samples and compare the results with those of sequence analysis. Molecular diagnostic methods were used to evaluate 127 samples, including tissues and blood samples from pigs suspected of PCMV infection. The detection rate for positive PCMV was 38.6% (n = 49), 23.6% (n = 30), and 12.6% (n = 16) in one-tube nested real-time PCR, nested PCR, and conventional PCR methods, respectively. All PCMV-positive samples in conventional PCR or nested PCR methods were also positive in the one-tube nested real-time PCR assay. All the PCR products in the three methods were checked for amplification of PCMV gene by PCR and subsequent direct sequencing. The results of one-tube nested real-time PCR were found to be consistent with those of sequence analysis for all the samples and showed good agreement (κ = 1). Our study found that the one-tube nested real-time PCR assay is more sensitive than the other two methods. This assay required approximately 1.5 h for completion. Therefore, we concluded that one-tube nested real-time PCR assay is a fast and reliable method for the characterizing pathogen responsible for PCMV infection.

Comparison of four serological tests for the diagnosis of swine brucellosis

The purpose of this article is to present results of a research in which the results of four serological tests were compared for the diagnosis of swine brucellosis in pigs from Brazilian herds in which the outbreak occurred in relation to other brucellosis-free. Rose Bengal (RBT), complement fixation (CFT), agglutination plus 2-mercaptoethanol test (TAT+ME), and commercial fluorescence polarization assay (FPA) were used to test 333 sera (271 sows and 62 finishing pigs) from a Brucella suis infected swine herd and 1,100 swine sera from brucellosis free pigs taken at a slaughterhouse. Considering infected all the 271 sows from the outbreak, and interpreting results of the FPA according to the manufacturer directions, sensitivities observed were 95.94% for FPA, 94.83% for RBT, 93.73% for CFT, and 92.25% for TAT+ME. Considering infected all the pigs from the infected herd with a positive result in at least one test, sensitivities observed were 98.92% for RBT, 97.13% for CFT, 96.06% for FPA, and 94.98% for TAT+ME. Specificities of the tests were CFT and TAT+ME, 100%; FPA, 99.55%; and RBT 99.27%. Results indicated a good performance of all the tests, and the agreement among them was almost perfect.

La crianza de cerdos en vida libre y pecaríes silvestres en zonas de transición de áreas protegidas del sureste de México

El estudio analizó los sistemas de producción de cerdos domésticos en vida libre y pecaríes silvestres para determinar los beneficios sociales, económicos y ambientales en comunidades ubicadas en la zona de transición de la Reserva de la Biósfera Calakmul, México. La información se obtuvo de 169 campesinos criadores de suinos, de un censo realizado en 13 comunidades. La información de la unidad de producción, los criadores, la piara de cerdos, la alimentación, los parámetros productivos y reproductivos, la comercialización, los costos e ingresos se registraron en un cuestionario. Los datos se analizaron con frecuencias, análisis de varianza, clúster, factorial y componentes principales con el paquete estadístico SAS. Se identificaron tres sistemas de crianza de cerdos domésticos en vida libre: traspatio (23,1%), itinerancia estacional (21,9%) e itinerante (55%). Los tres sistemas tuvieron pecarí de collar (Pecarí tajacu) como una forma de reproducción en cautiverio de esta especie silvestre. El sistema de crianza en traspatio fue el más intensivo en uso de mano de obra, forraje de corte, grano de maíz y la compra de alimento. El sistema de itinerancia estacional fue intermedio en el uso de recursos e insumos. La crianza itinerante tuvo mayor convivencia con el pecarí en cautiverio, con el uso de los ecosistemas naturales y una menor orientación al mercado. La crianza en traspatio y la itinerancia estacional se clasificaron como de transición a la producción en confinamiento, con una disminución de la crianza en libertad y se incrementó el uso de insumos para la alimentación.

Assessment of Influenza D Virus in Domestic Pigs and Wild Boars in France: Apparent Limited Spread within Swine Populations Despite Serological Evidence of Breeding Sow Exposure.

In order to assess influenza D virus (IDV) infections in swine in France, reference reagents were produced in specific pathogen free pigs to ensure serological and virological analyses. Hemagglutination inhibition (HI) assays were carried out on 2090 domestic pig sera collected in 2012–2018 in 102 farms. Only 31 sera from breeding sows sampled in 2014–2015 in six farrow-to-finish herds with respiratory disorders contained IDV-specific antibodies. In two of them, within-herd percentage of positive samples (73.3% and 13.3%, respectively) and HI titers (20–160) suggested IDV infections, but virus persistence was not confirmed following new sampling in 2017. All growing pigs tested seronegative, whatever their age and the sampling year. Moreover, PB1-gene RT-qPCR performed on 452 nasal swabs taken in 2015–2018 on pigs with acute respiratory syndrome (137 farms) gave negative results. In Corse, a Mediterranean island where pigs are mainly bred free-range, 2.3% of sera (n = 177) sampled on adult pigs in 2013–2014 obtained low HI titers. Finally, 0.5% of sera from wild boars hunted in 2009–2016 (n = 644) tested positive with low HI titers. These results provide the first serological evidence that sows were exposed to IDV in France but with a limited spread within the swine population.

Cross-protective efficacy of the O1 Manisa + O 3039 bivalent vaccine and the O 3039 monovalent vaccine against heterologous challenge with FMDV O/Jincheon/SKR/2014 in pig

After massive foot-and-mouth disease (FMD) outbreaks originated from Jincheon County from Dec. 2014 to Apr. 2015, the effectiveness of the previous FMD vaccine containing only the O1 Manisa as the O antigen, O1 Manisa + A Malaysia 97 + Asia 1 Sharmir trivalent vaccine, was questioned in South Korea, and a change in the O antigen in FMD vaccines was demanded to control the FMD caused by FMDV O/Jincheon/SKR/2014, the O Jincheon strain. Therefore, the efficacies of O1 Manisa + O 3039 bivalent vaccine and O 3039 monovalent vaccine were studied for cross-protection against heterologous challenge with the O Jincheon strain. In this study, the efficacy of the O1 Manisa + O 3039 bivalent vaccine was better than that of the O 3039 monovalent vaccine, even though the serological relationship (r1 value) between O Jincheon and O 3039 was matched according to the OIE Terrestrial Manual. According to serological test results from vaccinated specific pathogen free pigs, virus neutralization test titers against Jincheon were good estimates for predicting protection against challenge. A field trial of the O1 Manisa + O 3039 bivalent vaccine was performed to estimate the possibility of field application in conventional pig farms, especially due to concerns about the effect of maternally derived antibodies (MDA) in field application of the FMD vaccine. According to the result of the field trial, the O1 Manisa + O 3039 bivalent vaccine was considered to overcome MDA. The results of the efficacy and field trials indicated that the O1 Manisa + O3039 vaccine could be suitable to replace previous FMD vaccines to control the FMD field situation caused by O Jincheon FMDV.

Update on clinical trials results of kidney stone repositioning and preclinical results of stone breaking with one ultrasound system

Our goal is an office-based, handheld ultrasound system to target, detach, break, and/or expel stones and stone fragments from the urinary space to facilitate natural clearance. Repositioning of stones in humans (maximum 2.5 MPa, and 3-second bursts) and breaking of stones in a porcine model (maximum 50 cycles, 20 Hz repetition, 30 minutes, and 7 MPa peak negative pressure) have been demonstrated using the same 350-kHz probe. Repositioning in humans was conducted during surgery with a ureteroscope in the kidney to film stone movement. Independent video review confirmed stone movements (≥ 3 mm) in 14 of 15 kidneys (93%). No serious or unanticipated adverse events were reported. Experiments of burst wave lithotripsy (BWL) effectiveness on breaking human stones implanted in the porcine bladder and kidney demonstrated fragmentation of 4 of 4 stones on post mortem dissection. All clinical pathology, hematology, and urinalysis for a 1-week survival study with the BWL exposures in 10 specific pathogen free pigs were within normal limits. These results demonstrate that repositioning of stones with ultrasonic propulsion and breaking of stones with BWL are safe and effective. [Work supported by NIH P01 DK043881, K01 DK104854, and R44 DK109779.]

Islet Xenotransplantation for the Treatment of Type 1 Diabetes

Allogeneic islet transplantation has been established to prevent severe hypoglycemia for unstable type 1 diabetic patients. Recent phase 3 clinical trial clearly demonstrated the benefit of allogeneic islet transplantation. Severe hypoglycemia is serious issue not only for type 1 diabetic patients but also type 2 diabetic patients especially aged patients. Considering the possible demands for such patients, donor shortage will be the serious issue. To solve this issue, islet xenotransplantation using porcine islets is a promising remedy. There are several advantages of porcine islets comparing with human islets including unlimited and on demand supplies, higher quality of islets from healthy donor, safer with designated pathogen free (DPF) donor pigs and possible gene modifications. Comparing ES and iPS derived islets, porcine islets have more profound clinical experiences with evidences of safety and efficacy since 1990s. In addition, promising outcomes were published using porcine islets to restore normoglycemia in the non-human primate diabetes models. One of the major concerns of islet xenotransplantation is zoonotic infections. To prevent this possible complication, using DPF status donor pigs, and monitoring porcine endogenous retrovirus (PERV) are key elements. In addition, compliance with regulatory rule is critically important to prevent spreading the zoonotic infections. Recently, international xenotransplantation association updated their consensus statements to promote islet xenotransplantation and it emphasized the importance of the regulatory compliance. In 2014, a clinical trial of encapsulated porcine islet xenotransplantation for the treatment of type 1 diabetes under comprehensive regulation was published. This study demonstrated the safety of islet xenotransplantation and some efficacy. Recently, it was published the encapsulated porcine islet xenotransplantation provided clinical benefit for type 1 diabetic patients. Encapsulated porcine islet xenotransplantation seems promising, and several groups have demonstrated improved capsules which may solve the fibrosis issue of capsules. CRISPR/CAS9 system makes the gene editing technology stable and easy. In fact, it was demonstrated that PERVs were completely eliminated with CRISPR technology. Human compatible pig will be made with this technology. Eventually, human pancreas will be created in pig body with blastocyst complementation technology. These aggressive scientific advances will bring islet xenotransplantation as a standard therapy for diabetic patients.

ISOLATION AND IDENTIFICATION OF BRUCELLA SUIS IN PIGS AS ZOONOTIC DISEASE IN ENDEMIC AREAS OF EAST JAVA, INDONESIA

Background:Brucellosis in pigs at East Java Indonesia has not only cause great economic losses due to a decrease in productivity of livestock but also are zoonotic. Infection on free brucelosis pigs were initially begun with the infected pigs both male and female, or the use of superior male pigs together. The elimination of the disease either on a group or population is considered as the most effective way to prevent the spread of the disease in pigs. Prevention efforts mainly addressed to vaccination, sanitary maintenace and government policy. The purpose of this study was to isolated and identified Brucella suis as the causative agent.Material and Methods:The survey area were the pig farm owned by breeder farmers in the area of East Java Indonesia, at Kediri, Malang, Blitar and Probolinggo district. Blood samples obtained were tested with RBT. Pigs are suspected of being infected with Brucella if the RBT was positive that characterized with agglutination in the test results. If RBT was positive, bacteriological examination will be performed, with samples of visceral foetus organ, ie liver, spleen, placenta and amniotic fluid. Isolation and identification of Brucella suis were used Brucella Broth and Brucella Agar, and if the bacteri growthwill be continued with biochemical test ie H2S, urease, citrate, catalase and oxidase test. The positive results of Brucella suis showed positive urease, catalase andoxidase, but negative for citrate and H2S.Results:RBT and bacteriolgical examination showed that 1 sample was positive Brucella suis, and 19 negative. The positive results showed positive urease, catalase and oxidase, but negative for citrate and H2S.Conclusion:Based on RBT test and bacteriological examination, there was 1 positive sample of brucellla suis, that is sample coming from Kediri district.

ISOLATION AND IDENTIFICATION OF BRUCELLA SUIS IN PIGS AS ZOONOTIC DISEASE IN ENDEMIC AREAS OF EAST JAVA, INDONESIA

Brucellosis in pigs at East Java Indonesia has not only cause great economic losses due to a decrease in productivity of livestock but also are zoonotic. Infection on free brucelosis pigs were initially begun with the infected pigs both male and female, or the use of superior male pigs together. The elimination of the disease either on a group or population is considered as the most effective way to prevent the spread of the disease in pigs. Prevention efforts mainly addressed to vaccination, sanitary maintenace and government policy. The purpose of this study was to isolated and identified Brucella suis as the causative agent.The survey area were the pig farm owned by breeder farmers in the area of East Java Indonesia, at Kediri, Malang, Blitar and Probolinggo district. Blood samples obtained were tested with RBT. Pigs are suspected of being infected with Brucella if the RBT was positive that characterized with agglutination in the test results. If RBT was positive, bacteriological examination will be performed, with samples of visceral foetus organ, ie liver, spleen, placenta and amniotic fluid. Isolation and identification of Brucella suis were used Brucella Broth and Brucella Agar, and if the bacteri growthwill be continued with biochemical test ie H2S, urease, citrate, catalase and oxidase test. The positive results of Brucella suis showed positive urease, catalase andoxidase, but negative for citrate and H2S.RBT and bacteriolgical examination showed that 1 sample was positive Brucella suis, and 19 negative. The positive results showed positive urease, catalase and oxidase, but negative for citrate and H2S.Based on RBT test and bacteriological examination, there was 1 positive sample of brucellla suis, that is sample coming from Kediri district.

Current Status of Prevalence, Possible Control and Risk Factors Associated with Porcine Cysticercosis from Endemic Countries in Africa

Porcine cysticercosis (PC) caused by Taenia solium larvae is continuing being an important zoonotic neglected disease in many developing countries, is responsible for severe health disorders such as seizures and death in human and it poses a serious public health risk. In general estimated prevalence for porcine cysticercosis by Ag-ELISA was 29.7% (95% CI [4.8 - 58.2]), by EITB was 24.7% (95% CI [9.2 - 38.2]), by Tongue examination was 9.4% (95% CI [0.2 - 13.2]), and by postmortem examination was 15% (95% CI [0.2 - 27.4]). Average seroprevalence of human cysticercosis by circulating antibody detection (Ab-ELISA) was ranged from 1.3% to 45.3%. Average seroprevalence of human cysticercosis by circulating T. solium antigen detection (Ag-ELISA) was ranged from 4.6% to 11.9%. Average seroprevalence of human cysticercosis by EITB was ranged from 6.9% to 16.7%. Average prevalence of human cysticercosis by CT scan was ranged from 23.2% to 54.6%. A fundamental factor in the occurrence of infections in most of the article was lack of health education to the community on T. solium, cysticercosis and taeniasis complex as the key towards control and eradication. The major causal factors of the disease occurrence were free roaming pigs and poor sanitary conditions.

Different susceptibility and pathogenesis of rabbit genotype 3 hepatitis E virus (HEV-3) and human HEV-3 (JRC-HE3) in SPF rabbits

Hepatitis E virus (HEV) is an increasingly important zoonotic infection in humans with HEV genotypes 3 and 4 being recognized as zoonotic pathogens. The relatively recently isolated genotype 3 rabbit HEV (rHEV-3) and the more well known genotype 3 isolates from humans and swine (hsHEV-3) have all been confirmed experimentally to be capable of infecting both non-human primates and specific-pathogen free (SPF) pigs. In a previous study rHEV-3 was shown to cause acute hepatitis in experimentally infected rabbits. However, whether hsHEV-3 can productively infect rabbits remained unclear. The objective of this study was to investigate the experimental infection of rabbits with human HEV-3 (hHEV-3, JRC-HE3), to compare it to that with rHEV-3 (CHN-BJ-rb14) and to further characterise the pathogenesis of the two isolates. All animals inoculated with rHEV-3 (CHN-BJ-rb14) became infected, exhibiting an intermittent viremia, elevated liver enzymes, and persistent fecal virus shedding throughout the 15 week study period. Liver histopathology showed acute inflammation and both positive- and negative-stranded viral RNA was detected in various tissues from necropsied rabbits. By contrast, neither sero-conversion nor alanine aminotransferase (ALT) elevation was observed in most rabbits inoculated with hHEV-3 (JRC-HE3). In addition, rHEV-3 (CHN-BJ-rb14) but not hHEV-3 (JRC-HE3) recovered from primary infected rabbits was transmissible to naive rabbits. These results showed that SPF rabbits are readily susceptible to infection with rHEV-3 (CHN-BJ-rb14) but not hHEV-3 (JRC-HE3), which might indicate the influence of viral genomic organization on its pathogenicity.