Search Results (160)

Variants in the human PAX4 gene are associated with both monogenic and complex forms of diabetes, yet their pathogenic effects remain difficult to define in models that accurately mimic human islet architecture and neonatal metabolic transitions. Here, we created a porcine PAX4 loss-of-function model using CRISPR/Cas9 cytidine deaminase base editing to introduce a premature stop codon in the PAX4 coding sequence. PAX4 knockout piglets developed severe hyperglycemia within 24 h of birth, followed by rapid postnatal clinical deterioration and uniform death by day 3. Biochemical analysis showed significant diabetic decompensation, including electrolyte imbalances, hyperosmolality, azotemia, dyslipidemia, and metabolic acidosis. Gross and histological examinations revealed notable pancreatic hypoplasia with preservation of exocrine tissue. Single-nucleus RNA sequencing and immunohistochemistry demonstrated an almost complete loss of insulin- and somatostatin-producing β- and δ-cells, respectively, with relative preservation of glucagon-expressing α-cells. Overall, these results establish PAX4 as a crucial factor in pancreatic endocrine development and postnatal glucose regulation in a large-animal model. This platform offers a human-relevant system for studying diabetes-associated PAX4 variants and for testing regenerative and gene-based therapies for insulin-deficient diabetes. Full article

Newborn piglets in intensive pig farms are often prone to pulmonary diseases due to underdeveloped fetal lung neural systems, and ATRA is often regarded as an important morphogen that displays pleiotropic functions during embryonic development. However, information about the effect of maternal administration with ATRA on the peripheral neural system of fetal lungs is still scare. Fifteen pregnant sows were assigned to the ATRA0 (0 mg/kg diet), ATRA4, ATRA8, ATRA16 and ATRA32 treatment groups and offered their own feed from d12 to d95 after artificial insemination; then two neonatal pigs with birth weights similar to the average birth weight were taken out from each litter for the collection of lung samples, and samples were subjected to immunofluorescence staining, RNA-seq and RT-qPCR assays. Results indicated that compared to newborn piglets from the ATRA0 treatment group, newborn piglets from the ATRA4 treatment group had higher percentages of GFAP-positive astrocyte cells (p < 0.05) and GFAP-TUBB3 colocalization (p < 0.05) in the lungs. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis showed that differential expressed genes are mainly enriched in the pathways of neuroactive ligand–receptor interaction, GABAergic synapses and cell adhesion molecules. Addition of ATRA at 4 mg/kg to the diet of pregnant sows can enhance the healthy development of the pulmonary nervous systems of fetal pigs. Full article

The family Caliciviridae encompasses a diverse group of non-enveloped, positive-sense RNA viruses that are significant pathogens in veterinary medicine. This narrative review summarizes the current state of knowledge regarding the clinical, molecular, and epidemiological features of the three calicivirus genera identified in bovine hosts: Norovirus, Nebovirus, and Vesivirus. Bovine noroviruses and neboviruses are neglected enteric pathogens, frequently detected in association with neonatal calf diarrhea and often present in co-infections with other enteric agents. Clinical presentations for these enteric viruses range from severe, watery diarrhea to asymptomatic shedding, with distinct pathogenic profiles observed between norovirus genotypes GIII.1 and GIII.2. In contrast, the genus Vesivirus exhibits a broad host range, and bovine vesivirus strains are phylogenetically linked to vesiviruses identified in pigs and marine animals. Bovine vesivirus infections are associated with a broader spectrum of clinical manifestations, including respiratory disease, vesicular lesions, and abortion. Serological and virological surveys indicate that exposure to these viruses is ubiquitous in cattle populations globally. While direct evidence of human infection by bovine noroviruses and neboviruses remains limited, vesiviruses possess a confirmed capacity for cross-species transmission to humans. Significant knowledge gaps remain, particularly regarding in vitro culture systems, necessitating further research to facilitate vaccine development and clarify transmission dynamics. Full article

Background/Objectives: Porcine epidemic diarrhea (PED) is a highly contagious enteric disease caused by porcine epidemic diarrhea virus (PEDV) and is associated with severe clinical signs and high mortality in neonatal piglets. Vaccination is an important strategy for PED control through the induction of humoral immunity. This study aimed to compare immune responses induced by inactivated and live-attenuated PEDV vaccines and to evaluate a heterologous prime-boost vaccination strategy in PEDV-naïve replacement gilts. Methods: Twenty-four PEDV-naïve replacement gilts were randomly assigned to four groups: unvaccinated control, inactivated vaccine administered twice (K/K), live-attenuated vaccine administered twice (L/L), and live-attenuated priming followed by an inactivated booster (L/K). Pigs received two intramuscular vaccinations at 16 weeks of age and two weeks later. Serum samples collected up to 42 days post-vaccination were analyzed for PEDV-specific IgG and IgA antibodies by ELISA, and serum-neutralizing antibody titers were determined using a serum neutralization test. Results: The L/K regimen induced the highest PEDV-specific IgG responses, with peak levels at day 28 post-vaccination that were significantly higher than those in the K/K and control groups. Serum-neutralizing antibody titers were significantly higher in the L/K and L/L groups than in the K/K and control groups. Serum IgA responses were low and transient across all vaccination groups. Conclusions: A heterologous prime-boost vaccination strategy using a live-attenuated PEDV vaccine followed by an inactivated booster induces strong systemic humoral immune responses in replacement gilts and represents a promising approach for PEDV vaccination programs. Full article

Senecavirus A (SVA) is a picornavirus that causes vesicular disease in swine and has been associated with increased neonatal mortality. Although SVA had only been detected sporadically in the United States since the 1980s, there was a sharp increase in cases in the United States and around the world starting in 2015. The cause of this shift in SVA epidemiology remains unknown; however, changes in the virus that have resulted in enhanced infectivity may have contributed. The aim of this research was to establish the infectious dose of a post-2015 SVA isolate in neonatal pigs and compare its infectivity to previous work with a 2011 SVA isolate. A 2018 SVA isolate (SVA/KS/2018) was serially 10-fold diluted to generate six inoculums. Animals were individually housed with four pigs inoculated with 2 mL orally per dilution. Detection of SVA RNA in serum and swabs, as well as the presence of neutralizing antibodies, were used to classify the infection status of animals. The minimum infectious dose for SVA/KS/2018 in neonates was 10^2.5 TCID₅₀/mL (2 × 10^2.5 or 632 TCID₅₀/pig). This value is similar to the infectious dose determined for SVA/CAN/2011, thus providing evidence that the increase in SVA detections was not due to increases in infectivity of contemporary isolates. Neonatal mortality has not been experimentally reproduced; however, pigs inoculated with higher doses of SVA/KS/2018 developed diarrhea and mortality, suggesting increased virulence, which should be investigated further. Full article

Introduction: Gestational hypoxia (GH) increases the risk of cardiovascular diseases by inducing oxidative stress and vascular dysfunction. This study investigates whether prenatal melatonin can mitigate these effects in guinea pigs. Methods: Pregnant guinea pigs were exposed to normoxia or hypoxia and treated with melatonin (1 mg/kg/day). Echocardiography, vascular reactivity, and molecular assays were used to assess cardiovascular structure, function, and redox balance in neonates. Results: GH reduced neonatal birth weight and altered left ventricular (LV) development, resulting in increased LV systolic function and aortic blood flow velocity. Melatonin treatment reversed these effects, restoring endothelial-dependent vasodilation and decreasing oxidative stress in the LV and thoracic aorta. Catalase antioxidant enzyme activity was elevated in melatonin-treated hypoxic neonates. Unexpectedly, melatonin treatment altered cardiac structure in normoxic pregnancies, increasing LV length and decreasing LV myocardial nuclei density. Conclusions: Prenatal melatonin partially modulates GH-induced endothelial dysfunction and oxidative stress, offering potential therapeutic value. However, its effects under normoxic conditions deserve caution, emphasizing the need for targeted use only in pregnancies with evident hypoxic and oxidative stress conditions. Full article

Bump feeding is a nutritional management strategy in swine production that involves increasing feed allowance and/or dietary nutrient density during the final weeks of gestation, usually from day 90 to farrowing, to support rapid fetal growth and prepare sows for lactation. This strategy is widely applied to improve piglet birth weight, neonatal viability, and subsequent reproductive performance. This review synthesizes current evidence on the effects of increased maternal feed intake during late gestation on sow body condition and feeding-related behavioral responses, and farrowing outcomes. Available studies suggest that increasing feed allowance during late gestation can influence litter characteristics, piglet survival at birth, and sow energy reserves, as reflected by changes in backfat thickness (BFT) and body condition score (BCS). The nutritional composition of bump-feeding diets, including dietary energy and amino acid balance, is critically evaluated in relation to pregnancy maintenance, farrowing duration, and early lactation performance. In addition, the roles of parity and feeding behavior during late gestation are examined, with particular emphasis on their associations with sow activity patterns, restlessness around parturition, and farrowing efficiency. Despite these reported effects, findings across studies remain inconsistent, particularly regarding the balance between improved reproductive outcomes and the risk of excessive fat deposition in sows. This review highlights key knowledge gaps and underscores the need for optimized, parity-specific bump-feeding strategies that integrate nutritional management with feeding behavior to enhance farrowing performance, piglet survival, sow welfare, and economic sustainability in modern pig production. Full article

Porcine epidemic diarrhea virus (PEDV) is an enteric alphacoronavirus that causes severe diarrhea and high mortality in neonatal pigs, leading to substantial economic loss in the porcine industry. Previous studies have primarily focused on the spike protein because of its role in viral entry and induction of neutralizing antibody responses. However, accumulating evidence indicates that other viral components also contribute to host immune modulation and pathogenesis. This review summarizes the current knowledge on PEDV structural proteins, with an emphasis on membrane proteins as regulators of porcine innate immune responses. The molecular characteristics and intracellular localization of membrane proteins were described, and the reported effects on interferon signaling, inflammatory pathways, and cellular stress responses were examined. Findings from related coronaviruses were incorporated to highlight the conserved features and virus-specific differences in membrane protein-mediated host modulation. Available evidence suggests that membrane protein-associated interference with innate immune signaling may contribute to intestinal immune dysregulation and disease severity in neonatal piglets. The implications of these observations on PEDV pathogenesis and intervention strategies are also discussed. By shifting attention from spike-centered frameworks to structural protein-driven host interactions, this review highlights membrane proteins as an underexplored but biologically relevant factor in porcine coronavirus research. Full article

Epileptic seizures in the neonatal brain induce oxidative stress and disrupt the blood–brain barrier (BBB), leading to long-term cerebrovascular and neurodevelopmental deficits. This study examined the protective effects of selective head cooling and NADPH oxidase (NOX) inhibition on BBB integrity following seizures. Neonatal seizures were induced in newborn pigs with bicuculline under normothermic or selective head cooling conditions. BBB disruption was assessed by Evans Blue extravasation and quantification of circulating brain-derived endothelial cells (CD45⁻/CD146⁺/GluT1⁺). Seizures under normothermia caused marked BBB leakage, cerebrovascular apoptosis, and elevated endothelial biomarkers, whereas selective head cooling (cortical temperature ~25 °C, body ~35 °C) significantly reduced these effects. Pharmacological inhibition of NOX with setanaxib (5 mg/kg) or sulforaphane (0.4 mg/kg) also prevented BBB disruption during normothermia. In vitro, primary porcine and human brain endothelial cells exposed to glutamate or TNF-α showed increased NOX activity, ROS production, apoptosis, and barrier leakage, all attenuated by NOX inhibitors or moderate hypothermia (<30 °C). These findings identify endothelial NOX as a key mediator of seizure-induced BBB injury and demonstrate that both NOX inhibition and selective head cooling effectively preserve cerebrovascular integrity. Combined hypothermic and antioxidant therapy may offer a promising strategy to prevent cerebrovascular injury and BBB damage in neonatal epilepsy. Full article

(1) Background: How to reduce respiratory diseases in animals is a matter of great concern, and this study aimed to investigate the effect of maternal administration with all-trans retinoic acid (ATRA) on the lung health of neonatal pigs. (2) Methods: Fifteen sows were randomly allotted to one of five ATRA groups (0, 4, 8, 16, and 32 mg/kg diet), respectively; on average, from days 12 to 95 after insemination, lung samples of neonatal pigs were harvested for HE staining, 16S DNA sequencing, and transcriptomic analysis. (3) Results: Compared to neonatal pigs from the ATRA0 group, neonatal pigs from the ATRA4 group had an increased (p > 0.05) alveolar number and the thinner (p < 0.05) alveolar septum, higher (p < 0.05) abundance of Akkermansia and unidentified_Mitochondria, and a lower (p < 0.05) abundance of Acinetobacter, Cutibacterium, Stenotrophomonas, Enterobacter, Saccharomonospora, and Alistipes in the lungs. Maternal supplementation of ATRA at 4 mg/kg diet had the benefits of reducing pathogen virulence and drug resistance, but it poses the risk of increasing the resistance to β-lactam antibiotics. In addition, it reduced (p < 0.05) the virulence, drug resistance, and stress tolerant capacity of potential pathogens (Proteobacteria, Acinetobacter, Cupriavidus, and Pseudomonas), enriched the highest number of genes in neuroactive ligand–receptor interaction pathway, and decreased gene expression of CCL11 and IL9 in the asthma pathway. (4) Conclusions: Maternal supplementation of ATRA at 4 mg/kg diet can strengthen the lung health of neonatal pigs by improving alveolar development, decreasing the number and virulence of pathogens, and down-regulating the expression of asthma-related genes. Full article

Neonatal hypoxic–ischemic encephalopathy (HIE) remains a major cause of neonatal mortality and long-term disability, despite therapeutic hypothermia (TH) treatment, underscoring the need for further preclinical research. In the present study, we compared the neuroprotection afforded by TH and inhaled molecular hydrogen (H₂) treatment in a translational newborn pig HIE model. Following 20 min of asphyxia induced by a hypoxic/hypercapnic gas mixture, piglets were reoxygenated and monitored for 48 h. Animals were randomly assigned to normothermia, continuous H₂ ventilation (2.1%), or TH (33.5 °C for 37 h followed by slow rewarming) groups. Physiological parameters, electroencephalography (EEG), visual evoked potentials (VEPs), and neuropathology were assessed. TH eliminated post-asphyxia seizures and improved VEP latency, while H₂ delayed seizure onset and increased quantitative EEG markers of signal complexity. Neuropathology revealed severe thalamic injury in normothermic controls, which was significantly attenuated by both H₂ and TH, while neocortical, hippocampal, and basal ganglia injury was less extensive and not significantly altered by either of the neuroprotective interventions. These findings demonstrate that continuous H₂ inhalation provides neuroprotection in HIE comparable to TH, particularly in the thalamus. H₂ also exerts distinct electrophysiological effects, suggesting its therapeutic potential as a treatment for neonatal HIE. Full article

Background/Objectives: Respiratory syncytial virus (RSV) causes severe infection in neonates and infants. However, a suitable RSV vaccine for children is yet to be approved. The development of KD-409 is focused on creating an effective and safe RSV vaccine for newborns and children. The safety and efficacy of the RSV FG chimeric protein KD-409 were evaluated in several rodent models. Methods/Results: The effect of vaccine-induced antibody transfer was verified in a guinea pig model. Next, the exacerbation of infection was evaluated in a BALB/c mouse model of passive immunity designed to mimic the vaccination of pregnant women. KD-409 did not exacerbate infection when administered with alum, unlike pre-F with alum. Our active immunization model of BALB/c mice, which involved stimulating vaccination with a pediatric vaccine, suggested that KD-409 with alum was less likely to exacerbate inflammation than FI-RSV or pre-F with alum. The efficacy was evaluated in a cotton rat model, in which KD-409 demonstrated greater protection against infection than pre-F without adjuvant, the only currently approved formulation for immunizing pregnant women. Conclusions: KD-409 eliminated concerns about vaccine-enhanced disease in pediatric vaccination and demonstrated superior efficacy to current vaccines in rodent models. The safety in mice during passive and active immunization, and efficacy in cotton rats demonstrate the high potential of KD-409 as a safe and effective next-generation RSV vaccine candidate that can cover the neonatal-to-pediatric age range. Full article

Porcine epidemic diarrhea (PED), a severe and highly contagious disease induced by porcine epidemic diarrhea virus (PEDV), impacts pigs across all age groups but has a particularly high lethality in neonatal piglets, with mortality rates reaching 80 to 100%, leading to substantial economic losses in the swine industry. In this investigation, 128 intestinal samples obtained from 65 large-scale pig farms in eight prefectures of Guangdong Province were screened by RT-qPCR between 2022 and 2024. Of these, 50 samples (39.06%) tested positive for Porcine Epidemic Diarrhea Virus (PEDV). The complete S1 genes of 31 representative strains were sequenced. Phylogenetic analysis revealed G2c as the exclusive dominant lineage (29/31, 93.6%), with single representatives of G2a and G2d. Nucleotide identity among the local strains ranged from 88.9 to 100% and 88.1 to 93.5% to prototype CV777 and from 91.2 to 99.1% to vaccine strain AJ1102. The COE neutralizing epitope (aa 499–638) carried 26 substitutions versus AJ1102; T499I/S, A520S/L, F539L, K566N and F615L were most prevalent. The SS2 epitope was fully conserved, whereas SS6 showed three low-frequency changes (S766P, S769F, G770V). Six distinct N-glycosylation patterns were identified relative to AJ1102. The predominance of G2c, accompanied by marked epitope drift and altered glycosylation, indicates the need for further investigation into vaccine efficacy. Continuous surveillance and the careful evaluation of G2c-based vaccine candidates are warranted. Full article

Objectives: This study utilized IPEC-J2, a neonatal pig jejunum-derived cell line, to assess how iron deficiency (ID) and excess (IE) alter enterocyte metabolism and the transcription of inflammatory markers. Methods: Cells were treated with deferiprone (DFP) or ferric ammonium citrate (FAC) to induce ID or IE, respectively. The study evaluated: (1) transcriptional changes in iron-regulatory genes over 96 h under ID or IE; (2) the interaction between iron imbalance and lipopolysaccharide (LPS) exposure on mRNA expression of inflammation markers and iron transporters; and (3) cellular metabolic responses to ID, IE, and iron repletion using untargeted metabolomics. Results: ID triggered dynamic transcriptional changes in iron regulatory genes and suppressed cellular proliferation via impaired DNA replication. IE resulted in a persistent reduction in TFRC expression. LPS increased CYBRD1 (p < 0.001) and IL8 (p = 0.004) and tended to elevate TLR4 and TNF expression (p ≤ 0.07), while iron deficiency upregulated IL8 expression (p < 0.001). ID disrupted the TCA cycle, reduced glucuronic acid synthesis, and elevated glycolysis for energy production, whereas IE increased cholesterol biosynthesis and decreased alpha-tocopherol levels. Repletion of iron partially reversed ID-induced metabolic changes. Conclusions: ID impaired enterocyte proliferation and profoundly disrupted cellular metabolism, whereas IE enhanced cholesterol synthesis and depleted alpha-tocopherol levels. Restoration of cellular metabolism following iron repletion was observed, highlighting the resilience of enterocytes. Full article

Despite lower birth weight, Meishan piglets exhibit a notably higher pre-weaning survival rate compared to Western commercial breeds. This study aimed to evaluate the effect of low birth weight (LBW) on intestinal barrier function in Meishan neonates. Six pairs of neonatal piglets (one normal birth weight, NBW: 0.85 ± 0.06 kg; one LBW: 0.65 ± 0.02 kg) from the same sow were euthanized at birth prior to suckling. Morphological parameters, goblet cell density, antioxidant enzyme activities, cytokine gene expression, and tight junction protein levels in the small intestine (SI) were assessed. Results showed that LBW piglets had a significantly higher SI length-to-body weight ratio (p < 0.05), along with reduced villus height, villus height-to-crypt depth ratio, and villus surface area in the jejunum and ileum (p < 0.01). Notably, microvillus structure remained intact despite the presence of mitochondrial swelling. LBW piglets also exhibited decreased goblet cell numbers, lower antioxidant capacity, dysregulated expression of cytokines (CD8, IFNγ, IL4, IL2), and reduced levels of mucin 2, ZO-1, and occludin (p < 0.05). In conclusion, although LBW Meishan piglets showed impairments in multiple aspects of intestinal barrier function, the structural integrity of the microvillus was preserved, which may contribute to their higher survival rate and represents a key adaptive advantage over commercial pig breeds. Full article