Newborn Piglets Research Articles

Novel Porcine Getah Virus from Diarrheal Piglets in Jiangxi Province, China: Prevalence, Genome Sequence, and Pathogenicity.

Getah virus (GETV) is a mosquito-borne virus belonging to the genus Alphavirus in the family Togaviridae. Its infection poses an increasing threat to animals and public health in China. In this study, an epidemiological survey of GETV on 46 pig farms in Jiangxi Province, China, was performed; GETV isolation and characterization were carried out, including a complete sequence determination and phylogenetic analysis; and pathogenicity of the GETV was experimentally investigated by inoculating newborn piglets with the isolated GETV strain. Epidemiological studies conducted on the organs of infected pigs, aborted piglets, and the blood of aborted sows sampled from pig farms in Jiangxi Province, China, demonstrated that 44 out of the 46 pig farms were positive for GETV, which is a positivity rate of 95.65% (44/46). Of the 411 samples tested, 47.93% (197/411) were found positive for GETV. A GETV strain called GETV-JX-CHN-22 was obtained, which showed stable proliferation in Vero cells. One-step growth curve results showed that the GETV-JX-CHN-22-P7 (passage 7) isolate reached a peak titer of 108.3 TCID50/mL at 24 hpi. An analysis of the whole-genome sequencing results showed that GETV-JX-CHN-22 (prototype) and GETV-JX-CHN-22-P7 shared nucleotide sequence similarities of 95.3% to 99.6% with 73 reference strains of GETV in GenBank. Genetic evolution analysis revealed that GETV-JX-CHN-22 and GETV-JX-CHN-22-P7 belonged to the GIII group, the same group members of most strains were reported in China. Animal inoculation experiments indicated that piglets exhibited typical symptoms and pathological changes of GETV infection after 24 h inoculation, which reproduced the pathogenicity of GETV field strain infections in piglets. To our knowledge, this study is the first report on the detection and isolation of porcine GETV associated with diarrhea from pig farms in Jiangxi Province, China. It is of great importance to study the infection spectrum, transmission mechanism, and public health significance of GETV. The results provide foundations for the genomic and biological (pathogenic) characteristics of the circulating GETV in Jiangxi Province, China.

Effects of organic and inorganic selenium mixes in pregnant sows on piglet growth, selenium levels in serum and milk, and selenium deposition in newborn piglet tissues.

This study was conducted to evaluate the effects of organic and inorganic selenium mixes in pregnant sows on piglet growth, selenium levels in serum and milk, and selenium deposition in newborn piglet tissues. A total of 44 multiparous sows (Yorkshire×Landrace) with average body weight (BW), backfat thickness, and parity were assigned to one of the three treatments with 14 or 15 sows per treatment in a completely randomized design. The treatments were as follows: i) Control, corn-soybean meal-based diet with no addition of selenium premix; ii) ISOS(mixed inorganic selenium and organic selenium) 30, a basal diet supplemented with 0.15 ppm of inorganic Se and 0.15 ppm of organic Se; iii) ISOS50, a basal diet supplemented with 0.25 ppm of inorganic Se and 0.25 ppm of organic Se. At day 21 of lactation, supplementing a high level of mixed Se at 0.50 ppm resulted in higher piglet BW and weight gain than adding a low level of mixed Se at 0.30 ppm (p<0.05). Selenium concentration of colostrum in sows fed ISOS50 diet was significantly higher than those in sows fed ISOS30 diet (p<0.05). Selenium concentrations in the serum at days 90 and 110 of gestation and 24 hours postpartum were highest when sows were fed ISOS50 diet (p<0.05). Additionally, increasing levels of mixed Se led to an increase in piglet serum Se concentration at 24 hours postpartum (p<0.05). Before ingesting colostrum, piglets from sows fed a mixed selenium (Se) diet had significantly higher kidney Se concentrations compared to those from the control group, with the ISOS50 treatment showing the most significant difference (p<0.05). Supplementation of the gestation diet with 0.5 ppm of mixed Se may improve piglet growth performance, increase Se concentrations in milk, and enhance Se status in the serum of sows, as well as in the serum and tissues of their offspring.

Towards automatic farrowing monitoring-A Noisy Student approach for improving detection performance of newborn piglets.

Nowadays, video monitoring of farrowing and automatic video evaluation using Deep Learning have become increasingly important in farm animal science research and open up new possibilities for addressing specific research questions like the determination of husbandry relevant indicators. A robust detection performance of newborn piglets is essential for reliably monitoring the farrowing process and to access important information about the welfare status of the sow and piglets. Although object detection algorithms are increasingly being used in various scenarios in the field of livestock farming, their usability for detecting newborn piglets has so far been limited. Challenges such as frequent animal occlusions, high overlapping rates or strong heterogeneous animal postures increase the complexity and place new demands on the detection model. Typically, new data is manually annotated to improve model performance, but the annotation effort is expensive and time-consuming. To address this problem, we propose a Noisy Student approach to automatically generate annotation information and train an improved piglet detection model. By using a teacher-student model relationship we transform the image structure and generate pseudo-labels for the object classes piglet and tail. As a result, we improve the initial detection performance of the teacher model from 0.561, 0.838, 0.672 to 0.901, 0.944, 0.922 for the performance metrics Recall, Precision and F1-score, respectively. The results of this study can be used in two ways. Firstly, the results contribute directly to the improvement of piglet detection in the context of birth monitoring systems and the evaluation of the farrowing progress. Secondly, the approach presented can be transferred to other research questions and species, thereby reducing the problem of cost-intensive annotation processes and increase training efficiency. In addition, we provide a unique dataset for the detection and evaluation of newborn piglets and sow body parts to support researchers in the task of monitoring the farrowing process.

No Short-Term Effect of Low-Dose Nicotine on Inflammation after Global Hypoxia in Newborn Piglets

Introduction: Perinatal asphyxia initiates cytokine release and complement activation with risk of brain damage. We assessed the effect of nicotine on innate immunity and hypothesized that nicotine infusion in a newborn piglet model of asphyxia would decrease the immune response and be neuroprotective. Methods: Newborn piglets (n = 41) were randomized to one of three groups after hypoxia: two groups receiving nicotine, (1) 18 µg/kg/h (n = 17), (2) 46 µg/kg/h (n = 15), and (3) control group receiving saline (n = 9). C3a, IL-6, TNF, and IL-10 were measured in plasma and IL-6 and IL-8 in microdialysis fluid from cerebral periventricular white matter, using immuno-assays. Results: Plasma C3a and IL-6 increased significantly from start to end hypoxia (mean 4.4 ± 0.55 to 5.6 ± 0.71 ng/mL and 1.66 ± 1.04 to 2.68 ± 0.71 pg/mL, respectively), while IL-10 and TNF increased significantly after 4 h (mean 1.4 ± 1.08 to 2.9 ± 1.87 and 3.3 ± 0.67 to 4.0 ± 0.58 pg/mL, respectively) (p < 0.001 for all). IL-6 increased significantly (p < 0.001) in microdialysis samples from end hypoxia to end experiment (mean 0.65 ± 0.88 to 2.78 ± 1.84 ng/mL). No significant differences were observed between the nicotine groups and the control group neither in plasma nor in microdialysis samples. Conclusion: Hypoxia leads to rapid release of cytokines in plasma and cerebral microdialysis fluid, and complement activation measured on C3a. However, low-dose nicotine administration did not affect the immune response.

Deletion of maternal CD163 PSTII-domain-coding exon 13 protects fetuses from infection with porcine reproductive and respiratory syndrome virus (PRRSV)

Following infection of a porcine dam with PRRSV around 90 days of gestation, the virus crosses the placenta and starts to infect fetuses. This can lead to consequences such as abortions, stillbirths, and respiratory issues in newborn piglets. CD163 is an essential cellular viral entry receptor for porcine reproductive and respiratory syndrome virus (PRRSV). CD163 contains nine scavenger receptor cysteine-rich (SRCR) and two proline-serine-threonine (PST) domains. Gene-edited pigs possessing a complete deletion of CD163 are resistant to PRRSV infection. Recently, we demonstrated that pigs harboring a clean deletion of CD163 exon 13 (ΔExon13 CD163 pigs) which encodes the first 12 amino acids of the CD163 PSTII domain were not susceptible to PRRSV infection. In this study, ΔExon13 CD163 (−/−) gilts were bred with wildtype CD163 (+/+) boars producing heterozygous, CD163 (+/−) fetuses. We found that fetuses with a wildtype CD163, recovered between day 103 of gestation or 17 days after the maternal infection with PRRSV, were fully protected from PRRSV in dams containing a clean deletion of CD163 exon 13. These findings suggest a feasible approach for eliminating PRRSV-related reproductive illness, which is a significant cause of economic losses in agriculture.

Endoplasmic Reticulum Stress Contributes to Intestinal Injury in Intrauterine Growth Restriction Newborn Piglets.

Intrauterine growth retardation (IUGR) in piglets is associated with a high rate of morbidity and mortality after birth due to gut dysfunction, and the underlying mechanisms remain poorly understood. This study selected six pairs of IUGR newborn male piglets and normal birth weight newborn piglets (Large White × Landrace) to investigate differences in intestinal structure and digestive functions, intestinal ERS and apoptosis, intestinal barrier function, and inflammatory response. The results showed that IUGR significantly reduced the jejunal villi height (p < 0.05) and the ratio of villus-height-to-crypt-depth (p = 0.05) in neonatal piglets. Additionally, the microvilli in the jejunum of IUGR neonatal piglets were shorter than those in normal-weight piglets, and swelling of the mitochondria and expansion of the endoplasmic reticulum were observed. IUGR also significantly reduced serum glucose and lactase levels (p < 0.05) while significantly increasing mRNA levels of jejunal IRE1α, EIF2α, CHOP, Bax, Caspase9, Mucin2, Claudin-1, Occludin, ZO-1, Bcl-2, IL-6, and IFN-γ (p < 0.05), as well as GRP78 protein levels in neonatal piglets (p < 0.05). These findings suggest that IUGR impairs intestinal structure and barrier function in newborn piglets by enhancing intestinal inflammatory responses, activating intestinal ERS and the signaling pathways related to the unfolded protein response, thereby inducing ERS-related apoptosis.

108 Early colostrum uptake by the newborn requires prepartum initiation of lactogenesis in ungulate species - Sponsored by EAAP

Abstract Due to anatomical characteristics of ungulate placenta types, immunoglobulins (IG) cannot be transferred via the placenta from the maternal to the fetal circulation during pregnancy. Therefore, passive immunization of the newborn ungulate immediately after birth is crucial for its survival. Colostrum, the first milk produced contains immunoglobulins (IG) as well as greater concentrations of several nutrients and bioactive components to provide specific immunity for the first weeks of life as well as an activation of the digestive tract and metabolism through colostral hormones, growth factors, vitamins, and special nutrients such as oligosaccharides. The uptake of colostrum by newborn calves, piglets, or foals within the first few hours after birth requires the availability of colostrum in a sufficiently liquid form. The formation of pre-colostrum starts days or even weeks before parturition, which allows concentrations of IG and other factors far beyond the concentrations in maternal blood circulation. Pre-colostrum components are partially secreted into the alveolar lumen and partially stored in the lactocytes of the mammary gland. These secretions are characterized by increased viscosity and density, which are difficult to harvest by the newborn. Through lactogenesis the secretion of milk components such as lactose is initiated, which causes a considerable water influx into the mammary secretion through an osmotic gradient between maternal blood and mammary secretion. The typical high concentrations of gestagens, mostly progesterone, inhibit the initiation of lactogenesis during pregnancy. A declining gestagen activity is the key signal to initiate a lactogenic prolactin increase and hence milk secretion. Mechanisms and timing of gestagen withdrawal differ considerably among mammalian species. Most species initiate both parturition and lactation through luteolysis (cattle, goat, pig, cat, dog, rabbit, mouse rat). In some species the placenta takes over the main portion of gestagen production (human, sheep, horse). In human, IG are transferred to the fetus during pregnancy, and the start of colostrum secretion only several days after parturition is no problem. In sheep and horses, gestagen secretion during late pregnancy is provided by the placenta. Both of these species are ungulates, and a delayed start of lactogenesis would be critical for the survival of the newborn. Therefore, the placental steroid secretion is subject to characteristic changes in both species towards the end of pregnancy to allow a timely availability of colostrum immediately after birth. At the end of pregnancy, the placenta of sheep produces estrogens at the expense of progesterone, and in horses the less potent dihydroprogesterone is produced instead of progesterone. A considerable variation in the resulting colostrum composition is likely due to the existing wide range of the interval between the commencement of lactogenesis and parturition or due to local regulatory effects at the level of individual glands.

Can immunocrit be used as a monitoring tool for swine vaccination and infection studies?

BackgroundThe immunocrit is a cost-effective and straightforward technique traditionally used to assess passive immunity transfer to newborn piglets. However, it has not been previously used for monitoring the effect of vaccination and/or infections. Therefore, this study aimed to evaluate the usefulness of the immunocrit technique as an immunological monitoring tool in a vaccination and challenge scenario, using porcine circovirus 2 (PCV-2) as pathogen model. The immunocrit ratio was monitored in PCV-2 vaccinated (V) and non-vaccinated (NV) 3-week-old piglets (study day 0, SD0) that were subsequently challenged with this virus at SD21 and followed up to SD42. Additional techniques (PCV-2 IgG ELISA, optical refractometry, and proteinogram) were performed to further characterize the results of the immunocrit analysis.ResultsImmunocrit, γ-globulin concentration and PCV-2 S/P values followed similar dynamics: descending after PCV-2 vaccination but ascending after an experimental PCV-2 inoculation. However, statistically significant differences between V and NV animals were only found with the PCV-2 ELISA. In this case, V animals had significantly higher (p < 0.05) S/P values (S/P ratio = 0.74) than NV (S/P ratio = 0.39) pigs only after challenge at SD42. On the other hand, serum total protein obtained by refractometer (STPr) were maintained from SD0 to SD21 and increased in both groups from SD21 to SD42. Correlations between techniques were low to moderate, being the most robust ones found between immunocrit and optical refractometry (ρ = 0.41) and immunocrit with γ-globulins (ρ = 0.39). In a subset of sera, the proteinogram technique was applied to the whole serum and the supernatant of the immunocrit, with the objective to characterize indirectly the immunocrit fraction. The latter one included all protein types detectable through the proteinogram, with percentages varying between 64.3% (γ-globulins) and 82% (β-globulins).ConclusionThe immunocrit technique represented a fraction of the total serum proteins, with low to moderate correlation with all the complementary techniques measured in this study. Its determination at different time points did not allow monitoring the effect of vaccination and/or infection using PCV-2 as a pathogen model.

Molecular and phylogenetic analysis of transmissible gastroenteritis virus strain VET-16, isolated from piglets in Vietnam.

Porcine transmissible gastroenteritis virus (TGEV) is a major pathogen that causes viral enteritis and severe diarrhea in newborn piglets. TGEV strains have been isolated in the USA, Europe, and China, and their molecular characteristics are well known. However, there have been few reports of molecular analysis of TGEV strains isolated in Southeast Asia. In 2016, we isolated TGEV strain VET-16 from fecal samples collected from piglets in Vietnam and determined its complete genome sequence by Sanger sequencing. We found that, while the full genome of the VET-16 strain was 92.4-99.9% identical to those of other TGEV strains, the ORF3 gene showed very little sequence similarity. Phylogenetic analysis suggested that the VET-16 strain belongs to the Purdue subgroup. Comparison of the predicted amino acid (aa) sequence of the spike protein of strain VET-16 with those of other TGEV strains revealed three aa substitutions (V378L, S379T, and D380N) and a 3-aa insertion (F383_F387insWEK) in antigenic site D of the VET-16 strain. Also, a single aa deletion (∆F1413) was found in the transmembrane domain of the spike gene of VET-16. Like the ORF3 gene from the TGEV Miller M60 vaccine strain, the VET-16 strain has a large deletion (∆725 nt) in the ORF3 gene. Previous studies have suggested that these mutations in the spike and ORF3 genes might be associated with a reduction in pathogenicity. The data from this study will facilitate further genetic analysis and research into the evolution of TGEV in pigs in Vietnam.

Mucociliary clearance is impaired in small airways of cystic fibrosis pigs.

Cystic fibrosis (CF) is a genetic disorder characterized by recurrent airway infections, inflammation, impaired mucociliary clearance, and progressive decline in lung function. The disease may start in the small airways; however, this is difficult to prove due to the limited accessibility of the small airways with the current single-photon mucociliary clearance assay. Here, we developed a dynamic positron emission tomography assay with high spatial and temporal resolution. We tested that mucociliary clearance is abnormal in the small airways of newborn cystic fibrosis pigs. Clearance of [68Ga]-tagged macroaggregated albumin from small airways started immediately after delivery and continued for the duration of the study. Initial clearance was fast but slowed down a few minutes after delivery. Cystic fibrosis pigs' small airways cleared significantly less than non-CF pigs' small airways (non-CF 25.1 ± 3.1% vs. CF 14.6 ± 0.1%). Stimulation of the cystic fibrosis airways with the purinergic secretagogue uridine-5'-triphosphate (UTP) further impaired clearance (non-CF with UTP 20.9 ± 0.3% vs. CF with UTP 13.0 ± 1.8%). None of the cystic fibrosis pigs treated with UTP (n = 6) cleared more than 20% of the delivered dose. These data indicate that mucociliary clearance in the small airways is fast and can easily be missed if the assay is not sensitive enough. The data also indicate that mucociliary clearance is impaired in the small airways of cystic fibrosis pigs. This defect is exacerbated by stimulation of mucus secretions with purinergic agonists.NEW & NOTEWORTHY We developed a novel positron emission tomography scan assay with unprecedented temporal and spatial resolution to measure mucociliary clearance in the small airways. We proved a long-standing but unproven assertion that mucociliary clearance is inherently abnormal in the small airways of newborn cystic fibrosis piglets that are otherwise free of infection or inflammation. This technique can be easily extended to other airway diseases such as asthma, idiopathic pulmonary fibrosis, or chronic obstructive pulmonary disease.

Comparison of the Fatty Acid Profiles of Sow and Goat Colostrum.

Currently, the utilization of hyperprolific sows has stimulated the search for supplements aimed at enhancing piglet survival, as these sows yield more offspring than they can adequately feed with their colostrum production. In contrast, intensive goat farming often yields surplus colostrum, thus necessitating its removal, since kids are exclusively fed colostrum through lactation solely within the initial day of birth. The objective of this study was to examine and compare the fatty acid (FA) profiles of colostrum from sows and goats, together with possible influencing factors such as sow parity and the postpartum day of the goat, for possible use as an energy supplement for neonatal piglets. Swine colostrum was collected from sows with a 0-5 parity. In addition, samples of goat colostrum were collected on their first (D1) and second (D2) days of postpartum milking. The FA profiles of the colostrum were analyzed via gas chromatography. The parity value of the sows did not affect (p > 0.05) the FA colostrum composition. High proportions of palmitic, oleic, and linoleic acids were found in both types of colostrum. Levels of palmitic, oleic, and linoleic acids were significantly higher in D1 goat colostrum, whereas saturated FAs of less than 14 carbons (4:0, 6:0, 8:0, 10:0, and 12:0) were found in higher proportions in D2. These FAs play an important role in colostrum as they are a readily available source of energy and have also been attributed strong antibacterial activity. Therefore, goat colostrum, especially D2, could be used as an alternative energy supplement for newborn piglets, in particular for the weakest and smallest of the litter, which are the most in need.

Lipopolysaccharide induced systemic inflammation and heart rate variability in a term newborn piglet model.

Early biomarkers are needed to improve diagnosis and support antibiotic stewardship in neonatal sepsis. Heart rate variability (HRV) is proposed as such a biomarker. However, there is a lack of studies in term newborns. Infusion of lipopolysaccharide (LPS) from Escherichia coli induces systemic inflammation comparable to sepsis in newborns. We aimed to study the effect of systemic LPS induced inflammation on HRV in term newborn piglets. Baseline HRV was recorded for 1 h. This control period was compared to the hourly HRV for each piglet (n = 9) during 4 h of LPS infusion. For comparison, we used a mixed-effects regression model. Systemic inflammation induced by LPS was found to reduce HRV. Compared to baseline, most measures of HRV decreased to lower values compared to baseline at 2 h, 3 h, and 4 h after initiation of LPS infusion. Heart rate (HR) was increased at 2 h, 3 h, and 4 h. When adjusting for HR in the mixed-effects regression model all reductions in HRV were explained by the increase in HR. Reduced HRV may be an early biomarker of neonatal sepsis. However, an increase in HR alone could be an already available, more accessible, and interpretable biomarker of sepsis in term neonates. In a term newborn piglet model, systemic inflammation induced by lipopolysaccharide from Escherichia coli reduced heart rate variability measures and increased heart rate. All reductions in heart rate variability were mediated by heart rate. While heart rate variability may be a biomarker of sepsis in term newborns, changes in heart rate alone could be a more readily available biomarker.

Colostrum intake and neonatal characteristics in piglets experiencing varying lengths of expulsion phase

The objective of this study was to investigate how the cumulative expulsion interval, litter size, and birth order influence the characteristics of newborn piglets from hyperprolific sows, as well as their intake of colostrum. Analysis was conducted on the temporal aspects of parturition in 254 sows, including 3920 piglets. Newborn piglet characteristics examined encompassed delivery time, birth order, birth status, umbilical cord rupture, meconium staining, delivery ease, birth weight, blood oxygen saturation, and heart rate. From the raw data, colostrum intake, birth interval, and cumulative birth interval of each piglet were computed. Piglets were sorted based on the cumulative expulsion interval into nine categories: 0–30, 31–60, 61–90, 91–120, 121–150, 151–180, 181–210, 211–240, and >240 min. Litter sizes were categorized into three groups: 10–13, 14–17, and ≥18 piglets per litter. The birth-order ranking referred to the piglet's position within a litter and was categorized into four quartiles: 1, 2, 3, and 4. On average, the expulsion interval, cumulative expulsion interval, and colostrum consumption were 14.8 ± 21.8 min, 138.6 ± 155.7 min, and 407.6 ± 162.1 g, respectively. In litters with ≥18 piglets, the percentage of piglets with inadequate colostrum intake (<300 g) was higher compared to litters with 10–13 piglets (33.1 % vs. 12.9 %, P < 0.001) and 14–17 piglets (23.9 %, P < 0.001). The occurrence of stillbirths in the fourth quartile of the litter was higher than in the first (16.0 % vs. 2.2 %, P < 0.001), second (4.0 %, P < 0.001), and third (7.8 %, P < 0.001) quartiles. Among litters with ≥18 piglets, those born in the third quartile exhibited lower colostrum consumption compared to those born in the first quartile (346.0 ± 9.9 vs. 391.4 ± 8.6 g, P = 0.028). Colostrum intake decreased with longer cumulative expulsion intervals, evident from a notable decline starting at 90 min post-farrowing compared to piglets born within the first 30 min (P = 0.004). Interestingly, colostrum consumption exhibited a negative correlation with the cumulative expulsion interval (P < 0.001), but not with individual piglet expulsion intervals (P = 0.460). Piglets born with meconium staining were most prevalent among those with a cumulative expulsion interval of >240 min. In summary, both the cumulative expulsion interval and birth-order ranking significantly impact the physiological characteristics of newborn piglets and their colostrum intake. Colostrum consumption decreases and newborn piglet characteristics change with higher cumulative expulsion intervals. Identifying piglets at high risk of hypoxia and implementing appropriate treatments and management strategies within an optimal timeframe could aid pig producers in reducing pre-weaning mortality among piglets born to hyperprolific sows.

FlagT4G Vaccine Prevents Transplacental Transmission of Highly Virulent Classical Swine Fever Virus after Single Vaccination in Pregnant Sows.

The transplacental transmission of CSFV and the resulting persistent congenital infection in newborn piglets have been abundantly discussed in pregnant sows suffering from virus infection. Importantly, the availability of safe commercial vaccines with proven efficacy to prevent the generation of congenital and postnatal persistent infections in pregnant sows are critical tools for controlling the disease in CSF endemic areas. Here, we demonstrate the high efficacy of a single dose of the recombinant FlagT4G vaccine to provide solid protection in pregnant sows against transplacental transmission of a highly virulent CSFV. Pregnant sows vaccinated with FlagT4G at 44 days of gestation elicited a strong CSFV-specific antibody response, with neutralizing antibody levels above those required for protection against CSFV. Importantly, after the challenge with a highly virulent CSFV, all foetuses from FlagT4G-vaccinated sows lacked CSF macroscopic lesions and showed a complete absence of the challenge virus in their internal organs at day 79 of gestation. Therefore, pregnant sows safely vaccinated with FlagT4G without affecting reproductive efficacy are efficaciously protected, along with their foetuses, against the infection and disease caused by a CSFV virulent field strain.

Integrated Metagenomic and Metabolomics Profiling Reveals Key Gut Microbiota and Metabolites Associated with Weaning Stress in Piglets.

(1) Background: Weaning is a challenging and stressful event in the pig's life, which disrupts physiological balance and induces oxidative stress. Microbiota play a significant role during the weaning process in piglets. Therefore, this study aimed to investigate key gut microbiota and metabolites associated with weaning stress in piglets. (2) Methods: A total of ten newborn piglet littermates were randomly assigned to two groups: S (suckling normally) and W (weaned at 21 d; all euthanized at 23 d). Specimens of the cecum were dehydrated with ethanol, cleared with xylene, embedded in paraffin, and cut into 4 mm thick serial sections. After deparaffinization, the sections were stained with hematoxylin and eosin (H&E) for morphometric analysis. Cecal metagenomic and liver LC-MS-based metabolomics were employed in this study. Statistical comparisons were performed by a two-tailed Student's t-test, and p < 0.05 indicated statistical significance. (3) Results: The results showed that weaning led to intestinal morphological damage in piglets. The intestinal villi of suckling piglets were intact, closely arranged in an orderly manner, and finger-shaped, with clear contours of columnar epithelial cells. In contrast, the intestines of weaned piglets showed villous atrophy and shedding, as well as mucosal bleeding. Metagenomics and metabolomics analyses showed significant differences in composition and function between suckling and weaned piglets. The W piglets showed a decrease and increase in the relative abundance of Bacteroidetes and Proteobacteria (p < 0.05), respectively. The core cecal flora in W piglets were Campylobacter and Clostridium, while those in S piglets were Prevotella and Lactobacillus. At the phylum level, the relative abundance of Bacteroidetes significantly decreased (p < 0.05) in weaned piglets, while Proteobacteria significantly increased (p < 0.05). Significant inter-group differences were observed in pathways and glycoside hydrolases in databases, such as the KEGG and CAZymes, including fructose and mannose metabolism, salmonella infection, antifolate resistance, GH135, GH16, GH32, and GH84. We identified 757 differential metabolites between the groups through metabolomic analyses-350 upregulated and 407 downregulated (screened in positive ion mode). In negative ion mode, 541 differential metabolites were identified, with 270 upregulated and 271 downregulated. Major differential metabolites included glycerophospholipids, histidine, nitrogen metabolism, glycine, serine, threonine, β-alanine, and primary bile acid biosynthesis. The significant differences in glycine, serine, and threonine metabolites may be potentially related to dysbiosis caused by weaning stress. Taken together, the identification of microbiome and metabolome signatures of suckling and weaned piglets has paved the way for developing health-promoting nutritional strategies, focusing on enhancing bacterial metabolite production in early life stages.

Morphometric traits to estimate brain and liver weight and their ratio for the diagnosis of intrauterine growth restriction in newborn piglets

Intrauterine growth restriction (IUGR) is defined as inadequate foetal growth during gestation. In response to placenta insufficiency, IUGR piglets prioritise brain development as a survival mechanism. This adaptation leads to a higher brain-to-liver weight ratio (BrW/LW) at birth. This study assessed the potential of using morphometric traits to estimate brain (BrW) and liver (LW) weights, enabling non-invasive diagnosis of IUGR in newborn piglets. At birth, body weight (BtW) of individual piglets (n = 144) was recorded. One day (± 1) after birth, BrW and LW were measured with computed tomography (n = 94) or by weighing the organs after natural death or euthanasia (n = 50). Additionally, 20 morphometric traits were captured from images of each piglet and correlated with the BrW and LW. The morphometric traits that showed a r ≥ 0.70 in linear correlation with the BrW or LW were selected. Each selected trait was combined as an independent variable with BtW to develop multiple linear regression models to predict the BrW and LW. Six models were chosen based on the highest adjusted R2 value: three for estimating BrW and three for LW. The dataset was then randomly divided into a training (75% of the data) and a testing (remaining 25%) subsets. Within the training subset, three equations to predict the BrW and three to predict the LW were extrapolated from the six selected models. The equations were then applied to the testing subset. The accuracy of the equations in predicting organ weight was assessed by calculating mean absolute and mean absolute percentage error (MAE and MAPE) between predicted and actual BrW and LW. To predict the BrW/LW, an equation including BtW and the two morphometric traits which better predicted BrW and LW was used. In the testing dataset, the equation combining ear distance and BtW better estimated the BrW. The equation performed with a MAE of 1.95 and a MAPE of 0.06 between the true and estimated weight of the brain. For the liver, the equation combining the abdominal area delimited by a square and BtW displayed the best performance, with a MAE of 9.29 and a MAPE of 0.17 between the true and estimated weight. Finally, the MAE and MAPE between the actual and estimated BrW/LW were 0.14 and 0.17, respectively. These findings suggest that specific morphometric traits can be used to estimate brain and liver weights, facilitating accurate and non-invasive identification of IUGR in newborn piglets.

Butyrate induces higher host transcriptional changes to inhibit porcine epidemic diarrhea virus strain CV777 infection in porcine intestine epithelial cells

Newborn piglets’ health is seriously threatened by the porcine epidemic diarrhea virus (PEDV), which also has a significant effect on the pig industry. The gut microbiota produces butyrate, an abundant metabolite that modulates intestinal function through many methods to improve immunological and intestinal barrier function. The objective of this investigation was to ascertain how elevated butyrate concentrations impacted the host transcriptional profile of PEDV CV777 strain infection. Our findings showed that higher concentrations of butyrate have a stronger inhibitory effect on PEDV CV777 strain infection. According to RNA-seq data, higher concentrations of butyrate induced more significant transcriptional changes in IPEC-J2 cells, and signaling pathways such as PI3K-AKT may play a role in the inhibition of PEDV CV777 strain by high concentrations of butyrate. Ultimately, we offer a theoretical and experimental framework for future research and development of novel approaches to harness butyrate’s antiviral infection properties.

Disrupted Microbiota of Colon Results in Worse Immunity and Metabolism in Low-Birth-Weight Jinhua Newborn Piglets.

The Jinhua pig is well known in China due to its delicious meat. However, because of large litter size, low birth weight always happens. This experiment used this breed as a model to research bacterial evidence leading to growth restriction and provide a possible solution linked to probiotics. In this experiment, the differences in organs indexes, colonic morphology, short chain fatty acid (SCFA) concentrations, microbiome, and transcriptome were detected between piglets in the standard-birth-weight group (SG) and low-birth-weight group (LG) to find potential evidence leading to low birth weight. We found that LG piglets had a lower liver index (p < 0.05), deeper colonic crypt depth (p < 0.05), fewer goblet cells (p < 0.05), and more inflammatory factor infiltration. In addition, differentially expressed genes (DEGs) were mainly enriched in B-cell immunity and glucose metabolism, and LG piglets had lower concentrations of SCFAs, especially butyrate and isobutyrate (p < 0.05). Finally, most of the significantly differentially abundant microbes were fewer in LG piglets, which affected DEG expressions and SCFA concentrations further resulting in worse energy metabolism and immunity. In conclusion, colonic disrupted microbiota may cause worse glucose metabolism, immunity, and SCFA production in LG piglets, and beneficial microbes colonized in SG piglets may benefit these harmful changes.

Probiotic Lactobacillus rhamnosus GG improves insulin sensitivity and offspring survival via modulation of gut microbiota and serum metabolite in a sow model

BackgroundSows commonly experience insulin resistance in late gestation and lactation, causing lower feed intake and milk production, which can lead to higher mortality rates in newborn piglets. The probiotic Lactobacillus rhamnosus GG (LGG) is known to improve insulin resistance. However, whether supplementing LGG can improve insulin sensitivity in sows and enhance lactation performance, particularly the early survival of offspring remains unclear. Hence, we explored the effects and mechanisms of supplementing LGG during late gestation and lactation on sow insulin sensitivity, lactation performance, and offspring survival. In total, 20 sows were randomly allocated to an LGG (n = 10) and control group (n = 10).ResultsIn sows, LGG supplementation significantly improved insulin sensitivity during late gestation and lactation, increased feed intake, milk production and colostrum lactose levels in early lactation, and enhanced newborn piglet survival. Moreover, LGG treatment significantly reshaped the gut microbiota in sows, notably increasing microbiota diversity and enriching the relative abundance of insulin sensitivity-associated probiotics such as Lactobacillus, Bifidobacterium, and Bacteroides. Serum metabolite and amino acid profiling in late-gestation sows also revealed decreased branched-chain amino acid and kynurenine serum levels following LGG supplementation. Further analyses highlighted a correlation between mitigated insulin resistance in late pregnancy and lactation by LGG and gut microbiota reshaping and changes in serum amino acid metabolism. Furthermore, maternal LGG enhanced immunity in newborn piglets, reduced inflammation, and facilitated the establishment of a gut microbiota.ConclusionsWe provide the first evidence that LGG mitigates insulin resistance in sows and enhances offspring survival by modulating the gut microbiota and amino acid metabolism.

Surface display of the COE antigen of porcine epidemic diarrhoea virus on Bacillus subtilis spores.

Porcine epidemic diarrhoea virus (PEDV) infects pigs of all ages by invading small intestine, causing acute diarrhoea, vomiting, and dehydration with high morbidity and mortality among newborn piglets. However, current PEDV vaccines are not effective to protect the pigs from field epidemic strains because of poor mucosal immune response and strain variation. Therefore, it is indispensable to develop a novel oral vaccine based on epidemic strains. Bacillus subtilis spores are attractive delivery vehicles for oral vaccination on account of the safety, high stability, and low cost. In this study, a chimeric gene CotC-Linker-COE (CLE), comprising of the B. subtilis spore coat gene cotC fused to the core neutralizing epitope CO-26 K equivalent (COE) of the epidemic strain PEDV-AJ1102 spike protein gene, was constructed. Then recombinant B. subtilis displaying the CLE on the spore surface was developed by homologous recombination. Mice were immunized by oral route with B. subtilis 168-CLE, B. subtilis 168, or phosphate-buffered saline (PBS) as control. Results showed that the IgG antibodies and cytokine (IL-4, IFN-γ) levels in the B. subtilis 168-CLE group were significantly higher than the control groups. This study demonstrates that B. subtilis 168-CLE can generate specific systemic immune and mucosal immune responses and is a potential vaccine candidate against PEDV infection.