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Abstract Background Rotavirus B (RVB) causes diarrhea in humans and pigs. Although various RVB strains were identified in humans and various animals globally, little is known about the epidemiology RVB infection in Africa. In this study, we attempted to examine the prevalence of RVB infection in pig populations in Zambia. Methods Metagenomic analyses were conducted on pig feces collected in Zambia to detect double stranded RNA viruses, including RVB. To clarify the prevalence of RVB infection in pig populations in Zambia, 147 fecal samples were screened for the RVB detection by RT-qPCR. Full genome sequence of a detected RVB was determined by Sanger sequencing and genetically analyzed. Results The metagenomic analyses revealed that RVB sequence reads and contigs of RVB were detected from one fecal sample collected from pigs in Zambia. RT-qPCR screening detected RVB genomes in 36.7% (54/147) of fecal samples. Among 54 positive samples, 13 were positive in non-diarrheal samples (n = 48, 27.1%) and 41 in diarrheal samples (n = 99, 41.4%). Genetic analyses demonstrated that all the segments of ZP18-18, except for VP4, had high nucleotide sequence identities (80.6–92.6%) with all other known RVB strains detected in pigs. In contrast, the VP4 sequence of ZP18-18 was highly divergent from other RVB strains (

Biosecurity measures, coupled with effective on-farm management, encompass all aspects aimed at preventing the entry of pathogens into pig farms and their subsequent spread within the facility. This study assessed biosecurity practices on commercial pig farms in Serbia by surveying farm veterinarians/managers. To evaluate biosecurity on these farms, a Biocheck online survey (https://biocheckgent.com) was conducted across ten commercial pig farms in Northern Serbia, where the highest concentration of industrial pig production occurs. The average overall biosecurity score was 65.56 ±11.65. External biosecurity scored higher (75.56 ±9.39), whereas internal biosecurity scored lower (54.89 ±16.92). Within external biosecurity, the weakest scores were observed in categories such as feed, water, and equipment supply (47 ±25.48), showing considerable variation among farms, particularly in feed, water, equipment supply (54.22), and farm location (57.65). Regarding internal biosecurity, the lowest scores were noted during the farrowing and suckling period (37.22 ±15.08) and the finishing unit (37.66 ±24.12). This study reveals that biosecurity measures in commercial pig farms in Serbia are slightly below the global average. Given the predominance of extensive production systems in Serbia, urgent improvements in biosecurity practices on industrial farms are essential. Identification and implementation of appropriate biosecurity measures tailored to existing animal production systems are urgently needed.

Abstract Melatonin supplementation during in vitro maturation (IVM) improves porcine oocyte maturation and embryonic development by exerting antioxidative effects. Nevertheless, the mechanism by which melatonin prevents polyspermy after in vitro fertilization (IVF) remains unclear. Here, we examined the effects of melatonin on cytoplasmic maturation and the incidence of polyspermic penetration in porcine oocytes. No statistically significant difference was observed in the rate of first polar body formation between the groups (Control, Melatonin, Melatonin + Luzindole, and Melatonin + 4-P-PDOT). Interestingly, melatonin supplementation significantly improved the cytoplasmic maturation of porcine oocytes by enhancing the normal distribution of organelles (Golgi apparatus, endoplasmic reticulum and mitochondria) and upregulating organelle-related gene expressions (P

This study aims to elucidate the mechanism of lipid metabolism disorder in intrauterine growth retardation (IUGR) pigs and the potential alleviating effects of dimethylglycine sodium salt (DMG-Na). A total of 60 male newborn piglets were selected for this study. Within each litter, one normal birth weight (NBW) male piglet (1.53 ± 0.04 kg) and two IUGR male piglets (0.76 ± 0.06 kg) were chosen based on their birth weight. The piglets were divided into three groups for the study: NBW pigs received a PBS gavage and a common basal diet (NBW-C group), IUGR pigs received the same PBS gavage and common basal diet (IUGR-C group), and IUGR pigs received a 70-mg DMG-Na gavage along with a common basal diet supplemented with 0.1% DMG-Na (IUGR-D group). At 150 d of age, all piglets underwent euthanasia by exsanguination following electrical stunning, after which plasma, liver, and longissimus dorsi (LM) samples were promptly collected. The IUGR-D group demonstrated improvements in plasma parameters (P < 0.05), with lower triglyceride and free fatty acid (FFA) values, and hormone levels (P < 0.05), with lower growth hormone, insulin, and homeostasis model assessment of insulin resistance values. Restoration of lipid metabolism was observed (P < 0.05), with lower triglyceride and FFA, and higher hepatic lipase and total lipase values in the liver, and lower triglyceride and FFA values in the LM. Mitochondrial ETC complexes showed increased levels (P < 0.05), including higher complex III values in the liver, and higher complex I, complex III, and complex V values in the LM. Enhanced levels of energy metabolites were noted (P < 0.05), with higher NAD+, NAD+/NADH, adenosine triphosphate, and mtDNA values, and lower NADH values in the liver and LM. Additionally, meat quality parameters showed improvement (P < 0.05), with higher pH 24 h and a∗ values, and lower drip loss 48 h, L∗, and b∗ values. The expressions of lipid metabolism and mitochondrial function-related genes and proteins were upregulated (P < 0.05) compared to the IUGR-C group. In conclusion, it was indicated that IUGR pigs experienced lipid metabolism disorders and diminished performance. However, supplementation with DMG-Na showed promise in mitigating these adverse physiological effects by safeguarding body tissues and modulating energy metabolism.

The porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) belonging to the Arteriviridae family is the cause of PRRS disease. After being discovered for the first time in the United States in 1987, this illness quickly expanded to Canada. The disease was initially discovered in late 1990 in Germany, from where it quickly spread throughout Europe. The consequences of PRRSV lead to a number of epidemiological issues, including a sickness with a delayed immune response that permits extended viremia, which facilitates viral transmission. The virus penetrates the nasal epithelium, tonsils, lung macrophages, and uterine endometrium through the oronasal and genital pathways. Abortions performed late in pregnancy and premature or delayed deliveries resulting in dead and mummified fetuses, stillborn pigs, and weakly born piglets are indicative of reproductive syndrome. In the meanwhile, dyspnea, fever, anorexia, and lethargic behavior are signs of respiratory syndrome. The virus can be isolated from the tissue or serum of animals that have been infected to confirm the diagnosis. Pig movements and potential airborne dissemination are two ways that the virus can enter new herds and propagate through nose-to-nose contact or aerosols. Various supportive therapies may enhance infant survival, and antibiotics may or may not lessen the impact of secondary bacterial infections. The absence of simple diagnostic tests, the virus's airborne transmission, the occurrence of subclinical infections, and the virus's persistence in infected populations have all contributed to the failure of control efforts for PRRS. [Open Vet J 2024; 14(9.000): 2138-2152]

The significance of pork meat quality extends far beyond mere consumer satisfaction, encompassing crucial aspects such as health and nutrition, economic impact, reputation and branding, food safety, and sustainability within the global food system. Influenced by a multitude of factors, each playing a pivotal role in shaping its sensory attributes and consumer appeal, pork meat quality stands as a cornerstone of the meat industry. Thus, understanding these factors are imperative for ensuring consistent high-quality pork production, aligning with consumer preferences, and elevating overall satisfaction levels. In this review, we provide a comprehensive overview of the diverse factors affecting pork meat quality, including genetic characteristics, rearing systems, feed composition, gender differences, pre-slaughter handling, and meat aging processes.

This study was to investigate effects of different phytogenic feed additives (PFA) in grower finishing pigs with stressed by high stocking density. A total of 84 growing pigs ([Landrace × Yorkshire] × Duroc) with initial body weight (BW) of 28.23 ± 0.21 kg were used for 10 weeks (4 replicate pens with 3 pigs per pen). The dietary treatment consisted of basal diets in animal welfare density (positive control [PC]), basal diet in high stocking density (negative control [NC]), NC + 0.04% bitter citrus extract (PT1), NC + 0.01% microencapsulated blend of thymol & carvacrol (PT2), NC + 0.10% mixture of 40% bitter citrus extract and 10% microencapsulated blend of thymol and carvacrol (PT3), NC + 0.04% premixture of grape seed and grape marc extract, green tea and hops (PT4), and NC + 0.10% fenugreek seed powder (PT5). The reduction of space allowance significantly decreased (p < 0.05) growth performance (average daily gain, average daily feed intake, feed efficiency) and nutrient digestibility (dry matter, crude protein). Also, the fecal score of NC group increased (p < 0.05) compared with other groups. In blood profiles, lymphocyte decreased (p < 0.05), and neutrophil, cortisol, TNF-α increased (p < 0.05) when pigs were in high stocking density. Basic behaviors (feed intake, standing, lying) were inactive (p < 0.05) and singularity behavior (biting) were increased (p < 0.05) under high stocking density. However, PFA groups alleviated the negative effects such as reducing growth performance, nutrient digestibility, increasing stress indicators in blood and animal behavior. In conclusion, PFA groups improved the health of pigs with stressed by high stocking density and PT3 is the most effective.

Abstract Background The porcine gall bladder and cystic duct gained attention in experimental research aimed at improving human clinical care. While the common bile duct has been investigated before, there is almost no data on the porcine cystic duct. Its relevance for research originates from its potential use in xenotransplantation. Methods We included 10 consecutive pigs (1♀, 9♂) that had been sacrificed after participation in another study. At necropsy, the distance from the biliary papilla to the gall bladder (gall bladder distance) was measured. The cystic duct was then subjected to linear traction. Ethical approval for the study was granted. Associations between force measurements and anatomical parameters were tested by correlation analysis. Results The pigs had a mean body weight of 21.9 kg (standard deviation 2 kg), a mean liver weight of 590 g (standard deviation 88 g), and a mean crown rump length of 67.3 cm (standard deviation 3 cm). The gall bladder distance was 8.2 cm (95% confidence interval: 7.6–8.7 cm). The cystic duct withstood mean linear traction forces of 4.8 N (95% confidence interval: 3.7–5.8 N) and could be elongated by a mean of 6 mm (95% confidence interval: 3.9–8 mm). Linear breaking strength was neither correlated to gall bladder distance (R = 0.3, 95% confidence interval: − 0.41 to 0.78, P = 0.406) or crown rump length (R = 0.42, 95% confidence interval: − 0.28 to 0.83, P = 0.222) nor liver weight (R = 0.02, 95% confidence interval: − 0.62 to 0.64, P = 0.954) or body weight (R = 0.36, 95% confidence interval: − 0.35 to 0.81, P = 0.304). This was also true for gall bladder distance. Conclusions Our study indicates that allometric parameters were not associated with the gall bladder distance or its resistance to traction forces. Consequently, substantial variation of these parameters can be expected in these surgically important parameters, which cannot be derived from easily accessible anatomical parameters.

Background and Aim: Pig industries are currently facing a crisis in terms of protein and energy costs. Proteases were used to increase protein digestibility and metabolizable energy (ME) in diets. This study evaluated the effects of protease supplementation on in vitro protein digestibility and productive performance in starter-to-finisher pigs. Materials and Methods: A total of 691 starter pigs were randomly allocated into three dietary treatments using a randomized complete block design. Diets were provided in three phases according to body weight (BW): Starter, grower, and finisher phases. Each phase was fed for 30, 60, and 24 days of treatment diets as T1: basal diet and T2 and T3: the basal diet supplemented with 240 ppm protease reduced by 50 kcal/kg ME plus 1% crude protein (CP) and by 100 kcal/kg ME plus 2% CP, respectively. Protease and in vitro protein digestibility were measured. BW and feed intake were recorded to calculate the average daily gain (ADG), average daily feed intake (ADFI), feed-to-gain (F:G), and gain-to-feed (G:F) ratios. Results: There were no significant differences (p > 0.05) in the percentage of in vitro protein digestibility between the groups with and without protease supplementation. In the finisher phase, T2 had lower (p < 0.05) ADFI and F:G than T1 and T3. Overall, T3 had lower (p < 0.05) ADG, ADFI, and F:G than T1 and T2. Conclusion: Protease supplementation significantly affects protein digestibility. Supplementing basal diets with 240 ppm protease reduced ME by 50 kcal/kg and CP by 1% without affecting ADG, ADFI, F:G, and G:F ratios for starter-to-finisher pigs. Keywords:

Porcine reproductive and respiratory syndrome virus (PRRSV) causes abortions, stillbirths, and dummy pregnancies. Previous studies found that PRRSV can promote secondary bacterial infections and elevate bacterial endotoxin levels, further increasing the abortion rate in sows. However, the pathways by which bacterial endotoxins invade the bodies of PRRSV(+) sows and aggravate their clinical symptoms are unknown. In this study, we established a model of PRRSV and lipopolysaccharide (LPS) working together on porcine endometrial epithelial cells (PEECs). We speculate that PRRSV and LPS affect PEECs through viral protein interaction with cytokines and cytokine receptors, natural killer cell-mediated cytotoxicity, and regulation of actin cytoskeleton signaling pathways by analyzing seq-RNA. The PRRSV proteins act on inflammatory factors and their receptors to activate chemokines-5 (CCL5), chemokines-4 (CCL4), and chemokines-8 (CCL8) mRNA expression, causing severe inflammatory reactions. In addition, the elevation of MEK1/2 factors and the integrins acting on NK cells promote the upregulation of VAV1/Tiam1, RAC, and IRSp53, leading to increased expression of Arp2/3 and F-actin in PEECs in the PRRSV + LPS(+) groups. However, the highly expressed cell microfilaments and cytoskeleton disrupt the original network structure, causing changes in the original physiological function of the PEECs. In summary, the PRRSV protein interacts with cytokines and cytokine receptors of PEECs, thereby enhancing virus-mediated chemokine factors and their receptor activity, accelerating bacterial endotoxin entry into the body and the invasion of cells. They destroy the cytoskeletal structure of the cells and increase damage to uterine tissue.

Compared with the widely used rodents, pigs are anatomically, physiologically, and genetically more similar to humans, making them high-quality models for the study of liver diseases. Here, we review the latest research progress on pigs as a model of human liver disease, including methods for establishing them and their advantages in studying cystic fibrosis liver disease, acute liver failure, liver regeneration, non-alcoholic fatty liver disease, liver tumors, and xenotransplantation. We also emphasize the importance of genetic engineering techniques, mainly the CRISPR/Cas9 system, which has greatly enhanced the utility of porcine models as a tool for substantially advancing liver disease research. Genetic engineering is expected to propel the pig as one of the irreplaceable animal models for future biomedical research.

Abstract Background In patients requiring extracorporeal cardiopulmonary resuscitation (ECPR), there is a need for studies to assess the potential benefits of therapeutic interventions to improve survival and reduce hypoxic-ischemic brain injuries. However, conducting human studies may be challenging. This study aimed to describe two experimental models developed in our laboratory and to conduct a systematic review of existing animal models of ECPR reported in the literature. Results In our experiments, pigs were subjected to 12 min (model 1) or 5 min (model 2) of untreated ventricular fibrillation, followed by 18 min (model 1) or 25 min (model 2) of conventional cardiopulmonary resuscitation. Results showed severe distributive shock, decreased brain oxygen pressure and increased intracranial pressure, with model 1 displaying more pronounced brain perfusion impairment. A systematic review of 52 studies, mostly conducted on pigs, revealed heterogeneity in cardiac arrest induction methods, cardiopulmonary resuscitation strategies, and evaluated outcomes. Conclusions This review emphasizes the significant impact of no-flow and low-flow durations on brain injury severity following ECPR. However, the diversity in experimental models hinders direct comparisons, urging the standardization of ECPR models to enhance consistency and comparability across studies.

Abstract Background There is a higher risk for nematode infections associated with outdoor-reared pigs. Next to Ascaris suum, Oesophagostomum dentatum and Trichuris suis, there is the potential of infections with other nodular worm species, Hyostrongylus rubidus, Stongyloides ransomi and Metastrongylus spp. lungworms. Next-generation sequencing methods describing the nemabiome have not yet been established for porcine nematodes. Methods FLOTAC was used for faecal egg counts of porcine gastrointestinal nematodes and lungworms in piglets, fatteners and adults individually. A nemabiome analyses based on ITS-2 gene region metabarcoding was used to differentiate strongyle species. Additionally, questionnaire data was analysed using mixed-effect regression to identify potential risk factors associated with parasite occurrences and egg shedding intensity. Results On 15 of 17 farms nematode eggs were detected. Ascaris suum, strongyles and T. suis were detected on 82%, 70% and 35% of the 17 farms, respectively. Lungworms were detected on one out of four farms with access to pasture. Strongyloides ransomi was not detected. 32% (CI 28–36%), 27% (24–31%), 5% (4–7%) and 3% (0.9–8%) of the samples where tested positive for strongyles, A. suum, T. suis and lungworms, respectively. The nemabiome analysis revealed three different strongyle species, with O. dentatum being the most common (mean 93.9%), followed by O. quadrispinulatum (5.9%) and the hookworm Globocephalus urosubulatus (0.1%). The bivariate and multivariate risk factor analyses showed among others that cleaning once a week compared to twice a week increased the odds significantly for being infected with A. suum (OR 78.60) and strongyles (2077.59). Access to pasture was associated with higher odds for A. suum (43.83) and strongyles (14.21). Compared to shallow litter systems, deep litter and free range systems resulted in significant higher odds for strongyles (85.74, 215.59, respectively) and T. suis (200.33, 623.08). Conclusions Infections with A. suum, O. dentatum, O. quadrispinulatum, T. suis, Metastrongylus spp. and G. urosubulatus are present in German outdoor-reared pigs. This is the first report of G. urosubulatus in domestic pigs in Europe. Metabarcoding based on the ITS-2 region is a suitable tool to analyse the porcine nemabiome. Furthermore, management practices have the potential of reducing the risk of parasite infections.

Abstract Background Infertility is a growing global health concern affecting millions of couples worldwide. Among several factors, an extreme body weight adversely affects reproductive functions. Leptin is a well-known adipokine that serves as an endocrine signal between adiposity and fertility. However, the exact mechanisms underlying the effects of high leptin level on female reproduction remain unclear. Methods Transgenic pigs overexpressing leptin (♀) were produced by backcrossing and screened for leptin overexpression. The growth curve, fat deposition, reproductive performance, apoptosis, serum hormones and cholesterol production, RNA sequencing, and single-nucleus RNA sequencing (snRNA-seq) of the leptin-overexpressing pigs and wild-type group were evaluated. Results Transgenic pigs overexpressing leptin (♀) were obtained, which exhibited significantly reduced body weight, body size, and back fat thickness. These pigs manifested a late onset of puberty (330 ± 54.3 vs. 155 ± 14.7 days), irregular estrous behavior characterized by increased inter-estrous interval (29.2 ± 0 vs. 21.3 ± 0.7 days), and more number of matings until pregnancy (at least 3 times). This reproductive impairment in leptin pigs was related to hormonal imbalances characterized by increased levels of FSH, LH, prolactin, E2, P4, and TSH, altered steroidogenesis such as increased levels of serum cholesterol esters along with steroidogenic markers (StAR, CYP19A), and ovarian dysfunctions manifested by neutrophilic infiltration and low expression of caspase-3 positive cells in the ovaries. Moreover, bulk RNA sequencing of the ovaries also revealed neutrophilic infiltration followed by upregulation of inflammation-related genes. Furthermore, snRNA-seq reflected that leptin overexpression triggered immune response, suppressed follicle development and luteinization, resulting in metabolic dysfunction and hormone imbalance in the ovary. Conclusions Low body weight in leptin overexpressing pigs adversely affects the reproductive performance, causing delayed puberty, irregular estrous cycles, and reduced breeding efficiency. This is linked to metabolic imbalances, an increased immune response, and altered ovarian functions. This study provides a theoretical basis for the complex mechanisms underlying leptin, and infertility by employing leptin-overexpressing female pigs.

Abstract Background Swine dysentery, caused by Brachyspira hyodysenteriae, is a severe pig disease. Resistance to tylosins is common and resistance to tiamulin has been reported since the 1990s. Still, dysentery is not notifiable to authorities. The disease therefore escapes control from an overall population perspective. In Sweden, a program that aimed to control dysentery at national level was initiated in 2020, mainly due to the unexpected diagnosis of tiamulin resistant Brachyspira hyodysenteriae in 2016. Results Through joint efforts of a network including farmers, government, animal health organisations and abattoirs it was concluded that outbreaks of dysentery had taken place in 25 herds between 2016 and 2019. By 1 January 2020, nine of these herds were still not declared free from the disease. From that date, the network decided that Brachyspira hyodysenteriae was to be cultured whenever dysentery could be suspected. Thus, 148, 157 and 124 herds were scrutinised for Brachyspira hyodysenteriae in 2020, 2021 and 2022, respectively, whereof five, three and two new herds were confirmed positive. By 31 December 2022, four herds were judged as impossible to sanitise. However, they posed no problem since they were identified by the network, pigs to and from these enterprises could be transported without jeopardising other herds. When Brachyspira hyodysenteriae was diagnosed in fattening herds purchasing growers, Brachyspira hyodysenteriae could not be detected in the delivering herds. That result, together with other observations, indicated that Brachyspira hyodysenteriae ought to be regarded as ubiquitous, although at a low level in healthy pigs. Conclusions Eradication of dysentery contributed to substantial welfare and financial improvements in affected herds. Dysentery was controlled successfully at national level through the united efforts from competing stake holders, such as different abattoirs and animal health organisations. However, as Brachyspira hyodysenteriae was assumed to be ubiquitous, although at a low level in healthy pigs, the duration of the successful control of dysentery was concluded to only be transient. Without permanent monitoring for Brachyspira hyodysenteriae, the knowledge of the national status will rapidly decline to the level prior to the initiation of the control program.

Abstract Capacitation is an essential post-testicular maturation event endowing spermatozoa with fertilizing capacity within the female reproductive tract, significant for fertility, reproductive health, and contraception. By using a human-relevant large animal model, the domestic boar, this study focuses on furthering our understanding of the involvement of the ubiquitin-proteasome system (UPS) in sperm capacitation. The UPS is a universal, evolutionarily conserved, cellular proteome-wide degradation and recycling machinery, that has been shown to play a significant role in reproduction during the past two decades. Herein, we have used a bottom-up proteomic approach to (i) monitor the capacitation-related changes in the sperm protein levels, and (ii) identify the targets of UPS regulation during sperm capacitation. Spermatozoa were capacitated under proteasomal activity-permissive and inhibiting conditions and extracted sperm proteins were subjected to high-resolution mass spectrometry. We report that 401 individual proteins differed at least two-fold in abundance (P

Abstract Background The establishment of stable porcine embryonic stem cells (pESCs) can contribute to basic and biomedical research, including comparative developmental biology, as well as assessing the safety of stem cell-based therapies. Despite these advantages, most pESCs obtained from in vitro blastocysts require complex media and feeder layers, making routine use, genetic modification, and differentiation into specific cell types difficult. We aimed to establish pESCs with a single cell-passage ability, high proliferative potency, and stable in long-term culture from in vitro-derived blastocysts using a simplified serum-free medium. Methods We evaluated the establishment efficiency of pESCs from in vitro blastocysts using various basal media (DMEM/F10 (1:1), DMEM/F12, and a-MEM) and factors (FGF2, IWR-1, CHIR99021, and WH-4-023). The pluripotency and self-renewal capacity of the established pESCs were analyzed under feeder or feeder-free conditions. Ultimately, we developed a simplified culture medium (FIW) composed of FGF2, IWR-1, and WH-4-023 under serum-free conditions. Results The pESC-FIW lines were capable of single-cell passaging with short cell doubling times and expressed the pluripotency markers POU5F1, SOX2, and NANOG, as well as cell surface markers SSEA1, SSEA4, and TRA-1-60. pESC-FIW showed a stable proliferation rate and normal karyotype, even after 50 passages. Transcriptome analysis revealed that pESC-FIW were similar to reported pESC maintained in complex media and showed gastrulating epiblast cell characteristics. pESC-FIW were maintained for multiple passages under feeder-free conditions on fibronectin-coated plates using mTeSR™, a commercial medium used for feeder-free culture, exhibiting characteristics similar to those observed under feeder conditions. Conclusions These results indicated that inhibition of WNT and SRC was sufficient to establish pESCs capable of single-cell passaging and feeder-free expansion under serum-free conditions. The easy maintenance of pESCs facilitates their application in gene editing technology for agriculture and biomedicine, as well as lineage commitment studies.

Abstract Background Leptospiraceae comprise a diverse family of spirochetal bacteria, of which many are involved in infectious diseases of animals and humans. Local leptospiral diversity in domestic animals is often poorly understood. Here we describe the incidental detection of Leptospira (L.) licerasiae in an Austrian pig. Case presentation During an experiment to characterize the pathogenesis of L. interrogans serovar Icterohaemorrhagiae in pigs, cultivation of a urine sample from a non-challenged contact pig resulted in growth of a spirochetal bacterium that tested negative for pathogenic Leptospira (LipL32 gene). PCR, Sanger sequencing and standard serotyping further confirmed that the recovered isolate was clearly different from the challenge strain L. interrogans serovar Icterohaemorrhagiae used in the animal experiment. Whole genome sequencing revealed that the isolate belongs to the species L. licerasiae, a tropical member of the Leptospiraceae, with no prior record of detection in Europe. Conclusions This is the first report describing the occurrence of L. licerasiae in Europe. Since L. licerasiae is considered to have intermediate pathogenicity, it will be important to follow the geographical distribution of this species and its pathogenic and zoonotic potential in more detail.

Influenza D virus was isolated from pigs on a mixed pig and beef farm in France. Investigation suggested bull-to-pig transmission and spread among pigs. The swine influenza D virus recovered was a reassortant of D/660 and D/OK lineages. Reported mutations in the receptor binding site might be related to swine host adaptation.

Indonesia has a long history of livestock brucellosis, but the overall pooled prevalence remains unclear. This study aims to determine the pooled estimated prevalence of livestock brucellosis in Indonesia using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Five databases were searched and screened using predefined inclusion and exclusion criteria. Data from included studies were extracted and analyzed using a random effects model in R 4.3.0 for pooled estimated prevalence, subgroup meta-analysis, and meta-regression. Publication bias and sensitivity tests were conducted using Egger’s test, funnel plot, trim and fill plot, and leave-one-out. The screening process identified 46 included studies, representing 47,057 samples for brucellosis testing. The pooled estimated prevalence for livestock brucellosis was 3.25% (95% CI, 1.81%–5.78%) with high heterogeneity (Q = 2130.91, p = 0, I2=98%). Subgroup meta-analysis indicated no significant difference in the prevalence of livestock brucellosis across the main islands in Indonesia (p = 0.44) and across provinces in Sulawesi Island (p = 0.83), but significant differences were found among provinces in Java (p

AbstractAs COVID-19 has shown, pandemics and outbreaks of emerging infections such as Zika, Nipah, monkeypox and antimicrobial-resistant pathogens, especially emerging zoonotic diseases, continue to occur and may even be increasing in Southeast Asia. In addition, these infections often result from environmental changes and human behaviour. Overall, public health surveillance to identify gaps in the literature and early warning signs are essential in this region. A systematic review investigated the prevalence of emerging zoonotic diseases over 11 years from 2011 to 2022 in Southeast Asia to understand the status of emerging zoonotic diseases, as well as to provide necessary actions for disease control and prevention in the region. During the 2011–2022 period, studies on pigs, poultry, ruminants, companion animals and wildlife in Southeast Asia were reviewed thoroughly to assess the quality of reporting items for inclusion in the systematic review. The review was performed on 26 studies of pigs, 6 studies of poultry, 21 studies of ruminants, 28 studies of companion animals and 25 studies of wildlife in Southeast Asia, which provide a snapshot of the prevalence of the emerging zoonotic disease across the country. The findings from the review showed that emerging zoonotic diseases were prevalent across the region and identified a few zoonotic diseases associated with poultry, mainly stemming from Cambodia and Vietnam, as high priority in Southeast Asia.Clinical relevance: Appropriate prevention and control measures should be taken to mitigate the emerging zoonotic diseases in Southeast Asia.

The enteric nervous system (ENS) interacts bidirectionally with the local immune system, responding to inflammation within the gastrointestinal (GI) tract. In a previous study using the same samples, several gene targets were identified as being differentially expressed in the inflamed colonic tissue of pigs challenged with dextran sodium sulphate (DSS). Additionally, animals in the basal DSS group, exhibited reduced growth and increased fecal and pathology scores, while the relative abundance of beneficial taxa was reduced and harmful bacteria increased. While changes in the innate immune response and barrier function are widely cited regarding inflammatory bowel disease (IBD), the effects of inflammation on the local structures of the enteric nervous system (ENS) are less well understood. Hence, the objectives of this study were to: (1) evaluate the expression of a range of functionally diverse neuroactive receptors, transporters and neurotrophic factors in RNA derived from the colonic tissue from the same pigs; (2) examine associations with these neuroactive components and inflammatory, barrier function and matrix remodeling targets. Mature pigs were split into two experimental groups: (1) basal diet (n = 10); (2) basal diet + DSS (n = 11). The pigs were orally challenged with DSS once daily for four days and sacrificed humanely. Colonic tissue was collected for gene expression analysis. Most of the targets evaluated in this study were present at low levels or in some cases were undetectable by QPCR, including the dopamine receptor DRD5 and the serotonin receptor HTR3A. The dopamine receptors (DRD1, DRD3, DRD4), serotonin receptor (HTR4), and other selected neuroactive receptors (GRM7, GABRA2) were down-regulated in the DSS-challenged animals relative to the basal group (p < 0.05). Most notably, DRD2, was up-regulated four-fold, suggesting an active process involving this receptor (p < 0.05). Relationships with (previously published) gene expression data from the same samples suggest that DRD1 and DRD2 are influenced by different pathways and may also be interlinked with matrix remodeling and, more specifically, genes relevant to the epithelial to mesenchymal transition (CDH1, CDH2, IL6, IL13, IL10, MMP1, MMP2) an important fibrotic process in the pathogenesis of IBD.

Different dietary patterns affect the gut microbial compositions and diversity. Consistently, microbiome alterations are linked to digestion, immunity, and productivity. Sasa quelpaertensis Nakai (SQ) is a perennial bamboo species rich in proteins and fiber. Previous studies have confirmed the health benefits of SQ; however, the effects of SQ supplementation on gut microbiome and production performance are unclear. Herein, Landrace pigs were supplemented with SQ extract (SQE) and gut microbial compositions as opposed to the control group were assessed using 16S rRNA sequencing. Additionally, the influences of SQE supplementation on average daily gain (ADG) and backfat thickness (BF) were assessed after slaughter. In the SQE group, Firmicutes and Actinobacteria phyla increased significantly, whereas Bacteroidetes and Spirochaetes phyla markedly decreased (p < 0.05). The expression level of Bifidobacterium and Lactobacillus genera increased, whereas that of Treponema, Prevotella, and Turicibacter decreased (p < 0.05). The microbial richness was similar between groups; however, microbial diversity decreased in the SQE supplementation group. Additionally, the SQE supplementation in pigs resulted in a slight increase in ADG. In contrast, BF in the SQE group decreased notably (p < 0.05). These results underscore the significant influence of SQE supplementation on the gut microbiota and demonstrate the potential of SQ as a valuable feed resource for enhancing animal productivity.

Different dietary patterns affect the gut microbial compositions and diversity. Consistently, microbiome alterations are linked to digestion, immunity, and productivity. Sasa quelpaertensis Nakai (SQ) is a perennial bamboo species rich in proteins and fiber. Previous studies have confirmed the health benefits of SQ; however, the effects of SQ supplementation on gut microbiome and production performance are unclear. Herein, Landrace pigs were supplemented with SQ extract (SQE) and gut microbial compositions as opposed to the control group were assessed using 16S rRNA sequencing. Additionally, the influences of SQE supplementation on average daily gain (ADG) and backfat thickness (BF) were assessed after slaughter. In the SQE group, Firmicutes and Actinobacteria phyla increased significantly, whereas Bacteroidetes and Spirochaetes phyla markedly decreased (p < 0.05). The expression level of Bifidobacterium and Lactobacillus genera increased, whereas that of Treponema, Prevotella, and Turicibacter decreased (p < 0.05). The microbial richness was similar between groups; however, microbial diversity decreased in the SQE supplementation group. Additionally, the SQE supplementation in pigs resulted in a slight increase in ADG. In contrast, BF in the SQE group decreased notably (p < 0.05). These results underscore the significant influence of SQE supplementation on the gut microbiota and demonstrate the potential of SQ as a valuable feed resource for enhancing animal productivity.

Abstract Monitoring the use of antimicrobials and the emergence of resistance in animals and people is important for the control of antimicrobial resistance, and for establishing sustainable and effective disease management practices. In this study, we used Enterococcus spp. and Escherichia coli as indicator species to investigate antimicrobial susceptibility patterns and how these change over time, on ten Swedish pig farms. Indoor environmental sock sampling was performed once a month during the entire production cycle of one batch of pigs on each farm, resulting in 60 samples collected in total. Selective culture for E. coli and Enterococcus spp. resulted in 122 isolates of E. coli, 74 isolates of E. faecium, but no isolates of E. faecalis. Microdilution was used to determine minimum inhibitory concentrations for twelve antimicrobial substances in E. coli and fifteen substances in E. faecium. The overall prevalence of resistance was low. Among the E. coli isolates, the proportions non-wild type (resistant, NWT) isolates were as follows: azithromycin and amikacin 1% (n = 1), trimethoprim and sulfamethoxazole 2% (n = 3), ampicillin 6% (n = 7) and tetracycline 9% (n = 11). Among the E. faecium isolates, the NWT proportions were: teicoplanin, linezolid and gentamicin 1% (n = 1), daptomycin 3% (n = 2), erythromycin 26% (n = 19), tetracycline 27% (n = 20), quinupristin/dalfopristin 58% (n = 42). The resistance patterns differed between the farms, likely due to different antimicrobial use, biosecurity measures and source of the animals. The NWT prevalence among E. coli decreased over time, whereas no similar trend could be observed in E. faecium. The results of the current study illustrate the complex factors affecting the antimicrobial resistance patterns observed on each farm, indicating that specific practices and risk factors have an impact on the prevalence and type of antimicrobial resistance. Further studies of the farm environments in combination with antimicrobial use and other risk factor data are needed to elucidate the multifaceted drivers of antimicrobial resistance development on livestock farms.

Abstract Background Integration of high throughput DNA genotyping and RNA-sequencing data enables the discovery of genomic regions that regulate gene expression, known as expression quantitative trait loci (eQTL). In pigs, efforts to date have been mainly focused on purebred lines for traits with commercial relevance as such growth and meat quality. However, little is known on genetic variants and mechanisms associated with the robustness of an animal, thus its overall health status. Here, the liver, lung, spleen, and muscle transcriptomes of 100 three-way crossbred female finishers were studied, with the aim of identifying novel eQTL regulatory regions and transcription factors (TFs) associated with regulation of porcine metabolism and health-related traits. Results An expression genome-wide association study with 535,896 genotypes and the expression of 12,680 genes in liver, 13,310 genes in lung, 12,650 genes in spleen, and 12,595 genes in muscle resulted in 4,293, 10,630, 4,533, and 6,871 eQTL regions for each of these tissues, respectively. Although only a small fraction of the eQTLs were annotated as cis-eQTLs, these presented a higher number of polymorphisms per region and significantly stronger associations with their target gene compared to trans-eQTLs. Between 20 and 115 eQTL hotspots were identified across the four tissues. Interestingly, these were all enriched for immune-related biological processes. In spleen, two TFs were identified: ERF and ZNF45, with key roles in regulation of gene expression. Conclusions This study provides a comprehensive analysis with more than 26,000 eQTL regions identified that are now publicly available. The genomic regions and their variants were mostly associated with tissue-specific regulatory roles. However, some shared regions provide new insights into the complex regulation of genes and their interactions that are involved with important traits related to metabolism and immunity.

Abstract International interest in loose-housed farrowing is growing and there are ongoing discussions within the European Union (EU) on new legal requirements. However, there is a lack of empirical data on loose-housed farrowing pen sizes and sow dimensions in commercial production. The aim of this study was to map and describe sow size and loose-housing farrowing pen size on commercial piglet-producing farms in Sweden. The study included 146 sows and 51 pen types on 35 medium sized to large Swedish piglet-producing farms (ranging from 106 to 1300 sows in production). Sow length ranged from 129 to 238 cm (mean ± SD 191.3 ± 19.3 cm) and sow height from 74 to 133 cm (86.7 ± 7.7 cm). Floor space occupied by the sow when lying down (length x height) ranged from 1.0 to 3.2 m2 (1.7 ± 0.3 m2). Pen length ranged from 259 to 415 cm (315.1 ± 24.3 cm), pen width from 188 to 245 cm (207.0 ± 10.7 cm), total pen area from 5.7 to 8.9 m2 (6.5 ± 0.5 m2), piglet corner area from 0.5 to 1.8 m2 (1.1 ± 0.4 m2) and area available for the sow (total area - piglet corner area) from 3.9 to 6.4 m2 (5.4 ± 0.6 m2). These results show that there is substantial variation in sow, pen and piglet corner size on commercial piglet-producing farms in Sweden. This poses a risk of mismatches between sow and pen size (pens too short in relation to sow dimensions), especially for older sows. These findings are of practical significance for animal welfare and production and emphasise the importance of designing loose-housed pens adapted to future sow, litter and piglet size.

Abstract Background Long-term natural and artificial selection has resulted in many genetic footprints within the genomes of pig breeds across distinct agroecological zones. Nevertheless, the mechanisms by which these signatures contribute to phenotypic diversity and facilitate environmental adaptation remain unclear. Results Here, we leveraged whole-genome sequencing data from 82 individuals from 6 domestic pig breeds originating in tropical, high-altitude, and frigid regions. Population genetic analysis suggested that habitat isolation significantly shaped the genetic diversity and contributed to population stratification in local Chinese pig breeds. Analysis of selection signals revealed regions under selection for adaptation in tropical (55.5 Mb), high-altitude (43.6 Mb), and frigid (17.72 Mb) regions. The potential functions of the selective sweep regions were linked to certain complex traits that might play critical roles in different geographic environments, including fat coverage in frigid environments and blood indicators in tropical and high-altitude environments. Candidate genes under selection were significantly enriched in biological pathways involved in environmental adaptation. These pathways included blood circulation, protein degradation, and inflammation for adaptation to tropical environments; heart and lung development, hypoxia response, and DNA damage repair for high-altitude adaptation; and thermogenesis, cold-induced vasodilation (CIVD), and the cell cycle for adaptation to frigid environments. By examining the chromatin state of the selection signatures, we identified the lung and ileum as two candidate functional tissues for environmental adaptation. Finally, we identified a mutation (chr1: G246,175,129A) in the cis-regulatory region of ABCA1 as a plausible promising variant for adaptation to tropical environments. Conclusions In this study, we conducted a genome-wide exploration of the genetic mechanisms underlying the adaptability of local Chinese pig breeds to tropical, high-altitude, and frigid environments. Our findings shed light on the prominent role of cis-regulatory elements in environmental adaptation in pigs and may serve as a valuable biological model of human plateau-related disorders and cardiovascular diseases.

Abstract Background The pituitary belongs to the most important endocrine glands involved in regulating reproductive functions. The proper functioning of this gland ensures the undisturbed course of the oestrous cycle and affects the female’s reproductive potential. It is believed that visfatin, a hormone belonging to the adipokine family, may regulate reproductive functions in response to the female’s metabolic state. Herein we verified the hypothesis that suggests a modulatory effect of visfatin on the anterior pituitary transcriptome during the mid-luteal phase of the oestrous cycle. Results RNA-seq analysis of the porcine anterior pituitary cells revealed changes in the expression of 202 genes (95 up-regulated and 107 down-regulated in the presence of visfatin, when compared to the non-treated controls), assigned to 318 gene ontology terms. We revealed changes in the frequency of alternative splicing events (235 cases), as well as long noncoding RNA expression (79 cases) in the presence of the adipokine. The identified genes were associated, among others, with reproductive system development, epithelial cell proliferation, positive regulation of cell development, gland morphogenesis and cell chemotaxis. Conclusions The obtained results indicate a modulatory influence of visfatin on the regulation of the porcine transcriptome and, in consequence, pituitary physiology during the mid-luteal phase of the oestrous cycle.

Background: To overcome organ shortage during transplantation, interspecies organ generation via blastocyst complementation has been proposed, although not yet in evolutionarily distant species. To establish high levels of chimerism, low chimerism is required early in development, followed by high chimerism, to effectively complement the organ niche. Very few human cells are expected to contribute to chimerism in heterologous animals. Previous studies had demonstrated increased donor chimerism in both intra- and interspecies chimeras in rodents, using insulin-like growth factor 1 receptor (Igf1r) knockout (KO) mice; deletion of the Igf1r gene in the mouse host embryo created a cell-competitive niche. The current study aimed to generate IGF1R–KO pigs and evaluate whether they have the same phenotype as Igf1r-KO mice. Methods: To generate IGF1R–KO pigs, genome-editing molecules were injected into the cytoplasm of pig zygotes. The fetuses were evaluated at 104 days of gestation. Results: IGF1R–KO pigs were generated successfully. Their phenotypes were almost identical to those of Igf1r-KO mice, including small lungs and enlarged endodermal organs in fetuses, and they were highly reproducible. Conclusions: Pigs may allow the generation of organs using blastocyst complementation with developmentally-compatible xenogeneic pluripotent stem cells over a large evolutionary distance.

For the intracytoplasmic sperm injection (ICSI) procedure in pigs, an electrical pulse (EP) has been used as an effective method for oocyte stimulation, but unlike sperm, EP is unable to induce Ca2+ oscillations. In this study, we investigated the effects of generating artificial Ca2+ oscillations with phospholipase Cζ (PLCζ) mRNA, a candidate sperm factor, on fertilization, embryonic development, and gene expression after ICSI. Firstly, the concentration of PLCζ mRNA of a fixed volume (1.0 pl) that would induce a pattern of Ca2+ rise similar to that of in vitro fertilized (IVF) sperm was examined and determined to be 300 ng/μl. Secondly, the effects of oocyte stimulation methods on fertilization and embryonic development were investigated. ICSI-oocytes were activated by EP (EP group) or by PLCζ mRNA (PLCζ group). Furthermore, IVF-oocytes (IVF group) and ICSI-oocytes with and without an injection of buffer (buffer and untreated groups, respectively) were used as controls. It was found that the rates of normal fertilization in the PLCζ and EP groups were significantly higher than those in the buffer and untreated groups. The blastocyst formation rates did not differ among the groups. The embryo quality in the EP group was inferior to those in the PLCζ and IVF groups. Additionally, the expression level of a proapoptosis-related gene (Caspase-3) in the EP group was significantly higher than those in the PLCζ and IVF groups. Our data suggest that oocyte activation by PLCζ mRNA has the effect of improving embryo quality.

Pseudomonas aeruginosa poses a significant threat to both immunocompetent and immunocompromised individuals, often resulting in life-threatening infections. With increasing antimicrobial resistance, novel therapeutic strategies are urgently needed. Although animal models are crucial for preclinical studies, limited data are available for porcine models, more specifically for P. aeruginosa complicated skin and soft tissue infections (cSSTIs). This study presents a novel porcine model inducing and sustaining cSSTI for 14 days. Six pigs (120 wounds) were used for the development of infections, and within this group, two pigs (40 wounds) were used to evaluate the progression of the cSSTI infection. The model demonstrated bacterial loads of more than 107 CFU/gram of tissue or higher. The cSSTI fully developed within three days and remained well above these levels until day 14 post-infection. Due to the immunocompetence of this model, all the immunological processes associated with the response to the presence of infection and the wound healing process are preserved.

Abstract Background Swine dysentery (SD) is a severe mucohaemorrhagic colitis in pigs caused classically by Brachyspira hyodysenteriae. Although several aspects of B. hyodysenteriae infection dynamic are already described, further research in the early stage of this infection is required. In this study, 7-week-old pigs were orally challenged with B. hyodysenteriae to obtain information about faecal shedding, macro and microscopic intestinal lesions and serum acute phase proteins in pigs at the onset of B. hyodysenteriae shedding (early infection group, n = 8), in pigs with mucohaemorrhagic diarrhoea (acute infection group, n = 8) and in non-infected controls (n = 16). Results First B. hyodysenteriae detection by q-PCR and first loose stools with blood and mucus occurred both at 8 days post-inoculation. The lapse between a positive q-PCR and observation of mucohaemorrhagic diarrhoea ranged from 0 to 3 days, except in a single pig in which this period lasted 5 days. Macroscopic lesions were observed in the large intestine from both infected groups although more frequent and severe in acute infection group. Microscopic observation of the apex mucosa revealed that in early infection only higher ulceration values were observed compared to healthy controls. In contrast, the acute infection group exhibited higher ulceration, neutrophils infiltration and increased mucosal thickness compared to the other two groups. Among the serum biomarkers tested, only haptoglobin, C-reactive protein, and creatine kinase showed a significant increase in pigs in the acute infection period compared to controls, whereas haptoglobin was the only factor with a significant increase at the early infection compared to non-infected animals. Conclusions This study provides new insights about SD and remarks the complex and limited options to perform an early detection of infected animals beyond PCR diagnosis.

Stomach ulcers are relatively common in pigs raised in modern large-scale farms. They can result in reduced daily weight gain, reduced feed intake, and sudden death, resulting in significant economic losses. They also belong to the most common findings in the stomach of pigs at slaughterhouses and are among the most important causes of peracute mortality, associated with bleeding into the digestive system of pigs. The aim of this article was to provide an overview of the most important etiological factors, and pathogenesis of gastric ulcers in pigs.

Abstract Visfatin (VIS) is a hormone belonging to the adipokines’ group secreted mainly by the adipose tissue. VIS plays a crucial role in the control of energy homeostasis, inflammation, cell differentiation, and angiogenesis. VIS expression was confirmed in the hypothalamic–pituitary–gonadal (HPG) axis structures, as well as in the uterus, placenta, and conceptuses. We hypothesised that VIS may affect the abundance of proteins involved in the regulation of key processes occurring in the corpus luteum (CL) during the implantation process in pigs. In the present study, we performed the high-throughput proteomic analysis (liquid chromatography with tandem mass spectrometry, LC–MS/MS) to examine the in vitro influence of VIS (100 ng/mL) on differentially regulated proteins (DRPs) in the porcine luteal cells (LCs) on days 15–16 of pregnancy (implantation period). We have identified 511 DRPs, 276 of them were up-regulated, and 235 down-regulated in the presence of VIS. Revealed DRPs were assigned to 162 gene ontology terms. Western blot analysis of five chosen DRPs, ADAM metallopeptidase with thrombospondin type 1 motif 1 (ADAMTS1), lanosterol 14-α demethylase (CYP51A1), inhibin subunit beta A (INHBA), notch receptor 3 (NOTCH3), and prostaglandin E synthase 2 (mPGES2) confirmed the veracity and accuracy of LC–MS/MS method. We indicated that VIS modulates the expression of proteins connected with the regulation of lipogenesis and cholesterologenesis, and, in consequence, may be involved in the synthesis of steroid hormones, as well as prostaglandins’ metabolism. Moreover, we revealed that VIS affects the abundance of protein associated with ovarian cell proliferation, differentiation, and apoptosis, as well as CL new vessel formation and tissue remodelling. Our results suggest important roles for VIS in the regulation of ovarian functions during the peri-implantation period.

Abstract Background Neurodevelopmental disorders (NDD), such as autism spectrum disorders (ASD) and intellectual disorders (ID), are highly debilitating childhood psychiatric conditions. Genetic factors are recognized as playing a major role in NDD, with a multitude of genes and genomic regions implicated. While the functional validation of NDD-associated genes has predominantly been carried out using mouse models, the significant differences in brain structure and gene function between mice and humans have limited the effectiveness of mouse models in exploring the underlying mechanisms of NDD. Therefore, it is important to establish alternative animal models that are more evolutionarily aligned with humans. Results In this study, we employed CRISPR/Cas9 and somatic cell nuclear transplantation technologies to successfully generate a knockout miniature pig model of the MIR137 gene, which encodes the neuropsychiatric disorder-associated microRNA miR-137. The homozygous knockout of MIR137 (MIR137 –/– ) effectively suppressed the expression of mature miR-137 and led to the birth of stillborn or short-lived piglets. Transcriptomic analysis revealed significant changes in genes associated with neurodevelopment and synaptic signaling in the brains of MIR137 –/– miniature pig, mirroring findings from human ASD transcriptomic data. In comparison to miR-137-deficient mouse and human induced pluripotent stem cell (hiPSC)-derived neuron models, the miniature pig model exhibited more consistent changes in critical neuronal genes relevant to humans following the loss of miR-137. Furthermore, a comparative analysis identified differentially expressed genes associated with ASD and ID risk genes in both miniature pig and hiPSC-derived neurons. Notably, human-specific miR-137 targets, such as CAMK2A, known to be linked to cognitive impairments and NDD, exhibited dysregulation in MIR137 –/– miniature pigs. These findings suggest that the loss of miR-137 in miniature pigs affects genes crucial for neurodevelopment, potentially contributing to the development of NDD. Conclusions Our study highlights the impact of miR-137 loss on critical genes involved in neurodevelopment and related disorders in MIR137 –/– miniature pigs. It establishes the miniature pig model as a valuable tool for investigating neurodevelopmental disorders, providing valuable insights for potential applications in human research.

This study aimed to investigate the impact of intradermal (ID) and intramuscular (IM) vaccination with a porcine reproductive and respiratory syndrome virus (PRRSV)-modified live vaccine (MLV) in piglets on serum copeptin levels and hematological profile. This study included 104 suckling piglets (2 weeks of age) from a commercial farrow-to-finish pig farm suffering from positive unstable PRRSV status. Animals were assigned to four groups, with two replicates (13 piglets/group/replicate); group A: IM vaccination with a PRRSV MLV vaccine, group B: ID vaccination with the same vaccine, group C: ID of Diluvac Forte, and group D: IM of Diluvac Forte. Blood samples were collected from the same three pigs/group/replicate at 4, 7, and 10 weeks of age. Blood samples were used for the performance of the complete blood count, and they were also examined by PCR for PRRSV and by ELISA for copeptin. No significant differences in serum copeptin levels and the number of blood cell counts (packed cell volume—PCV, numbers of white blood cells—WBCs, and platelets number—PLTs) were noticed in the same group over time and among groups. In conclusion, it seems that the vaccination against PRRSV does not affect the levels of the released copeptin. Based on our results, the measurement of serum copeptin could not be proposed as a potential stress biomarker in pigs.

Abstract Feed efficiency (FE) is essential for pig production, has been reported to be partially explained by gut microbiota. Despite an extensive body of research literature to this topic, studies regarding the regulation of feed efficiency by gut microbiota remain fragmented and mostly confined to disorganized or semi-structured unrestricted texts. Meanwhile, structured databases for microbiota analysis are available, yet they often lack a comprehensive understanding of the associated biological processes. Therefore, we have devised an approach to construct a comprehensive knowledge graph by combining unstructured textual intelligence with structured database information and applied it to investigate the relationship between pig gut microbes and FE. Firstly, we created the pgmReading knowledge base and the domain ontology of pig gut microbiota by annotating, extracting, and integrating semantic information from 157 scientific publications. Secondly, we created the pgmPubtator by utilizing PubTator to expand the semantic information related to microbiota. Thirdly, we created the pgmDatabase by mapping and combining the ADDAGMA, gutMGene, and KEGG databases based on the ontology. These three knowledge bases were integrated to form the Pig Gut Microbial Knowledge Graph (PGMKG). Additionally, we created five biological query cases to validate the performance of PGMKG. These cases not only allow us to identify microbes with the most significant impact on FE but also provide insights into the metabolites produced by these microbes and the associated metabolic pathways. This study introduces PGMKG, mapping key microbes in pig feed efficiency and guiding microbiota-targeted optimization.

Abstract At present, liquid storage is the most efficient method for pig semen preservation. This approach relies upon reducing sperm metabolism, allowing for the maintenance of cell lifespan. In this context, the study of proteins that could protect sperm during liquid storage is of high relevance. The 70 kDa Heat Shock Protein (HSP70) is an anti-apoptotic protein that has been reported to be relevant to sperm survival. Thus, we explored the role of HSP70 during prolonged storage of pig semen at 17 °C. Six semen pools were incubated with YM-1 (0, 0.05, 0.1 and 0.2 μM), an HSP70 inhibitor, and stored at 17 °C for 21 days. On days 0, 4, 10, 14 and 21, sperm quality and function were evaluated through flow cytometry and Computer-Assisted Sperm Analysis (CASA), and HSP70 activity and chromatin condensation were also determined. While inhibition of HSP70 increased progressive motility, Ca2+ and Reactive Oxygen Species (ROS) levels, and mitochondrial activity during the first 10 days of storage, it had a detrimental effect on sperm motility after 14 and 21 days. In spite of this, sperm viability was not altered. We can conclude that HSP70 contributes to the liquid storage of pig semen because it keeps mitochondrial activity low, which is needed for the maintenance of sperm function.

Abstract Background Mammalian intestine harbors a mass of phages that play important roles in maintaining gut microbial ecosystem and host health. Pig has become a common model for biomedical research and provides a large amount of meat for human consumption. However, the knowledge of gut phages in pigs is still limited. Results Here, we investigated the gut phageome in 112 pigs from seven pig breeds using PhaBOX strategy based on the metagenomic data. A total of 174,897 non-redundant gut phage genomes were assembled from 112 metagenomes. A total of 33,487 gut phage genomes were classified and these phages mainly belonged to phage families such as Ackermannviridae, Straboviridae, Peduoviridae, Zierdtviridae, Drexlerviridae, and Herelleviridae. The gut phages in seven pig breeds exhibited distinct communities and the gut phage communities changed with the age of pig. These gut phages were predicted to infect a broad range of 212 genera of prokaryotes, such as Candidatus Hamiltonella, Mycoplasma, Colwellia, and Lactobacillus. The data indicated that broad KEGG and CAZy functions were also enriched in gut phages of pigs. The gut phages also carried the antimicrobial resistance genes (ARGs) and the most abundant antimicrobial resistance genotype was diaminopyrimidine resistance. Conclusions Our research delineates a landscape for gut phages in seven pig breeds and reveals that gut phages serve as a key reservoir of ARGs in pigs. Video Abstract

Abstract Background Pork quality is affected by the type of muscle fibers, which is closely related to meat color, tenderness and juiciness. Exosomes are tiny vesicles with a diameter of approximately 30–150 nm that are secreted by cells and taken up by recipient cells to mediate communication. Exosome-mediated muscle-fat tissue crosstalk is a newly discovered mechanism that may have an important effect on intramuscular fat deposition and with that on meat quality. Various of adipose tissue-derived exosomes have been discovered and identified, but the identification and function of muscle exosomes, especially porcine fast/slow myotube exosomes, remain unclear. Here, we first isolated and identified exosomes secreted from porcine extensor digitorum longus (EDL) and soleus (SOL), which represent fast and slow muscle, respectively, and further explored their effects on lipid accumulation in longissimus dorsi adipocytes. Results Porcine SOL-derived exosomes (SOL-EXO) and EDL-derived exosomes (EDL-EXO) were first identified and their average particle sizes were approximately 84 nm with double-membrane disc- shapes as observed via transmission electron microscopy and scanning electron microscopy. Moreover, the intramuscular fat content of the SOL was greater than that of the EDL at 180 days of age, because SOL intramuscular adipocytes had a stronger lipid-accumulating capacity than those of the EDL. Raman spectral analysis revealed that SOL-EXO protein content was much greater than that of EDL-EXO. Proteomic sequencing identified 72 proteins that were significantly differentially expressed between SOL-EXO and EDL-EXO, 31 of which were downregulated and 41 of which were upregulated in SOL-EXO. Conclusions Our findings suggest that muscle-fat tissue interactions occur partly via SOL-EXO promoting adipogenic activity of intramuscular adipocytes.

In vitro model provides alternatives to the use of live animals in research. In pig nutrition, there has been a tremendous increase in in vivo research over the decades. Proper utilization of in vitro models could provide a screening tool to reduce the needs of in vivo studies, research duration, cost, and the use of animals and feeds. This study aimed to develop a multi-step porcine in vitro system to simulate nutrient digestion and intestinal epithelial immune responses affected by feedstuffs and feed additives. Seven feedstuffs (corn, corn distillers dried grains with solubles [corn DDGS], barley, wheat, soybean meal, soy protein concentrates, and Corynebacterium glutamicum cell mass [CGCM]), feed enzymes (xylanase and phytase), and supplemental amino acids (arginine, methionine, and tryptophan), were used in this in vitro evaluation for their efficacy on digestibility, digesta characteristics, and intestinal health compared with the results from previously published in vivo studies. All in vitro evaluations were triplicated. Data were analyzed using Mixed procedure of SAS9.4. Evaluations included (1) nutrient digestibility of feedstuffs, (2) the effects of feed enzymes, xylanase and phytase, on digestibility of feedstuffs and specific substrates, and (3) the effects of amino acids, arginine, tryptophan, and methionine, on anti-inflammatory, anti-oxidative, and anti-heat stress statuses showing their effects (P

Glutamine, one of the most abundant amino acids in the body, has been shown to exert various beneficial effects in pigs. However, knowledge regarding the role of dietary glutamine in low-protein diet-fed piglets remains scarce. The present study aimed to investigate the effects of different levels of L-glutamine on growth performance, serum biochemistry parameters, redox status, amino acids, and fecal microbiota in low-protein diet-fed piglets. A total of 128 healthy crossbred piglets (Landrace × Yorkshire) were randomly allocated into 4 groups of 4 replicate pens, with 8 piglets per pen. Piglets in the 4 groups were fed with corn and soybean meal-based low-protein diets (crude protein level, 17%) that contained 0%, 1%, 2%, and 3% L-glutamine, respectively, for 28 d. Pigs administered 1% L-glutamine had greater body weight on d 28 and average daily gain (ADG, P

The reduced nutrient digestibility of low-protein (LP) diets has been shown to be caused by the weakened fermentative capacity of the post-gut flora. The dynamic regulation of dietary protein contents on post-gut microbial population and fermentative metabolism is unclear. Twelve growing barrows (19.9 ± 0.8 kg) fitted with a T-cannula at the blind end of the cecum were randomly administered a high-protein (HP, 21.5% crude protein [CP]) diet or an LP (15.5% CP) diet for 28 d. The cecal content and feces were collected at d 1, 14, and 28 of the experiment for microflora structures and metabolite concentrations analysis. The nutrient digestibility coefficient and plasma biochemical parameters were also determined. Compared with the HP treatment, the LP treatment showed decreased plasma urea nitrogen concentration and apparent total tract digestibility of dry matter, gross energy, and CP (P

【Objective】The study aimed to isolate Escherichia coli without virulence genes and antibiotic resistance from pig intestines and to analyze their growth performance and genetic evolution, with a view to laying a foundation for the development of microecologics or oral vaccines for pigs.【Method】Fecal samples of 11 and 15 healthy adult pigs were collected from two pig farms in Guangdong Province, respectively. First, E. coli strains were isolated by McConkey plate and identified by 16S rRNA gene sequencing. Then, PCR was used to verify whether the selected E. coli contained 13 common virulence genes of pig pathogenic E. coli. Antimicrobial susceptibility tests were performed with 7 classes (14 kinds) of antimicrobial agents. Finally, the growth curves of selected E. coli strains were analyzed, and a phylogenetic tree was constructed based on the extracted genome sequences.【Result】Among 260 suspected E. coli strains, 206 strains were identified as E. coli by 16S rRNA gene sequencing analysis. 107 strains of E. coli did not contain 13 common virulence genes, of which 25 strains were sensitive to 7 classes (14 kinds) of antibiotics. Among the 25 E. coli strains, 23 non-pathogenic and non-antibiotic resistant E. coli strains (numbered as 2-9, 3-2, 3-4, 5-1, 6-1, 6-2, 6-9, 6-10, 8-2, 8-9, 10-5, B-4, B-6, B-7, B-10, D-10, E-2, E-10, J-1, J-4, K-6, L-6 and O-9) showed similar growth performance to E. coli Nissle 1917 and MG1655, and 2 E. coli strains (numbered as 6-4 and 5-2) showed higher growth rates than those of E. coli Nissle 1917 and MG1655. Among them, 10 strains (numbered as E-10, E-2, 6-4, 6-9, 8-2, O-9, 3-2, 6-2, J-1, K-6) had good acid tolerance and may be suitable for long-term colonization in the gastrointestinal tract of pigs. The phylogenetic tree analysis based on SNP core genome showed that 9 strains (numbered as 3-2, 6-10, 10-5, 6-1, 8-9, 5-2, L-6, O-9 and K-6) of the 25 strains were in the same group at the beginning of the phylogenetic tree, which were most likely to belong to the ancestral group of pig native E. coli.【Conclusion】Based on the comprehensive analysis of antibiotic sensitivity, growth performance, genome-wide evolutionary tree and gastric environment tolerance, one strain (numbered as O-9) was dominant in these four aspects. The growth kinetics of this strain is similar to or even better than that of E. coli Nissle 1977 and MG1655, which means that O-9 strain of E. coli isolates was likely to be high-quality pig native E. coli strain, with great research potential.

The aim of this experiment was to evaluate the effects of low crude protein (CP) level with essential amino acids (AA) addition on growth performance, nutrient digestibility, microbiota, and volatile fatty acid composition in growing pigs. A total of 160 growing pigs (Landrace × Yorkshire × Duroc [LYD]; average initial body weight 16.68 ± 0.12 kg) were randomly allotted to one of the four treatments on the basis of initial body weight. A randomized complete block design was used to conduct this experiment in the Research Center of Animal Life Sciences at Kangwon National University. There were ten pigs/replicate with four replicates in each treatment. The treatments include; CON (Control, 17.2% dietary CP level), low protein (LP)-1.10 (15.7% dietary CP level + 1.10% lysine level), LP-1.15 (15.7% dietary CP level + 1.15% lysine level), LP1.2 (15.7% dietary CP level + 1.20% lysine level). The pigs fed CON and LP-1.2 diet showed greater final body weight than that of LP-1.1 diet (p < 0.05). Although average daily gain, average daily feed intake, and feed efficiency did not show any difference in phase 2 and 3, average daily gain and feed efficiency was significantly greater in CON and LP-1.20 in phase 1. However, the average daily feed intake did not show any difference during the experimental period. Isobutyric acid and isovaleric acid composition of LP treatments were lower than CON treatment in phase 2. Total branched chain fatty acid composition was significantly lower in LP treatment in phases 1 and 2. However, there was no significant difference among treatments in phase 3. The results of this study underscore the importance of AA supplementation when implementing a low-protein diet during the early growth phase (16–50 kg) in pigs.

Abstract Background Skeletal muscle development and fat deposition have important effects on meat quality. The study of regulating skeletal muscle development and fat deposition is of great significance in improving the quality of carcass and meat. In the present study, whole transcriptome sequencing (including RNA-Seq and miRNA-Seq) was performed on the longissimus dorsi muscle (LDM) of Jinfen White pigs at 1, 90, and 180 days of age. Results The results showed that a total of 245 differentially expressed miRNAs were screened in any two comparisons, which may be involved in the regulation of myogenesis. Among them, compared with 1-day-old group, miR-22-5p was significantly up-regulated in 90-day-old group and 180-day-old group. Functional studies demonstrated that miR-22-5p inhibited the proliferation and differentiation of porcine skeletal muscle satellite cells (PSCs). Pearson correlation coefficient analysis showed that long non-coding RNA (lncRNA) LOC106505926 and CXXC5 gene had strong negative correlations with miR-22-5p. The LOC106505926 and CXXC5 were proven to promote the proliferation and differentiation of PSCs, as opposed to miR-22-5p. In terms of mechanism, LOC106505926 functions as a molecular sponge of miR-22-5p to modulate the expression of CXXC5, thereby inhibits the differentiation of PSCs. In addition, LOC106505926 regulates the differentiation of porcine preadipocytes through direct binding with FASN. Conclusions Collectively, our results highlight the multifaceted regulatory role of LOC106505926 in controlling skeletal muscle and adipose tissue development in pigs and provide new targets for improving the quality of livestock products by regulating skeletal muscle development and fat deposition.

Abstract Intramuscular fat (IMF) in pork holds significant importance for economic performance within the pig industry and dietary calcium supplementation enhances the accumulation of intramuscular fat. Additionally, calcium ions inhibit translation and reduce protein synthesis. However, the mechanism by which calcium regulates IMF deposition in muscle through translation remains largely unknown. In this study, we compared the ribosome profiles of the longissimus dorsi muscles of Duroc $$\times$$ × Landrace $$\times$$ × Large white pigs from the normal calcium (NC) group or calcium supplement (HC) group by Ribo-seq, and RNA-seq. By integrating multiple-omics analysis, we further discovered 437 genes that were transcriptionally unchanged but translationally altered and these genes were significantly enriched in the oxidative phosphorylation signaling pathway. Furthermore, experimental data showed that inhibiting the expression of COX10 and mtND4L increased triglyceride accumulation in C2C12 cells, providing new targets for intramuscular fat deposition. Finally, this work links dietary calcium, translation regulation and IMF deposition, providing a new strategy for both meat quality and economic performance within the pig industry.

Abstract Porcine islet xenotransplantation is a promising therapy for severe diabetes mellitus. Maintenance of the quality and quantity of porcine islets is important for the success of this treatment. Here, we aimed to elucidate the influence of relatively short-term (14 days) culture on adult porcine islets isolated from three micro-minipigs (P111, P112 and P121). Morphological characteristics of islets changed little after 14 days of culture. The viability of cultured islets was also maintained at a high level (> 80%). Furthermore, cultured islets exhibited similar glucose-stimulated insulin secretion and insulin content at Day 14 were preserved comparing with Day 1, while the expressions of Ins, Gcg and Sst were attenuated at Day 14. Xenotransplantation using diabetic nude mice showed no normalization of blood glucose but increased levels of plasma porcine C-peptide after the transplantation of 14 day cultured porcine islets. Histological assessment revealed that relatively short-term cultured porcine islets were successfully engrafted 56 days following transplantation. These data show that relatively short-term culture did not impair the quality of adult porcine islets in regard to function, morphology, and viability. Prevention of impairment of gene correlated with endocrine hormone is warranted for further improvement.

Abstract Porcine reproductive and respiratory syndrome (PRRS), a highly contagious disease caused by Porcine reproductive and respiratory syndrome virus (PRRSV), results in huge economic losses to the world pig industry. MiRNAs have been reported to be involved in regulation of viral infection. In our study, miR-320 was one of 21 common differentially expressed miRNAs of Meishan, Pietrain, and Landrace pig breeds at 9-h post-infection (hpi). Bioinformatics and experiments found that PRRSV replication was inhibited by miR-320 through directly targeting PRRSV ORF6. In addition, the expression of CCAAT enhancer binding protein beta (CEBPB) was also inhibited by miR-320 by targeting the 3ʹ UTR of CEBPB, which significantly promotes PRRSV replication. Intramuscular injection of pEGFP-N1-miR-320 verified that miR-320 significantly inhibited the replication of PRRSV and alleviated the symptoms caused by PRRSV in piglets. Taken together, miR-320 have significant roles in the infection and may be promising therapeutic target for PRRS.