Oral phage therapy with microencapsulated phage A221 against Escherichia coli infections in weaned piglets.

BACKGROUND: Escherichia coli (E. coli) is a common pathogen that often causes diarrhea in piglets. Since bacteria are becoming more and more resistant to antibiotics, phages have become a promising alternative therapy. However, the therapy of oral phage often fails to achieve the desired effect. A novel phage named A221 was isolated by using E. coli GXXW-1103 as host strain, characterized by electron microscopy, genomic sequencing and analyzed by measuring lysis ability in vitro. RESULTS: Phage A221 was identified as a member of Ackermannviridae, Aglimvirinae, Agtrevirus with 153297 bp genome and effectively inhibited bacterial growth in vitro for 16 h. This study was conducted to evaluate the therapeutic effect of oral microencapsulated phage A221 on E. coli GXXW-1103 infections in weaned piglets. The protective effect of phage was evaluated by body weight analysis, bacterial load and histopathological changes. The results showed that with the treatment of phage A221, the body weight of piglets increased, the percentage of Enterobacteriaceae in duodenum decreased to 0.64%, the lesions in cecum and duodenum were alleviated, and the bacterial load in the jejunal lymph nodes, cecum and spleen were also significantly different with infected group (P < 0.001). CONCLUSIONS: The results showed that phage A221 significantly increased the daily weight gain of piglets, reduced the bacterial load of tissues and the intestinal lesions, achieved the same therapeutic effect as antibiotic Florfenicol. Taken together, oral microencapsulated phage A221 has a good therapeutic effect on bacterial diarrhea of weaned piglets, which provides guidance for the clinical application of phage therapy in the future.

In vivo assessment of bacteriophages specific to multidrug resistant Escherichia coli on fecal bacterial counts and microbiome in nursery pigs.

Escherichia coli is the most common cause of economic loss in swine industry. Nowadays, bacteriophages have been proven as good candidates for controlling bacterial infections. In this study, 6 phages were isolated and selected based on their high efficacy against 11 stains of E. coli isolated from diarrheal pigs. Six groups of weaned piglets were assigned (control, bacterial control (BC), two phage control (PC) and two phage treatment (PT) groups). Two titers (2 × 109 PFU/animal and 2 × 1010 PFU/animal) of phage cocktails consisting of these phages were tested in the PC and PT groups via oral gavage at 24, 48, and 72 h against an E. coli cocktail (2 × 109 CFU/animal) that was given to the piglets at 0, 12, 24, and 48 h of the trial. A significant reduction of fecal E. coli counts was observed in both PT groups from day 1 to 7 following the final phage dosage when compared to those of the BC group. Microbiomes in feces obtained 24 h after the final phage administration revealed phage therapy with both dosages could restore the gut's bacterial composition. Moreover, the given phage cocktails resulted in a significantly higher average daily gain of piglets during the first few weeks in both PC groups and the PT group receiving a higher phage dosage. These findings suggest that bacteriophages might be a potential alternative to antibiotics in the treatment of pathogens. In addition, they could also be utilized to improve pig growth performance.

Diagnosis and management of obstructive urolithiasis in miniature pigs.

This article provides information to assist practitioners in the diagnosis, medical, and surgical management of obstructive urolithiasis in miniature pigs. The article focuses primarily on pigs managed as pets because production swine rarely undergo intensive treatment for urolithiasis. As much as possible, the information in this article is based on published literature, but given the limited number of publications addressing obstructive urolithiasis in pigs, some information is based on the authors' clinical experience. Medical and surgical management of obstructive urolithiasis of pigs is often similar to management in ruminant and small animal species, but differences in swine anatomy, handling, and temperament create unique challenges. Pigs tend to be more difficult to restrain and examine than other livestock and often require heavy sedation or anesthesia to facilitate physical examination and basic diagnostic procedures. Because pigs are monogastrics, some oral treatments used in small animals may be used effectively in pigs. Clinicians should follow AMDUCA and consult the Food Animal Residue Avoidance Databank prior to extralabel drug use because pet pigs are still considered a major food-producing species in the United States.

Phage Treatment Trial to Eradicate LA-MRSA from Healthy Carrier Pigs.

The increase of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) causes a threat to human health. LA-MRSA can be transmitted from animals to animal caretakers, which may further spread MRSA to communities and health care facilities. The objective of this work was to study the efficacy of phage treatment in the eradication of LA-MRSA from healthy carrier pigs. A total of 19 MRSA -positive weanling pigs were assigned to a test (n = 10) and a control group (n = 9). A phage cocktail containing three Staphylococcus phages, or a control buffer was administered to the nares and skin of the pigs three times every two days, after which the phage and MRSA levels in nasal and skin swab samples were monitored for a three-week period. The sensitivity of the strains isolated during the follow-up period to the phage cocktail and each phage individually was analyzed and the pig sera were tested for antibodies against the phages used in the cocktail. The phage treatment did not cause any side effects to the pigs. Phages were found in the skin and nasal samples on the days following the phage applications, but there was no reduction in the MRSA levels in the sampled animals. Phage-resistant strains or phage-specific antibodies were not detected during the experiment. The MRSA load in these healthy carrier animals was only 10-100 CFU/swab or nasal sample, which was likely below the replication threshold of phages. The effectiveness of phage treatment to eradicate MRSA from the pigs could thus not be (reliably) determined.

Efficient influence of ssDNA virus PCV2 replication by CRISPR/Cas9 targeting of the viral genome.

Porcine circovirus type 2 (PCV2), a ubiquitous pathogen that primary cause of postweaning multisystemic wasting syndrome (PMWS), had caused significant morbidity and mortality in swine populations with huge economic losses in the worldwide swine industry. Currently, looking for effective antiviral drugs for PCV2 infection remains an important works. In our study, CRISPR/Cas9 system was used to further detected the key sites of PCV2 replication. We designed 8 single guide RNAs (sgRNA) by targeting essential genes across the genome of PCV2. Western-blot(WB), Cell counting kit-8 for high-throughput sgRNA screening were applied to detect PCV2 replication levels. The results showed that Oc8, O13, O134, NQT and NPS sgRNAs can edit the PCV2 genome efficiently and inhibit PCV2 replication in PK-15 cell; H3 sgRNA cannot edit the PCV2 genome successfully; NAT sgRNA can edit the PCV2 genome efficiently to improve the PCV2 replication in PK-15 cell; O26 sgRNA can edit the PCV2 genome successfully but it is not known yet of its effect on PCV2 replication, besides the Cas9 expression had no effect on cell viability. These data suggest that CRISPR/Cas9 system targeting PCV2 essential genes may serve as a novel therapeutic agent against PCV2 infection in the future.

Field evaluation of a sing-dose bivalent vaccine of porcine circovirus type 2b and Mycoplasma hyopneumoniae.

BACKGROUND: The field efficacy of a bivalent vaccine containing porcine circovirus type 2b (PCV2b) and Mycoplasma hyopneumoniae was evaluated on three pig farms. METHODS: Three pig farms were used, two of which had a history of subclinical PCV2 and clinical M. hyopneumoniae infections between 84 and 126 days of age while concurrent porcine circovirus-associated disease and clinical M. hyopneumoniae infection between 70 and 105 days of age. Each farm vaccinated pigs with a single dose of a bivalent vaccine at 10 days of age while unvaccinated pigs were administered a single dose of phosphate buffered-saline at the same age. RESULTS: Vaccination improved growth performance and reduced clinical scores significantly (p < .05) when compared with unvaccinated animals. The amount of PCV2d loads in blood and M. hyopneumoniae loads in nasal swabs of vaccinated animals were also significantly lower (p < .05) when compared with unvaccinated animals. Immunologically, vaccinated groups elicited a significantly higher (p < .05) level of protective immunity against PCV2d such as neutralizing antibodies and interferon-γ secreting cells (IFN-γ-SC), as well as protective immunity against M. hyopneumoniae such as IFN-γ-SC when compared with unvaccinated animals. Pathologically, vaccination significantly lowered (p < .05) the scores of M. hyopneumoniae-induced pneumonia and PCV2-associated lymphoid lesions when compared with unvaccinated animals. CONCLUSIONS: The evaluated bivalent vaccine provided good protection against PCV2d and M. hyopneumoniae infection under field conditions.

Managing respiratory disease in finisher pigs: Combining quantitative assessments of clinical signs and the prevalence of lung lesions at slaughter.

This study aimed to assess the relationship between quantitative assessments of clinical signs of respiratory disease (recorded manually and automatically) and the prevalence of lung lesions at slaughter to validate the use of both in the management of respiratory disease on farm. This was an observational study where pigs (n = 1573) were monitored from 25 ± 5.3 kg (week 12) to slaughter at 114 ± 15.4 kg (week 24). Pigs were housed in eight rooms divided into six pens on a wean-to-finish farm. A manual pen-based coughing (CF) and sneezing (SF) frequency was recorded weekly, for ten consecutive weeks, and a SOMO box (SoundTalks®) was installed in each room, issuing a daily respiratory distress index (RDI) for 13 weeks. Lungs were individually scored for pneumonia, scarring and dorsocaudal (DC) and cranial (CP) pleurisy lesions at slaughter. Relationship between prevalence of lung lesions and weekly RDI and CF and SF was assessed using Spearman's rank correlations and multivariable linear and logit-normal models. Both coughing and lung lesions were largely pen-specific, which fit the disease presentation of Mycoplasma hyopneumoniae. Results showed agreement between RDI and CF (rs = 0.5, P < 0.001), measuring higher levels of coughing at the beginning (weeks 13-14) and end (weeks 21-24, and weeks 21-22, respectively) of the finisher period. Positive associations were found between the prevalence of pneumonia and CF on week 21 and 22 (P < 0.001 and P = 0.011, respectively) and RDI on week 21-24 (rs > 0.70; P < 0.050); the prevalence of DC and CP, and CF on week 22 (P < 0.001); and prevalence of scar lesions and CF on week 17 and 21 (P = 0.013 and P = 0.004, respectively), and RDI on week 21-24 (rs > 0.70; P < 0.050). In the earlier weeks of the finisher stage, coughing was recorded but was not reflected in a higher prevalence of lung lesions at slaughter. These findings highlight the benefit of including measurements of coughing frequency to complement post mortem findings, to improve the management of respiratory disease on farm.

Clinical management and outcomes for goats, sheep, and pigs hospitalized for treatment of burn injuries sustained in wildfires: 28 cases (2006, 2015, and 2018).

OBJECTIVE: To characterize injuries and describe medical management and clinical outcomes of goats, sheep, and pigs treated at a veterinary medical teaching hospital for burn injuries sustained during wildfires. ANIMALS: Goats (n = 9), sheep (12), and pigs (7) that sustained burn injuries from wildfires. PROCEDURES: Medical records were searched to identify goats, sheep, and pigs that had burn injuries associated with California wildfires in 2006, 2015, and 2018. Data regarding signalment, physical examination findings, treatments, clinical outcomes, time to discharge from the hospital, and reasons for death or euthanasia were recorded. RESULTS: The eyes, ears, nose, mouth, hooves, perineum, and ventral aspect of the abdomen were most commonly affected in both goats and sheep. In pigs, the ventral aspect of the abdomen, distal limb extremities, ears, and tail were most commonly affected. The median (range) time to discharge from the hospital for goats and pigs was 11 (3 to 90) and 85.5 (54 to 117) days, respectively. One of 9 goats, 12 of 12 sheep, and 5 of 7 pigs died or were euthanized. Laminitis and devitalization of distal limb extremities were common complications (13/28 animals) and a common reason for considering euthanasia in sheep and pigs. CONCLUSIONS AND CLINICAL RELEVANCE: Burn injuries in small ruminants and pigs required prolonged treatment in some cases. Results suggested prognosis for survival may be more guarded for sheep and pigs with burn injuries than for goats; however, further research is needed to confirm these findings.

Curing piglets from diarrhea and preparation of a healthy microbiome with Bacillus treatment for industrial animal breeding.

High-throughput farming of animals for an essential purpose such as large scale health and production of hogs is a challenge for the food industry in the modern world. The problem is that the breeding of livestock for fast growth or high yields of meat is often associated with illness and microbial infection that develop under the breeding conditions. Piglet diarrhea is most common pig disease, leading to heavy mortality and thereby economic loss. We proved that chemical drugs can relieve the symptoms of diarrhea in ill piglets, but they do not treat the underlying cause, i.e. significantly altered bacterial gut flora. Using Illumina sequencing of fecal DNA, we showed that the bacterial gut flora of piglets treated with antibiotics remain close to the ill conditions. However, using Illumina sequencing of fecal DNA from piglets treated with a specific Bacillus (Bacillus subtilis Y-15, B. amyloliquefaciens DN6502 and B. licheniformis SDZD02) demonstrated the efficiency of natural bioproducts not only on curing diarrhea, but also on beneficial bacteria to re-establish in the piglet gut. We therefore propose a new natural "medicine" to be explored by the world farm animal agriculture industry, particularly for sustainable improvement of swine livestock production and health.

Dietary osteopontin-enriched algal protein as nutritional support in weaned pigs infected with F18-fimbriated enterotoxigenic Escherichia coli.

This study investigated the effects of dietary osteopontin (OPN)-enriched algal protein on growth, immune status, and fecal fermentation profiles of weaned pigs challenged with a live infection of F18-fimbriated enterotoxigenic E. coli (ETEC). At 21 d of age, 54 pigs (5.95 ± 0.28 kg BW; blocked by BW) were allotted to 1 of 3 experimental groups combining dietary and health statuses. A control diet, containing 1% wild-type algal protein, was fed to both sham-inoculated (NC) and ETEC-inoculated (PC) pigs, while the test diet contained 1% OPN-enriched algal protein as fed only to ETEC-inoculated pigs (OA). All pigs received their assigned dietary treatment starting at study initiation to permit a 10-d acclimation period prior to inoculation. Growth performance, fecal dry matter, as well as hematological, histopathological, immune, and microbiota outcomes were analyzed by ANOVA, where treatment and time were considered as fixed effects and pig as a random effect; significance was accepted at P < 0.05. Overall, ETEC-inoculated pigs (PC and OA) exhibited decreased (P < 0.05) ADG and G:F, as well as increased (P < 0.05) peripheral blood helper T-cells and total leukocyte counts, compared with NC pigs during the postinoculation period. The OA treatment also elicited the highest (P < 0.05) concentrations of circulating tumor necrosis factor-α and volatile fatty acid concentrations in luminal contents at various postinoculation time-points, compared with other treatments. A principal coordinate analysis based on Unifrac weighted distances indicated that NC and OA groups had similar overall bacterial community structures, while PC pigs exhibited greater diversity, but infection status had no impact on α-diversity. Osteopontin-specific effects on microbial community structure included enrichment within Streptococcus and Blautia genera and decreased abundance of 12 other genera as compared with PC pigs. Overall, ETEC-infected pigs receiving 1% OPN-enriched algal protein exhibited changes immunity, inflammatory status, and colonic microbial community structure that may benefit weanling pigs experiencing F18 ETEC infection.

Increasing dietary tryptophan in conjunction with decreasing other large neutral amino acids increases weight gain and feed intake in weaner pigs regardless of experimental infection with enterotoxigenic Escherichia coli.

Dietary tryptophan (Trp) is a precursor for serotonin, a neuromediator involved in stress responses. Tryptophan competes with other large neutral amino acids (LNAA: tyrosine, isoleucine, leucine, valine, and phenylalanine) to cross the blood-brain barrier; therefore, the regulation of circulating LNAA can influence Trp availability in the cortex and serotonin biosynthesis. The hypothesis examined in this study was that increased supplementation of dietary Trp and a reduction in LNAA for weaned pigs experimentally infected with enterotoxigenic Escherichia coli (ETEC; F4) will increase Trp availability in plasma and reduce indices of the stress response, which will translate to reduced production losses. At 21 ± 3 d of age (mean ± SEM), 96 male pigs (Large White × Landrace) weighing 6.3 ± 0.98 kg (mean ± SEM) were individually penned and allocated to a 4 × 2 factorial arrangement of treatments, with respective factors being 1) four dietary standardized ileal digestible (SID) Trp and LNAA contents, being HTrpHLNAA (Low Trp-High LNAA; 0.24% SID Trp: 5.4% SID LNAA), HTrpHLNAA (Low Trp-Low LNAA; 0.24% SID Trp: 4.6% SID LNAA), HTrpHLNAA (High Trp-High LNAA; 0.34% SID Trp: 5.4% SID LNAA), and HTrpHLNAA (High Trp-Low LNAA; 0.34% SID Trp: 4.6% SID LNAA), and 2) without/with ETEC infection. Pigs were orally infected with 0.8 mL (3.6 × 109 CFU/mL) ETEC at days 7 and 8 after weaning. Pigs fed diets high in Trp irrespective of the level of LNAA (HTrpHLNAA and HTrpLLNAA) had higher plasma Trp concentrations (P < 0.001) and a Trp:LNAA ratio (P < 0.001) before infection and 6 d after infection. Following infection, noninfected pigs had higher plasma Trp (P = 0.03) and a Trp:LNAA ratio (P = 0.004) compared with pigs infected with ETEC. Plasma cortisol levels after infection were higher in ETEC-infected pigs (P = 0.05) and altering dietary Trp and LNAA concentrations did not influence (P > 0.05) plasma cortisol. Pigs fed diet HTrpLLNAA had higher serum serotonin levels 24 h after infection (P = 0.02) compared with pigs fed diets LTrpLLNAA and HTrpHLNAA. Similarly, pigs fed diet HTrpLLNAA had a higher (P = 0.02) average daily gain during the 3-wk study. Overall, average daily feed intake tended to be higher in pigs fed an HTrpLLNAA diet compared with the other diets (P = 0.08). These results suggest that the increased supplementation of dietary Trp with reduced LNAA increased circulating Trp levels that, in turn, likely caused higher serum serotonin levels, irrespective of infection with ETEC, and improved aspects of post-weaning performance.

Practical oxygen therapy for newborn piglets.

Aims: To evaluate the effect of a novel method of practical oxygen therapy on physiological parameters related to survival, weaning weight and preweaning mortality of neonatal piglets under commercial farm conditions. Methods: Piglets from hyperprolific sows born with signs of asphyxia, (n = 109; <6 on a score of respiration, meconium staining and activity) or very low birth weight (VLBW; n = 112; <1.05 kg) were selected for the study. Approximately half of each group (n = 55 VLBW piglets and n = 57 piglets with asphyxia) received 100% oxygen immediately after birth using a specially designed facemask for 45 seconds (VLBW) or 1 minute (asphyxiated). Physiological parameters (peripheral blood oxygen saturation (SpO2) blood glucose concentration and rectal temperature) were measured before oxygen treatment 5 minutes after birth (SpO2) and 24 hours later (SpO2, blood glucose concentration, temperature). Weight at birth, at 24 hours and at 21 days of age, preweaning mortality, and estimated colostrum intake were also recorded. Results: A significant treatment effect on SpO2 was observed (p = 0.013 and p < 0.001 for VLBW and asphyxiated piglets respectively). VLBW and asphyxiated piglets that received oxygen treatment had higher SpO2 after treatment (measured 5 minutes after birth, 97.7 and 97.8% respectively) compared to immediately after birth (93.3 and 86.8% respectively) while untreated piglets showed no variation. Blood glucose concentrations increased in all piglets between birth and 24 hours of age (p = 0.003 and p < 0.001 for asphyxiated and VLBW piglets respectively) and this was higher in asphyxiated piglets that received oxygen than those that did not (5.6 (SE 0.2) mmol/L; p < 0.05). Estimated colostrum intake was higher in asphyxiated (401.6 (SD 24.4) g/kg) and VLBW (374.9 (SE 23.4 g/kg) piglets that received oxygen than those that did not (273.2 (SE 24.1) g/kg; p < 0.001 and 249.0 (SE 22.5) g/kg; p < 0.001 respectively). Similarly weight at weaning was higher in asphyxiated (5.8 (SE 0.2) kg) and VLBW (4.9 (SE 0.2) kg) piglets that received oxygen therapy than control animals (4.9 (SE 0.2) kg; = 0.005 and 4.1 (SE 0.2) kg; p = 0.008 respectively). Furthermore, oxygen treatment markedly reduced preweaning mortality from 9/52 (17%) untreated to 1/57 (1.7%) oxygen-treated piglets suffering asphyxia at birth (p = 0.006). Conclusions: Oxygen therapy improves physiological and productive parameters in piglets born with signs of asphyxia or VLBW. The incorporation of this strategy as part of the farrowing routine enhances the advantages of rearing hyperprolific sows.

[Field trial for eradication of methicillin- resistant Staphylococcus aureus (MRSA) in a pig breeding farm by bacteriophages]. / Feldversuch zur Eradikation von Methicillin-resistenten Staphylococcus aureus (MRSA) mittels Bakteriophagen in einem Schweinezuchtbetrieb.

INTRODUCTION: In recent years, Livestock Associated Methicillin-Resistant Staphylococcus aureus (LA-MRSA) are found frequently in pigs. The colonization of the care staff with LA-MRSA is strongly associated with the intensity and duration of animal contact and LA-MRSA herd prevalence. In human medicine, staphylococcal infections have been controlled successfully by topical or systemic administration of Staphylococcus - associated bacteriophages. Therefore, the present study investigated the effect of a bacteriophage cocktail on skin and mucosal colonization of pigs with MRSA in a pig farm with high MRSA prevalence. In a first experiment, the sows were washed with a bacteriophage cocktail and nose, mouth and vagina were rinsed before the sows were admitted to the farrowing house. Then, 10 ml of the bacteriophage cocktail was administered daily to the sows over the feed until weaning. The suckling piglets were sprayed and sampled twice a week during the suckling period and treated with the bacteriophage cocktail over the feed during the weaning period. In further experiments, the weaning room was nebulized three times a day with a bacteriophage cocktail and different concentrations of bacteriophages were added to the drinking water via Dosatron®. None of the experiments, however, showed an eradication of MRSA neither in nose nor in feces.

INTRODUCTION: Au cours des dernières années, des Staphylococcus aureus résistant à la méthicilline associé au bétail (LA-MRSA) ont été fréquemment trouvés chez les porcs. La colonisation du personnel soignant par des LA-MRSA est fortement corrélée à l'intensité et à la durée du contact avec les animaux et à la prévalence de LA-MRSA dans le troupeau. En médecine humaine, les infections à staphylocoques ont été contrôlées avec succès par l'administration topique ou systémique de bactériophages associés à Staphylococcus. C'est pourquoi la présente étude a étudié l'effet d'un cocktail de bactériophages sur la colonisation de la peau et des muqueuses des porcs atteints de SARM dans une exploitation porcine à forte prévalence de SARM. Dans une première expérience, les truies ont été lavées avec un cocktail de bactériophages et le nez, la bouche et le vagin ont été rincés avant leur admission dans le local de mise bas. Ensuite, 10 ml du cocktail de bactériophages ont été administrés quotidiennement aux truies sur l'aliment jusqu'au sevrage. Les porcelets allaitants ont été pulvérisés et échantillonnés deux fois par semaine pendant la période d'allaitement et traités avec le cocktail de bactériophages sur l'aliment pendant la période de sevrage. Dans d'autres expériences, le local de sevrage a été nébulisée trois fois par jour avec un cocktail de bactériophages et différentes concentrations de bactériophages ont été ajoutées à l'eau de boisson via Dosatron®. Cependant, aucune des expériences n'a montré d'éradication du SARM ni dans le nez ni dans les fèces.

The role of viral particle integrity in the serological assessment of foot-and-mouth disease virus vaccine-induced immunity in swine.

The efficacy of foot-and-mouth disease virus (FMDV) inactivated vaccines is mainly dependent on the integrity of the whole (146S) viral particles. If the intact capsids disassemble to 12S subunits, antibodies against internal-not protective epitopes, may be induced. Serological correlates with protection may be hampered if antibodies against internal epitopes are measured. Here we compared the performance of different ELISAs with the virus-neutralization test (VNT) that measures antibodies against exposed epitopes. Sera from pigs immunized with one dose of an expired commercial FMDV vaccine were used. This vaccine contained about 50% of O1/Campos and over 90% of A24/Cruzeiro strains total antigen as whole 146S particles. Specific-total antibodies were measured with the standard liquid-phase blocking ELISA (LPBE). We also developed an indirect ELISA (IE) using sucrose gradient purified 146S particles as capture antigen to titrate total antibodies, IgM, IgG1 and IgG2. A good correlation was found between VNT titers and IgG-ELISAs for A24/Cruzeiro, with the lowest correlation coefficient estimated for IgG2 titers. For O1/Campos, however, the presence of antibodies against epitopes different from those of the whole capsid, elicited by the presence of 12S particles in the vaccine, hampered the correlation between LPBE and VNT, which was improved by using purified O1/Campos 146S-particles for the liquid-phase of the LPBE. Interestingly, 146S particles but not 12S were efficiently bound to the ELISA plates, confirming the efficiency of the IE to detect antibodies against exposed epitopes. Our results indicate that any serological test assessing total antibodies or IgG1 against epitopes exposed in intact 146S-particles correlate with the levels of serum neutralizing antibodies in vaccinated pigs, and might potentially replace the VNT, upon validation. We recommend that antigen used for serological assays aimed to measure protective antibodies against FMDV should be controlled to ensure the preservation of 146S viral particles.

Capsulized faecal microbiota transplantation ameliorates post-weaning diarrhoea by modulating the gut microbiota in piglets.

Early weaning-induced stress causes diarrhoea, thereby reducing the growth performance of piglets. Gut bacterial dysbiosis has emerged as a leading cause of post-weaning diarrhoea. The present study aimed to investigate the effect of capsulized faecal microbiota transplantation (FMT) on the gut bacterial community, immune response and gut barrier function of piglets. Thirty-two weaned barrows were randomly divided into two groups. The recipient group was inoculated orally with capsulized faecal microbiota of healthy Tibetan pigs during the whole period of the trial, while the control group was given an empty capsule. The feed-to-gain ratio, diarrhoea ratio, and histological damage score of recipient piglets were significantly decreased. FMT treatment significantly increased the colon length of piglets. Furthermore, the relative abundances of Firmicutes, Euryarchaeota, Tenericutes, Lactobacillus, and Methanobrevibacter in the colon of recipient piglets were increased, and the relative abundances of Campylobacter and Proteobacteria were significantly decreased compared with those in the control group. CD4+ lymphocytes and CD4+/CD8+ ratio in the peripheral blood of recipient piglets were significantly increased. FMT treatment increased the IL-4 and IL-10 levels and decreased the TNF-α and INF-γ levels in the colonic tissue of piglets. The recipient piglets' mRNA expression of TLR2, TLR8, NF-κB, and iNOS was significantly regulated. In addition, FMT significantly enhanced the gene expression of ZO-1. Overall, treatment with capsulized FMT ameliorated diarrhoea in piglets, with significant effects on limiting colon inflammatory responses, downregulating the TLR signalling pathway and the gene expression of iNOS, and strengthening intestinal barrier function by modulating the constituents of the gut microbiota.

In vitro probiotic properties of Pediococcus pentosaceus L1 and its effects on enterotoxigenic Escherichia coli-induced inflammatory responses in porcine intestinal epithelial cells.

This study aimed to evaluate in vitro probiotic characteristics of Pediococcus pentosaceus strain L1 from pickled radish and investigate its impacts on inflammatory responses in porcine intestinal epithelial cells (IEC) to enterotoxigenic Escherichia coli (ETEC) F4+. The abilities of P. pentosaceus L1 to tolerate gastrointestinal conditions and to antagonize ETEC F4+ growth were determined. Adhesion of P. pentosaceus L1 and its effect on ETEC F4+ adhesion to porcine IPEC-J2 IEC were evaluated. Furthermore, the effects of this strain on proinflammatory gene expression and cytokines/chemokine production in porcine IPEC-J2 IEC induced by ETEC F4+ were determined. P. pentosaceus L1 showed good tolerance to the medium adjusted at pH 2.5 and consequently supplemented with 0.3% oxgall. Reduction of ETEC F4+ growth in co-culture with L1 was found. Effective adhesion of L1 to porcine. IPEC-J2 IEC was observed under these conditions. P. pentosaceus L1 decreased the adhesion of ETEC F4+ to IPEC-J2 IEC and the extent of inhibition of ETEC F4+ adhesion depended on the timing of L1 addition. Further analysis revealed down-regulation of expression of ETEC F4+-induced proinflammatory genes encoding interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-8 (IL-8) in IPEC-J2 IEC. Expression of the genes involved in NF-κB pathway, including RELA and NFKB1, were also repressed, as was production of IL-6, TNF-α, and IL-8. These results indicate that P. pentosaceus L1 may have potential as a probiotic for control of ETEC infection in pigs.

Time course determination of the effects of rapid and gradual cooling after acute hyperthermia on body temperature and intestinal integrity in pigs.

Rapid cooling after acute hyperthermia may cause a sustained increase in body temperature and exacerbate intestinal damage in pigs. Therefore, the study objective was to evaluate the temporal effects of rapid and gradual cooling on body temperature response and intestinal integrity after acute hyperthermia in pigs. In three repetitions, 54 pigs [83.3 ± 6.7 kg initial body weight (BW)], balanced by sex were exposed to thermoneutral conditions for 6 h (TN; n = 6 pigs/repetition; 21.1 ± 2.0°C), or heat stress conditions (HS; 39.3 ± 1.6°C) for 3 h, followed by a 3 h recovery period of gradual cooling [HSGC; n = 6 pigs/repetition; gradual decrease from HS to TN conditions] or rapid cooling [HSRC; n = 6 pigs/repetition; rapid TN exposure and cold water (4.0°C) dousing every 30 min for 1.5 h]. Feed was withheld throughout the entire 6 h period, but water was provided ad libitum. Gastrointestinal (TGI) and rectal (TR) temperatures were recorded every 15 min during the HS and recovery periods. Six pigs per repetition (n = 2/treatment) were euthanized and jejunal and ileal samples were collected for histology immediately after (d 0), 2 d after, and 4 d after the recovery period. Data were analyzed using PROC MIXED in SAS 9.4. Overall, rapid cooling reduced TR and TGI (P < 0.01; 0.95°C and 0.74°C, respectively) compared to gradual cooling. Jejunal villus height was reduced overall (P = 0.02; 14.01%) in HSGC compared to HSRC and TN pigs. Jejunal villus height-to-crypt depth ratio was reduced overall (P = 0.05; 16.76%) in HSGC compared to TN pigs. Ileal villus height was reduced overall (P < 0.01; 16.95%) in HSGC compared to HSRC and TN pigs. No other intestinal morphology differences were detected. In summary, HSRC did not cause a sustained increase in body temperature and did not negatively impact biomarkers of intestinal integrity in pigs.

Lactobacillus johnsonii L531 ameliorates enteritis via elimination of damaged mitochondria and suppression of SQSTM1-dependent mitophagy in a Salmonella infantis model of piglet diarrhea.

Newly weaned piglets challenged with Salmonella infantis were particularly susceptible, whereas oral preadministration of Lactobacillus johnsonii L531 alleviated enteritis and promoted intestinal secretory IgA production. Salmonella infantis-induced activation of NLRC4 and NLRP3 inflammasomes and (nuclear factor kappa B) NF-κB signaling in the small intestine was also inhibited by L. johnsonii L531 pretreatment, thus limiting inflammation. An IPEC-J2 cell model of S. infantis infection yielded similar results. Salmonella infantis infection also resulted in mitochondrial damage and impaired mitophagy in the ileum and IPEC-J2 cells, as demonstrated by immunofluorescence colocalization of mitochondria with microtubule-binding protein light chain 3 (LC3) and high expression of autophagy-related proteins PTEN-induced putative kinase 1 (PINK1), sequestosome 1 (SQSTM1/p62), optineurin (OPTN), and LC3 by Western blotting analysis. However, L. johnsonii L531 pretreatment reduced both the extent of mitochondrial damage and autophagy-related protein expression. Our findings suggest that the amelioration of S. infantis-associated enteritis by L. johnsonii L531 is associated with regulation of NLRC4 and NLRP3 inflammasomes and NF-κB signaling pathway activation and suppression of mitochondrial damage. Amelioration of impaired mitophagy by L. johnsonii L531 could involve eliminating damaged mitochondria and regulating S. infantis-induced activation of the NF-κB-SQSTM1mitophagy signaling pathway in host cells to prevent the further mitochondrial damage and S. infantis dissemination.