Weaning diet supplemented with health-promoting feed additives influences microbiota and immune response in piglets challenged with Salmonella.

The aim of this study was to evaluate the potential of micronutrients and feed additives to modulate intestinal microbiota and systemic and mucosal immune responses in weaned pigs infected with Salmonella. At weaning, 32 litters of 12 piglets each were allocated to four dietary treatments: 1) control diet (CTRL), 2) CTRL supplemented with chlortetracycline (ATB), 3) CTRL supplemented with a cocktail of feed additives (CKTL); and 4) CKTL diet containing bovine colostrum in replacement of spray-dry animal plasma (CKTL+COL). The CKTL supplement included cranberry extract, encapsulated carvacrol and yeast-derived products and an enriched selenium and vitamin premix. Three weeks after weaning, four pigs per litter were orally inoculated with Salmonella Typhimurium DT104. Half of them were euthanized 3 days post-infection (dpi) and the other half, 7 dpi. The expression of IL6, TNF, IL8, monocyte chemoattractant protein 1 (MCP1), IFNG, cyclooxygenase 2 (COX2), glutathione peroxidase 2 (GPX2) and ß-defensin 2 (DEFB2) showed a peaked response at 3 dpi (P < 0.05). Results also revealed that DEFB2 expression was higher at 3 dpi in CTRL and CKTL groups than in ATB (P = 0.01 and 0.06, respectively) while GPX2 gene was markedly increased at 3 and 7 dpi in pigs fed CKTL or CKTL+COL diet compared to CTRL pigs (P < 0.05). In piglets fed CKTL or CKTL+COL diet, intestinal changes in microbial communities were less pronounced after exposure to Salmonella compared to CTRL and progressed faster toward the status before Salmonella challenge (AMOVA P < 0.01). Furthermore, the relative abundance of several families was either up- or down-regulated in pigs fed CKTL or CKTL+COL diet after Salmonella challenge. In conclusion, weaning diet enriched with bovine colostrum, vitamins and mixture of feed additives mitigated the influence of Salmonella infection on intestinal microbial populations and modulate systemic and intestinal immune defences.

Prevalence of Mycoplasma synoviae and Its Impact on Productivity in Commercial Poultry Farms in Quebec, Canada.

Mycoplasma synoviae (MS) is associated with upper respiratory disease, joint, and reproductive system disease in poultry. Economic losses are due to stunting, increased mortality, lower egg production, and higher slaughterhouse condemnations. The seroprevalence of MS is increasing worldwide, and more pathogenic strains have emerged over the past few years. Where this increase is noted, the economic consequences are considerable, even when there are no obvious clinical signs. The best control strategy is to maintain mycoplasma-free flocks. Since 2014 in Quebec, Canada, MS has been isolated with greater frequency in poultry farms and at times, as a primary pathogenic agent. The aim of this study was to evaluate the prevalence and impact of MS in commercial poultry farms in Quebec because the poultry industry was considering an insurance program that would cover losses in case of an outbreak. MS was shown to be present in all types of commercial production, although egg layers were principally affected with over 50% of flocks sampled being MS-positive in all producing regions of the province. On the basis of vlhA gene sequencing, several strains were identified with the most prevalent ones being type E, followed by Qc-1, a strain specific to Quebec. On average, the impact of MS on production parameters were not significant for any of the different types of commercial poultry production.

Prevalencia de Mycoplasma synoviae y su impacto en la productividad en granjas avícolas comerciales en Quebec, Canadá. Mycoplasma synoviae (MS) se asocia con enfermedades de las vías respiratorias superiores, problemas articulares y del sistema reproductivo en la avicultura comercial. Las pérdidas económicas se deben al retraso en el crecimiento, aumento de la mortalidad, menor producción de huevo y mayores decomisos en plantas de procesamiento. La seroprevalencia de M. synoviae está aumentando en todo el mundo y en los últimos años han surgido cepas más patógenas. En los lugares donde se ha observado este aumento, las consecuencias económicas han sido considerables, incluso cuando no se presentan signos clínicos evidentes. La mejor estrategia de control es mantener las parvadas libres de micoplasmas. Desde 2014 en Quebec, Canadá, M. synoviae se ha aislado con mayor frecuencia en granjas avícolas y en ocasiones como agente patógeno primario. El objetivo de este estudio fue evaluar la prevalencia y el impacto de M. synoviae en granjas avícolas comerciales en Quebec porque la industria avícola estaba considerando un programa de seguros que cubriría las pérdidas en caso de un brote. Se demostró que M. synoviae estaba presente en todos los tipos de producción comercial, aunque las aves de postura de huevo se vieron afectadas principalmente, con más del 50% de las parvadas muestreadas positivas a M. synoviae en todas las regiones productoras de la provincia. Con base en la secuenciación del gene vlhA, se identificaron varias cepas, siendo las más prevalentes el tipo E, seguidas de la cepa QC-1, que es específica de Quebec. En promedio, el impacto de M. synoviae en los parámetros de producción no fue significativo para ninguno de los diferentes tipos de producción avícola comerciales.

Salmonella contamination in a network of 10 pig farms interconnected within the same cooperative.

OBJECTIVE: To evaluate whether pig farms interconnected within the same cooperative share similar Salmonella contamination patterns. SETTING: Ten finishing pig farms within a 100 km radius of a common slaughterhouse were selected. Their inclusion was based on their association to the same cooperative and the sharing of common resources: piglets, feed, swine transporters, slaughterhouse, technicians and veterinarians. PROCEDURE: Each farm was visited three times over a 10-month period. Pig faeces, the barn front door handle, the feed pipeline, mobile objects (shovel, balance and pig board), the landing stage, the concrete slab of the feed bins, the tire tracks left on the pathways by the animal feed truck, the pig delivery truck and the carcase knacker truck and the mudguards and cabin carpets of the veterinarian and technician vehicles on their arrival at the farm were all analysed for the presence of Salmonella. RESULTS: All farms were not equally contaminated with Salmonella. Whereas some farms yielded up to 12 Salmonella isolates, other farms were Salmonella free. Some locations, most notably the landing stage, were more contaminated than others. Salmonella contamination was dynamic in time. Some contaminations seen on farms, on specific locations on the first visit, had disappeared on the second and third visits, but new contaminations were detected on different locations. CONCLUSIONS: Contamination with Salmonella was not disseminated through the network of the 10 pig farms interconnected within the same cooperative but was rather most often restricted in time to specific locations on specific farms.

Impact of zinc and arginine on antioxidant status of weanling piglets raised under commercial conditions.

The effects of dietary zinc and L-arginine supplements on the weight gain, feed efficiency, antioxidant capacity and oxidative status of weanling piglets raised under commercial conditions were examined. A total of 288 piglets aged 21 d were fed for 15 d a diet supplemented or not with 2,500 mg/kg of zinc (provided as zinc oxide) and 1% L-arginine·HCl. The 4 treatments were distributed in a randomized complete block design with 6 initial body weight categories (12 animals per pen). Access to feed and water was ad libitum. Data were analyzed as a 2 × 2 factorial experiment using the SAS MIXED procedure, with zinc and arginine as the main independent variables. Blood collection day (d 8 and 15, samples were collected from the same 2 piglets in each pen before the morning feeding) was included as a third factor. The zinc supplement increased the average daily gain (ADG) from d 0 to 7, d 8 to 15 and d 0 to 15 (0.289 vs. 0.217 kg/d), average daily feed intake (ADFI) from d 8 to 15 and d 0 to 15 (0.338 vs. 0.279 kg/d) and the gain to feed (G:F) ratio from d 0 to 7 and d 0 to 15 (0.86 vs. 0.77) (P < 0.001). Both supplements significantly decreased the malondialdehyde concentration (zinc: 4.37 vs. 3.91 µmol/L, P = 0.005; arginine: 4.38 vs. 3.89 µmol/L, P = 0.002). Total antioxidant capacity and reduced glutathione (GSH) increased from d 8 to 15 (0.953 vs. 1.391 µmol/L, 2.22 vs. 3.37 µmol/L, P < 0.05) regardless of dietary treatment. Total and oxidized GSH concentrations on d 8 were higher in response to the combined supplements (zinc × arginine interaction, P < 0.05). Piglets fed either Zn-supplemented diet had a lower haptoglobin serum concentration (509 vs. 1,417 mg/L; P < 0.001). In conclusion, the zinc supplement improved piglet growth performance (ADG and ADFI) and oxidative status (based on malondialdehyde concentration). The arginine supplement had a limited effect on growth performance and oxidative status under these conditions.

Overlooked sources of Salmonella contamination in the pig production network: Slaughterhouse yard pathways and mudguards and carpets from transport trucks.

This report describes various Salmonella serovars which were found on often overlooked locations in a pig farm/slaughterhouse interface. These include slaughterhouse yard pathways and mudguards and carpets of transport trucks arriving at and departing from production sites.

Sources négligées de contamination par Salmonella dans un réseau de production de porcs: les voies de circulation de l'abattoir et les garde-boues et les tapis de cabine des camions de transport. Nous montrons ici que Salmonella, l'agent causal de la salmonellose, peut être trouvé sur des sites très inhabituels et négligés dans l'interface ferme porcine/abattoir: les voies de circulation de la cour d'abattoir, et les garde-boues et tapis des camions de transport qui arrivent et partent vers les sites de production.(Traduit par les auteurs).

The fecal presence of enterotoxin and F4 genes as an indicator of efficacy of treatment with colistin sulfate in pigs.

BACKGROUND: Enterotoxigenic Escherichia coli (ETEC) strains producing multiple enterotoxins are important causes of post-weaning diarrhea (PWD) in pigs. The aim of the present study was to investigate the fecal presence of ETEC enterotoxin as well as F4 and F18 genes as an indicator of colistin sulfate (CS) efficacy for treatment of PWD in pigs. Forty-eight piglets were weaned at the age of 21 days, and were divided into four groups: challenged treated, challenged untreated, unchallenged treated, and unchallenged untreated. Challenge was performed using 109 CFU of an ETEC: F4 strain, and treatment was conducted using oral CS at the dose of 50,000 IU/kg. The fecal presence of genes encoding for STa, STb, LT, F4 and F18 was detected using PCR. RESULTS: The PCR amplification of ETEC virulence genes showed that nearly 100% of pigs excreted genes encoding for STa and STb toxins in the feces before the challenge. These genes, in the absence of the gene encoding F4, were considered as a marker for F4-negative ETEC. One day after ETEC: F4 oral challenge pigs in the two challenged groups excreted the genes encoding LT and F4 in the feces. These genes were considered as a marker for F4-positive ETEC. However, the gene encoding F18 was not detected in any fecal samples of the 4 groups throughout the experiment. After only 3 days of successive oral treatment with CS, a significant reduction in both the F4-positive and negative ETEC populations was observed in the challenged treated group compared to the challenged untreated group (p < 0.0001). CONCLUSIONS: Our study is among the first to report that under controlled farming conditions, oral CS treatment had a significant effect on both fecal F4-positive and F4-negative ETEC in pigs. However, CS clinical efficiency was correlated with non-detection of F4-positive ETEC in the feces. Furthermore the fecal presence of F4-negative ETEC was not associated with clinical symptoms of post-weaning diarrhea in pigs.

Feed supplementation with arginine and zinc on antioxidant status and inflammatory response in challenged weanling piglets.

Although supplementing the diet with zinc oxide and arginine is known to improve growth in weanling piglets, the mechanism of action is not well understood. We measured the antioxidant status and inflammatory response in 48 weanling castrated male piglets fed diets supplemented with or without zinc oxide (2,500 mg Zn oxide per kg) and arginine (1%) starting at the age of 20 days. The animals were injected with lipopolysaccharide (100 µg/kg) on day 5. Half of them received another injection on day 12. Blood samples were taken just before and 6, 24 and 48 h after injection and the mucosa lining the ileum was recovered following euthanizing on days 7 and 14. Zinc supplementation increased reduced and total glutathione (GSH) (reduced and total) during days 5 to 7 and arginine decreased oxidized GSH measured on days 5 and 12 and the ratio of total antioxidant capacity to total oxidative status during days 12 to 14. Zinc decreased plasma malondialdehyde measured on days 5 and 12 and serum haptoglobin measured on day 12 and increased both metallothionein-1 expression and total antioxidant capacity measured in the ileal mucosa on day 14. Tumour necrosis factor α concentration decreased from days 5 to 12 (all effects were significant at P < 0.05). This study shows that the zinc supplement reduced lipid oxidation and lipopolysaccharide-induced inflammation during the post-weaning period, while the arginine supplementation had only a limited effect.

In vivo therapeutic efficacy and pharmacokinetics of colistin sulfate in an experimental model of enterotoxigenic Escherichia coli infection in weaned pigs.

Enterotoxigenic Escherichia coli (ETEC: F4) associated with post-weaning diarrhea (PWD) in pigs has developed resistance against several antimicrobial families, leading to increased use of colistin sulfate (CS) for the treatment of this disease. The objective of this study was to determine the efficacy of oral CS treatment in experimental PWD due to ETEC: F4 challenge and determine the effect of this challenge on CS intestinal absorption. In this study, 96 pigs were divided into two trials based on CS dose (100 000 or 50 000 IU/kg). Fecal shedding of ETEC: F4, total E. coli, and CS-resistant E. coli, diarrhea scores, and weight changes were evaluated. Colistin sulfate plasma concentrations were determined by HPLC-MS/MS. Regardless of the dose, CS treatment resulted in a reduction of fecal ETEC: F4 and total E. coli shedding, and in diarrhea scores but only during the treatment period. However, CS treatment resulted in a slight increase in fecal shedding of CS resistant E. coli and did not prevent weight loss in challenged pigs. In addition, challenge with ETEC: F4 resulted in an increase of CS intestinal absorption. Our study is among the first to demonstrate that under controlled conditions, CS was effective in reducing fecal shedding of ETEC: F4 and total E. coli in experimental PWD. However, CS treatment was associated with a slight selection pressure on E. coli and did not prevent pig weight loss. Further studies are needed in field conditions, to better characterize CS therapeutic regimen efficacy and bacterial resistance dissemination.

Gastric stability and oral bioavailability of colistin sulfate in pigs challenged or not with Escherichia coli O149: F4 (K88).

The aim of the present study was to investigate the in vitro gastric stability of colistin sulfate (CS) and its antimicrobial activity against Escherichia coli and to study the impact of ETEC O149: F4 (K88) infection in pigs on CS intestinal absorption. The stability profile of CS was evaluated in a simulated gastric fluid (SGF). Antimicrobial activity of CS and its degradation products were examined in a 96-well polystyrene microplate model. The effect of experimental infection with ETEC O149: F4 on CS intestinal absorption was determined by quantification of CS systemic concentration using a validated LC-MS/MS method. A rapid degradation of CS accompanied by an increase in CS antimicrobial activity by comparison with non-degraded CS (P<0.0001) was observed in SGF. Additionally, CS levels were not quantifiable in systemic circulation using a highly sensitive method and concurrent oral challenge did not affect CS absorption in an induction model of subclinical post-weaning diarrhea (PWD).

Characterization of Salmonella Typhimurium isolates associated with septicemia in swine.

Salmonella Typhimurium is frequently isolated from pigs and may also cause enteric disease in humans. In this study, 33 isolates of S. Typhimurium associated with septicemia in swine (CS) were compared to 33 isolates recovered from healthy animals at slaughter (WCS). The isolates were characterized using phenotyping and genotyping methods. For each isolate, the phage type, antimicrobial resistance, and pulsed-field gel electrophoresis (PFGE) DNA profiles were determined. In addition, the protein profiles of each isolate grown in different conditions were studied by Coomassie Blue-stained sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot. Various phage types were identified. The phage type PT 104 represented 36.4% of all isolates from septicemic pigs. Resistance to as many as 12 antimicrobial agents, including some natural resistances, was found in isolates from CS and WCS. Many genetic profiles were identified among the PT 104 phage types. Although it was not possible to associate one particular protein with septicemic isolates, several highly immunogenic proteins, present in all virulent isolates and in most isolates from clinically healthy animals, were identified. These results indicated that strains associated with septicemia belong to various genetic lineages that can also be recovered from asymptomatic animals at the time of slaughter.

Interaction between host cells and septicemic Salmonella enterica serovar typhimurium isolates from pigs.

Salmonella enterica serovar Typhimurium is an important pathogen in swine and is also a frequently reported zoonotic agent. The objective of this study was to characterize isolates of S. enterica serovar Typhimurium associated with septicemia in swine and to compare them to isolates recovered from clinically healthy pigs. We were particularly interested in comparing the two groups of isolates for their ability to adhere to and invade host cells, to be phagocytized and survive in monocyte cells, to induce apoptosis, and to adhere to intestinal mucus. Their surface properties were also evaluated by interactions with solvents. The isolates recovered from diseased animals were shown to invade intestinal epithelial cell lines at a higher rate (P = 0.003) than isolates from healthy pigs. Septicemic isolates were phagocytized by human monocytes at a higher rate than isolates from healthy pigs (P = 0.009). The mean percentages of phagocytosis were significantly lower for human monocytes than for porcine monocytes (P = 0.02 and P = 0.008, respectively) for isolates from both diseased and healthy animals. Healthy animal isolates were phagocytized more by porcine monocytes at 15 min (P = 0.02) than septicemic isolates. No difference between isolates from septicemic pigs and isolates from healthy pigs was detected for other tested parameters. These results suggest that septicemic isolates have a particular pattern of invasion.