The vehicle of administration, feed or water, and prandial state influence the oral bioavailability of amoxicillin in piglets.

Feed and water components may interact with drugs and affect their dissolution and bioavailability. The impact of the vehicle of administration (feed and water) and the prandial condition of weaner piglets on amoxicillin´s oral bioavailability was evaluated. First, amoxicillin's in vitro dissolution and stability in purified, soft, and hard water, as well as release kinetics from feed in simulated gastric and intestinal media were assessed. Then, pharmacokinetic parameters and bioavailability were determined in fasted and fed pigs using soft water, hard water, or feed as vehicles of administration following a balanced incomplete block design. Amoxicillin showed similar dissolution profiles in soft and hard water, distinct from the dissolution profile obtained with purified water. Complete dissolution was only achieved in purified water, and merely reached 50% in soft or hard water. Once dissolved, antibiotic concentrations decreased by around 20% after 24 h in all solutions. Korsmeyer-Peppas model best described amoxicillin release from feed in simulated gastric and intestinal media. Feed considerably reduced antibiotic dissolution in both simulated media. In vivo, amoxicillin exhibited significantly higher bioavailability when delivered via water to fasted than to fed animals, while in-feed administration yielded the lowest values. All treatments showed a similar rate of drug absorption. In conclusion, we demonstrated that water and feed components, as well as feed present in gastrointestinal tract of piglets decrease amoxicillin´s oral bioavailability. Therefore, the use of oral amoxicillin as a broad-spectrum antibiotic to treat systemic infections in pigs should be thoroughly revised.

Fosfomycin modifies the replication kinetics of bovine alphaherpesvirus-1 and reduces the timing of its protein expression on bovine (MDBK) and human (SH-SY5Y) cell lines.

Bovine alphaherpesvirus 1 (BoAHV-1) predisposes cattle to respiratory secondary bacterial infections, which can be treated with the broad-spectrum antibiotic fosfomycin. This drug also suppresses NF-kB activity and pro-inflammatory responses. Therefore, cattle may be exposed to an interaction between the virus and the antibiotic which may have effects on it. The aim of this study was to determine the effect of calcium fosfomycin (580 µg/mL) on BoAHV-1 (moi = 0.1) replication. Two cell lines (MDBK and SH-SY5Y) were used in this study. Our results show that fosfomycin has novel properties. By MTT assay we have shown that it is non-cytotoxic for any of the cell lines. Extracellular and intracellular viral titers demonstrated that fosfomycin has a cell-type and time-dependent effect on BoAHV-1 replication. By direct immunofluorescence it was shown that it reduces the timing of BoAHV-1 protein expression, and by qPCR, we found that its effect on NF-kB mRNA expression depends on the cell type.

Citrullinemia is a suitable biomarker for post weaning performance in piglets under intensive farming.

OBJECTIVE: To describe citrullinemia profiles during the weaning transition and correlate citrulline production with stress and growth in a commercial pig farm. ANIMALS: 240 healthy piglets of homogenous weight, weaned from second and third parity sows, were selected at weaning and subjected to the farm's routine management practices in May to July 2020 and May to July 2021. PROCEDURES: Piglets were weighed at weaning, then 15 and 49 days later in order to calculate daily weight gain during the first 15 and 49 days after weaning. Blood samples were collected from each piglet to determine citrulline and cortisol profiles during the early postweaning period. RESULTS: Citrullinemia decreased dramatically during the first week postweaning and then increased progressively to reach preweaning values by 15 days postweaning. Citrulline production during the first 2 weeks postweaning was negatively correlated with cortisol production (r: -0.2949) and positively correlated with mean daily weight gain during the first 15 (ρ: 0.5450) and 49 (ρ: 0.6603) days postweaning. CLINICAL RELEVANCE: Citrullinemia profile of piglets during the early postweaning period showed a temporal negative impact of stress (assessed by plasmatic cortisol levels) on intestinal enterocytes' mass and function, which resulted in a lower average daily weight gain. We demonstrated that a single biomarker, plasmatic citrulline, is useful to describe intestinal metabolism during the early postweaning period and that the greater the citrulline production during the first days after weaning, the higher the weight gain during the entire postweaning period.

Effect of Dietary Oregano (Lippia origanoides) and Clover (Eugenia caryophillata) Essential Oils' Formulations on Intestinal Health and Performance of Pigs.

The incorporation of natural essential oils to the pigs' diet in intensive production systems is a potential tool to improve gut health and prevent infections without using antibiotics. Nevertheless, different products, even containing the same compounds, coming from the same botanical species, may exert dissimilar biological effects due differences in the technological processes by which they are produced and preserved. For this reason, suitability of a given product based on natural extracts, intended for swine production must be thoroughly evaluated. In the present study, we assessed the effects of three additives containing oregano (Lippia origanoides) essential oil, alone or in combination with clover (Eugenia caryophillata) essential oil, with or without being microencapsulated, on gastrointestinal health and on some performance parameters in a commercial pig production farm. Recently weaned piglets were randomly divided in four groups, and basal diet or essential oil-supplemented diet (OCE; MOCE; MOE) was randomly assigned to each of the groups from weaning to finishing. Blood samples were collected at pre-established days after weaning. Intestinal sampling took place at 42 and 72 days of age. Pigs consuming the supplemented diets showed higher intestinal metabolic activity during the post-weaning period, decreasing the impact of weaning stress on enterocytes' metabolism. Intestinal barrier function was not affected in pigs consuming microencapsulated products. All treated groups showed improved intestinal architecture, increased digestive enzymes activity and caecal VFA concentrations. The incorporation of the dietary essential oils products brought beneficial effects on gastrointestinal health that were reflected in improved performance parameters.

Zearalenone Adsorbent Based on a Lyophilized Indigenous Bacterial Lactobacillus plantarum Strain as Feed Additive for Pigs: A Preliminary Study In Vivo.

Feed contamination with naturally occurring mycotoxins is an unavoidable condition of significant concern in intensive productions. The presence of high concentrations of zearalenone >1 ppm in the diet can cause major reproductive disorders, particularly in swine. In order to reduce the consequences of intoxication, mycotoxin adsorbents are incorporated into feed. In the present study, zearalenone adsorption capacity of a lyophilized indigenous strain of Lactobacillus plantarum (L4; previously isolated from pig's rectal swabs) was first evaluated in vitro. A preliminary study in vivo was then performed in which the indigenous Lactobacillus plantarum strain was lyophilized and the powder obtained (L-L4) was incorporated into the diet of gilts two gilts received basal diet (control) and two received basal diet containing 2 g/kg L-L4 (treated). After an adaptation period, all the feed was contaminated with zearalenone at a dose of 0.93 mg ZEA/kg. Results from in vitro assay showed that L-L4 adsorbed 87.9% (SD 3.97) of zearalenone in 0.9% NaCl solution. In the in vivo exploratory study, higher daily weight gain and lower vulva area were observed in gilts that incorporated L-L4 to the diet. Additionally, higher zearalenone concentrations were eliminated in faeces from treated animals. The use of a product based on a lyophilized indigenous Lactobacillus strain to protect gilts from detrimental effects of zearalenone consumption has shown promising results so far. However, further studies are required in order to accurately assess its impact and evaluate doses according to different degrees of mycotoxins contamination.

Potential interactions between an oral fosfomycin formulation and feed or drinking water: Impact on bioavailability in piglets.

Feed and drinking water are the most frequently used vehicles for administration of antibiotics in intensive pig production. Interactions of drugs with feed and water components may affect dissolution and bioavailability. Therefore, antibiotic formulations should be tested in order to assure their suitability for oral use. In this study, an oral fosfomycin (FOS) formulation was evaluated considering dissolution in water (soft and hard), release kinetics from feed in simulated gastrointestinal fluids and bioavailability after oral administration blended into feed or dissolved in water (soft and hard), to fed and fasted piglets. FOS reached immediate dissolution in soft and hard water. The presence of feed significantly decreased antibiotic dissolution in simulated intestinal medium. Bioavailability was lower when feed was used as a vehicle for FOS administration than when the drug was dissolved in water (soft or hard). The fed or fasted condition of piglets did not affect bioavailability. Probably, FOS interactions with feed components alter its dissolution in the gastrointestinal tract, and only a fraction of the dose would be available for absorption. This information must be considered to support decisions on eligibility of antibiotic pharmaceutical formulations and the vehicle for their administration in order to pursue a responsible use of antibiotics.

Impact of water hardness on oxytetracycline oral bioavailability in fed and fasted piglets.

Water hardness is a critical factor that affects oxytetracycline dissolution by chelation with cations. These interactions may lead to impaired dosing and consequently decrease absorption. Moreover, feed present in gastrointestinal tract may interact with antibiotic and alter pharmacokinetic parameters. In the present study, dissolution profiles of an oxytetracycline veterinary formulation were assessed in purified, soft and hard water. Furthermore, oxytetracycline absolute bioavailability, after oral administration of the drug dissolved in soft or hard water, was evaluated in fed and fasted piglets. A maximum dissolution of 86% and 80% was obtained in soft and hard water, respectively, while in purified water dissolution was complete. Results from in vivo study reconfirmed oxytetracycline's very low oral bioavailability. The greatest values were attained when antibiotic was dissolved in soft water and in fasted animals. Statistically significant lower absolute bioavailability was achieved when hard water was used and/or animals were fed. Moreover, Cmax attained in all treatments was lower than MIC90 of most important swine pathogens. For these reasons, the oral use of OTC formulations, that have demonstrated low oral bioavailability, should be avoided to treat systemic diseases in pigs.

Effect of fosfomycin, Cynara scolymus extract, deoxynivalenol and their combinations on intestinal health of weaned piglets.

Weaning is a challenging stage of pig farming. Animals undergo environmental, social and dietary changes leading to weaning stress syndrome. In order to compensate for the detrimental effects of weaning stress, antibiotics and natural extracts are used as feed additives, sometimes without fully understanding the interactions between them or even with low concentrations of mycotoxins that are frequently present in feed. The aim of this study was to evaluate the effect of fosfomycin (FOS), Cynara scolymus extract (CSE), deoxynivalenol (DON) and their combined administration on intestinal health of weaned piglets. The experiment was designed as a 2 × 2 × 2 factorial arrangement with 3 factors (FOS, CSE and DON treatments), 2 levels each (presence and absence) and 3 repeats. Weaned piglets (n = 24) were randomly divided in groups to receive the different treatments, namely DON administered in diet (50 µg/kg BW), FOS administered into the drinking water (30 mg/kg BW), CSE administered in diet (15 mg/kg BW) and all their combinations. After 15 d, the animals were euthanized and gastrointestinal tract samples were immediately taken to evaluate gastrointestinal pH, Enterobacteriaceae to lactic acid bacteria (E:L) ratio, volatile fatty acid (VFA) concentrations, disaccharidase (lactase, sucrase and maltase) activity, histology (intestinal absorptive area [IAA] and goblet cells count) and mucus ability to adhere pathogenic Escherichia coli. From our results, FOS and CSE treatments, individually or combined, produced a lower E:L ratio, an enhanced production of butyrate, increased disaccharidase activity (particularly maltase), and a greater IAA and goblet cells count along with an increase in pathogenic bacteria adherence to intestinal mucus. Deoxynivalenol did not show interactions with the other factors and its administration produced decreases on VFA, disaccharidase activity and goblet cells count. In conclusion, weaning piglets receiving diets containing FOS, CSE or both exhibited evident beneficial intestinal effects compared to animals receiving diets free from these compounds. On the contrary, the presence of DON at sub-toxic concentrations produced detrimental effects on intestinal health. The knowledge of the physiological and pathological gut changes produced by these compounds contributes to understand their potential productive consequences.

Ex vivo penetration of fosfomycin into healthy and Lawsonia intracellularis-colonized swine intestinal mucosa.

Fosfomycin (FOS) is an antibiotic used, mostly in Latin America, for the treatment of lung and enteric infections of pigs. Intracellular fluids of enterocytes can act as biophase for Lawsonia intracellularis, the causative agent of porcine proliferative enteropathy (PPE). The aim of this study was to determine whether the presence of L. intracellularis in the enterocytes modifies FOS penetration. Eight healthy pigs in growth-finishing stage were used to produce healthy (group A) and L. intracellularis-colonized (group B) intestinal explants. For both groups, treatment consisted of a 580 µg/ml concentration of calcium FOS, which was added to each explant (0.5-6 hr). For group B, the Enterisol Ileitis® vaccine was used as source of the micro-organism. Previously to the assay, the time necessary for L. intracellularis to colonize the enterocytes was defined. Also, a PCR protocol was optimized to determine the presence of the pathogen in the explants. There were nonstatistical differences for the penetration of the antibiotic into healthy and L. intracellularis-colonized enterocytes. MIC90 of FOS for L. intracellularis is unknown; nevertheless, MIC90 of various antibiotics ranges between 0.125 and 128 µg/ml. FOS reaches inside the enterocyte concentrations which surpass the MICs90 of other antibiotics that also act by the inhibition of cell wall synthesis; however, further studies should be carried out to determine fosfomycin MIC90 for L. intracellularis to discern the usefulness of this antibiotic in the treatment of PPE.

Natural Contamination with Mycotoxins Produced by Fusarium graminearum and Fusarium poae in Malting Barley in Argentina.

Two of the most common species of toxin-producing Fusarium contaminating small cereal grains are Fusarium graminearum and F. poae; with both elaborating diverse toxins, especially deoxynivalenol (DON) and nivalenol (NIV), respectively. The objective of our work during the 2012-2014 growing seasons was to screen crops for the most commonly isolated Fusarium species and to quantify DON and NIV toxins in natural malting-barley samples from different producing areas of Argentina. We identified 1180 Fusarium isolates in the 119 samples analyzed, with 51.2% being F. graminearum, 26.2% F. poae and 22.6% other species. We found high concentrations of mycotoxins, at maximum values of 12 µg/g of DON and 7.71 µg/g of NIV. Of the samples, 23% exhibited DON at an average of 2.36 µg/g, with 44% exceeding the maximum limits (average of 5.24 µg/g); 29% contained NIV at an average of 2.36 µg/g; 7% contained both DON and NIV; and 55% were without DON or NIV. Finally, we report the mycotoxin contamination of the grain samples produced by F. graminearum and F. poae, those being the most frequent Fusarium species present. We identified the main Fusarium species affecting natural malting-barley grains in Argentina and documented the presence of many samples with elevated concentrations of DON and NIV. To our knowledge, the investigation reported here was the first to quantify the contamination by Fusarium and its toxins in natural samples of malting barley in Argentina.

Zearalenone adsorption capacity of lactic acid bacteria isolated from pigs

ABSTRACT The ability to adsorb zearalenone by five strain of lactic acid bacteria was evaluated: four strains of Lactobacillus spp. isolated from pig rectal swabs and one commercial strain (Lactobacillus rhamnosus). Several factors affecting the adsorption capacity were evaluated in order to improve the adsorption of the mycotoxin by bacteria. The stability of the zearalenone-bacteria complex was analyzed. In every case, bacterial adsorption capacity was higher than 40.0%. The strain showing the highest adsorption (68.2%) was selected for the following steps of this research. The adsorption percentages obtained after processing 6.5 and 7.5 mL MRS broth were 57.40% + 3.53 and 64.46% + 0.76, respectively. The stability of zearalenone-bacteria complex was evaluated by successively rinsing. In the first rinsing step 42.26% + 0.414 was still bound. In the second rinsing step 25.12% + 0.664 was still bound, whereas 15.82% + 0.675 remained in the pellet after the third rinse. Results obtained demonstrated that Lactic Acid Bacteria has capacity to adsorb zearalenone. Finally adsorption was increased using a higher volume of initial broth. These results could be used to design a new lyophilized powder for detoxification, using lactic acid bacteria as potential zearalenone adsorbents.

Zearalenone adsorption capacity of lactic acid bacteria isolated from pigs.

The ability to adsorb zearalenone by five strain of lactic acid bacteria was evaluated: four strains of Lactobacillus spp. isolated from pig rectal swabs and one commercial strain (Lactobacillus rhamnosus). Several factors affecting the adsorption capacity were evaluated in order to improve the adsorption of the mycotoxin by bacteria. The stability of the zearalenone-bacteria complex was analyzed. In every case, bacterial adsorption capacity was higher than 40.0%. The strain showing the highest adsorption (68.2%) was selected for the following steps of this research. The adsorption percentages obtained after processing 6.5 and 7.5mL MRS broth were 57.40%+3.53 and 64.46%+0.76, respectively. The stability of zearalenone-bacteria complex was evaluated by successively rinsing. In the first rinsing step 42.26%+0.414 was still bound. In the second rinsing step 25.12%+0.664 was still bound, whereas 15.82%+0.675 remained in the pellet after the third rinse. Results obtained demonstrated that Lactic Acid Bacteria has capacity to adsorb zearalenone. Finally adsorption was increased using a higher volume of initial broth. These results could be used to design a new lyophilized powder for detoxification, using lactic acid bacteria as potential zearalenone adsorbents.

Effect of the combination of crude extracts of Penicillium griseofulvum and Fusarium graminearum containing patulin and zearalenone on rumen microbial fermentation and on their metabolism in continuous culture fermenters.

Six single-flow continuous cultures were used to study the effects of the mycotoxins patulin (PAT) and zearalenone (ZEN) alone or in combination on rumen microbial fermentation. In each of the four 7-d periods, the fermenters were supplemented in a 2 × 3 factorial arrangement with two levels of PAT (0 and 20 mg/l) and three levels of ZEN (0, 5 and 10 mg/l). The treatments did not affect the apparent and true digestibility of organic matter. PAT alone decreased the digestibility of neutral detergent fibre (NDF) and acid detergent fibre (ADF) (p < 0.01), but in the presence of 5 or 10 mg/l of ZEN, there were no effects of PAT. In contrast, the digestibility of NDF and ADF was decreased at 10 mg/l of ZEN in the absence of PAT (p < 0.05). The pH of the fermenters increased after 2 and 3 d of PAT treatment (p < 0.01). PAT decreased the concentration of total volatile acids (VFA), the molar proportion of acetate and the acetate:proportionate ratio (p < 0.01). The molar concentrations of other VFA were unchanged. Ammonia N (NH3-N) flow increased (p < 0.05) and there was a tendency to a higher NH3-N concentration (p < 0.1) in fermenters with PAT. Total N, non-ammonia N and bacterial N as well as efficiency of microbial protein synthesis and efficiency of N utilisation were not affected by treatments. PAT was nearly completely degraded during incubation. The mean recovery of ZEN, α-zearalenol and ß-zearalenol expressed as a proportion of administered ZEN was less than 50% in effluents from fermenters receiving only ZEN and ZEN plus PAT, respectively. With exception of fibre digestion, the co-administration of PAT and ZEN did not elicit interaction effects on most measured parameters of rumen metabolism.