In Ovo Administration of Silver Nanoparticles and/or Amino Acids Influence Metabolism and Immune Gene Expression in Chicken Embryos.

Due to their physicochemical and biological properties, silver nanoparticles (NanoAg) have a wide range of applications. In the present study, their roles as a carrier of nutrients and an immunomodulator were tested in chicken embryos. Cysteine (Cys)+NanoAg injected embryos had smaller livers but heavier breasts on the 19th day of embryogenesis. Cys injected embryos had lower oxygen consumption compared to threonine (Thr) or NanoAg injected embryos. The energy expenditure in Thr+NanoAg, or NanoAg injected embryos was higher than Cys or Cys+NanoAg but was not different from uninjected control embryos. Relative expression of the hepatic insulin-like growth factor-I (IGF-I) gene was higher in Cys or NanoAg injected embryos after lipopolysaccharide (LPS) induction. The gene expression of hepatic tumour necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) did not differ among amino acids, NanoAg and uninjected controls in the non-LPS groups, but increased by many folds in the LPS treated NanoAg, Cys and Cys+NanoAg groups. In LPS treated spleens, TNF-α expression was also up-regulated by NanoAg, amino acids and their combinations, but interleukin-10 (IL-10) expression was down-regulated in Thr, Cys or Thr+NanoAg injected embryos. Toll like receptor-2 (TLR2) expression did not differ in NanoAg or amino acids injected embryos; however, toll like receptor-4 (TLR4) expression was higher in all treated embryos, except for Cys+NanoAg, than in uninjected control embryos. We concluded that NanoAg either alone or in combination with amino acids did not affect embryonic growth but improved immunocompetence, indicating that NanoAg and amino acid complexes can act as potential agents for the enhancement of innate and adaptive immunity in chicken.

Effect of an organic acid blend as an antibiotic alternative on growth performance, antioxidant capacity, intestinal barrier function, and fecal microbiota in weaned piglets.

This study was conducted to evaluate the effects of dietary organic acid blend on growth performance, antioxidant capacity, intestinal barrier function, and fecal microbiota in weaned piglets compared to antibiotic growth promoters (AGP). A total of 90 weaned crossbred barrows (24 ± 1 days of age) with an initial body weight of 7.40 kg were allocated into 3 experimental treatments. Each treatment consisted of 6 replicate pens, with 5 piglets housed in each pen. The dietary treatments included the basal diet (NC), the basal diet supplemented with antibiotics (PC), and the basal diet supplemented with organic acid blend (OA). On day 42, one piglet per pen was randomly selected for plasma and small intestinal sample collection. The results showed that dietary AGP significantly improved growth performance and reduced diarrhea incidence compared to the NC group (P < 0.05). Dietary OA tended to increase body weight on day 42 (P = 0.07) and average daily gain from day 0 to 42 (P = 0.06) and reduce diarrhea incidence (P = 0.05). Dietary OA significantly increased plasma catalase (CAT) activity and decreased the plasma concentration of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), interleukin (IL)-8, and IL-6, which were accompanied by upregulated the relative mRNA abundance of superoxide dismutase 1 (SOD1), glutathione peroxidase 1 (GPX1) and nuclear factor erythroid 2-related factor 2 (NRF2) in comparison to that in the NC group (P < 0.05). Moreover, pigs fed the OA diet significantly increased the ratio of villus height to crypt depth and upregulated the relative expression of zonula occludens-1 (ZO-1) and Claudin1 gene in the jejunum compared to the NC group (P < 0.05). Interestingly, dietary AGP or OA did not affect the fecal microbiota structure or volatile fatty acid content (P > 0.05). In conclusion, our results suggested that dietary OA supplementation could improve growth performance and antioxidant capacity and protect the intestinal barrier of weaned piglets, therefore it has the potential to be consideredas an alternative to AGP in the pig industry.

Nano-nutrition of chicken embryos. The effect of silver nanoparticles and ATP on expression of chosen genes involved in myogenesis.

It has been suggested that the quantity and quality of nutrients stored in the egg might not be optimal for the fast rate of chicken embryo development in modern broilers, and embryos could be supplemented with nutrients by in ovo injection. Recent experiments showed that in ovo feeding reduces post-hatch mortality and skeletal disorders and increases muscle growth and breast meat yield. Adenosine triphosphate (ATP) is a "ready for use" energetic molecule, while nanoparticles of silver (Nano-Ag) may penetrate tissues as well as cells and localise inside cells. In this investigation, we hypothesised that silver nanoparticles could be used as a protective carrier for ATP as well as an active agent. ATP and/or an ATP complex with Nano-Ag would be delivered to the muscle cells as a gene expression regulator and promoter of growth and development of embryo breast muscle. A collection of 160 broiler eggs was randomly divided into a Control group without injection and injected groups with hydrocolloids of Nano-Ag, ATP or a complex of Nano-Ag and ATP (Nano-Ag/ATP). The embryos were evaluated on day 20 of incubation. The results indicate that the application of ATP to chicken embryos increases expression of fibroblast growth factor 2 (FGF2), vascular endothelial growth factor (VEGF) and Na(+)/K(+) transporting ATPase (ATP1A1), which may indicate that an extra energy source can enhance molecular mechanisms of muscle cell proliferation. Nano-Ag also up-regulated expression of FGF2, VEGF, ATP1A1 and, also up-regulated expression of myogenic differentiation 1(MyoD1), affecting cell differentiation. The results indicate that ATP and Nano-Ag may accelerate growth and maturation of muscle cells.

Effect of supplementing hydroxy trace minerals (Cu, Zn, and Mn) on egg quality and performance of laying hens under tropical conditions.

OBJECTIVE: A pivotal study was designed to investigate the effect of Hydroxy (HYC) Cu, Zn, and Mn on egg quality and laying performance of chickens under tropical conditions. METHODS: A total of 1,260 Babcock White laying hens (20-wk-old) were randomly assigned to one of 4 treatments with 15 replicates of 21 hens each in a Randomized Complete Block Design. The birds were reared for 16 weeks and were fed the corn-soybean meal diets supplemented with one of the following mineral treatments: T1, inorganic (INO, 15 ppm CuSO4, 80ppm MnSO4 and 80 ppm ZnO); T2, Hydroxy-nutritional level (HYC-Nut, 15 ppm Cu, 80 ppm Mn, 80 ppm Zn from Hydroxy); T3, Hydroxy-Low (HYC-Low, 15 ppm Cu, 60 ppm Mn, 60 ppm Zn from Hydroxy); T4, Hydroxy plus inorganic (HYC+INO, 7.5 ppm HYC Cu+7.5 ppm CuSO4, 40 ppm HYC ZnO+40 ppm ZnSO4, 40 ppm HYC Mn+40 ppm MnSO4). The egg production was recorded daily, while the feed consumption, feed conversion ratio (FCR) and egg mass were determined at the end of each laying period. The egg quality parameters were assayed in eggs collected over 48 h in each laying period. RESULTS: Overall, no significant effect of treatments was observed on percent egg production, egg weight and FCR (p>0.05). Feed intake was significantly lower in birds fed Hydroxy plus inorganic (p<0.05) diet. The supplementation of HYC-Low significantly increased the egg mass compared to the other treatments (p<0.05). HYC supplementation alone or in combination with INO elicited a positive effect on shell thickness, shell weight, shell weight per unit surface area, yolk colour, albumen and yolk index for a certain period (p<0.05), but not throughout the whole laying period. CONCLUSION: Dietary supplementation of HYC-Low (15-60-60 mg/kg) showed similar effects on production performance and egg quality characteristics in laying hens as compared to 15-80-80 mg/kg of Cu-Zn-Mn from inorganic sources. This indicates that sulphate based inorganic trace minerals can effectively be substituted by lower concentration of hydroxyl minerals.

Sows fed with synergistic blend of short- and medium chain organic acid has a carryover effect on post-weaning growth rate.

This study investigated the effect of a synergistic blend of free and buffered organic acid (FMP) on the performance of piglets born to sows supplemented with a blend of short- and medium-chain organic acids (SGG) during the late gestation and lactation period. A total of 150 multiparous sows (n = 50/treatment, Landrace × Yorkshire) were blocked (2.4 parity) and assigned to 1 of 3 dietary treatments: CON - corn-soybean meal-based basal diet, SGG-Low - CON+ 1.5 kg/ton SGG, and SGG-High - CON + 3kg/ton SGG. During weaning, 600 piglets (6.72 ± 0.5kg) which weaned from sows supplemented with 3 levels of SGG were allocated to 2 weaner diets (Control and FMP - 3kg/ton) following 3 × 2 factorial arrangement. Supplemental effects on performance were measured at d0-d21 and d 21-42, and the entire period. Pigs fed with FMP and born to sows supplemented with SGG-High gained more weight and ate more (p < 0.05) compared with those in the CON group in both phases, and with SGG-Low in the second phase. Over the entire post-weaning period, piglets born to sows supplemented with SGG-Low and SGG-High had a higher average daily gain (ADG) and body weight (BW) (p < 0.05). Regardless of sow treatment, pigs fed with an FMP diet had higher ADG (p < 0.001), BW (p = 0.045), and a lower feed conversion ratio (p = 0.033). Also, feeding FMP diets reduced the fecal Escherichia coli and Clostridium perfringens counts at d42. The current study indicates that sows fed SGG supplement had a positive carry-over effect on the post-weaning growth rate, and FMP supplement enhances the growth performance and reduced the number of C. perfringens and E. coli. Thus, the application of 3 kg/ton of SGG in sows' diet and subsequent feeding of piglets with FMP would be an effective strategy to improve growth rate and reduce pathogenic bacteria in post-weaned piglets.

Organic acid blends improve intestinal integrity, modulate short-chain fatty acids profiles and alter microbiota of broilers under necrotic enteritis challenge.

Controlling enteric diseases of broilers is crucial. Among many additives, organic acids (OA) and their blends are gaining attention to combat diseases in the post-antibiotic era. The current study evaluated the potentials of short-chain fatty acids (SCFA) and medium-chain fatty acids (MCFA) blends and/or phenolic compounds on intestinal integrity, intestinal pH, caecal microbiota, and caecal SCFA profiles of broilers under necrotic enteritis (NE) challenge. The additives used were: (A) a blend of SCFA, MCFA, and a phenolic compound (SMP), (B) a blend of free and buffered SCFA with MCFA (SMF), and (C) a blend of free and buffered SCFA with a high concentration of MCFA (SHM). A total of 1,404 male parental chicks of Ross 308 broilers were randomly allocated to 78 floor pens on hatching day with 6 treatments replicated 13 times with 18 birds per pen. The treatments were: UCC, unchallenged control; CHC, challenged control; BAC, challenged group plus zinc bacitracin; SMP, challenged group plus additive SMP; SMF, challenged group plus additive SMF; SHM, challenged group plus additive SHM. Birds were challenged with field-strain Eimeria spp. on d 9 and Clostridium perfringens on d 14. Birds challenged with NE increased fluorescein isothiocyanate dextran (FITC-d) concentration in serum, reduced acetate and butyrate concentrations, and increased Bacteroides and C. perfringens load in the caeca (P < 0.05). Birds fed additives decreased FITC-d from gut to serum, reduced Bacteroides (d 16, P < 0.05) and numerically reduced C. perfringens load compared to CHC group. Birds fed additive SHM had higher concentrations of acetate and butyrate (d 21, P < 0.05) than CHC group but were not different from SMP and SMF groups. All the additives exhibited similar intestinal protection against NE compared to the BAC group indicated by FITC-d concentration in serum, acetate, propionate and butyrate concentrations in the caeca, and caecal bacterial loads except for the C. perfringens (P > 0.05). The SMP group had a higher load compared to BAC (P < 0.05). These findings suggest the promising effects of OA blends as alternatives to BAC to ameliorate the impact of NE challenge of broilers as indicated by improved intestinal health.

Impacts of feeding organic acid-based feed additives on diarrhea, performance, and fecal microbiome characteristics of pigs after weaning challenged with an enterotoxigenic strain of Escherichia coli.

Post weaning diarrhea (PWD) caused by enterotoxigenic strains of E. coli (ETEC) remains a major problem in the industry, causing decreases in performance and survival of weaned pigs. Traditionally, antimicrobials have been used for its mitigation/control. This study tested the hypothesis that a combination of two organic acid (OA)-based commercial feed additives, Presan FX [an OA, medium-chain fatty acid (MCFA) and phenolic compound-based product] and Fysal MP (free and buffered OA based on formic acid), would reduce PWD and improve post-weaning performance in pigs challenged with an F4-ETEC. This combination was assessed against a Negative control diet without any feed additives and a diet containing amoxicillin. Combined with a reduction in temperature during the infection period, inoculation with F4-ETEC resulted in 81% of pigs developing diarrhea, but with no differences between treatments (P > 0.05). However, between days 14 to 20 of the study and due to colonization by Salmonella serovars, pigs fed the combination of Presan FX and Fysal MP showed less (P = 0.014) diarrhea commensurate with a lower (P = 0.018) proportion of Salmonella numbers relative to total bacterial numbers. This caused less (P = 0.049) therapeutic antibiotic administrations relative to the diet with amoxicillin during this time. The diversity of bacteria within amoxicillin-treated pigs was lower (P = 0.004) than the diversity in control or Presan FX + Fysal MP-treated pigs (P = 0.01). Pair-wise comparisons showed that amoxicillin-treated pigs had altered (P < 0.001) fecal microbial communities relative to both Presan FX + Fysal MP-treated pigs and control pigs. Amoxicillin-treated pigs were characterized by an increased abundance of bacterial families generally linked to inflammation and dysbiosis in the gastrointestinal tract (GIT), whereas Presan FX + Fysal MP-treated pigs had an increased abundance of bacterial families considered beneficial commensals for the GIT. Control pigs were characterized by an increased abundance of Spirochaetaceae associated with healthy piglets, as well as bacterial families associated with reduced feed intake and appetite. The combination of two OA-based feed additives did not reduce the incidence of F4 ETEC-associated diarrhea nor enhance performance. However, the combination markedly reduced diarrhea caused by Salmonella that occurred following the ETEC infection, commensurate with less therapeutic administrations relative to the diet with amoxicillin.

The Efficacy of a Smectite-Based Mycotoxin Binder in Reducing Aflatoxin B1 Toxicity on Performance, Health and Histopathology of Broiler Chickens.

The aim of the experiment was to investigate the efficacy of a smectite-based clay binder (Toxo-MX) in reducing the toxicological effects of aflatoxin B1 (AFB1) in commercial broiler chickens. A total of 450 one-day old male broiler chickens were randomly allocated into three treatment groups with ten replicates of 15 birds each in a 42-day feeding experiment. The dietary treatments included a negative control (NC, a basal diet with no AFB1 and binder), a positive control (PC, a basal diet contaminated with 500 ppb of AFB1) and a smectite-based mycotoxin binder(Toxo-MX, PC with smectite clay binder). AFB1 challenge resulted in 14 to 24% depression in growth performance, elevated levels of aspartate aminotransferase (AST), and gamma-glutamyl transferase (GGT), organ enlargement and immuno-suppression.As compared to PC, feeding of Toxo-MX improved the final weight (15%; p < 0.0001), average daily gain (ADG) (15%; p < 0.001) and feed efficiency of broilers (13%; p < 0.0003) but did not have any effects on liver enzyme activities. Supplementation of smectite claysignificantly increased serum globulin levels and reduced the weight of the liver (p < 0.05) as compared to AFB1-fed broiler chickens. The severity of lesions (inflammatory and degenerative changes) observed in the liver, kidney, heart, pancreas, and lymphoid organs in PC birds was reduced by feeding smectite clay. The immuno-suppression caused by AFB1 was moderately ameliorated in Toxo-MX groupby stimulating the production of antibodies against IBD at day 42 (p < 0.05). In conclusion, dietary supplementation of a smectite-based mycotoxin binder to the diet containing AFB1 improved growth performance, reduced toxicological effects in liver and improved humoral immune response in broilers, suggesting its protective effect against aflatoxicosis.

Potential of blended organic acids to improve performance and health of broilers infected with necrotic enteritis.

Organic acids (OA) and their blends have been shown to positively affect performance and health of broilers. However, the data in the literature are not consistent. This study examined the potential of blended short-chain fatty acids (SCFA) with medium-chain fatty acids (MCFA) as alternatives to antibiotic growth promoters (AGP) on performance, health and welfare of broilers infected with necrotic enteritis (NE). The additives used were: A) a blend of SCFA, MCFA, and a phenolic compound (SMP); B) a blend of free and buffered SCFA with MCFA (SMF); C) a blend of free and buffered SCFA with a high concentration of MCFA (SHM). A total of 1,404 Ross 308 one-day-old male parental chicks were randomly distributed into 78-floor pens with 13 replicates of 18 birds each. Six treatments were the following: T1, unchallenged control (UCC); T2, challenged control (CHC); T3, challenged group plus zinc bacitracin (BAC); T4, challenged group plus additive SMP; T5, challenged group plus additive SMF; T6, challenged group plus additive SHM. Challenged birds were gavaged with Eimeria spp. on d 9 and Clostridium perfringens EHE-NE18 on d 14. Post NE challenge and cumulatively, BWG, FCR, and nutrient digestibility of birds were compromised (P < 0.05) by NE challenge indicating a successful induction of sub-clinical NE. Additive SHM had higher BWG compared to CHC and BAC groups (P = 0.001; d 10 to 24) but not different from SMP and SMF groups (P > 0.05). All the 3 additive groups had lower FCR compared to CHC (P = 0.001; d 0 to 35), and exhibited similar jejunal lesions (d 16) compared to BAC and apparent ileal protein digestibility (d 21) compared to UCC and BAC groups (P > 0.05). Birds in additive SHM group had a higher concentration of serum IgA compared to all groups (P = 0.001) except additive SMF (P > 0.05; d 21). All the additive groups had lower footpad dermatitis and hock burns compared to CHC (P < 0.05). The findings suggest the potential of blended OA as alternatives to BAC to protect broilers from NE indicated by improved FCR, immunity, digestibility, and bird welfare.

The effects of synergistic blend of organic acid or antibiotic growth promoter on performance and antimicrobial resistance of bacteria in grow-finish pigs.

This study was carried out to evaluate the effect of Selacid Green Growth (GG) or antibiotic growth promoter (AGP) on the performance and economics of grow-finish (GF) pigs. The Selacid GG is a blend of short-chain fatty acids (formic acid, acetic acid, lactic acid, propionic acid, citric acid, and sorbic acid), buffered organic acid (ammonium formate), and a combination of medium-chain fatty acids (C8, C10, and C12). A total of 312 grower pigs (Yorkshire × Landrace × Duroc) with initial body weight (BW) of 26.5 ± 0.92 kg were used in a 90-d feeding trial. The pigs were allocated randomly to three treatments consisting of eight replicate pens with 13 pigs each. The treatments tested included a 1) negative control (control): basal diet without colistin and Selacid GG, 2) positive control (AGP): basal diet with colistin (20 g/ton), and 3) Feed additive (Selacid GG): basal diet with Selacid GG (2 kg/ton). The results showed that, over the entire period of the experiment, the dietary supplementation of Selacid GG elicited a similar effect as AGP on feed cost and on all growth parameters measured (P > 0.05). In relation to the control group, Selacid GG significantly improved the final BW (+3.4 kg or 3.6%), average daily gain (+39 g/pig or 5.3%), and gain:feed (+30 g or 8.1%) of pigs (P < 0.05). In addition, the feeding of Selacid GG reduced feed cost (-0.078 USD) per kilogram BW gain. The average daily feed intake was not affected by dietary treatments (P > 0.05). Escherichia coli was prevalent in 46 out of 48 fecal samples tested. All E. coli isolates were resistant to colistin, amox-colistin, ciprofloxacin, and enrofloxacin. The number of E. coli isolates resistant to amoxicillin/clavulanic, cefotaxime, ceftiofur, and norfloxacin was significantly reduced, whereas the inhibitory zones of amocxillin/clavulanic acid were increased; and the susceptibility of E. coli to amoxcillin/clavulanic, cefotaxime, ceftiofur, ciprofloxacin, nofloxacin, and flumequin was increased when Selacid GG was added in the feeds (P < 0.05). The findings of the study suggest that Selacid GG is a cost-effective product with the same efficacy as AGP in promoting the growth and economic performance of GF pigs. The product is safe and can be added to the diet of GF pigs without developing resistance to selected antibiotics.

Silver nanoparticles administered to chicken affect VEGFA and FGF2 gene expression in breast muscle and heart.

Nanoparticles of colloidal silver (AgNano) can influence gene expression. Concerning trials of AgNano application in poultry nutrition, it is useful to reveal whether they affect the expression of genes crucial for bird development. AgNano were administered to broiler chickens as a water solution in two concentrations (10 and 20 ppm). After dissection of the birds, breast muscles and hearts were collected. Gene expression of FGF2 and VEGFA on the mRNA and protein levels were evaluated using quantitative polymerase chain reaction and enzyme-linked immunosorbent assay methods. The results for gene expression in the breast muscle revealed changes on the mRNA level (FGF2 was up-regulated, P < 0.05) but not on the protein level. In the heart, 20 ppm of silver nanoparticles in drinking water increased the expression of VEGFA (P < 0.05), at the same time decreasing FGF2 expression both on the transcriptional and translational levels. Changes in the expression of these genes may lead to histological changes, but this needs to be proven using histological and immunohistochemical examination of tissues. In general, we showed that AgNano application in poultry feeding influences the expression of FGF2 and VEGFA genes on the mRNA and protein levels in growing chicken.

Effect of silver nanoparticles on growth performance, metabolism and microbial profile of broiler chickens.

This study evaluated the potential of silver nanoparticles (AgNano) as an antimicrobial growth-promoting supplement for broiler chickens. One hundred forty-four seven-day-old broiler chicks were distributed randomly to AgNano treatments at 0, 10 and 20 mg/kg (Control, Group AgNano10, and Group AgNano20, respectively) provided via the drinking water from day 7 to 36 post-hatching. Body weight and feed consumption were measured weekly. In addition, balance and respiration experiments were carried out to determine nitrogen (N) utilisation and energy retention. At days 22 and 36, blood samples and intestinal content were collected to evaluate the effects of AgNano on plasma concentration of immunoglobulins and the intestinal microflora, respectively. The provision of water solutions containing different concentrations of AgNano had no effect on postnatal growth performance and the energy metabolism of broiler chickens. However, in Group AgNano10 N intake (p = 0.05) and retention (p = 0.03) was increased, but N excretion and efficiency of utilisation was not affected. The populations of bacteria in the intestinal samples were not affected by AgNano supplementation. The concentration of immunoglobulin (IgG) in the blood plasma of broilers supplemented with AgNano decreased at day 36 (p = 0.012). The results demonstrated that AgNano affects N utilisation and plasma IgG concentration; however, it does not influence the microbial populations in the digestive tract, the energy metabolism and growth performance of chickens.