An Immobilized Form of a Blend of Essential Oils Improves the Density of Beneficial Bacteria, in Addition to Suppressing Pathogens in the Gut and Also Improves the Performance of Chicken Breeding.

Antimicrobial growth promoters (AGP) are used in chicken production to suppress pathogens in the gut and improve performance, but such products tend to suppress beneficial bacteria while favoring the development and spread of antimicrobial resistance. A green alternative to AGP with the ability to suppress pathogens, but with an additional ability to spare beneficial gut bacteria and improve breeding performance is urgently required. We investigated the effect of supplementation of a blend of select essential oils (cinnamon oil, carvacrol, and thyme oil, henceforth referred to as EO; at two doses: 200 g/t and 400 g/t feed) exhibiting an ability to spare Lactobacillus while exhibiting strong E. coli inhibition ability under in vitro tests and immobilized in a sunflower oil and calcium alginate matrix, to broiler chickens and compared the effects with those of a probiotic yeast (Y), an AGP virginiamycin (V), and a negative control (C). qPCR analysis of metagenomic DNA from the gut content of experimental chickens indicated a significantly (p < 0.05) lower density of E. coli in the EO groups as compared to other groups. Amplicon sequence data of the gut microbiome indicated that all the additives had specific significant effects (DESeq2) on the gut microbiome, such as enrichment of uncultured Clostridia in the V and Y groups and uncultured Ruminococcaceae in the EO groups, as compared to the control. LEfSe analysis of the sequence data indicated a high abundance of beneficial bacteria Ruminococcaceae in the EO groups, Faecalibacterium in the Y group, and Blautia in the V group. Supplementation of the immobilized EO at the dose rate of 400 g/ton feed improved body weight gain (by 64 g/bird), feed efficiency (by 5 points), and cellular immunity (skin thickness response to phytoheamagglutinin lectin from Phaseolus vulgaris by 58%) significantly (p < 0.05), whereas neither yeast nor virginiamycin showed a significant effect on performance parameters. Expression of genes associated with gut barrier and immunity function such as CLAUDIN1, IL6, IFNG, TLR2A, and NOD1 were significantly higher in the EO groups. This study showed that the encapsulated EO mixture can improve the density of beneficial microbes in the gut significantly, with concomitant suppression of potential pathogens such as E.coli and improved performance and immunity, and hence, has a high potential to be used as an effective alternative to AGP in poultry.

Effects of Dietary Antimicrobial Growth Promoters on Performance Parameters and Abundance and Diversity of Broiler Chicken Gut Microbiome and Selection of Antibiotic Resistance Genes.

Antimicrobial growth promoters (AGPs) are commonly used in broiler production. There is a huge societal concern around their use and their contribution to the proliferation of antimicrobial resistance (AMR) in food-producing animals and dissemination to humans or the environment. However, there is a paucity of comprehensive experimental data on their impact on poultry production and the AMR resistome. Here, we investigated the effect of five antimicrobial growth promoters (virginiamycin, chlortetracycline, bacitracin methyl disalicylate, lincomycin, and tylosin) used in the commercial broiler production in the Indian subcontinent and in the different parts of the world for three consecutive production cycles on performance variables and also the impact on gut bacteria, bacteriophage, and resistome profile using culture-independent approaches. There was no significant effect of AGPs on the cumulative growth or feed efficiency parameters at the end of the production cycles and cumulative mortality rates were also similar across groups. Many antibiotic resistance genes (ARGs) were ubiquitous in the chicken gut irrespective of AGP supplementation. In total, 62 ARGs from 15 antimicrobial classes were detected. Supplementation of AGPs influenced the selection of several classes of ARGs; however, this was not correlated necessarily with genes relevant to the AGP drug class; some AGPs favored the selection of ARGs related to antimicrobials not structurally related to the AGP. AGPs did not impact the gut bacterial community structure, including alpha or beta diversity significantly, with only 16-20 operational taxonomic units (OTUs) of bacteria being altered significantly. However, several AGPs significantly reduced the population density of some of the potential pathogenic genera of bacteria, such as Escherichia coli. Chlortetracycline increased the abundance of Escherichia phage, whereas other AGPs did not influence the abundance of bacteriophage significantly. Considering the evidence that AGPs used in poultry production can select for resistance to more than one class of antimicrobial resistance, and the fact that their effect on performance is not significant, their use needs to be reduced and there is a need to monitor the spread of ARGs in broiler chicken farms.

Supplementation of sulfur and folic acid improves performance of broiler chicken fed suboptimal concentrations of dietary methionine.

BACKGROUND: Supplementation of sulfur (S) and folic acid (FA) are known to spare the dietary requirement of methionine (Met) in chicken. An experiment was conducted to determine the effects of feeding graded concentrations of FA (0, 2, 4, 6, 8 mg kg-1 ) and two concentrations of S (0 and 2 g kg-1 ) in 5 × 2 factorial design. Diet without FA and S was considered as negative control (NC). Another diet with the recommended concentration of Met was considered as the positive control (PC). Each diet was offered ad libitum from day 1 to 42 to 10 pens of 20 male chicks each. RESULTS: Broilers fed the NC diet had lower body weight gain (BWG), high feed conversion ratio, lower ready-to-cook yield, higher lipid peroxidation and reduced concentrations of protein and albumin in serum. BWG improved with FA at day 21 and showed a trend of improvement at day 42. Similarly, BWG improved with S supplementation at day 42. The FA and S interaction indicated a significant reduction in lipid peroxidation with S supplementation at the majority of FA levels. Serum protein fractions increased with increased dietary FA content and increased further with S supplementation. CONCLUSION: Based on the results, it is concluded that the combination of FA (4 mg kg-1 ) and S (2 g kg-1 ) improved BWG. Similarly, the feed efficiency with 6 mg FA alone or with S was similar to the PC group. The improvement was probably due to the increase in protein accretion and reduction in lipid peroxidation with FA and S supplementation. © 2022 Society of Chemical Industry.

Effect of methyl donors supplementation on performance, immune responses and anti-oxidant variables in broiler chicken fed diet without supplemental methionine.

OBJECTIVE: Methionine (Met) is involved in methyl group transfer besides protein synthesis. As the availability is limited and cost is high for synthetic Met, reductions in its inclusion in broiler diet may be possible by supplementing the low Met diets with methyl donors (MD) like betaine (Bet), folic acid (FA), vitamin B12 (B12), and biotin (Bio). An experiment was conducted to study the effects of supplementing the MD on performance (average daily gain [ADG], daily feed intake, feed efficiency [FE]), anti-oxidant variables, immune responses and serum protein concentration in broilers fed sub-optimal concentrations of dietary Met. METHODS: Maize-soybean meal diet was used as control (CD). Different MD like Bet (0.2%), B12 (0.1 mg), FA (4 mg), or Bio (1.5 mg/kg) were supplemented to basal diet (BD) having no supplemental Met. The BD without MD was kept for comparison. Each diet was fed ad libitum to 10 replicates of 25 chicks in each from 1 to 42 d of age. RESULTS: At the end of experiment, the ADG in MD group was higher than BD and lower than CD. The FE improved with FA or Bet compared to the BD. Breast meat weight was higher in Bet compared to the BD, while it was intermediate between BD and CD in other groups. The lipid peroxidation reduced with Bio, B12, or Bet, while the glutathione peroxidase activity improved with Bio or B12 compared to the BD. Lymphocyte proliferation improved with Bet compared to the BD. The serum protein concentrations increased with FA, Bio, or Bet compared to those fed BD. CONCLUSION: It can be concluded that the ADG can be improved partially with supplementation of MD while the FE improved with FA or Bet. Some MD also reduced the stress indices and improved immune responses compared to the BD fed broilers.

Gut Microbial Composition Differs Extensively among Indian Native Chicken Breeds Originated in Different Geographical Locations and a Commercial Broiler Line, but Breed-Specific, as Well as Across-Breed Core Microbiomes, Are Found.

Gut microbiota plays an important role in the health and performance of the host. Characterizations of gut microbiota, core microbiomes, and microbial networks in different chicken breeds are expected to provide clues for pathogen exclusion, improving performance or feed efficiency. Here, we characterized the gut microbiota of "finishing" chickens (at the end of production life) of indigenous Indian Nicobari, Ghagus, and Aseel breeds, originating from the Nicobari island, coastal India, and the Indian mainland, respectively, as well as a global commercial broiler line, VenCobb 400, using 16S rDNA amplicon sequencing. We found that diversity, as well as richness of microbiota, was higher in indigenous breeds than in the broiler line. Beta diversity analysis indicated the highest overlap between Ghagus and Nicobari breeds and a very low overlap between the broiler line and all indigenous breeds. Linear discriminant analysis effect size (LEfSe) revealed 82 breed- or line-specific phylotype operational taxonomic unit (OTU) level biomarkers. We confirm the presence of breed specific and across-breed core microbiomes. Additionally, we show the existence of breed specific complex microbial networks in all groups. This study provides the first (and comprehensive) insight into the gut microbiota of three indigenous breeds and one commercial broiler line of chickens reared without antimicrobials, and underscores the need to study microbial diversity in other indigenous breeds.

Effect of Dietary Supplementation of Organic Trace Minerals at Reduced Concentrations on Performance, Bone Mineralization, and Antioxidant Variables in Broiler Chicken Reared in Two Different Seasons in a Tropical Region.

Two experiments were conducted to study the performance, antioxidant activity, and bone mineral variables in broilers fed organic trace minerals (OTM) at lowered concentrations in the diet. In experiment 1, a total of 1500 day-old broiler male chicks were randomly distributed into six groups with 10 replicates of 25 chicks each and housed in floor pens (1.90 × 1.22 m). One group was fed the maize-soybean meal-based control diet, supplemented with inorganic trace mineral (ITM) premix containing Mn, Zn, Fe, Cu, Se, and Cr at 50, 45, 40, 7.5, 0.30, and 0.25 mg/kg, respectively. The remaining groups of chicks were fed the diets, where the ITM premix was replaced with OTM mix so as to provide the respective minerals at graded levels in the diet (100, 75, 50, 40, and 30% of the control diet). Similarly, in experiment II, a total of 1350 day-old male broiler chicks were divided at random into six groups with 9 replicates of 25 chicks each and fed the maize-soybean meal-based control diet having ITM at levels similar to those of experiment I or the test diets, where the ITM was replaced with OTM so as to provide the minerals at 100, 80, 60, 40, and 20% of the ITM-based control diet. Each diet was fed ad libitum from 0 to 42 days of age. The body weight gain, feed intake, and feed efficiency were not affected by supplementing OTM at 30 and 20% in experiments I and II, respectively. Slaughter variables, activities of glutathione peroxidase and super oxide dismutase, and lipid peroxidation and ferric reducing activity in serum were not affected by supplementing OTM at the lowest level of 20% in the diet. Similarly, bone-breaking strength, ash, and Mn contents were not affected by supplementing OTM at 20% of the mineral concentration in control diet containing ITM. Deposition of Ca, P, Cu, and Fe in tibia ash increased with increased concentration of OTM in the diet. However, Zn content in tibia ash reduced with reduction in OTM level in the diet. Based on the results, it is concluded that trace mineral supplementation in organic form even at 20% of the concentration recommended for ITM may be sufficient in maize-soybean meal-based diet to support the optimum growth, bone ash, bone strength, and serum antioxidant status in commercial broilers.

Effect of nutrient density on production performance, egg quality and humoral immune response of brown laying (Dahlem Red) hens in the tropics.

A study was conducted to evaluate the effect of various concentrations of metabolizable energy (ME) with graded incremental levels of crude protein (CP) and essential amino acids (lysine and methionine) on production performance, egg quality and humoral immune response of Dahlem Red laying hens. Four experimental diets based on maize-soybean meal-deoiled rice bran were prepared. Diet 1 was fed as a control diet containing 2,600 kcal ME/kg, 15% CP, 0.75% Lys and 0.36% Met, and in the other three diets (D2, D3 and D4), concentrations of the above nutrients were increased by 2.5%, 5.0% and 7.5%, respectively. The levels of Ca (3.5%) and available P (0.32) were constant in all the diets. Each diet was offered ad libitum from 28 to 40 weeks of age to eight replicates containing six birds in each replicate. The egg production, egg weight and egg mass (in grams of egg per hen per day) were not affected by increasing the nutrient density up to 7.5% (2,795 kcal ME/kg diet) compared to the control group (2,600 ME/kg diet). However, feed consumption and feed efficiency (in grams of egg per gram of feed) were influenced by the variation in the nutrient density of diets. As the nutrient density increased by 5% (2,730 ME/kg diet), birds consumed significantly (P < 0.001) less feed. The birds in the 7.5% higher density group produced significantly (P < 0.05) higher egg mass per unit feed consumption compared to the control diet. Increasing nutrient density up to 7.5% had no effect on relative weight of albumen, yolk or shell. The Haugh unit, yolk colour and shell thickness were also not affected due to variation in the nutrient density. The humoral immune response measured at 34 and 40 weeks was progressively improved by increasing the nutrient density up to 5%. Increasing the nutrient density beyond 5% in the diet had no further influence on the humoral immune response. Based on the results of the present study, it can be concluded that Dahlem Red laying hens required 2,795 kcal/kg ME, 16% CP, 0.8% lysine and 0.4% methionine for eliciting optimum performance and immune response during 28 to 40 weeks of age.