Thymol, alpha tocopherol, and ascorbyl palmitate as a natural feed strategy to modulate immune variables and Heterophil to Lymphocyte ratio in broiler chickens

Name: Thymol, alpha tocopherol, and ascorbyl palmitate as a natural feed strategy to modulate immune variables and Heterophil to Lymphocyte ratio in broiler chickens
License: https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Keywords: Immunity

Datos de investigación

Thymol, alpha tocopherol, and ascorbyl palmitate as a natural feed strategy to modulate immune variables and Heterophil to Lymphocyte ratio in broiler chickens

Autores: Videla, Emiliano Ariel Icon

; Luna, Agustin Icon

; Picco, Romina Paula Andrea Icon

; Marin, Raul Hector Icon

; Nazar, Franco Nicolas Icon

; Caliva, Jorge Martín Icon

Publicador: Consejo Nacional de Investigaciones Científicas y Técnicas

Fecha de depósito: 02/08/2024

Fecha de recolección: 15/03/2016-15/10/2016

Clasificación temática:

Ciencias Veterinarias

Resumen

Immune response observed on broiler chickens fed different natural supplements (BHT, Thymol and a mix of Ascorbyl palmitate and alpha tocopherol

Información Técnica

2.1 Animals and Husbandry One-day-old Cobb-500 male broiler chicks with similar weight (43.0 ± 1.0) g were obtained from a local hatchery (INDACOR S.A.), and located in a commercial farm operated by INDACOR S.A. Upon arrival, birds were identified (leg banded for 2 weeks and wing banded after that) and randomly housed in groups of 40 individuals in one of 24 pens measuring 2.4×1.2 m. Water and feed were provided ad libitum throughout the experiment. Appropriate amounts of dietary supplements (see below) were dissolved in soybean oil (normally included in feed’s formulation), sprayed to ground basal diet and mixed until homogenization before extrusion. 2.2 Treatment procedures All birds within each pen were randomly assigned to 1 of 6 dietary treatments. There were 4 replicates (pens) of each dietary treatment. Each replica initially consisted of 40 birds. Dietary treatments were characterized as follows: 1) Basal (no feed supplements added), 2) Promotor (Basal + 6.26 µmol flavomycin/kg feed), 3) BHT (Basal + 1.33 mmol of BHT/kg feed), 4) Prom-BHT (Basal + a mix of 6.26 µmol flavomycin/kg feed and 1.33 mmol of BHT/kg feed), 5) THY (Basal + 1.33 mmoles of THY/kg feed), and 6) TOC-AP (Basal + 0.665 mmoles of TOC + 0.665 mmoles of AP/kg feed). The Basal, Promotor (Flavomycin 8%, Hugestone Enterprise Co., China), BHT (FlukaAG, Buchs SG, Switzerland), and Prom-BHT, were used as controls. THY (SAFCR©, ≥99%, FCC, USA) and TOC-AP (GRINDOX 497 DuPONT, Danisco Argentina) were considered the experimental groups. When birds were 21 days (d) old, stocking density in each pen was randomly reduced to 32 birds per pen. Doses were selected based on previous reports28 and the current promoter and BHT doses used in the farm. All supplements but flavomycin (commercial doses as growth promoter) were chosen to achieve the same final (1.33 mol/kg) concentration. The feeding regimen consisted of a starter (1–10 d), grower (11–26 d), and finisher (27–42 d) diet. The ingredients and chemical composition of the basal diets are detailed elsewhere13. Birds were raised following a standard husbandry program for slaughter weight between 2.5-3.0 kg (Cobb-vantress; Broiler Management Guide 2017). 2.3 Sampling procedures On d 35 of age (one week before slaughtering) blood samples were taken from the left brachial vein of 10 broilers from each of the 24 pens (40 birds in total from each dietary treatment). Blood withdrawal was done with ethylenediaminetetraacetic-acid (EDTA) to avoid blood coagulation; anaesthesia was not used in line with guidance in the National Centre for the Replacement Refinement and Reduction of Animals in Research (www.nc3rs.org.uk/general-principles). The sampling procedure was randomly performed between treatments. One blood drop was used for blood smears, and the rest was centrifuged at 2500 g for 15 minutes to obtain plasma, pellets were discarded. Plasma samples were stored at -80ºC and then used for microagglutination assay to determine humoral response against SRBC. 2.4 Immunological determinations To assess the inflammatory response, the day before blood samples were taken, the peripheral zone of the right-wing web was measured with a digital caliper and then intradermally injected with 0,1 ml of PHA-P (Sigma Chemical, St Louis, USA) solution in phosphate saline buffer (PBS, pH= 7, 1 mg.ml-1) (Vinkler et al., 2010). Twenty-four hours later the same region previously injected was measured to determine the percentage of inflammation, calculated using the following formulae: percentage of inflammation = [(inflammation previous 24hs) / (inflammation post 24hs)] x 100. For the microagglutination assay, a 30 µl aliquot of plasma was used as a pure sample. Another 30 µl aliquot of plasma was serially diluted in 30µl of PBS, across 10 wells of a 96 U well plate. Then, 30 µl of a 2% solution of SRBC were added to each well, except for the blanks. The 11 and the 12 wells were used as blank (serum diluted with PBS only) and negative controls (PBS with SRBC), respectively. Plates were incubated for 45 minutes at 40ºC. Antibody titers were reported as the Log2 of the highest dilution yielding significant agglutination. Blood smears previously made were stained with May-Grünwald Giemsa. Using a light optical microscope (1000x), 100 white cells per smear were counted and differentiated in Heterophils, Basophils, Eosinophils, Monocytes and Lymphocytes. The Heterophil to Lymphocyte ratio was calculated for each individual. H/L ratio was calculated using the following formulae: H/L ratio = (number of heterophils) / (number of lymphocytes) (Nazar et al., 2018). 2.5 Statistical analysis Generalized Linear Mixed Models (GLMM) were used to compare differences between groups on each response variable. According to the data distribution, the percentage of inflammation, titers against SRBC and H/L ratio variables were respectively analyzed using a Gamma, Gamma, and Lognormal function. Dietary treatment was defined as a fixed effect, considering the pen as an experimental unit and as a random effect. DGC test was performed for post-hoc analysis29. A P value < 0.05 was considered to represent significant differences. Statistical analyses were performed through an ‘R’ (The R Foundation for Statistical Computing) user-friendly interface implemented in InfoStat (Di Rienzo et el., 2016).

Palabras clave: Immunity, Welfare, Poultry, Supplementation

Alcance geográfico

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Identificador del recurso

URI: http://hdl.handle.net/11336/241615

Colecciones

Datos de Investigación(CIBICI)
Datos de Investigación de CENTRO DE INV.EN BIOQUI.CLINICA E INMUNOLOGIA

Datos de Investigación(IIBYT)
Datos de Investigación de INSTITUTO DE INVESTIGACIONES BIOLOGICAS Y TECNOLOGICAS

Citación

Videla, Emiliano Ariel; Luna, Agustin; Picco, Romina Paula Andrea; Marin, Raul Hector; Nazar, Franco Nicolas; Caliva, Jorge Martín; (2024): Thymol, alpha tocopherol, and ascorbyl palmitate as a natural feed strategy to modulate immune variables and Heterophil to Lymphocyte ratio in broiler chickens. Consejo Nacional de Investigaciones Científicas y Técnicas. (dataset). http://hdl.handle.net/11336/241615

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