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Essential oils, extracts, their components, and other non-antibiotic beneficial additives are becoming increasingly popular as naturally occurring immunomodulatory agents. A study was conducted to evaluate the effects of a blend (hereafter refers to as ImmunoMax (IMX)) of peppermint, coneflower, thyme, propolis, and prebiotic on performance, immune response, hematological indices, and serum biochemistry of broiler chickens. A total of 300 day-old broiler male chicks were randomly assigned to 4 treatments: control (A) (not treated), 0.25 mL/L (B), 0.5 mL/L (C), and 1 mL/L (D) supplementing drinking water with IMX on alternate days, starting at day 21. The results demonstrated a higher body weight gain (BWG) and feed intake (FI) in groups B, C, and D (P < 0.05) with no significant difference in FCR. The results also revealed a superior immune response to NDV, IB, and IBD (P < 0.05), albeit no constant linear response. The largest portion of the heterophil, the smallest amount of the lymphocyte, and therefore the highest heterophil-to-lymphocyte ratio (H:L) were observed in the control group, while the lowest ratio belonged to groups B and C (P < 0.05). Hemoglobin content, hematocrit, total number of red blood cells (RBC), and white blood cells (WBC) were also affected by IMX, showing a significant rise compared to the control group (P < 0.05). Administering 0.25 mL/L of IMX resulted a higher thigh and breast yield as well as the smallest amount of abdominal fat (P < 0.05). The serum content of total protein (TP) and albumin (ALB) was superior in groups receiving 0.25 and 0.5 mL/L of IMX; these treatments also yielded an inferior content of glucose (GLU), triglycerides (TG), cholesterol (CHOL), LDL, and VLDL, in addition to causing the highest content of HDL (P < 0.05). The results obtained in this study revealed the immunomodulatory efficacy of a standardized natural multidrug, IMX, thereby improving the performance and carcass yield of modern broiler chickens with the optimal dosages of 0.25 and 0.5 mL/L.

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Marek's disease (MD) is a highly contagious, lymphoproliferative poultry disease caused by the oncogenic herpesvirus, serotype 1 Marek's disease virus (MDV-1), or Gallid herpesvirus 2 (GaHV-2). MDV strains have shown a continued evolution of virulence leading to immune failure, and MD cases continue to occur or surge. Meq, the major MDV-1 oncoprotein, induces T-cell neoplastic transformation through several mechanisms including inhibition of apoptosis, cell cycle regulation, and serum-anchorage independent growth. There is no current information on the MDV serotypes and pathotypes circulating in vaccinated commercial farms in Iran, where the birds are vaccinated at the hatchery with GaHV-2 and Meleagrid herpesvirus 1 (MeHV-1) vaccines. This study reports the molecular characterization of a GaHV-2 strain detected in 19 flocks of Iranian layer farms exhibiting MDV-1-like clinical signs and visceral lymphomas. Based on sequencing and phylogenetic analysis of the Meq gene, the Iranian GaHV-2 isolates could be divided into two separate clades regarding molecular features. The clade containing strains was closely related to Italian, Indian, and Hungarian virulent isolates, and the clade was related to American very virulent plus (vv+) isolates. For the first time, the MDV-1 virus was characterized by an outbreak in poultry flocks in Iran. Although MDV-1 strains obtained in Iran's present outbreak are presumably related to virulent (v) and vv+ pathotypes based on nucleotide, amino acid, and phylogenetic analysis of the viruses, they are not confirmed so far. Thus, it is highly recommended to perform further analyses to demonstrate the pathotype characteristics in vivo.

Caracterización molecular y análisis filogenético del virus de la enfermedad de Marek en Irán. La enfermedad de Marek (MD) es una enfermedad altamente contagiosa linfoproliferativa en la avicultura causada por el herpesvirus oncogénico, el virus de la enfermedad de Marek de serotipo 1 (MDV-1) o Gallid herpesvirus 2 (GaHV-2). Las cepas del virus de Marek han mostrado una evolución continua de virulencia que conduce a una falla inmunológica, y los casos de Marek continúan ocurriendo o aumentando. El gene Meq, codifica la principal oncoproteína de MDV-1, induce la transformación neoplásica de células T a través de varios mecanismos que incluyen la inhibición de la apoptosis, la regulación del ciclo celular y el crecimiento independiente del anclaje sérico. No hay información actual sobre los serotipos y patotipos del virus de Marek que circulan en las granjas comerciales vacunadas en Irán, donde las aves se vacunan en la planta de incubación con las vacunas GaHV-2 y Meleagrid herpesvirus 1 (MeHV-1). Este estudio reporta la caracterización molecular de una cepa del Gallid herpesvirus 2 detectada en 19 lotes de granjas de aves de postura iraníes que presentaron signos clínicos sugestivos del serotipo 1 del virus de la enfermedad de Marek y linfomas viscerales. Según la secuenciación y el análisis filogenético del gene Meq, los aislamientos iraníes de GaHV-2 podrían dividirse en dos clados separados con respecto a las características moleculares. El clado que contenía las cepas estaba estrechamente relacionado con los aislados virulentos de Italia, India y de Hungria y el clado estaba relacionado con los aislados americanos muy virulentos plus (vv+). Por primera vez, el serotipo 1 del virus de la enfermedad de Marek se caracterizó por un brote en parvadas avícolas en Irán. Aunque las cepas del virus de Marek, serotipo 1 obtenidas en el brote actual de Irán están presuntamente relacionadas con patotipos virulentos (v) y muy virulentos plus según el análisis de nucleótidos, aminoácidos y filogenético de los virus, hasta el momento no se han confirmado. Por lo tanto, se recomienda realizar más análisis para demostrar las características del patotipo in vivo.

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The unprecedented rate of global warming requires more immaculate strategies to fight the heat stress and its detrimental effects on poultry sector. Nutritional strategies, particularly herbal extracts, play a crucial importance in this scenario. Therefore, this study was conducted to evaluate the effects of a novel selected mixture (hereafter refers to as Stress-Bio-Max, SBM) comprising St John's wort, lavender, Melissa officinalis extracts, betaine, and Vit C on heat-stressed broilers. A total of 300 male broiler chicks were allocated to 4 treatments: T1, Control (no SBM in drinking water); T2, 0.25 mL/L of SBM in drinking water; T3, 0.5 and T4, 1 mL/L of SBM, which applied since d 32. The birds underwent heat stress (40 ± 1) daily from d 35 to 42 of the experiment (2 pm-8 pm). The results show the superior weight gain of T2 and T3 from d 7 to 42 compared to other groups, 72.5 and 72.7 g/d, respectively (P < 0.05). Corticosterone level was higher on d 35 in T1 (4.674 ng/mL, P < 0.05) and lower in T2 and T3 (2.64 and 2.952 ng/mL, respectively, P < 0.05); T2 and T3 also caused the lowest concentrations of corticosterone on d 40 (5.198 and 6.458 ng/mL, respectively, P < 0.05). The superior triiodothyronine levels belonged to T2 (0.935 ng/mL, P < 0.05) on d 35 and T2 and T3 on d 40 (0.699 and 0.582 ng/mL, respectively, P < 0.05). T2, T3, and T4 caused a rise in glutathione peroxidase (GPX) and superoxide dismutase (SOD) activities (P < 0.05). A significant higher amount of total antioxidant capacity (TAC) belonged to the groups treating with SBM (P < 0.05). The SBM in T2 and T3 resulted in the lowest levels of malondialdehyde (MDA), aspartate transaminase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). Results from this study indicate that SBM may alleviate the negative impacts associated with heat stress in broiler chickens.

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[This corrects the article DOI: 10.1371/journal.pone.0229009.].

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Broiler chickens experience an acute-phase response (APR) through vaccination, which reflects the innate immunity and stress related to immunization. It is also considered that APR can modulate adaptive immunity and response to infection. As biomarkers for APR, assessing the acute-phase proteins (APPs) function and their levels in response to immunization is of great value for vaccine design, development and administration. In this study, the heterophils/lymphocyte (H/L) ratio and the level of APPs was evaluated in broilers with three different Newcastle disease (ND) vaccination regimens. Inactivated ND vaccine (IND) was administered by the intramuscular route. Live attenuated strains, Lasota and Vitapest, was administered by ocular routes. H/L ratio, serum amyloid A (SAA) and alpha-1 acid glycoprotein (AGP) were measured before and after two rounds of vaccination on days 10 and 21. In a comparison between the three vaccines, H/L ratio in IND group significantly increased to 3 fold (1.48 ± 0.41) after the first vaccination while the Lasota and Vitapest showed a milder response. The concentration of SAA increased after 24h by 1.8-fold in IND group (0.116 ± 0.015 mg/L) and 2-fold in Lasota group (0.14 ± 0.002 mg/L). Significant changes were found in Vitapest group after 48h post vaccination (0.113 ± 0.016 mg/L). Elevation pattern of AGP, 24 hours after first vaccination in IND (3.5-fold) and Vitapest (2.5-fold) was different from Lasota in which the peak was reached after 48 hours (2.9-fold). Except for IND group, no significant changes in SAA and AGP concentrations were detected after the second vaccination. A significant positive correlation between SAA values at day 22 and HI titers at day 28 (r = 0.998, P≤0. 0.005) was found. According to these results, different types of ND vaccines can cause different patterns of acute phase responses. Assessment of stress and level of acute-phase proteins can be used for prediction of immune response outcomes in vaccine design and development.

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