Sentinel serosurveillance of backyard hens proved West Nile virus circulation in the western provinces of Turkey.

West Nile virus (WNV) is a mosquito-borne virus of a re-emergence importance with a wide range of vertebrate hosts. Granted, it causes asymptomatic infection, but fatal cases and neurologic disorders were also recorded, especially in humans, horses and some exposed birds. The virus is globally spread and birds are considered an amplifying and reservoir host of WNV, helping to spread the disease due to their close contact with main hosts. In this study, we aimed to detect the presence of antibodies against WNV in backyard hens that were reared in the western Anatolian part of Turkey. A total of 480 chicken sera were randomly collected from six provinces in the west of Turkey (Mugla, Izmir, Aydin, Afyonkarahisar, Kutahya and Manisa) with 80 samples from each province (40 in spring and 40 in fall seasons). They were tested by using a competitive ELISA method to identify the specific avian antibodies of IgG that produced against the WNV envelope proteins (pr-E). Twelve of 480 (2.5%) sera were found seropositive, three of these positive sera were detected from the Izmir province (3.75%) collected in the spring session and the other nine positive sera were detected from the Mugla province (11.25%) collected in the fall session. Both of these provinces are located seaside and have suitable climate conditions for vectors of infection. The results indicated that WNV infection is in circulation in these provinces, and that may put the other susceptible vertebrates under risk of infection.

Evaluation of adjuvant activity of Astragaloside VII and its combination with different immunostimulating agents in Newcastle Disease vaccine.

Astragaloside VII (AST-VII), a major cycloartane saponin isolated from Turkish Astragalus species, turned out to be one of the most active metabolites demonstrating Th1/Th2 balanced immune response. As Quillaja saponins are extensively used in adjuvant systems, this study made an attempt to improve AST-VII based adjuvant systems by using different immunostimulatory/delivery agents (monophosphoryllipid A (MPL), Astragalus polysaccharide (APS) and squalene) and to induce cellular and humoral immune response against a viral vaccine. For this purpose, Newcastle Disease vaccine (NDV) was chosen as a model vaccine. Swiss albino mice were immunized subcutaneously with LaSota vaccines in the presence/absence of AST-VII or developed adjuvant systems. AST-VII administration both in live/inactivated LaSota vaccines induced neutralizing and NDV specific IgG, IgG1 and IgG2b antibodies response as well as IL-2 and IL-4 production. APS based delivery systems enhanced the production of neutralizing antibody and the minor augmentation of IFN-γ and IL-2 levels. Squalene emulsion (SE) alone or combined with AST-VII were effective in NDV restimulated splenocyte proliferation. As a conclusion, AST-VII and AST-VII containing adjuvant systems demonstrated Th1/Th2 balanced antibody and cellular immune responses in NDV vaccines. Thus, these systems could be developed as vaccine adjuvants in viral vaccines as alternative to saponin-based adjuvants.

Adjuvant potency of Astragaloside VII embedded cholesterol nanoparticles for H3N2 influenza vaccine.

Adjuvants are substances that increase the immune response to a given antigen. In the development of novel vaccine adjuvants/systems, saponins are one of the most attractive molecules due to their altered immunomodulatory activities. In this study, we tried to develop PEG (polyethylene glycol)/cholesterol-based lipid nanoparticles (LNPs) to deliver the Astragaloside VII (AST-VII) and potentiate adjuvant properties of AST-VII for the influenza vaccine. In the formation of PEG/cholesterol/AST-VII-based LNPs (PEG300: Chol-AST-VII LNPs), 3 different primary solvents (acetone, ethanol, and chloroform) were evaluated, employing their effects on hydrodynamic particle size, distribution, surface chemistry, and colloidal stability. Prepared nanoparticles were simply admixtured with inactivated influenza antigen (H3N2) and applied to PMA (phorbol 12-myristate 13-acetate)-ionomycin treated human whole blood to evaluate their cytokine release profile. PEG300: Chol-AST-VII LNPs (80.2 ± 7.7 nm) were obtained using chloroform as a desolvation agent. Co-treatment of PMA-ionomycin with AST-VII and PEG300: Chol-AST-VII LNPs significantly increased the levels of IL-2 and IFN-g, compared to PMA-ionomycin alone. In the presence of H3N2, AST-VII was able to augment IL-17A, while PEG300: Chol-AST-VII LNPs stimulated the production of IFN-g. Hemolysis was only observed in PEG300: Chol-AST-VII LNPs (250 µg/mL) treatment. AST-VII and AST-VII-integrated LNPs could be used as efficacious adjuvants for an inactivated H3N2 vaccine in vitro, and cytokine response through Th1/Th17 route was reported.

Correction to: First report of fowl aviadenovirus serotypes FAdV-8b and FAdV-11 associated with inclusion body hepatitis in commercial broiler and broiler-breeder flocks in Turkey.

The co-author names were found missed in the original publication of this article and the complete lists of authors were updated here. The original article has been corrected.

First report of fowl aviadenovirus serotypes FAdV-8b and FAdV-11 associated with inclusion body hepatitis in commercial broiler and broiler-breeder flocks in Turkey.

Inclusion body hepatitis (IBH), hydropericardium syndrome (HS), and gizzard erosion (GE) are all economically important diseases in the poultry industry worldwide and are all caused by fowl aviadenovirus (FAdV). It is important to identify the serotype of the virus to differentiate these diseases. In the present study, a total of six recent FAdV serotypes were isolated and identified in broiler and broiler-breeder flocks in Izmir, Manisa, and Aydin provinces of the Aegean region of Turkey between January and March 2019. The viruses were isolated from livers and pooled organs of chickens using primary chicken embryo kidney cell cultures (CEKC). Virus isolates were identified by PCR amplification of the loop 1 (L1) variable region of the hexon gene followed by Sanger sequencing. Sequence analysis revealed the presence of both FAdV-D (serotype 11) and FAdV-E (serotype 8b). The viruses that were isolated were associated with IBH, which is typically characterized by gross lesions such as enlarged and pale yellow liver with multiple petechial hemorrhages. Histopathological examination also showed necrotizing hepatitis with intranuclear inclusion bodies in hepatocytes. This study is the first report of the isolation and identification of FAdV serotypes associated with IBH in commercial broilers and broiler-breeder flocks in Turkey. The results of sequence analysis showed that FAdV-8b and FAdV-11 were the circulating serotypes that caused recent field outbreaks of IBH in the Aegean region between January and March, 2019.

Development of adjuvant nanocarrier systems for seasonal influenza A (H3N2) vaccine based on Astragaloside VII and gum tragacanth (APS).

Adjuvants are chemical/biological substances that are used in vaccines to increase the immunogenicity of antigens. A few adjuvants have been developed for use in human vaccines because of their limitations including lack of efficacy, unacceptable local or systemic toxicity, the difficulty of manufacturing, poor stability, and high cost. For that reasons, novel adjuvants/adjuvant systems are under search. Astragaloside VII (AST-VII), isolated from Astragalus trojanus, exhibited significant cellular and humoral immune responses. The polysaccharides (APS) obtained from the roots of Astragalus species have been used in traditional Chinese medicine and possess strong immunomodulatory properties. In the present study, the immunomodulatory effects of a newly developed nanocarrier system (APNS: APS containing carrier) and its AST-VII containing formulation (ANS: AST-VII + APNS), on seasonal influenza A (H3N2) vaccine were investigated. Inactivated H3N2 alone or its combinations with test compounds/formulations were intramuscularly injected into Swiss albino mice. Four weeks after immunization, the immune responses were evaluated in terms of antibody and cytokine responses as well as splenocyte proliferation. APNS demonstrated Th2 mediated response by increasing IgG1 antibody titers, whereas ANS showed response towards Th1/Th2 balance and Th17 by producing of IFN-γ, IL-17A and IgG2a. Based on these results, we propose that APNS and ANS are good candidates to be utilized in seasonal influenza A vaccines as adjuvants/carrier systems.

Effects of rearing systems on performance, egg characteristics and immune response in two layer hen genotype.

White (Lohmann LSL) and Brown (ATAK-S) laying hens, were reared under organic and conventional cage rearing systems, and the effects of the rearing system on performance parameters, egg production, egg characteristics, and immune response were investigated. For this purpose, a total of 832 laying hens of two commercial hybrids, i.e., 416 white (Lohmann LSL) and 416 Brown (ATAK-S) layers, were used. The experiment lasted between 23 and 70 wk of age. In this study, the white layers yielded more eggs as compared to the brown layers in both organic and conventional production systems. Egg weight exhibited a similar pattern to that of laying performance. However, the total hen-housed egg number for the white birds in the organic system was fewer than that of white birds in the conventional cage facility; conversely, a contradictory tendency was observed for the brown birds. Livability of the white layers in the organic system was remarkably lower (14%) than that of the brown line, whereas the white line survived better (3.42%) than their brown counterparts in conventional cages. The feed conversion ratio of the white hens was markedly inferior in the organic system as compared to that of the white hens in the conventional system, whereas relatively lower deterioration was reported in brown layers when reared in an organic system. The organic production system increased egg albumen height and the Haugh unit in eggs of the brown layers. The yolk color score of organic eggs was lower than that of conventional eggs for both brown and white hens. The egg yolk ratio of eggs from white layers was found to be higher in organic eggs as compared to those obtained in the conventional system. All organic eggs had heavier shells than those produced in the conventional system. Eggs from brown layers had more protein content than eggs from white layers. Neither housing systems nor genotype influenced egg yolk cholesterol concentration. When compared to conventional eggs, n-3 fatty acid content was lower in organic eggs, and the n-6:n-3 ratio was higher in organic eggs. In conclusion, two hen genotypes showed different responses in terms of performance and egg quality to two different rearing systems. A commercial white strain produced more eggs with higher egg quality as compared to a native brown strain. The brown strain was found to have adapted well to organic production conditions when survival and total egg number was taken into consideration.

Evidence for differing evolutionary dynamics of A/H5N1 viruses among countries applying or not applying avian influenza vaccination in poultry.

Highly pathogenic avian influenza (HPAI) H5N1 (clade 2.2) was introduced into Egypt in early 2006. Despite the control measures taken, including mass vaccination of poultry, the virus rapidly spread among commercial and backyard flocks. Since the initial outbreaks, the virus in Egypt has evolved into a third order clade (clade 2.2.1) and diverged into antigenically and genetically distinct subclades. To better understand the dynamics of HPAI H5N1 evolution in countries that differ in vaccination policy, we undertook an in-depth analysis of those virus strains circulating in Egypt between 2006 and 2010, and compared countries where vaccination was adopted (Egypt and Indonesia) to those where it was not (Nigeria, Turkey and Thailand). This study incorporated 751 sequences (Egypt n=309, Indonesia n=149, Nigeria n=106, Turkey n=87, Thailand n=100) of the complete haemagglutinin (HA) open reading frame, the major antigenic determinant of influenza A virus. Our analysis revealed that two main Egyptian subclades (termed A and B) have co-circulated in domestic poultry since late 2007 and exhibit different profiles of positively selected codons and rates of nucleotide substitution. The mean evolutionary rate of subclade A H5N1 viruses was 4.07×10(-3) nucleotide substitutions per site, per year (HPD 95%, 3.23-4.91), whereas subclade B possessed a markedly higher substitution rate (8.87×10(-3); 95% HPD 7.0-10.72×10(-3)) and a stronger signature of positive selection. Although the direct association between H5N1 vaccination and virus evolution is difficult to establish, we found evidence for a difference in the evolutionary dynamics of H5N1 viruses among countries where vaccination was or was not adopted. In particular, both evolutionary rates and the number of positively selected sites were higher in virus populations circulating in countries applying avian influenza vaccination for H5N1, compared to viruses circulating in countries which had never used vaccination. We therefore urge a greater consideration of the potential consequences of inadequate vaccination on viral evolution.