Preparation and evaluation of chicken embryo-adapted fowl adenovirus serotype 4 vaccine in broiler chickens

The current study was planned to develop an efficient vaccine against hydropericardium syndrome virus (HSV). Currently, formalin-inactivated liver organ vaccines failed to protect the Pakistan broiler industry from this destructive disease of economic importance. A field isolate of the pathogenic hydropericardium syndrome virus was adapted to chicken embryos after four blind passages. The chicken embryo-adapted virus was further serially passaged (12 times) to get complete attenuation. Groups of broiler chickens free from maternal antibodies against HSV at the age of 14 days were immunized either with 16th passage attenuated HSV vaccine or commercially formalized liver organ vaccine. The antibody response, measured by enzyme-linked immunosorbent assay was significantly higher (P < 0.05) in the group immunized with the 16th passage attenuated HSV vaccine compared to the group immunized with liver organ vaccine at 7, 14, and 21 days post-immunization. At 24 days of age, the broiler chickens in each group were challenged with 10^3.83 embryo infectious dose₅₀ of pathogenic HSV and were observed for 7 days post-challenge. Vaccination with the 16th passage attenuated HSV gave 94.73% protection as validated on the basis of clinical signs (5.26%), gross lesions in the liver and heart (5.26%), histopathological lesions in the liver (1.5 ± 0.20), and mortality (5.26%). The birds inoculated with liver organ vaccine showed significantly low (p < 0.05; 55%) protection estimated on the basis of clinical signs (40%), gross lesions in the liver and heart (45%), histopathological lesions in the liver (2.7 ± 0.72), and mortality (35%). Birds in the unvaccinated control group showed high morbidity (84%), mortality (70%), gross (85%), and histopathological lesions (3.79 ± 0.14) with only 10% protection. In conclusion, this newly developed HSV vaccine proved to be immunogenic and has potential for controlling HSV infections in chickens.     

Molecular study and genotyping of Cryptosporidium baileyi and Cryptosporidium parvum from free-range and commercial broiler chickens in Guilan province,Iran

Cryptosporidiosis acutely impacts the digestive and/or respiratory tract of the birds in many species of various orders. More importantly, it is also well known as a significant zoonotic disease, which can lead to diarrhea in humans and livestock. Regarding increasing demand for free-range products and increasing the number of free-range poultry farms, the present paper evaluated histopathological and molecular detection of Cryptosporidium baileyi and Cryptosporidium parvum in free-range and commercial broiler chickens in the north part of Iran. For this purpose, 100 fecal and tissue samples of the chickens in Guilan province were collected. After microscopic examination using Ziehl-Neelsen staining, molecular analyses of the fecal samples were processed by Nested-PCR targeting the 18S rRNA gene followed by sequencing of the amplicons and phylogenetic analyses. Eventually, the tissue samples were studied for histological lesions. Findings demonstrated the presence of Cryptosporidium baileyi and Cryptosporidium parvum in 6 % and 2 % of fecal samples, respectively. This is the first identification of C.parvum in avian hosts in Iran, and for the first time, C.baileyi and C.parvum are shown in native free-range chickens in Iran. All of the PCR positive birds with clinical symptoms showed gross lesions of respiratory infections. There was no significant difference between infection rate in free-range and commercial broiler chickens; however, the infection rate was significantly higher in chickens <25 days old. To conclude, we present here a notable Cryptosporidium infection rate in the free-range chicks in Iran, which notify the role of this host as a reservoir and should be more noted due to the economic and zoonotic importance.     

Effects of feeding dried broccoli floret residues on performance,ileal and total digestive tract nutrient digestibility,and selected microbial populations in broiler chickens

A study was conducted to determine the effects of feeding dried broccoli floret residues on growth performance, apparent ileal digestibility (AID) and apparent total tract nutrient digestibility (ATTD) coefficients, and intestinal microbial populations in broiler chickens. One thousand two hundred day-old male Ross 508 broilers were randomly allotted to 4 dietary treatments and were grown over a 35-d experimental period. Dietary treatments included 4 levels of dried broccoli floret residues: 0, 3, 6, and 9%. Results showed that inclusion of dried broccoli floret residues increased body weight gain (quadratic effect, P = 0.004) and feed conversion ratio (quadratic effect, P = 0.002) with no effect on feed intake. Apparent ileal crude protein (CP, quadratic effect, P = 0.003) and dry matter (DM, quadratic effect, P = 0.002) digestibility for younger birds (25 d of age) increased as the level of dried broccoli floret residues in the diet increased. However, apparent ileal CP (linear effect, P = 0.022), DM (linear effect, P = <0.001) and gross energy (linear effect, P = 0.001) digestibility for older birds (35 d of age) decreased as a result of dried broccoli residue inclusion. Nitrogen (N) corrected apparent metabolizable energy decreased (linear effect, P < 0.001) as the level of dried broccoli floret residues in the diet increased. However, N retention was not influenced by dried broccoli floret residue inclusion. It was concluded that incorporation of dried broccoli floret residues in broiler diet at moderate levels (i.e., 3 to 6%) may improve the growth of broiler chickens with no detrimental effects on nutrient digestibility and retention. However, at high levels (i.e., 9%), dietary dried broccoli floret residues may compromise ileal and total tract nutrient digestibility.     

Pathogenicity of serotype 1 fowl adenovirus in commercial broiler chickens.

The pathogenicity of a serotype 1 fowl adenovirus (FAV-99ZH), isolated from broiler chickens exhibiting gizzard erosion, was investigated in commercial broiler chickens. Five-, 3-, and 1-wk-old commercial broiler chickens were inoculated with FAV-99ZH by both oral and ocular routes. In the 5-wk-old chickens (trial 1), none of which had the maternal antibody to FAV-99ZH, severe gizzard erosions were observed on days 5, 7, and 10 postinoculation (PI). Among the 3-wk-old chickens (trial 2), which were separated into a control group and three treatment groups according to their maternal antibody titer levels, some chickens showed clinical signs such as depression and anorexia. Compared with the control group, all the treatment groups showed decreased weight gain. One treatment group, moreover, showed significantly decreased (P < 0.05) weight gain on day 10 PI. Severe gizzard lesions, such as erosion or ulcers, were observed from day 4 PI in all treatment groups regardless of their maternal antibody levels. The 1-wk-old chickens (trial 3) were separated into a control group and two treatment groups according to their titer levels of the inoculated virus. In spite of high maternal antibody levels, severe gizzard lesions were observed in both treatment groups, which also showed decreased weight gain. One treatment group, inoculated with the higher dose, showed significantly decreased (P < 0.05) weight gain on days 10 and 14 PI compared with the control group. Fowl adenovirus was recovered mainly from gizzard and rectal (including feces) samples from inoculated chickens but was not recovered from liver samples in any of the trials or in any of the control chickens. Although the reproduced disease was similar to that described in a previous report of experimental infection of specific-pathogen-free (SPF) white leghorn chickens with fowl adenovirus, the pathogenicity of FAV-99ZH in commercial broiler chickens was more severe than that in the SPF white leghorn chickens. The results of the present study indicate that FAV-99ZH isolated from gizzard erosion had pathogenicity and produced severe lesions in the gizzards of broiler chickens and that FAV-99ZH could infect and produce illness in broiler chickens with maternal antibodies against this virus.     

Inclusion body hepatitis in broiler chickens in Iraq

Eighteen outbreaks of inclusion body hepatitis (IBH) were identified in broiler chickens in Baghdad in 1977 and 1978. The disease was seen mainly in 4-to-6-week-old broiler chickens. The mortality rate did not exceed 1% in any outbreak investigated. The most common gross findings were stellate or punctiform hemorrhagic areas in markedly fatty livers. Noticed in many cases were enlarged and pale kidneys, hemorrhagic lesions in the skeletal muscles, and pale fattened bone marrow. Histological examination of tissues revealed fat droplets and intranuclear inclusion bodies in degenerated liver cells. Eosinophilic inclusion bodies were seen in all cases. Only five cases had basophilic inclusions along with the eosinophilic ones. The etiologic agent was isolated on chorioallantoic membrane (CAM) of 12-day-old embryonated chicken eggs. The disease was produced experimentally in 4-week-old chicks using infected CAM suspension.     

Efficacy of VP2 protein expressed in E. coli for protection against highly virulent infectious bursal disease virus

The ability of a heat-inactivated whole virus from a highly virulent infectious bursal disease virus (hvIBDV) and VP2 protein from hvIBDV expressed in E. coli provided protection against a hvIBDV challenge in specific-pathogen-free (SPF) chickens. Six out of seven chickens that were injected three times with crude VP2 protein developed significant antibody titer against IBDV. However, only four out of the seven chickens survived the hvIBDV challenge. Despite showing low antibody titer profiles, all chickens immunized with the heat-inactivated whole virus also survived the challenged with hvIBDV. However, all of these chickens had bursal atrophy and mild to moderate depletion of lymphocytes. Thus, antibodies raised against IBDV VP2 protein expressed in E. coli and denatured IBDV proteins induced some degree of protection against mortality but not against bursal damage following challenge with hvIBDV.     

Immunoreactivity and morphological changes of bursal follicles in chickens infected with vaccine or wild-type strains of the infectious bursal disease virus

Infectious bursal disease (IBD) is characterized by immunosuppression due to the depletion of lymphocytes in the atrophied bursa of Fabricius (BF). We have sometimes encountered contradictory findings: chickens infected with the vaccine IBD virus (IBDV) strain have sometimes exhibited a highly atrophied BF, but not immunosuppression. In this study, chickens administered vaccine or wild-type strains of IBDV were later vaccinated with the B1 strain of the Newcastle disease virus (NDV). Bursal changes were examined histologically with a focus on the bursal follicle. The immunoreactivity to NDV was also evaluated with the hemagglutination inhibition test. In gross examination, we observed a few chickens with a severely atrophied BF in vaccine strain-administered groups (vaccine groups), and the level of severity was the same as that in the wild-type strain-administered group (wild-type group). However, these chickens retained humoral antibody responses to NDV and were revealed to possess a higher number of bursal follicles than those of the wild-type group. These results indicated that macroscopic evaluation dose not accurately reflect the immunoreactivity and degree of bursal damage in IBDV-administered chickens. We also found non-immunosuppressed chickens in the wild-type group. These non-immunosuppressed chickens retained a significantly higher number of normal follicles and total follicles according to our statistical analysis. Furthermore, a high correlation coefficient between the NDV-HI titer and the number of normal follicles was found in the wild-type group. These results implied that the retained number of normal follicles is important for the immunoreactivity of chickens infected with IBDV.     

霉变饲料中添加复方霉菌毒素吸附剂对肉鸡人工感染发生的影响

本试验旨在研究霉变饲料和／或添加复方霉菌毒素吸附剂对肉鸡人工感染发生的影响。选取140羽1日龄健康的AA肉鸡随机分成7个组．每组4个重复。第1组为空白对照组,饲喂基础日粮;第2组饲喂霉变饲料;第3～7组以混合感染病鸡病变组织匀浆饮水制作人工感染疾病模型;第3～7组分别饲喂基础日粮、霉变饲料、霉变饲料＋复方霉菌毒素吸附剂、霉变饲料＋复方霉菌毒素吸附剂＋抗氧化剂、霉变饲料＋复方霉菌毒素吸附剂＋抗氧化剂＋中药增免剂。试验期35d。结果发现,肉鸡摄入霉变饲料与摄入正常饲料相比,提高肉鸡料重比和死淘率,显著降低肉鸡增重、胸腺指数、法氏囊指数,显著降低血清IBDV、H9N1v和NDV抗体水平（P〈0．05）。给人工感染肉鸡饲喂霉变饲料与饲喂正常饲料相比．显著提高料重比和死淘率,显著降低肉鸡增重、胸腺指数和脾脏指数,显著降低血清IFN-1、IL-2、IL-4和IL-12含量以及LTR,显著降低血清IBDV、H9N1V和NDV抗体水平（P〈0．05）。霉变饲料中添加复方霉菌毒素吸附剂能显著提高混合感染肉鸡增重,并降低料重比和死淘率（P〈0．05）,显著提高胸腺指数、法氏囊指数（P〈0．05）,显著提高血清IBDV、H9N1V和NDV抗体水平（P〈0．05）。可见,霉变饲料可导致肉鸡免疫功能抑制,加重肉鸡混合感染病情;复方霉菌毒素吸附剂能有效缓解霉变饲料对人工感染肉鸡生产性能、免疫功能和病情的不利影响,抗氧化剂和免疫功能增强剂可减轻霉变饲料的毒性作用。     

Effects of salinomycin and Bacillus subtilis on growth performance and immune responses in broiler chickens

The present study was undertaken to compare the effect of salinomycin and Bacillus subtilis on growth performance, serum antibody levels against Clostridium spp. and Eimeria spp., and cytokine mRNA expression levels in broiler chickens raised in the used litter. Broiler chickens fed a diet containing salinomycin showed lower (P < 0.05) body weights compared with the control diet-fed counterparts. Serum nitric oxide levels were significantly (P < 0.05) elevated in chickens fed the B. subtilis-enriched diet compared with those on either the salinomycin-fed or control diet-fed chickens. None of the dietary treatments affected (P > 0.05) serum antibody levels against Clostridium perfringens toxins. Both salinomycin and B. subtilis significantly lowered (P < 0.05) the serum levels of Eimeria-specific antibodies compared with the control group. Salinomycin, but not B. subtilis, significantly modulated (P < 0.05) the expression of cytokines encoding interferon-γ (IFN-γ), interleukin10 (IL-10) and tumor necrosis factor superfamily 15 (TNFSF15) compared with the control group. In conclusion, dietary salinomycin and B. subtilis affected serum anticoccidial antibody and intestinal cytokine expression, but failed to improve growth performance in broiler chickens. Further study is warranted to investigate the mode of action of salinomycin on host immune response and growth performance in broiler chickens.     

Immunostimulant effect of a mixed herbal extract on infectious bursal disease virus (IBDV) vaccinated chickens in the context of a co-infection model of avian influenza virus H9N2 and IBDV

This study was conducted to assess the comparative effects of a mixed herbal extract (MHE) containing Ocimum sanctum, Withania somnifera, Emblica officinalis, Tinospora cordifolia, Mangifera indica, and Asphaltum (shilajit) on infectious bursal disease virus (IBDV)-vaccinated (VAC) chickens infected with IBDV and avian influenza virus (AIV) H9N2. The experiment included three groups (G1-G3): G1, the negative control group; G2, the VAC + challenged (Ch) group; and G3, the VAC + Ch + MHE group. MHE was orally administered continuously for 5 weeks post-vaccination (PV) with IBDV at 12 days of age, and the chicks were simultaneously challenged with virulent IBDV (intraocularly) and AIV H9N2 (intranasally) at 21 days PV. Blood and tissue samples as well as tracheal and cloacal swabs were gathered at different times PV and post-challenge. Immunological and haematological parameters, histopathological lesions, relative organ weights and final live weights revealed significant differences (P ≤ 0.05) between G2 and G3 groups. Furthermore, in the G3 group, the protection rates, ELISA and HI titers and CD4+/CD8+ ratio were significantly increased, whereas viral shedding titers and the heterophil/lymphocyte ratio were decreased. In conclusion, the oral administration of the mixed herbal extract for 5 weeks can stimulate the immune response to IBDV vaccination and relieves the pathogenicity of an AIV H9N2 and IBDV co-infection in chickens.     

Proventriculitis in broiler chickens: effects of immunosuppression

Proventriculitis in broilers causes carcass condemnation when swollen proventriculi tear during evisceration. The cause of this proventriculitis is unknown, but several infectious agents have been associated with it. One such agent, infectious bursal disease virus (IBDV), has been implicated as a cause of proventriculitis, but a direct effect of this virus on the proventriculus has not been proven. The role of IBDV in proventriculitis may be indirect as a result of its ability to cause immunosuppression. The objective of this study was to understand how immunosuppression affects the incidence of proventriculitis in broiler chickens. Immunosuppression was induced in commercial and specific-pathogen-free broiler chickens using chemicals (cyclophosphamide and cyclosporin) or virus (IBDV). All groups were then exposed to a proventricular homogenate produced from diseased birds. At 7 and 14 days postinoculation, the incidence of proventriculitis in these groups was compared to that produced by homogenate exposure in immunocompetent broilers. All birds exposed to the proventricular homogenate from diseased birds developed proventriculitis. Cyclophosphamide and IBDV, both B cell suppressors, did not significantly affect the incidence or characteristics of the proventriculitis observed, although they did have an effect on the size of the proventriculus at 7 days postinoculation. Chickens immunosuppressed with cyclosporin, a T cell suppressor, developed more severe lesions and had a higher incidence of proventriculitis. These findings indicate that both B and T cells are involved in the immune response against proventriculitis, but cell-mediated immunity appears to have a more important role in controlling the disease. IBDV affects both humoral and cellular immunity in the chicken, so although under experimental conditions it didn't have a major effect on proventriculitis, it may explain why control of IBDV in the field seems to reduce the incidence of proventriculitis.     

Effect of dietary supplementation of medium-chain fatty acids on growth performance and prevalence of carcass defects in broiler chickens raised in different stocking densities

A 3 × 3 factorial experiment was conducted to examine the effect of medium-chain fatty acids (MCFA) on performance and occurrence of carcass defects in broiler chickens raised in stocking densities of 14, 16, and 18 birds/m². A total number of 720 Ross 308 broiler chicks that were 1 d old were reared up to 49 d of age. Corn- and soybean meal (SBM)–based starter and grower diets were supplemented with 0, 1.5, and 2 g/kg Aromabiotic and provided to the birds ad libitum. Inclusion of 2 g/kg Aromabiotic in the diet significantly improved weight gain of the birds during 1 to 21 d of age compared with control birds (P< 0.001). A significant reduction in mortality (P= 0.004) and a significant increase in European performance efficiency index (EPEI;P= 0.036) were noted in the birds that received diets containing 2 g/kg Aromabiotic compared with control birds at 49 d of age. The birds grown in a placement density of 18 birds/m²had significantly improved FCR along with reduced feed intake (FI) during 1 to 49 d of age compared with those reared in a density of 14 birds/m². The mean EPEI was significantly greater in the birds raised in a density of 16 birds/m²(P< 0.001). At 48 d of age, litter caking was significantly increased in the birds raised in a density of 18 birds/m²and decreased in the birds that received dietary supplementation of Aromabiotic (P< 0.05). The influence of dietary Aromabiotic was significant related to reduced incidence and severity of footpad lesions and hock burns (P< 0.01). Frequency of lesions with a score of 2 to 5 on footpad and hock significantly increased for the birds raised in densities higher than 14 birds/m². It was concluded that dietary supplementation of 2 g/kg Aromabiotic improved early productive performance of broiler chickens and enhanced the PEI mainly due to decreased mortality. Dietary Aromabiotic improved broiler welfare as indicated by reduced foot defects in the birds raised in placement densities of 16 and 18 birds/m².     

Effects of dietary fiber and threonine on performance,intestinal morphology and immune responses in broiler chickens

The present experiment was conducted to investigate the effects of fiber and threonine (Thr) on growth performance, intestinal morphology and immune responses of broiler chickens. A total of 420 one-day-old mixed sex broiler chicks (Ross 308) were randomly divided into 6 experimental diets and 5 replicates of 14 chicks based on a factorial arrangement (2 × 3) from 1 to 42 d of age. Experimental factors included dietary supplemental fiber type (no supplemental fiber [NSF], 30 g/kg sugar beet pulp [SBP] or 30 g/kg rice hull [RH]) and Thr inclusion level (100% or 110% of Thr requirement recommended by breeder company [Ross 308]). Growth performance of broilers was assayed at different periods of the experiment. Intestinal morphometric features were measured at 21 d of age. Antibody titer against sheep red blood cells (SRBC), Newcastle and influenza disease viruses were measured on d 30 of trial. Dietary inclusion of SBP and RH significantly decreased feed intake and weight gain during the entire rearing period (P < 0.05). Interaction of fiber and Thr had no beneficial effects on the performance of chickens across the entire rearing phase. Dietary supplementation of 110% Thr required level improved jejunal morphometric features (P < 0.05), whereas its inclusion with fibrous materials failed to show the same effects. Dietary supplemental Thr together with SBP significantly increased antibody production against SRBC (P < 0.05). In conclusion, although supplemental 30 g/kg fibrous materials impaired growth performance, inclusion of SBP along with 110% Thr level improved the humoral immunity in broiler chickens.     

Subclinical infectious bursal disease in an integrated broiler production operation.

A field study was designed to determine the prevalence of subclinical infectious bursal disease (IBD) in broiler chickens from a commercial poultry company. Bursae of Fabricius (BF) from two vaccinated and three nonvaccinated broiler flocks were evaluated histologically, and antibody profiles of these broiler and matched parent breeder flocks were established. Lesions of IBD, including lymphoid necrosis, stromal edema, and infiltrates of heterophils and macrophages, were first detected in BF at 24 days of age in both vaccinated and nonvaccinated chickens. At 41 days, all BF had lesions characteristic of IBD, including severe lymphoid depletion, proliferation of epithelial cells, and mild fibroplasia. Although mean maternal antibody levels (measured by enzyme-linked immunosorbent assay) in broilers were apparently protective through day 12, IBD antibodies decreased to nonprotective levels (below 1,000) by day 16 or 20. Titers began to increase by day 28 or 32 because of field exposure. Sentinel birds, placed with broiler flocks, also developed IBD antibody titers. Broiler breeders had low and nonuniform antibody titers. Prevalence of field IBD exposure was high, and existing vaccination programs were not effective.     

Starch digestion rates in multiple samples of commonly used feed grains in diets for broiler chickens

In this study the starch digestion rates in broiler chickens from 18 samples of 5 commonly used feed grains (sorghum, wheat, maize, barley, triticale) were determined. The methodology to determine starch digestion rates in poultry is detailed herein. Starch digestion rates were not significantly different (P = 0.128) across the 18 feed grains, which reflects the wide variations that were observed within a given feedstuff. Nevertheless, starch digestion rates in broiler chickens offered wheat-based diets were significantly more rapid by 56.0% (0.117 versus 0.075 min⁻¹; P = 0.012) than their sorghum-based counterparts on the basis of a pair-wise comparison. In descending order, the following starch digestion rates were observed: wheat (0.117 min⁻¹), barley (0.104 min⁻¹), triticale (0.093 min⁻¹), maize (0.086 min⁻¹), sorghum (0.075 min⁻¹). The implications of these findings are discussed as they almost certainly have implications for poultry nutrition and the development of reduced crude protein diets for broiler chickens.     

Protein digestive dynamics of meat and bone meals in broiler chickens

This study determined the variations in protein digestibilities and digestion rates in broiler chickens offered diets containing 7 different meat and bone meals (MBM). A total of 252 male Ross 308 broiler chickens were offered 7 atypical diets largely based on maize and MBM from 24 to 28 d post-hatch. Each experimental diet was offered to 6 replicates with 6 birds per replicate cage. Excreta were collected in their entirety from 25 to 27 d post-hatch and on 28 d post-hatch. Digesta samples were collected from the proximal jejunum, distal jejunum, proximal ileum and distal ileum. Apparent digestibilities of protein were determined in each segment and apparent digestibilities of amino acids were measured in the distal ileum. There were significant differences in apparent protein digestibility coefficients in the proximal jejunum (P = 0.006), where broiler chickens offered the high ash beef meal (diet 7) generated the lowest protein digestibility in the proximal jejunum (0.318). Similarly, there were significant differences in apparent digestibility coefficients in the distal jejunum (P < 0.001) and distal ileum (P < 0.001) but not in the proximal ileum. More pronounced differences were found in the disappearance rate of protein and there were significant differences in all 4 segments of the small intestine (P < 0.001). Broiler chickens offered the high ash beef meal had the lowest protein disappearance rate (P < 0.001). No difference was observed in the predicted protein digestion rate (P = 0.486) but chickens offered the high ash beef meal had the lowest potential digestible protein (0.662, P = 0.034) whereas the highest potential digestible protein (0.739) was detected in diet 5 (containing a beef meal). This study contributed to the establishment of a preliminary database to include digestion rates of starch and protein into practical diet formulation.     

Depletion of glycogen reserves in fasting broiler chickens

1. Male broiler chickens were killed at various times up to 36 h after withdrawal of food; hepatic and muscle glycogen concentrations and pH were compared with those of control birds given continuous access to food and water. 2. Liver glycogen was reduced to negligible concentrations (less than 1 mg/g) within 6 h of food deprivation and the initial pH was elevated. 3. Withdrawal of food had no effect on glycogen concentrations in the m. pectoralis superficialis (PS) but reduced them in the m. biceps femoris (BF); the reduction was significant after 12 h. Ultimate pH was elevated by fasting in the BF, but not the PS.