Use of an enzyme-linked immunosorbent assay to measure antibody levels in turkey breeder hens, eggs, and progeny following natural infection or immunization with a commercial Bordetella avium bacterin

Twelve large white turkey hens were immunized with a commercially available Bordetella avium bacterin. Hens and eggs were tested using an enzyme-linked immunosorbent assay (ELISA) to determine the response to the bacterin. Three hundred poults were then obtained from two commercial flocks, the hens of one flock having been immunized with the same bacterin used on the group of 12 turkeys. Titers of the poults were monitored for 7 weeks, and poults were challenged by exposure to infected poults at 1, 7, 14, and 21 days post-hatch. Hens produced an antibody response following immunization, with a parallel antibody response being detected in eggs. Maternal antibodies were present in poults from immunized hens. Poult titers declined to near the level of poults from unimmunized hens by 14 days of age. Poults from immunized hens challenged at 1 and 7 days were resistant to development of clinical disease and gross lesions, whereas all poults from unimmunized hens exhibited clinical signs and gross lesions. After 14 days, the resistance of both groups to development of clinical disease, became near equal, neither group being affected as severely as the unimmunized hens challenged at days 1 and 7. Six commercial turkey breeding flocks and their progeny that had not been vaccinated for B. avium and had no history of B. avium infection were evaluated with the B. avium ELISA. There were variations between the flocks, with poult titers reflecting those found in the hens.     

Selected blood chemistry values in mobility-impaired broiler breeder hens with suspected calcium tetany using the i-STAT handheld clinical analyzer

Calcium tetany is a poorly defined disease of broiler breeder hens that results from acute hypocalcemia. It is characterized by impaired mobility, increased mortality, and absence of gross lesions that would explain the impaired mobility. To evaluate if hens with impaired mobility had calcium tetany or other abnormalities, blood values from normal and affected hens were determined using the i-STAT handheld clinical analyzer. Three flocks were evaluated weekly prior to peak production (range 25-30 wk of age) comparing normal hens to hens with clinically apparent calcium tetany. Calcium tetany suspect (CaTS) hens from four additional flocks were also evaluated. Significant hypocalcemia (P < 0.001) was observed in CaTS hens (average = 1.14 mmol/L ionized calcium [iCa]) compared to normal hens (average = 1.53 mmol/L iCa) in only one of three flocks sampled weekly. Clinically affected hens from one of the other four flocks also had hypocalcemia. Blood value abnormalities in mobility-impaired hens without hypocalcemia included hypernatremia. Findings in this study indicate calcium tetany is one cause of impaired mobility in breeder hens, but mobility impairment without hypocalcemia can also occur. Calcium tetany should be confirmed by finding significantly decreased levels of iCa in the blood, as diagnosis based on clinical presentation and necropsy results can be inaccurate. The i-STAT handheld clinical analyzer is an efficient, relatively low-cost method to determine iCa and other blood chemistry values that may be associated with impaired mobility in broiler breeder hens.     

Sudden death syndrome in turkey hens

A turkey breeder flock of 5600 31-week-old hens experienced an abrupt increase in daily mortality for a 2-day period. This higher mortality rate corresponded with the handling and moving of the hens 1 to 2 days before. No clinical illness was seen in the hens before carcasses were discovered. The four hens that were necropsied had small spleens, congested lungs, and enlarged livers. The most consistent histologic lesions were pulmonary hemorrhage and edema, and congestion of veins and capillaries in several visceral organs. One hen had subserosal hemorrhage of the oviduct and perirenal hemorrhage. The increased mortality pattern, the lack of clinical signs, and the histopathologic changes are consistent with a diagnosis of sudden death syndrome. The presence of concurrent inflammatory lesions in the lungs predisposed the hens to sudden death following the stress of handling and moving.     

Protection of turkey poults from Bordetella avium infection and disease by pili and bacterins

The role of pili in protection against Bordetella avium infection in turkey poults was studied. An isolate that produced the largest number of pili under growth conditions developed in our laboratory was used for preparation of pili and bacterin and for challenge. The pili were isolated, purified, examined by electron microscopy, and tested for purity by gel electrophoresis. Poults were vaccinated with oil-adjuvant pili, formaldehyde- or merthiolate-inactivated bacterins, or a commercial bacterin. Poults were vaccinated once or twice subcutaneously at different ages and challenged intranasally with a pathogenic B. avium isolate 5 days following the last vaccination. A few vaccinated birds had very mild clinical signs. B. avium was isolated from the sinuses of a few vaccinated birds, and growth was scanty. The mean colony counts from tracheal sections was significantly higher (P less than 0.1) in unvaccinated challenged poults than in vaccinated challenged poults. It is postulated that B. avium pili are important immunogens in turkey poults.     

Low pathogenicity avian influenza viruses infect chicken layers by different routes of inoculation

In order to develop better control measures against avian influenza, it is necessary to understand how the virus transmits in poultry. In a previous study in which the infectivity and transmissibility of the pandemic H1N1 influenza virus was examined in different poultry species, we found that no or minimal infection occurred in chicken and turkeys intranasally (IN) inoculated with the virus. However, we demonstrated that the virus can infect laying turkey hens by the intracloacal (IC) and intraoviduct (IO) routes, possibly explaining the drops in egg production observed in turkey breeder farms affected by the virus. Such novel routes of exposure have not been previously examined in chickens and could also explain outbreaks of low pathogenicity avian influenza (LPAI) that cause a decrease in egg production in chicken layers and breeders. In the present study, 46-wk-old specific-pathogen-free chicken layers were infected by the IN, IC, or IO routes with one of two LPAI viruses: a poultry origin virus, A/chicken/CA/1255/02 (H6N2), and a live bird market isolate, A/chicken/NJ/12220/97 (H9N2). Only hens IN inoculated with the H6N2 virus presented mild clinical signs consisting of depression and anorexia. However, a decrease in number of eggs laid was observed in all virus-inoculated groups when compared to control hens. Evidence of infection was found in all chickens inoculated with the H6N2 virus by any of the three routes and the virus transmitted to contact hens. On the other hand, only one or two hens from each of the groups inoculated with the H9N2 virus shed detectable levels of virus, or seroconverted and did not transmit the virus to contacts, regardless of the route of inoculation. In conclusion, LPAI viruses can also infect chickens through other routes besides the IN route, which is considered the natural route of exposure. However, as seen with the H9N2 virus, the infectivity of the virus did not increase when given by these alternate routes.     

A novel delivery of oxytetracycline in turkey breeder hens.

A novel product (SQ12) for subcutaneous (SQ) injectable delivery of oxytetracycline (OTC) has been developed for use in livestock. SQ12 employs microfluidic spheres encasing OTC crystals, which allows for longer release of the OTC compared with other injectable antibiotics. The objectives of the study were to determine serum and tissue levels of SQ12 in turkey breeder hens to 14 days postinjection and to evaluate effects of SQ12 on reproductive status. Thirty photostimulated hens were housed in litter floor pens and provided with 14.5 hr of light per day in a curtain-sided facility. Six hens served as untreated controls. Twelve hens per treatment group received SQ injections in the neck with SQ12 at 11.4 (L dose group) or 22.7 mg/kg (H dose group) to assess low and high doses, respectively. Serum samples were obtained from each hen at predose and 6, 12, 24, 48, 72, 96, 168, 240, and 336 hr postinjection. All hens were euthanatized at 14 and 15 days postinjection. One-half of the hens in each treatment group were sampled (liver, lung, kidneys, and breast muscle) for tissue residue levels of OTC. The control group had no detectable OTC in serum or tissues at any sample collection time. There were no detectable serum levels of OTC in either treatment group prior to injection. The average serum concentrations of the L and H dose groups showed similar depletion curves although the H dose group was 42% higher at maximum concentration than the L group. Average tissue concentration of OTC for all tissues sampled from the H dose group was twice that of the L dose group. All tissue levels were below the OTC residue tolerance limit. SQ12 provided an extended source of OTC in serum of turkey breeder hens with no effect on reproductive status. SQ12 may provide for a novel treatment of bacterial infection in turkey breeder hens with longer lasting serum levels compared with other single injectable OTC products.     

An enzyme-linked immunosorbent assay for detection of Bordetella avium infection in turkey flocks: sensitivity, specificity, and reproducibility.

An enzyme-linked immunosorbent assay (ELISA) for detection of Bordetella avium infection in turkey poults was developed. One-week-old poults challenged intratracheally with 10(12) colony-forming units of B. avium had detectable titers (greater than or equal to 11), with an average of 13.6% positive samples when the birds were 6 to 11 weeks old. The method was sensitive enough to detect maternal antibodies to B. avium in poults up to 3 weeks of age. The same poults challenged at 1 week of age had 100% tracheal infection up to 3 weeks of age, which dropped to 0% by 6 weeks. The method resulted in no false-positive samples (titer = 0) from birds not infected with B. avium and tested weekly between 4 and 11 weeks of age. Antibodies in turkey flocks infected with Newcastle disease virus, hemorrhagic enteritis virus, and Mycoplasma meleagridis, and birds infected with Escherichia coli had no apparent cross-reactivity to the B. avium antigens used in the ELISA. The percentages of B. avium-positive serum samples collected from different turkey flocks did not significantly differ (P greater than 0.05) when samples were tested by the developed ELISA at different times, an indication of the reproducibility of the method.     

Live Performance and Immune Responses of Straight-Run Broilers: Influences of Zinc Source in Broiler Breeder Hen and Progeny Diets and Ambient Temperature During the Broiler Production Period

Supplemental zinc in broiler diets has been reported to be more available to the chick when provided as a zinc-amino acid complex rather than an inorganic source. Previous work with broiler breeder and turkey hens has indicated that supplementing diets with zinc-amino acid complexes enhances immune status and livability of progeny. This study examined the effects of supplemental dietary zinc source [ZnSO₄ or Availa Zn 100 zinc-amino acid complex (ZnAA)] provided to broiler breeder hens and their progeny on live performance and immune status of broilers. Broiler breeder and progeny diets were supplemented with 160 and 140 ppm zinc, respectively. In addition to the dietary treatments, the broilers were subjected to either optimum or suboptimum temperatures during the first weeks of production. Supplementing broiler breeder hen and progeny diets with ZnAA and ZnAA + ZnSO₄, respectively, improved cumulative feed conversion (FC) of broilers. Providing diets supplemented with ZnSO₄ to hens or exposing broilers to optimum temperatures enhanced humoral immune response of progeny. Subjecting broilers to suboptimum brooding temperatures increased feed intake, but a higher incidence of mortality occurred primarily encompassing the first week of the growout. These data indicate that brooding temperature affected broiler performance, whereas the response to dietary zinc source was less pronounced.     

Vertical transmission of reticuloendotheliosis virus in breeder turkeys

Epizootiological studies were conducted on a commercial turkey breeder flock naturally infected with nondefective reticuloendotheliosis (RE) virus. Although RE virus was isolated from 27 (46%) of the 59 hens studied, only 4 of the 45 hens tested transmitted RE virus to progeny during a 6-week observation period and the overall transmission rate was 1.8%. The transmitter hens were of two types: three hens were consistently viremic and antigenemic and lacked antibody; one hen was viremic but lacked detectable viral antigen and possessed antibody. Toms appeared to play no role in vertical transmission of the infection. Of several tests evaluated for detection of transmitter hens, the direct enzyme-linked immunosorbent assay on albumen was probably best, since it detected three of four transmitter hens, detected relatively few nontransmitter hens, and had the best consistency of any test. No significant differences in hatchability were found between eggs from viremic and non-viremic hens. These findings can be utilized in the development of programs for eradication of RE virus from turkey breeding flocks.     

Effect of age and body weight on plasma concentrations of luteinizing hormone in turkey hens before and after photostimulation

The independent effects of age and body weight (BW) on photostimulatory response in turkey breeder hens were studied by measuring changes in plasma luteinizing hormone (LH; ng/mL) before and 3 d after photostimulation. The study was conducted with hens from two BW groups at 24–25, 27–28, and 31–32 wk of age. There was approximately a 1-kg difference in BW between groups within an age. Six hens per BW and age group were cannulated (jugular vein) and serially sampled during each of two 6-hr periods. Samples were collected at 10-min intervals. The two sampling periods were the last 6 hr of the short-day photoperiod (SD) and the same period during the third long day after photostimulation (LD). The photostimulatory response (PR) or difference between the SD and LD baseline LH concentrations was greatest in the 24–25-wk-old hens. The PR was unaffected by hen BW at any age. The baseline LH concentration during the SD photoperiod declined as hens aged. After photostimulation, baseline LH and LH peak amplitude concentrations were higher in 24–25-wk-old hens compared with the older ages. The number of LH peaks increased after photostimulation, but there were no significant effects attributable to age or BW within an age. In conclusion, the PR was affected by hen age but not hen BW or BW within a particular age.     

Effect of dietary manipulation and vaccination of turkey breeder hens on immunoglobulin levels of yolk,yolk sac and neonate poults

Two hundred turkey breeder hens and 24 viable toms of 30–35 weeks age of small white variety were distributed into two treatment groups having four replicates of 25 hens and three toms in each treatment. First four replicates were offered a turkey breeder diet (Diet A) (Nutrient requirements of poultry, 1994, National Academic Press, Washington, DC) and the rest four replicates were maintained on a higher plane of nutrition (Diet B) for 8‐week duration. After 6 weeks of experimental feeding, two replicates from each treatment groups were vaccinated with ND (R₂B) vaccine. Yolk sac of embryo from birds fed Diet B had a significantly higher (p < .05) IgG, IgM level and HI titre (log 2) than those fed Diet A. HI titre values of embryonic yolk sac from the vaccinated birds fed Diet B were significantly higher (p < .05) than that of the control groups. In addition, HI titre values were significantly higher (p < .05) in the day‐old poults of the birds fed Diet B than that of those fed Diet A. There was significantly (p < .01) positive correlation between serum IgG and IgM of the breeder birds and day‐old chicks. Similarly, there was significantly (p < .05) positive correlation between yolk IgG and IgM after 1‐month experimental feeding and yolk sac IgG and IgM. Positive correlation (p < .05) also existed between yolk sac IgM and day‐old chick serum IgM. Furthermore, the HI titres of breeder birds' serum at 14 days post‐vaccination were positively correlated with their egg yolk after 10 and 15 days post‐vaccination, yolk sac and day‐old chicks. Thus, the study envisaged that a higher immunity in neonate poults from turkey breeders maintained on a higher plane of nutrition may be elicited as there was maternal transfer of antibodies from the serum of breeder birds to their offsprings through their yolk sac.     

Influenza in commercial broiler breeders.

Influenza was detected in a flock of broiler breeders during routine serological monitoring. Although there were no clinical signs, egg production may have been affected in hens on one story of a two-story breeder house. Intensive measures were taken to avoid transmission to other farms. Two months after the flock was found to be serologically positive, sentinel hens were placed in the flock, and they became serologically positive 1 month later. In spite of this evidence for virus being present in the flock, no detectable transmission to any other farm occurred.     

Characteristics of oviposition patterns of turkey hens and the influence of different wavelengths of light

1. Characteristics of individual oviposition patterns of turkey breeder hens were determined for 160 d of photostimulation with fluorescent blue, green or red light and with incandescent light. 2. In all treatments most eggs were laid during the afternoon and evening. The maximum frequency occurred at 14.00 to 15.00 h (8 to 9 h after light onset); about 15% of all eggs were laid in the dark period. 3. The hourly oviposition distribution remained unchanged and was specific to individual hens throughout the study period. However, it was highly variable between hens, ranging from highly dispersed to tightly clustered. 4. The total number of clutches per hen did not differ between treatments and eggs were laid in relatively short clutches; 65% were of two eggs or fewer, 30% of three to 5 eggs and only 5% of 6 eggs or more. 5. The oviposition time of the first egg in a clutch was very variable; however, oviposition time became more clustered and uniform as the position of the egg in a clutch increased. 6. Mean time of lay, mean interval between eggs and total lag time were not influenced by light treatment. Mean interval was about 27 h for all eggs and was not influenced by clutch size. 7. Light quality at the intensity used in this study did not appear to play an important role in oviposition patterns of turkey hens.     

Organic matrix composition and ultrastructure of eggshell: a comparative study.

1. The avian eggshell is a biomineralised composite ceramic consisting of calcium carbonate embedded in an organic matrix. Matrix components are supposed to be involved in the control of mineralisation, crystallographic texture and biomechanical properties of eggshell. 2. The structure and eggshell matrix composition of various domesticated bird species were compared to gain insight into the universality of the eggshell mineralisation process. 3. The SDS-PAGE profiles of soluble eggshell matrix were specific within groups of birds (a: laying hen, breeder hen, quail, pheasant and possibly turkey; b: guinea fowl; c: duck and goose) but some of the protein bands were common to all groups. 4. Analogies between species were confirmed by Western blotting using hen protein antibodies. Ovocleidin-17 (OC-17) and ovalbumin were revealed in all species (except quail for OC-17). Lysozyme was present only in hen eggshell. Another egg white protein: ovotransferrin showed a positive signal in hens, turkey and quail. Osteopontin was observed in laying and breeder hens and quail. 5. Different proteoglycans were localised to discrete regions within the eggshell. Dermatan sulphate was observed within the matrix of the calcified shell of all species except quail which contained chondroitin-6-sulfate. Keratan sulphate was observed in mammillary bodies of breeder and laying hen, quail, pheasant and turkey while chondroitin sulphate was also present in guinea fowl and duck. 6. The general structural organisation of the different avian eggshells was similar but specific differences were observed in the ultrastructure of the mammillary layer. Species of the same taxonomic family could be grouped according to their structural analogies: breeder hen, turkey and pheasant resembled that of the domestic fowl. Guinea fowl was unique. Goose and duck were quite similar with large and confluent mammillary bodies. 7. Some matrix components are therefore common to eggshells of various species but more information is needed to relate differences in matrix composition between taxonomic groups with differences in ultrastructure.     

Degenerative myopathy in turkey breeder hens: a comparative study of normal and affected muscle

The pH of affected tissue within the green lesion characteristic of degenerative myopathy in the supracoracoideus muscle of turkey breeder hens was 7.4 to 7.8 and there was a high concentration of albumin. Histological examination showed that the striations of affected muscle fibres were more pronounced and curved than those of normal fibres. Examination of the affected fibres using the electron microscope showed that the curvature resulted from displacement of groups of myofibrils. The ultrastructure of these myofibrils differed markedly from that of normal muscle. The A‐band was granular and ill‐defined, while extensive degradation of the I‐band and Z‐line was observed. In addition, the sarcoplasmic reticulum was extensively disrupted and mitochondria had degenerated.     

Phenotypic and genotypic characterization of Salmonella arizonae from an integrated turkey operation

Fifty cases submitted between 2000 and 2002 were selected for retrospective analysis to evaluate possible relationships between Salmonella arizonae isolated from breeder flocks, hatching eggs, and meat bird flocks belonging to a single turkey integrator. In all the meat bird cases selected for this study, arizonosis was the primary diagnosis. In birds under 1 month of age, clinical signs and pathologic changes were observed in older birds. The Salmonella arizonae isolates were analyzed by antibiotic resistance pattern and serotype and genotyped by pulsed-field gel electrophoresis (PFGE). Serotyping and PFGE yielded similar results, but the antibiotic resistance patterns did not correspond to either serotyping or PFGE typing. The presence of common pulsed-field patterns in breeder flocks, eggs, and meat bird flocks suggested that S. arizonae was being transmitted vertically from the breeder flock.     

Effects of Bordetella avium toxin on turkey tracheal organ cultures as measured with a tetrazolium-reduction assay

Turkey tracheal organ cultures (TOCs) were exposed to one of the following Bordetella avium fractions or controls: live B. avium, formalin-killed B. avium, B. avium sonicate, heat-inactivated sonicate, culture supernatant, heat-inactivated culture supernatant, phosphate-buffered saline, or brain-heart infusion broth. After the TOCs were incubated for 2 hours with the bacterial fractions, the cellular metabolism of each TOC was evaluated using a tetrazolium chloride reduction assay, and cellular morphology was determined by light microscopy. Additionally, bacterial fractions and controls were injected into turkeys to test lethality. Although the bacterial sonicate was lethal for turkeys, neither the sonicate nor any other B. avium fraction significantly affected the metabolism or morphology of turkey TOCs.