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[Vaccination against infectious anemia of poultry (CAA)--results of field studies].

Infectious anemia of poultry is a disease of high economical significance. Connatal infection of chicks with the chicken anemia agent (CAA) via the embryonated egg causes anemia along with severe immunosuppression, thus rendering the chicken susceptible for secondary infections. In order to prevent infection of young chicks, it is necessary to induce immunity against CAA in parent flocks, with the aim to prevent connatal spread of the infection and provide maternal protection for baby chicks. In this publication, the efficacy and use of a live CAA vaccine is reported. From autumn 1986 until summer 1990, 3 experimental vaccine charges were applied in 85 broiler parent flocks with totally 3.1 million chickens. In addition, totally 293,000 broiler breeder and 171,000 layer breeder chicken were vaccinated in 1989/90. The vaccine was administered between the 13th and 19th week of life by drinking water without adverse effect to the birds. Chicken anemia symptoms were observed only at the begin of laying period in two parent flocks. These flocks had been vaccinated in the 17th and 19th week, respectively. The offsprings of all other vaccinated parent flocks remained free of chicken anemia. Day-old chicks derived from vaccinated parent flocks were protected against CAA challenge infection. It is emphasized, that vaccination should be performed within the 13th to 15th week of life, because according to our observations, this will lead to an immediate seroconversion.

Anemia↗

Experimental infection of chickens with Campylobacter jejuni: strains differ in their capacity to colonize the intestine.

Groups of broiler and layer type chickens (25 to 63 d.o.) were inoculated per os with separate isolates of 10 strains of Campylobacter jejuni. Nine of the 10 strains were originally isolated from chickens and one from a dog. The dog strain and five of the chicken isolates could be isolated after inoculation, but four strains were not recovered from cloacal swabs for up to 4 to 16 days after inoculation. However, it was possible to isolate C. jejuni from these birds, from cloacal swabs, when they were inoculated with organisms which had been previously shown to colonize other birds.

Journal Article↗

Localized binding of [3H]muscimol to synapses in chicken retina.

Binding sites for [3H]muscimol, an analogue of gamma-aminobutyric acid (GABA) were localized in the synaptic layers of chicken retina by light microscopic and electron microscopic autoradiography. Light microscopic autoradiography of cryostat sections incubated in [3H]muscimol or [3H]GABA revealed identical binding patterns: a band over the inner plexiform layer (IPL) and a band over the outer plexiform layer (OPL). This binding pattern differed from the uptake pattern for [3H]GABA: labeling over horizontal, amacrine, and ganglion cell bodies as well as very intense labeling over lamina 5 in the proximal IPL. Statistical analysis of electron microscopic autoradiography data from the IPL indicated that only amacrine synapses bind [3H]muscimol (i.e., make GABAergic synapses). Processes of amacrine, bipolar, or ganglion cells can be postsynaptic to these amacrine synapses. The highest concentration of synapses binding [3H]muscimol occurred in laminae 2 and 4 of the IPL and not in lamina 5 as might be expected from the density of [3H]GABA uptake. In the OPL, [3H]muscimol binding occurred over specialized junctions proximal to photoreceptor terminals. In cone receptor terminals, [3H]muscimol binding was suspected near horizontal cell dendrite/receptor terminal membranes lateral to the synaptic ribbon, supporting the hypothesis that horizontal cells are involved in a GABAergic feedback loop with cone terminals. We conclude that the synaptic binding pattern provides a more accurate concept of GABAergic synaptic interaction than does the uptake pattern for [3H]GABA because the two patterns in the IPL are not related.

Animals↗

Serological monitoring on layer farms with specific pathogen-free chickens.

To monitor the existence of avian pathogens in laying chicken flocks, specific pathogen-free (SPF) chickens were introduced into two layer farms and reared with laying hens for 12 months. SPF chickens were bled several times after their introduction and examined for their sero-conversion to avian pathogens. As a result, antibodies to eight or ten kinds of pathogens were detected in SPF chickens on each farm. Antibodies to infectious bronchitis virus (IBV), avian nephritis virus, Mycoplasma gallisepticum and M. synoviae were detected early within the first month. Antibody titer to IBV suggested that the laying chickens were infected with IBV repeatedly during the experiment on both farms. However, antibodies to infectious bursal disease virus and 6 pathogens were not detected.

Animals↗

Differential changes in inhibin A, activin A, and total alpha-subunit levels in granulosa and thecal layers of developing preovulatory follicles in the chicken.

Accumulating evidence implicates inhibins and activins as endocrine and local regulators of follicular development in mammals, and it was recently confirmed that inhibin/activin alpha and betaA genes are also expressed in the avian ovary. To investigate the potential involvement of these proteins in the chicken ovary, thecal and granulosa layers of the four largest follicles (F1-F4) and the most recent postovulatory follicle were collected from hens (10/group) killed 4, 12, and 20 h before the expected time of F1 ovulation. Inhibin A and activin A concentrations of tissue extracts (expressed per mg DNA) were measured using validated two-site enzyme-linked immunosorbent assays; total immunoreactive inhibin alpha-subunit (ir-alpha) was also measured by heterologous RIA (Monash assay). Inhibin A and ir-alpha were largely confined to the granulosa layer, whereas activin A was much more abundant in the thecal layer. Granulosa inhibin A contents were similar in F4 and F3, but increased approximately 40-fold from F3-F1 (P < 0.0001). As such, the F1 granulosa layer was by far the richest source of inhibin A in the chicken ovary, but contained very little activin A. Total ir-alpha in granulosa was much more abundant than inhibin A and increased only 3-fold from F4-F1 (P < 0.001). Activin A in both granulosa and theca showed little variation between F1 and F4 follicles (by ANOVA, P > 0.05). The inhibin A content of F1 granulosa was maximal 12 h before ovulation and had fallen approximately 6-fold (P < 0.0001) within 8 h, suggesting an inhibitory effect of the preovulatory LH surge on the F1 capacity to synthesize inhibin A. Inhibin A, activin A, and ir-alpha were all less in the postovulatory follicle compared with F1 before ovulation (P < 0.0001). In conclusion, application of the present two-site enzyme-linked immunosorbent assays to the chicken ovary revealed 1) divergent tissue distribution of inhibin A and activin A within preovulatory follicles, and 2) differential regulation of granulosa cell production of inhibin A and activin A dimers during preovulatory follicular development. These findings of dynamic changes in inhibin A, activin A, and total ir-alpha support the hypothesis that these proteins subserve regulatory roles during preovulatory follicular development in the hen.

Activins↗

Assessment by ELISA of passively acquired protection against infectious bursal disease virus in chickens.

The level of maternal antibody to infectious bursal disease (IBD) virus in the circulation of one-day-old layer strain chickens was found to be on average, 45% of the antibody titre in their respective dam, while the minimum ELISA titre which protected against a challenge of 1000 CID50 of virus was 400. Maternal antibody was found to disappear from the circulation of these crossbred chickens with a half-life of 6.7 days. From these data it is possible to estimate the ELISA titre necessary in vaccinated hens to provide the desired duration of passive protection; protection being assessed by an ELISA which measures IBD viral antigen in the bursa of Fabricius following challenge.

Animals↗

Differentiation of smooth muscle cells from undifferentiated cells of chicken gizzard occurs on the layer of fibroblast-like cells.

We examined the in vitro differentiation of smooth muscle cells from undifferentiated cells of embryonic chicken gizzard. We used the gizzard from 7-day-old embryos (Hamburger and Hamilton's stage 26-28) as a source of culture, since the expression of myosin is extremely low, and the gizzard consists of round cells, which are not stained by anti-smooth muscle myosin antiserum. When the dissections of the gizzard were cultured, they attached to the dish, and fibroblast-like cells migrated within 4 days after the culture. Then round cells migrated from the transplants over the layer of fibroblast-like cells. At 12-14 days after the culture, smooth muscle cells, which were ribbon-shaped and stained by anti-smooth muscle myosin antiserum, appeared in the layer of round cells. The dissected transplant itself was not stained by anti-smooth muscle myosin antiserum even after being cultured for 15 days. We concluded then that the smooth muscle cells were differentiated from the round cells, which spread on the layer of the fibroblast-like cells. We also observed the differentiation of smooth muscle cells when the cells separated from 7-day-old embryo cultured on the layer of cloned fibroblast cells. We suggest that the fibroblast-like cell may play an important role in the differentiation of smooth muscle cells.

Animals↗

Patterns of ribosomal gene variation in elite commercial chicken pure line populations.

The nucleolus organizer region (NOR) encodes the tandemly repeated 18S, 5.8S and 28S ribosomal (r) RNA genes. The NORs of broiler and layer commercial chicken pure lines were studied to establish the type and extent of genetic variation at this important locus. The parameters studied were gene copy number, repeat size, and diversity of NOR-types. The populations were organized into three groups for analysis including brown-egg broiler (13 lines), brown-egg layer (six lines), and white-egg layer (eight lines). The ribosomal gene copy number average of the white-egg layer populations was significantly lower (329 genes) than that of the brown-egg layers (372 genes); the brown-egg broiler ribosomal gene average was intermediate (350 genes). The white-egg layer populations exhibited a ribosomal repeat unit average size of 36 kb, significantly different from the brown-egg layer and brown-egg broiler average repeat unit size of 32.5 and 33.9 kb, respectively. NOR array size was similar among the three groups (6 mb). The brown-egg broiler populations exhibited polymorphic NOR patterns, intra- and interline, whereas the white-egg layer populations were essentially monomorphic for NOR-type; brown-egg layers exhibited an intermediate level of NOR diversity. Some NOR array characteristics may be a function of breed origin as brown-egg commercial populations, both broilers and layers, have similar breed origins and exhibited similarities for predominant repeat unit size as compared with white-egg layer populations. However, the finding that brown-egg broiler lines typically exhibit a greater number of segregating NOR-types than brown-egg layer lines suggests that the selection schemes of broiler vs. layer pure line populations may also have influenced the degree of variation at this gene complex.

Animals↗

A cross-sectional survey of Australian chicken farms to identify risk factors associated with seropositivity to Newcastle-disease virus.

Several outbreaks of virulent Newcastle-disease occurred in Australia in 1998-2000. We conducted a cross-sectional survey of 753 Australian chicken farms to identify risk factors associated with the seroprevalence of chicken flocks with Newcastle-disease virus (NDV). We had a 99.7% response rate to the survey and the overall prevalence of NDV seropositive farms was 39.8%. Associations were analysed for the layer, chicken-meat and breeder production sectors in sector-specific logistic-regression models using 187, 198 and 146 farms, respectively. In the layer sector, increased risk of seroprevalence was associated with increasing age of the chickens, and decreased risk when the nearest-neighbour poultry farm was >10 km distant (odds ratio (OR)=0.30). In the chicken-meat sector, increased risk of seroprevalence was associated with location in the Sydney basin (OR=13.67), eastern Victoria (OR=26.10) or western Victoria (OR=5.43), and decreased risk when the nearest-neighbour poultry farm was greater than 0.5 km distant (OR=0.34). In the breeder sector, increased risk of seroprevalence was associated with increasing age of the chickens, the presence of wild birds on the farm (OR=5.28) and location in eastern Victoria (OR=16.19). A conditional logistic-regression for 112 pairs of farms matched for age, survey region and production sector identified a distance of >1.0 km to the nearest-neighbour poultry farm (OR=0.24) and ownership by owner 2 (OR=0.02), owner 5 (OR=0.11) or owner 9 (OR=0.25) as significant in reducing the risk of NDV seroprevalence. Our survey found that high levels of biosecurity and hygiene practices had been adopted by most farms.

Age Factors↗

The proteome of chicken skeletal muscle: changes in soluble protein expression during growth in a layer strain.

The whole animal, and the pectoralis muscle in particular, grows at a greatly enhanced rate in chickens selected for meat production (broilers) when compared to those selected for egg production (layers). As part of an ongoing study to analyse muscle protein dynamics under conditions of rapid growth, we have embarked upon a preliminary characterisation of the proteome of layer chicken pectoralis muscle, at specified time-points from 1 to 27 days after hatching. Soluble extracts of muscle homogenates were separated by two-dimensional (2-D) gel electrophoresis and selected spots were analysed by in-gel tryptic digestion and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Of 90 spots, 51 gave mass spectra that matched to existing chicken proteins present in on-line databases, 12 matched equivalent proteins from non-avian species and 11 yielded good quality spectra but were unable to be matched against existing databases. For many of these proteins, growth over 27 days elicited dramatic changes in relative expression levels. Chicken skeletal muscle offers an excellent system for developmental proteomics.

Animals↗

Biological activities of recombinant chicken leptin C4S analog compared with unmodified leptins.

The chicken leptin sequence, in contrast to mammalian leptins, contains an unpaired Cys at position 3 of the original cDNA (AF012727). The presence of an extra Cys may confer a different structure and affect the leptin's biological activity. To address this, we studied the effects of wild-type and mutated (C4S) chicken leptins in vitro and in vivo and compared them with mammalian leptin prepared from ovine leptin cDNA. The prokaryotic expression vector pMON, encoding full-size A(-1) chicken leptin (AF012727), was mutated using a mutagenesis kit, yielding the C4S analog. Escherichia coli cells transformed with this vector overexpressed large amounts of chicken leptin C4S upon induction with nalidixic acid. The expressed protein, found in the inclusion bodies, was refolded and purified to homogeneity on a Q-Sepharose column, yielding three electrophoretically pure fractions, eluted from the column by 100, 125, and 150 mM NaCl, respectively. All three fractions showed a single band of the expected molecular mass (16 kDa) and were composed of >95% monomeric protein. Proper refolding was evidenced by comparing the circular dichroism spectrum of the analog with spectra of nonmutated chicken and ovine leptins. The biological activity of the C4S analog was evidenced by its ability to stimulate proliferation of leptin-sensitive BAF/3 cells transfected with a long form of human leptin receptor construct similar to its nonmutated counterpart, indicating that Cys4 plays no role in leptin activity. The in vitro activity of both wild-type and mutated chicken leptins was approximately 10-fold lower than that of ovine leptin. After intravenous or intraperitoneal injections, C4S analog and the nonmutated chicken and ovine leptins all lowered the food intake of starved 9-day-old broiler or 5-wk-old layer male chickens by 11-34%. Monitoring food behavior revealed that the attenuated food intake resulted not from a decreased number of approaches to the feeders but from a decrease in the average time spent eating during each approach.

Aging↗

Molecular changes of the fusion protein gene of chicken embryo fibroblast-adapted velogenic Newcastle disease virus: effect on its pathogenicity.

Molecular changes of cell culture-adapted Newcastle disease virus (NDV) were studied by adapting a velogenic NDV isolated from commercial layer chicken-to-chicken embryo fibroblast (CEF) cells. The isolate was passaged 50 times in CEF cells. At every 10th passage the virus was characterized conventionally by mean death time analysis, intracerebral pathogenicity index, and virus titration. As the passage level increased, a gradual reduction in the virulence of the virus was observed. Molecular characterization of the virus included cloning and sequencing of a portion of the fusion gene (1349 bp) encompassing the fusion protein cleavage site (FPCS), which was previously amplified by reverse transcription-polymerase chain reaction. Sequence analysis revealed a total of 134 nucleotide substitutions, which resulted in the change of 41 amino acids between the parent and the 50th passage virus. Pathogenicity studies conducted in 20-wk-old seronegative chickens revealed gross and histopathologic changes in the chickens injected with the parent virus and absence of the lesions in chickens injected with the adapted virus. The 50th passage cell culture virus was back-passaged five times in susceptible chickens and was subjected to virulence attribute analysis and sequence analysis of the FPCS region, with minor differences between them.

Adaptation, Biological↗

Thermoregulatory capacity and muscle membrane integrity are compromised in broilers compared with layers at the same age or body weight.

1. The effects of acute heat stress (2 h at 32 degrees C and 75% RH) on body temperature and indices of respiratory thermoregulation and skeletal muscle function were examined in two divergently selected male grandparent lines of broiler and layer-type chickens at two ages (35 and 63 d), or at a similar body weight (approximately 2.2 kg). 2. The two chicken lines exhibited markedly different baseline blood acid-base and skeletal muscle characteristics. At the same age or live weight, birds from the broiler line had significantly higher venous blood carbon dioxide tensions associated with lower blood pH. Plasma creatine kinase (CK) activities reflecting muscle membrane damage were also greatly elevated in the broiler line. 3. Exposure to acute heat stress caused an increase in deep body temperature, panting-induced acid-base disturbances and elevated plasma CK activity in both lines of chicken, an effect that increased with age. The extent of disturbances in acid-base regulation and heat-stress-induced myopathy were more pronounced in the broiler than the layer line at the same age or similar live weights. 4. It is suggested that genetic selection for high muscle growth in broiler lines has compromised their capacity to respond to an acute thermal challenge, leading to detrimental consequences for muscle function. This reduction in heat tolerance may have important implications for bird welfare and subsequent meat quality.

Aging↗

Chicken granulosa cells show differential expression of epidermal growth factor (EGF) and luteinizing hormone (LH) receptor messenger RNA and differential responsiveness to EGF and LH dependent upon location of granulosa cells to the germinal disc.

Granulosa cells in the chicken follicle exhibit different phenotypes according to their location relative to the germinal disc (GD). Granulosa cells proximal to the GD (referred to as proximal granulosa cells) are more proliferative, whereas granulosa cells distal to the GD (referred to as distal granulosa cells) are more differentiated. We have shown that epidermal growth factor (EGF) derived from the GD stimulated proliferation of granulosa cells proximal to the GD, whereas extraovarian LH promoted differentiation. We tested the hypothesis that phenotypic differences of granulosa cells are the result of differential responsiveness of granulosa cells to EGF and LH. We found that both granulosa and theca layers of chicken preovulatory follicles expressed mRNA for EGF receptor (EGFr) by polymerase chain reaction (PCR) analysis. However, only the granulosa layer showed differential expression of EGFr and LH receptor (LHr) mRNA. Competitive reverse transcription-PCR revealed that proximal granulosa cells expressed more EGFr mRNA but less LHr mRNA than distal granulosa cells. In addition, proximal granulosa cells proliferated more in response to EGF than their distal counterparts. We further demonstrated that EGF decreased LHr mRNA expression by granulosa cells in a dose-dependent manner, whereas EGF and LH had no effect on EGFr mRNA expression except at one dose of LH (15 ng/ml) that stimulated EGFr mRNA expression. Our findings suggest that EGF derived from the GD influences the phenotypes of granulosa cells. Granulosa cells proximal to the GD exhibit a proliferative phenotype possibly because they are exposed to and are more responsive to GD-derived EGF. Furthermore, GD-derived EGF decreases LHr mRNA expression by proximal granulosa cells and therefore results in less differentiated granulosa cell phenotype. In contrast, granulosa cells distal to the GD are not under the influence of EGF and exhibit a more differentiated phenotype.

Animals↗

A comparison of immune responses to infection with virulent infectious bursal disease virus (IBDV) between specific-pathogen-free chickens infected at 12 and 28 days of age.

Previous studies indicated that the pathogenesis of infectious bursal disease virus (IBDV) may vary depending on the age of the infected chicken. The reason for this difference is not clear, immunopathogenesis studies comparing different age groups are lacking. In the present study the IBDV-immunopathogenesis was compared in 12-day-old and 28-day-old specific-pathogen-free layer-type chickens. Birds were inoculated with the classic virulent strain IBDV-IM. Interestingly, the up-regulation of interferon (IFN)-gamma and interleukin-1beta expression varied between the two IBDV-infected age groups. The cytokine expression pattern was different between IBDV-infected age groups in magnitude and timing not only in bursal tissue and caecal tonsils but also in the spleen. No variations between the different age groups were observed in IBDV-induced bursa lesions, accumulation of intrabursal T cells and macrophages, and systemic stimulation of the release of nitric oxide inducing factors. Furthermore, virus-neutralizing antibody levels were comparable between age groups. Under these experimental conditions, age-related differences were found only for the cytokine patterns but interestingly this did not influence the outcome of the disease with IBDV-IM.

Age Factors↗

Effects of cholecystokinin on chicken cecal motility: mechanisms involved.

The aims of this work are to characterize the effects of cholecystokinin (CCK) on chicken ceca and to study in vitro the mechanisms through which such actions are mediated. Longitudinal and circular cecal strips kept in vitro in organ baths were responsive to CCK sulphated octapeptide (CCK-8s). On longitudinal strips the response consisted of a fast phasic contraction followed by a sustained increase in tone which was dose dependent and decreased markedly in the presence of tetrodotoxin (TTX). Ketanserin (10(-5) M) also caused a decrease in the CCK-8s response. CCK tetrapeptide (CCK-4) and CCK unsulphated octapeptide (CCK-8ns) induced slightly less contractile effects at concentrations of 2 x 10(-6) M only. L365,260 and L364,718 decreased the response of longitudinal strips to CCK-8s with similar efficacy. On circular strips CCK-8s caused rhythmic phasic contractions of dose dependent amplitude and frequency, and both effects were resistant to TTX. The EC50 for the amplitude was about 4 times higher than that for the frequency. CCK-8ns (2x 10(-6) M) also caused phasic contractions, whereas the same concentrations of CCK-4 did not elicit any motor effects. L365,260 and L364,718 showed different efficacy in decreasing amplitude or frequency of contraction. These results suggest that 1) Both muscularly and neurally located CCK receptors are present on the longitudinal layer of chicken ceca whereas only muscular receptors are present on the circular muscle. 2) 5HT2 receptors seem to be involved in the neurally mediated CCK-8s response observed in the longitudinal layer. 3) The different potency of CCK-8s, CCK-8ns and CCK4 to induce contractile effects and of the CCK-A and CCK-B antagonists to block such effects suggests the existence of two different CCK receptors on the circular layer.

Anesthesia↗

Clinical Mycoplasma gallisepticum infection in multiplier breeder and meat turkeys caused by F strain: identification by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, restriction endonuclease analysis, and the polymerase chain reaction.

In February 1991, a flock of North Carolina multiplier breeder turkeys experienced respiratory signs, sinusitis, airsacculitis, and increased mortality. Mycoplasma gallisepticum (MG) was isolated, and appropriate control measures were initiated. Ultimately, this outbreak involved several breeder flocks of an integrated turkey production company before the last infected flock was identified in May 1991. During this time, MG was also isolated from a flock of commercial layer-type chickens raised as pullets in close proximity to the index turkey flock. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and restriction endonuclease analysis indicated that these isolates were identical to each other and to examples of the vaccinal F strain. Additionally, MG isolates from the affected turkey breeder and layer flocks were identified as MG F strain by use of an F strain-specific DNA probe and polymerase chain reaction. A separate outbreak of MG disease in several meat-turkey flocks of a Midwest producer/processor yielded isolates identified as F strain by the polymerase chain reaction. These studies demonstrated: 1) the utility of newer technologies for disease outbreak investigations; and 2) the potential of MG F strain to cause disease in breeder and meat turkeys under field conditions.

Animals↗