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Fate mapping the neural plate and the intraembryonic mesoblast in the upper layer of the chicken blastoderm with xenografting and time-lapse videography.

The disposition of the Anlage fields of the neural plate and the intraembryonic mesoblast in the upper layer of the chicken blastoderm was studied at the primitive streak stage prior to the regression of Hensen's node (stages 5V to 6V, L. Vakaet (1970) Arch. Biol. 81, 387-426). Chicken blastoderms were cultured by New's technique on a mixture of thin egg white and agar. The anterior half of the deep layer was reflected with a tungsten needle. A circular fragment of the upper layer was punched out with a pulled out Pasteur pipette and discarded. It was replaced with an isotopic and isopolar piece of quail upper layer that was punched out with the same pipette. The deep layer was replaced and the chimeras were reincubated for 24 hours. The xenografts were followed with time-lapse videography. After fixation, the quail cells were located using Le Douarin's quail nucleolar marker technique. Integrating the observations with time-lapse videography and the results of Feulgen stained sections, we have drawn a new fate map of the disposition of the Anlage fields in the upper layer of the chicken blastoderm at stages prior to the regression of Hensen's node (stages 5V to 6V). The disposition of the neural plate and of the notochord, somites, nephrotome and lateral plates was therefore determined before the Anlage fields are morphologically discernible. The pathway of the fields in the upper layer towards their disposition was documented with time-lapse videography in chimeric chicken blastoderms that developed normally.

Animals↗

Role of cochlear integrity in cochlear nucleus glucose metabolism and neuron number after cochlea removal in aging broiler chickens.

In the chicken auditory system, cochlear nucleus (nucleus magnocellularis, NM) neurons receive their only excitatory input from the ipsilateral cochlea. Cochlea removal (CR) results in an immediate decrease in NM neuron electrical activity, followed by death of approximately 30% of NM neurons. Previous work showed a decrease in NM activity and subsequent loss of NM neurons in all chicks. Egg layer adults showed NM neuron loss after CR, while neuron number remained stable in broiler adults. This suggested that effects of CR on NM were age- and breed-dependent. We now know that most aging egg layer chickens maintain largely normal cochleae throughout adulthood. Some exhibit cochlear damage with age. The converse is true of broiler chickens. Most aging broiler chickens display cochlear degeneration, with some maintaining normal cochlear anatomy throughout adulthood. The presence of extensive cochlear damage may alter the effect of CR on NM, leading to the described differences. Here, we examine the effect of unilateral CR on NM glucose metabolism and neuron number in 2, 30, 39, and 52 week-old broiler chickens found to have normal cochleae. Chickens with damaged cochleae were excluded. Using 2-deoxyglucose uptake to evaluate bilateral NM glucose metabolism, we found significantly decreased uptake ipsilateral to CR at each age examined. Bilateral cell counts revealed significant neuron loss ipsilateral to CR at each age examined. This suggests that NM glucose metabolism decreases and subsequent neuron death occurs in aging broiler chickens when a normal cochlea is removed. The status of the cochlea must play a role in the effect of deafferentation on NM glucose metabolism and neuron survival. The effect of CR appears to be dependent upon neither age nor breed, but upon cochlear integrity instead.

Aging↗

Skeletal muscle protein turnover in broiler and layer chicks.

Dietary infusion of L-[U-14C]tyrosine was used to estimate the fractional protein synthesis rates (FSR) in broiler and layer chickens. Six 2-wk-old birds of each strain were placed in individual metabolism cages and given a purified diet in agar-gel containing 2 microCi L-[U-14C]tyrosine for 6 h. The birds were sacrificed and the pectoralis major (PM) and two combined leg (LM) muscles (gastrocnemius and peroneous longus) were removed for analysis. Subgroups of chickens were sacrificed 3 d before and 3 d after infusion to observe changes in muscle composition to calculate fractional protein accretion rates (FAR). Fractional protein breakdown rates (FBR) were calculated by difference (FBR = FSR-FAR). Protein, ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) concentrations were determined to observe relationships between these cellular constituents and FSR. Fractional whole body growth rate and FSR in PM was greater (P less than .05) in broiler than layer birds. The FSR in LM of the layer was not different (P greater than .05) from that of broilers, and from the FSR of the PM in each bird-type. The calculated FBR in the layer PM was at least 17% higher than that of the other muscles. Ratios of FSR to FBR indicated that 16% of the protein synthesized in the layer PM was retained, compared with 45% in the broiler PM. The RNA activity of the layer PM was less (P less than .05) than that of the other muscles investigated. Deoxyribonucleic acid activity was lower (P less than .05) in the PM than LM of either bird-type.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Status of Salmonella gallinarum-pullorum infections in poultry in Zambia.

Ten outbreaks of Salmonella gallinarum-pullorum infections on poultry farms in Zambia were investigated. Three cases were seen in day-old broiler chickens and were diagnosed by culture as S. gallinarum-pullorum and characterized as pullorum disease because the mortality was only in the first few weeks. Another case was diagnosed by culture from broiler parent stock. Day-old chicks from two of the three cases were supplied by a hatchery. Five cases in 5-to-18-month-old layer chickens were diagnosed by culture as S. gallinarum-pullorum and characterized as fowl typhoid because of the clinical disease appearing after 5 months of age and the typical lesions of fowl typhoid. The last case was in 5-month-old village-bred fowls and was diagnosed by culture and clinical manifestation as fowl typhoid. Outbreaks of S. gallinarum-pullorum are still manifest in Zambia. Clinically, both pullorum disease and fowl typhoid were observed, and it was indicated that hatchery infection plays an important role in the transmission of S. gallinarum-pullorum.

Animals↗

A rapid assay for detecting antibody against leucocytozoonosis in chickens with a latex agglutination test using recombinant R7 antigen.

A method for detecting antibody against leucocytozoonosis in chickens by latex agglutination (LA) was developed using latex beads coated with recombinant R7 (rR7), an outer membrane antigen of second-generation schizonts of Leucocytozoon caulleryi. Compared with the agar gel precipitation test, which is widely used in Japan, LA could detect antibody induced by L. caulleryi infection with greater sensitivity. No agglutination was detected with sera from specific pathogen free or layer chickens that had not been vaccinated with oil-adjuvanted rR7 antigen, or infected with L. caulleryi, nor with sera from chickens inoculated with other pathogens, establishing the specificity of the assay. The LA test was also able to be used to quantify serum antibody induced by vaccination, which is not possible with the agar gel precipitation test. This study has shown that LA using the rR7 antigen is a simple, quick, and useful method for detecting antibody against L. caulleryi.

Animals↗

A chicken leptin-specific radioimmunoassay.

Recombinant chicken leptin was used to produce an antiserum in order to develop a specific and sensitive radioimmunoassay (RIA) for chicken leptin in plasma and serum. We have used either murine or chicken leptin as tracer and competition curves were performed using recombinant chicken leptin. Variations in leptin plasma levels in different chicken strains and various nutritional states were correlated with the physiological status. Leptin plasma concentrations were regulated by the nutritional state with higher levels in the fed state as compared to the fasted state (3.36 +/- 0. 13 versus 2.78 +/- 0.11 ng/ml) and being dependent upon the age. Higher leptin levels were found in 22 week-old as compared to 15 week-old layer chickens (2.709 +/- 0.172 versus 1.478 +/- 0.102 ng/ml). We have also shown that the multispecies leptin RIA kit (LINCO Inc.) underestimated leptinemia compared to the chicken leptin- specific RIA reported here. In conclusion the RIA developed in the present study is specific to the chicken and thus may be considered as powerful tool for investigating the physiological significance of leptin in chickens.

Animals↗

Quantitative trait locus detection in commercial broiler lines using candidate regions.

A QTL that explained a large proportion of the phenotypic difference between broiler and layer chickens in an experimental cross was evaluated in a commercial broiler line. A three-generation design, consisting of 15 grandsires, 608 half-sib hens, and more than 50,000 third-generation offspring, was implemented within the existing breeding scheme of a broiler breeding company. Four markers from a candidate region on chicken chromosome 4 were selected for their informativeness in the grandsires and used to genotype the first two generations. Using half-sib analyses, linkage was studied between these markers and 13 growth and carcass traits. The QTL analyses confirmed the presence of significant QTL for body weight (P < 0.01) and residual feed intake (P < 0.05) on chicken chromosome 4. Furthermore, evidence was found for QTL affecting the relative weight of bone and muscle in the thigh. Four more markers were added to increase resolution of the QTL positions. This increased the significance of the QTL for body weight (P < 0.001) and residual feed intake (P < 0.01) and showed evidence (P < 0.05) for additional QTL affecting carcass weight and conformation score. This study showed for the first time that a QTL that explains differences between broilers and layers was segregating in lines that have been selected for body weight over 50 generations. A possible explanation could be a pleiotropic or closely linked effect on fitness-related traits that are not part of the present study. The results demonstrate the feasibility of QTL detection and the potential for marker-assisted selection within a commercial broiler line without altering the existing breeding scheme.

Animals↗

An attenuated Salmonella Enteritidis strain derivative of the main genotype circulating in Uruguay is an effective vaccine for chickens.

We have recently reported that Salmonella enterica serovar Enteritidis (S. Enteritidis) strains circulating in Uruguay, are unevenly distributed among different genetic subtypes, with a predominant genotype that is a common contaminant of poultry-derived food and that accounts for the vast majority of human cases of food-borne disease. Herein, we describe the construction of a genetically-defined aroC derivative (LVR02) of a local strain of S. Enteritidis belonging to the major genetic type. We demonstrated the attenuation and the immunogenicity of that strain in a mouse model, and evaluated it as a vaccine for commercial layer chickens. LVR02 proved to be stable, attenuated, innocuous, immunogenic and to induce protective immunity against a S. Enteritidis challenge when used for oral vaccination. A single oral dose of LVR02 administered to newly hatched chickens induced protection against oral challenge with the parental virulent strain, preventing systemic and persistent intestinal infection and significantly reducing the shedding of the challenge strain in birds' feces. A second vaccine dose at 15 days post-hatching boosted the immunogenicity of the vaccine, and strengthened the protection achieved with a single dose. This strain may represent the basis of a live vaccine to be included in national control programs to reduce circulation of this pathogen in the country.

Administration, Oral↗

Infectious bursal disease virus variant from commercial Leghorn pullets.

An infectious bursal disease virus (IBDV) was isolated from 39-to-43-day-old commercial leghorn pullets suspected of having infectious bursal disease (IBD). These chickens had been vaccinated with a commercial live IBDV vaccine at 28 and 35 days of age. An isolate designated IN was recovered using specific-pathogen-free (SPF) chickens and the BGM-70 established cell line. Experimental studies using SPF chickens vaccinated with either inactivated vaccines made from the vaccine strain used in the problem flock or a standard-type vaccine indicated no protection against the IN isolate. However, two variants and another standard-type vaccine induced protection against the IN isolate. Cross-neutralization tests indicated that the IN isolate differed antigenically from commercial vaccine strains and was related to the variant IBDV strains recently isolated from broilers. To our knowledge, this is the first report of a variant IBDV recovered from commercial layer chickens in the United States.

Animals↗

Effect of Campylobacter-specific maternal antibodies on Campylobacter jejuni colonization in young chickens.

Using laboratory challenge experiments, we examined whether Campylobacter-specific maternal antibody (MAB) plays a protective role in young chickens, which are usually free of Campylobacter under natural production conditions. Kinetics of C. jejuni colonization were compared by infecting 3-day-old broiler chicks, which were naturally positive for Campylobacter-specific MAB, and 21-day-old broilers, which were negative for Campylobacter-specific MAB. The onset of colonization occurred much sooner in birds challenged at the age of 21 days than it did in the birds inoculated at 3 days of age, which suggested a possible involvement of specific MAB in the delay of colonization. To further examine this possibility, specific-pathogen-free layer chickens were raised under laboratory conditions with or without Campylobacter infection, and their 3-day-old progenies with (MAB(+)) or without (MAB(-)) Campylobacter-specific MAB were orally challenged with C. jejuni. Significant decreases in the percentage of colonized chickens were observed in the MAB(+) group during the first week compared with the MAB(-) group. These results indicate that Campylobacter-specific MAB plays a partial role in protecting young chickens against colonization by C. jejuni. Presence of MAB in young chickens did not seem to affect the development of systemic immune response following infection with C. jejuni. However, active immune responses to Campylobacter occurred earlier and more strongly in birds infected at 21 days of age than those infected at 3 days of age. Clearance of Campylobacter infection was also observed in chickens infected at 21 days of age. Taken together, these findings (i) indicate that anti-Campylobacter MAB contributes to the lack of Campylobacter infection in young broiler chickens in natural environments and (ii) provide further evidence supporting the feasibility of development of immunization-based approaches for control of Campylobacter infection in poultry.

Animals↗

Growth and food intake responses to diets of different protein contents and a choice between diets containing two concentrations of protein in broiler and layer strains of chicken.

1. Food intake, protein intake and body weight gain were measured in male broiler and layer strains of chickens offered approximately isocaloric diets containing various concentrations of protein from 4 to 9 weeks of age. The carcases were analysed for protein, fat and ash. 2. In the first experiment 5 birds of each strain were given diets containing either 65, 115, 172, 225 or 280 g protein/kg fresh matter. The sixth group was given a choice between 65 and 280 g/kg. There was an approximately linear increase in protein deposition with dietary protein content up to 280 g/kg with broilers and 225 g/kg with layers. When a choice of diets was offered, birds of both strains grew at a rate not significantly different from that of birds on the diets with the lowest protein content which gave maximal growth given singly, by making an apparently appropriate choice from the two diets. 3. In the second experiment broilers were offered a choice of two diets in the following combinations: 65 and 115, 65 and 225, 115 and 225, 225 and 280 and 280, and 320 g/kg protein. They were able to differentiate successfully between two diets on the basis of their protein content and, where the two diets were on either side of the optimum, to change the proportions selected as they grew to match their changing requirement for dietary protein. When given two diets, both of which had protein contents lower than the single diets which gave maximal growth, birds ate predominantly from that closer to that optimal diet. When both diets had a higher protein content than the optimum birds ate mostly from that closer to optimum. 4. The results show that growing chickens can match their protein intake closely to their requirements when given a pair of diets that allows this; if both diets are on the same side of the optimum then the one closest to that required is predominantly chosen.

Animal Feed↗

Production and evaluation criteria of specific monoclonal antibodies to the hemagglutinin of the H7N2 subtype of avian influenza virus.

To enhance the rapidity in diagnosing the spread of avian influenza virus (AIV) in chicken layer flocks, studies were initiated to develop more sensitive and specific immunological and molecular methods for the detection of AIV. In this study, the purification of the hemagglutinin protein (H) from field isolates of H7N2, the production of monoclonal antibodies (MAbs), and their evaluation as diagnostic reagents are reported. Hybridomas were generated by fusion of SP2/0-Ag14 myelomas and spleen cells from immunized mice. Hybridomas secreting antibodies specific for the H protein were assayed by an ELISA and cloned using limiting dilution. The MAbs produced were characterized by hemagglutination inhibition (HI), immunohistochemistry (IHC), indirect fluorescent antibody assay (IFA), Western blots, and IFA flow cytometry using various AIV subtypes (i.e., H4N2, H5N3, H7N2). Of the various MAbs assayed, 6 had consistent and reproducible results in each of the assays used. The results obtained in this investigation enhanced the usage of the MAbs to viral H protein in the surveillance of AIV in chickens.

Animals↗

[Study on the possible presence of aflatoxin B 1 in liver and muscle tissue of slaughter animals].

Seventy-seven samples of liver (from pigs, bulls, cows, broiler chickens, layers), twenty-eight samples of white muscle (from pigs, broilers, layers), and twenty-four samples of red muscle (from pigs, broilers, layers) were examined by the modified Jacobson method (Jacobson et al. 1971) for the contingent presence of aflatoxin B1. The samples came from healthy animals, part of them from animals fed non-traditional feed containing pig excrements. Positive findings were only obtained in two samples of pork liver, out of the total number of 30 samples. One sample came from pigs fed a ration containing 5% of pig excrements, the other from a traditionally fed pig. The amounts of aflatoxin B1 found in these two samples can be treated as trace quantities, since the concentration did not exceed 0.1 microgram per kg. In the samples of muscle and liver of the other farm animals, the chromatogram did not show the presence of spots which might resemble, in colour and position, the spots of aflatoxin B1.

Aflatoxins↗

Biochemical changes in layer and broiler chickens when fed on a high-glucosinolate rapeseed meal.

Diets containing 500 g high-glucosinolate rapeseed meal/kg or an equivalent amount of soybean meal as the only protein supplement were fed to layer-type chickens and two broiler strains from 1 to 56 d of age. Additional groups of the former were maintained on the diets until they were 16 and 28 d old. The rapeseed meal produced thyroid hypertrophy in all strains but reduced the growth rate of only one of the broiler strains. The livers of chickens fed on rapeseed meal were enlarged and DNA analysis indicated hyperplasia, but no macroscopic lesions were found. The activities of aspartate transaminase, lactate dehydrogenase and alkaline phosphatase in the plasma were increased by rapeseed meal, suggesting liver damage. In all strains, feeding rapeseed meal increased plasma total protein, albumin and cholesterol and decreased urate. Hyperglycaemia accompanied by a decrease in plasma triglycerides occurred in the layer strain. Through its extra-thyroidal toxicity (-)5-vinyl-oxazolidine-2-thione (goitrin) was probably responsible for most of these changes.

Alkaline Phosphatase↗

Salmonella control programs in Denmark.

We describe Salmonella control programs of broiler chickens, layer hens, and pigs in Denmark. Major reductions in the incidence of foodborne human salmonellosis have occurred by integrated control of farms and food processing plants. Disease control has been achieved by monitoring the herds and flocks, eliminating infected animals, and diversifying animals (animals and products are processed differently depending on Salmonella status) and animal food products according to the determined risk. In 2001, the Danish society saved U.S.$25.5 million by controlling Salmonella. The total annual Salmonella control costs in year 2001 were U.S.$14.1 million (U.S.$0.075/kg of pork and U.S.$0.02/kg of broiler or egg). These costs are paid almost exclusively by the industry. The control principles described are applicable to most industrialized countries with modern intensive farming systems.

Animals↗

Production of rabies neutralizing antibody in hen's eggs using a part of the G protein expressed in Escherichia coli.

In an attempt to produce anti-rabies immunoglobulin affordable for people living in developing countries, we have immunized layer chickens with a part of the G protein of rabies virus expressed in Escherichia coli. Immunoglobulin (IgY) was purified from the yolks of eggs layed by immunized hens. It was revealed in vitro that the antibody specifically bound to virions as well as cells infected with rabies virus. Moreover, the antibody apparently neutralized rabies virus infectivity. Inoculation of the antibody into mice infected with rabies virus reduced the mortality caused by the virus, suggesting that IgY directed to the part of the G protein expressed in E. coli could serve as a possible alternative to currently available anti-rabies human or equine immunoglobulins.

Animals↗

A simple and sensitive liquid chromatography-mass spectrometry confirmatory method for analyzing sulfonamide antibacterials in milk and egg.

A simple and specific method able to identify and quantify traces of 14 sulfonamide antibacterials (SAs) in milk and eggs is presented. This method uses a single solid-phase extraction (SPE) cartridge for simultaneous extraction and purification of SAs in the above matrices. Milk and egg samples are passed through a Carbograph 4 sorption cartridge. After analyte desorption, an aliquot of the final extract is injected into a liquid chromatography-mass spectrometry (LC-MS) instrument equipped with an electrospray ion source (ESI) and a single quadrupole. MS data acquisition is performed in the positive-ion mode and by a time-scheduled multiple-ion selected ion monitoring program. Compared to two published methods, the present protocol extracted larger amounts of SAs from both milk and egg and decreased the analysis time by a factor of 3 with milk samples and by a factor of 2 with egg samples. Recovery of SAs in milk at the 5 ppb level ranged between 76 and 112% with relative standard deviations (RSDs) of <or=13%. Recovery of SAs in egg at the 50 ppb level ranged between 68 and 106% with RSDs of <or=12%. Estimated limits of quantification (S/N = 10) of the method were 1-6 ppb of SAs in whole milk and 5-13 ppb of SAs in eggs. Analyses of eggs from three layer chickens treated with sulfoquinoxaline revealed this antibacterial was still present at the 150 ppb level 1 week after withdrawal.

Animals↗

Interactions of dietary polyunsaturated fatty acids and vitamin E with regard to vitamin E status, fat composition and antibody responsiveness in layer hens.

1. Effects of dietary polyunsaturated fatty acids (PUFA) and vitamin E (VE) on an immune response may interact because VE may protect PUFA from in vivo oxidation. The present study was designed to study the presence of such an interaction in growing layer chickens. 2. Three dietary concentration of linoleic acid (LA, 3.3, 6.6 and 10%), in combination with 4 concentration of dietary VE (5, 20, 40 and 80 mg/kg) were used. Effects of LA and VE on circulating VE concentration, fatty acid composition of bursal and adipose fat, and antibody kinetics against keyhole limpet hemocyanin and Mycobacterim butyricum were established. 3. At high dietary LA concentration, bursal and adipose LA were higher but bursal arachidonic acid and long chain n-3 PUFA decreased. The dietary VE level did not consistently affect the deposition of PUFA in tissue. Plasma VE concentrations were affected by the dietary VE and LA content, but not by their interaction. Antibody responses before and 7 d after immunisation were affected by the dietary treatments. Antibody concentration were not affected by tissue fatty acid content. 4. In conclusion, the interaction effects of dietary PUFA and VE on fat deposition and immune responses are of minor importance compared to separate PUFA and VE effects. This implies that, within the studied range, adding extra VE to preserve or affect the effects of dietary PUFA on antibody responsiveness is unnecessary.

Adipose Tissue↗