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Anatomical parameters of cardiopulmonary system in three different lines of chickens: further evidence for involvement in ascites syndrome.

The present study was designed to compare the anatomical parameters of the cardiopulmonary system in three different lines of chickens with different susceptibility to ascites syndrome. Eggs from three different genetic lines-commercial broiler and layer lines and one native to Iran--were incubated and hatched, and 1-day-old chicks used. The relative heart and lung weights, the volumes of the heart, lung and thorax cavity, the incidence of ascites, and the related physiological parameters in these chickens were followed. Lung weight as a percentage of body weight, the relative lung and heart volume, and the volumes of the thorax cavity, before and after removing heart and lung tissues, were lower in fast-growing broiler chickens compared with the layer and native chickens. Additionally, most of these parameters (e.g. relative lung weight, lung volume and thorax cavity) were significantly decreased with age in broiler chickens but were increased in layer and native chickens, which was concomitant with the incidence of ascites in broiler chickens. Our data indicate that all cardiopulmonary parameters investigated are extremely unfavourable to broiler chickens and suggest a reduction in gas exchange area in broilers, and therefore higher susceptibility to pulmonary hypertension and ascites.

Aging↗

Altered colonizing ability for the ceca of broiler chicks by lipopolysaccharide-deficient mutants of Salmonella typhimurium.

Salmonella typhimurium strain 3333/O was used to assess the role of bacterial lipopolysaccharide (LPS) in intestinal colonization of broiler chicks by salmonellae. LPS-defective TnPhoA mutants of this strain were isolated. The sensitivities of the mutants to smooth and rough phages and LPS banding patterns in sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated a defect in the polysaccharide side chain of the LPS molecule. Colonization was determined by orally administering 10(8) cells each of the wild-type and/or the mutant strains per chick and counting the colony-forming units (CFU) from the ceca 1 to 3 weeks after gavage. CFU of chicks given the LPS-deficient strains either were not detected or were significantly lower than the CFU from chicks given the wild-type strain. The incidence of the wild-type strain in spleens was higher than incidence of the mutant strains. In vitro binding studies with LPS-deficient mutants derived in this study and from S. typhimurium LT2 suggest that LPS side-chain components may shield the bacterial cell from entrapment in the chicken mucus. The LPS layer appears to enhance persistence of Salmonella in the avian intestinal tract.

Animals↗

Genomic organisation of the chicken ghrelin gene and its single nucleotide polymorphisms detected by denaturing high-performance liquid chromatography.

(1) Ghrelin is a novel endogenous ligand for the growth hormone secretagogue receptor (GHS-R) and is expressed primarily in the stomach and hypothalamus with the probable function of stimulating GH secretion and food intake both in mammals and poultry. The complete sequences of ghrelin gene have been reported in humans and mice; however, that of chickens remains unclear. (2) Here, we report the complete sequence of chicken ghrelin gene (submitted to Genbank; accession number AY303688), which consists of 5 exons and 4 introns. As in mice, the first exon of chicken ghrelin gene does not encode any amino acid. (3) Scanning point mutations with denaturing high-performance liquid chromatography (DHPLC) using WAVE DNA Fragment Analysis Systems and confirmed with direct sequencing for polymerase chain reaction (PCR) products, we analysed the single nucleotide polymorphisms (SNPs) in the entire gene of chicken ghrelin. (4) Results showed that there were 19 SNPs in chicken ghrelin gene, and most of these SNPs were scattered in the 4 introns. In these SNPs, one mutation in exon 5 (A2355G) led to the change of amino acid from glutamine to arginine (Gln 113 Arg): as a result a different ghrelin precursor instead of a mature peptide was produced. In addition, one SNP in 5'UTR (C223G) determined the presence or absence of a potential binding site of transcription factor serum response factor (SRF), which might affect the expression of chicken ghrelin gene. Some of the SNPs detected in the present study could be used in quantitative trait loci (QTL) mapping for growth characters in chickens. (5) Because one SNP is located in a polymorphic site of restriction enzyme PagI of intron 4, it was possible to design a PCR-RFLP procedure and analyse the diversity of 10 chicken populations. Results showed the allelic frequencies of C2100T differ among these breeds, however, no significant difference was observed between imported breeds and Chinese native ones, nor between egg layers and meat type chickens. A chi-square test showed that 4 chicken populations of Beijing Fat, Xinghua, Recessive White and Silky followed the Hardy-Weinberg equilibrium.

Amino Acid Sequence↗

Simultaneous cell death and desquamation of the embryonic diffusion barrier during epidermal development.

The periderm is an epithelial layer covering the emerging epidermis in early embryogenesis of vertebrates. In the chicken embryo, an additional cellular layer, the subperiderm, occurs at later embryonic stages underneath the periderm. The questions arose what is the function of both epithelial layers and, as they are transitory structures, by which mechanism are they removed. By immunocytochemistry, the tight junction (TJ) proteins occludin and claudin-1 were localized in the periderm and in the subperiderm, and sites of close contact between adjacent cells were detected by electron microscopy. Using horseradish peroxidase (HRP) as tracer, these contacts were identified as tight junctions involved in the formation of the embryonic diffusion barrier. This barrier was lost by desquamation at the end of the embryonic period, when the cornified envelope of the emerging epidermis was formed. By TUNEL and DNA ladder assays, we detected simultaneous cell death in the periderm and the subperiderm shortly before hatching. The absence of caspases-3, -6, and -7 activity, key enzymes of apoptosis, and the lack of typical morphological criteria of apoptosis such as cell fragmentation or membrane blebbing point to a special form of programmed cell death (PCD) leading to the desquamation of the embryonic diffusion barrier.

Animals↗

[Substrate basis of the adhesion of the adenohypophyseal and diencephalic cell layers and cell anlagen in chick embryos].

At the 16-18th stages of development of the chicken embryos adhesion of cell layers of the adenohypophysis and diencephalon anlages is provided with a submicroscopically regulated complex of neutral and acid polysaccharides, bivalent kations and probably proteins. The mechanisms of cells contact in each of the layers with this complex are different.

Animals↗

Immunohistochemical characterization of monoclonal antibodies (PDs) as markers of the periderm in the developing chicken embryo.

The outermost surface cell layer of the developing embryo, the periderm, arises from the initial single layer of ectoderm and is eventually exfoliated from the stratified epidermis, which has the same ectodermal origin. In this study, monoclonal antibodies against chicken limb bud ectoderm were generated and screened for those which stained the periderm. Four separate antibodies termed PD2, 3, 7 and 9 were obtained from 180 mixed hybridomas. These PD antibodies stained the periderm selectively at all stages examined (stage 20-42). By correlating the results of immunohistochemistry with observations made by transmission electron microscopy, it was revealed that PD antibodies stained both the squamous periderm at an early stage and rounded bulging peridermal cells just before exfoliation. Therefore we feel that PD antibodies may be useful in further systematic investigations of the development and function of the chicken embryonic periderm.

Animals↗

Morphometry of the extremely thin pulmonary blood-gas barrier in the chicken lung.

The gas exchanging region in the avian lung, although proportionally smaller than that of the mammalian lung, efficiently manages respiration to meet the high energetic requirements of flapping flight. Gas exchange in the bird lung is enhanced, in part, by an extremely thin blood-gas barrier (BGB). We measured the arithmetic mean thickness of the different components (endothelium, interstitium, and epithelium) of the BGB in the domestic chicken lung and compared the results with three mammals. Morphometric analysis showed that the total BGB of the chicken lung was significantly thinner than that of the rabbit, dog, and horse (54, 66, and 70% thinner, respectively) and that all layers of the BGB were significantly thinner in the chicken compared with the mammals. The interstitial layer was strikingly thin in the chicken lung ( approximately 86% thinner than the dog and horse, and 75% thinner than rabbit) which is a paradox because the strength of the BGB is believed to come from the interstitium. In addition, the thickness of the interstitium was remarkably uniform, unlike the mammalian interstitium. The uniformity of the interstitial layer in the chicken is attributable to a lack of the supportive type I collagen cable that is found in mammalian alveolar lungs. We propose that the surrounding air capillaries provide additional structural support for the pulmonary capillaries in the bird lung, thus allowing the barrier to be both very thin and extremely uniform. The net result is to improve gas exchanging efficiency.

Animals↗

Plasminogen activator production by the granulosa layer is stimulated by factor(s) produced by the theca layer and inhibited by the luteinizing hormone surge in the chicken.

The stages of follicular maturation of a preovulatory follicle in the hen can be divided into an extended proliferative phase (prior to LH surge) and a brief ovulatory phase (after LH surge). Previous studies suggest involvement of plasminogen activator (PA) in both the proliferative and ovulatory phases. The goals of the present study were 1) to determine whether PA production by granulosa and theca is dependent upon interaction of the two cell layers; 2) to investigate whether the structural difference of the stigma (site of follicular rupture) and nonstigma regions of the theca layer affect PA production; 3) to determine whether there is a change in the ability of the granulosa layer and stigma or nonstigma regions of the theca layer to produce PA as the follicle makes the transition from the proliferative to the ovulatory phase; and 4) to characterize the type(s) of PA produced by the hen follicle. Equal proportions of the granulosa layer (10-mm diameter) and stigma or nonstigma regions of the theca layer (10 mg) obtained from the F1 preovulatory follicle 8 h before ovulation (before LH surge) or 2 h before ovulation (after LH surge) were incubated alone or in combination for 24 h. PA was measured in tissue homogenates and medium by use of the chromogenic substrate S-2251. The granulosa layer or stigma or nonstigma regions of the theca layer incubated alone and obtained either 8 h or 2 h before ovulation had very low amounts of PA activity in the medium and tissue homogenates.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Studies of avian urolithiasis associated with an infectious bronchitis virus.

Avian urolithiasis syndrome was diagnosed in 14-to-25-week-old chickens from a multiple-age caged-layer complex housing more than 2.5 million chickens. Losses from this syndrome ranged from 0.5 to 1.0% per week. Seven-to-14-week-old pullets from this facility had multifocal renal tubular necrosis leading to interstitial fibrosis, tophus formation, and tubular dilation. A coronavirus was isolated in embryos inoculated with pooled samples of trachea, kidney, and cecal tonsil of 4-week-old pullets. This virus, identified as 85-209, was related to infectious bronchitis virus strain Florida 88 by hemagglutination-inhibition assay. Day-old specific-pathogen-free chicks were inoculated with fifth-embryo-passage amnioallantoic fluid containing this virus. These chicks developed histologic lesions of tracheitis at 5 to 7 days postinoculation. Half the chicks inoculated by eyedrop developed renal tubular necrosis after 7 days. Urolithiasis in the flock investigated was attributed to renal damage by this strain of infectious bronchitis virus occurring in 4-to-7-week-old pullets and progressing to segmental atrophy, hyperplasia, and ureteral stone formation in 14-to-25-week-old chickens.

Animals↗

Preproenkephalin messenger RNA-containing amacrine cells in the chicken retina identified with in situ hybridization histochemistry.

Enkephalin peptides are present in the retina of several vertebrate species. In the avian retina, enkephalin immunoreactivity is primarily localized to a population of amacrine cells. In the present study, we determined the localization of cells expressing preproenkephalin (PPE) mRNA, which encodes the precursor of enkephalin peptides, in adult as well as in embryonic chicken retinas. The localization of PPE mRNA-expressing cells to the proximal inner nuclear layer (INL) in the adult chicken retina is similar to that of enkephalin-immunoreactive cells observed in previous studies, indicating that amacrine cells expressing PPE mRNA synthesize Met5- and Leu5-enkephalin peptides and related extended forms. Specific hybridization signal is absent in retinas at embryonic day (E) 11, but it is detected in retinas at E 15 and at hatching. PPE mRNA-expressing cells at these ages are located in the proximal INL, and they can be classified as amacrine cells on the basis of their soma size and laminar position. These findings confirm and extend previous observations on the presence of opioid peptides in amacrine cells of the chicken retina. The presence of PPE mRNA at embryonic ages, together with the evidence that enkephalins influence developmental processes, suggests that these peptides modulate retinal maturation in birds.

Animals↗

Occurrence of aflatoxins and aflatoxin-producing strains of Aspergillus spp. in soybeans.

Above average rainfall in Maryland during August, September, and October 1971 resulted in heavy mold growth in soybeans while still in the field. Of 28 samples of soybean seed, aflatoxins were found in 14, 2 of which had been used in poultry feed. Aflatoxins were identified by thin-layer chromatography, spectrophotometry, and chicken embryo bioassay. Aspergillus spp. were isolated from 11 samples, and 5 of these isolates produced aflatoxins when grown in liquid culture.

Aflatoxins↗

Form and function of lacunae in the ovary of the laying hen.

BACKGROUND: Spontaneous atresia of large, yolky, ovarian follicles occurs in the chicken and in other species when the stimulus for ovulation ceases. This means that as much as 40 ml of yolk must be resorbed by the ovary if normal ovarian morphology is to be regained. This study examines the process of yolk resorption in the ovarian stroma following bursting atresia. METHOD: Light, transmission, and scanning electron microscopy were used to study the morphology of lacuna spaces and their changes in ovaries of broiler and layer breeds of domestic chicken, both in actively laying birds and in birds undergoing spontaneous or induced follicular atresia. A comparative study was made of the ultrastructural differences between lacuna-lining cells, blood-vessel endothelium, and germinal epithelium of the ovary. RESULTS: Transformation of lacuna-lining cells and peritoneal cells into macrophages occurred after bursting atresia and exposure of these cells to released yolk. Complete atresia of large yolky follicles occurred very quickly. CONCLUSION: The resorption of this large volume of yolk, at least in part, seems to be mediated by the transformed lacuna-lining cells.

Animals↗

Serovars of Listeria monocytogenes and Listeria innocua from food.

The increase of listeriosis outbreaks in recent years has mainly been due to the consumption of contaminated dairy products. This prompted a revision of prevention and control measures. Cheese and other foods (meat, poultry meat) were investigated. For isolation, samples were prepared and investigated according to the recommendations of the FDA with some modifications including, in particular, sub-enrichment. Suspicious colonies were tested biochemically followed by serological identification with factor sera. Out of 100 chicken (95 broilers, 5 layers) Listeria monocytogenes was isolated from 85 carcases. The contamination rate for minced beef and pork (n = 76) was approximately 30%. That for selected ripened soft cheeses (n = 89) was 9%. The counts of Listeria in red meat were usually very low (approximately 10/g). Serovar 1/2a was predominant (78 out of 102 strains). Further serovars detected were: 1/2b, 1/2c, 3a, 4a, 4b, 4d. For comparison, isolated strains of L. monocytogenes and Listeria innocua were inoculated into chicken embryos. In contrast to the former, L. innocua did not kill the embryos.

Animals↗

Serological response of chickens to Salmonella enteritidis infection.

Fifty-eight sera, from 29 chickens originating from two layer flocks known to be naturally infected with Salmonella enteritidis phage type (PT) 4, were examined for antibodies to S. enteritidis. Using the techniques of immunoblotting and ELISA, antibodies to the lipopolysaccharide (LPS) of S. enteritidis were detected in 43 of 58 sera. Antibodies were of the IgG class and bound to the S. enteritidis LPS antigen O = 12. Bacterial agglutination reactions using whole-cell preparations of S. enteritidis and S. pullorum, correlated with anti-LPS antibody reactions as detected by immunoblotting and ELISA. A rapid means of screening chicken sera for antibodies to the LPS of S. enteritidis as an indicator of infection is discussed.

Agglutination Tests↗

Short-term effects of food protein content on subsequent diet selection by chickens and the consequences of alternate feeding of high- and low-protein foods.

1. Growing male chickens of broiler and layer strains were allowed to eat either a high-protein food (HP) or a low-protein food (LP) for 10 min after an overnight fast and then offered a choice between HP and LP. During the next hour they ate significantly more of the food other than the initial meal. Similar results were obtained when there was a gap of 45 min without food between the initial meal and the choice period. 2. When the initial meal was given by tube into the crop, followed by 45 min without food, there was no significant effect on subsequent diet selection. These results show that it is necessary for the birds to eat the food themselves for it to exert its effect on subsequent choice. 3. Birds with experience of eating two foods identical in nutrient composition but different in colour did not select either food in preference to the other following an initial meal of one alone. This is evidence against the possibility that neophagia accounts for the results of the first two experiments. When an initial meal of low protein food was followed by a choice of high and low protein foods, birds ate a greater proportion of the high protein when it was placed on the opposite side of the cage from that in which the initial meal was given. Thus they are not choosing a food simply by its position within the cage. 4. To see the extent to which broilers can extend their ability to compensate for protein content over several weeks, male broilers were given one of 4 treatments from 5 to 10 weeks of age: (A) HP and LP concurrently; (B) HP and LP on alternate days; (C) HP in the morning and LP in the afternoon, (D) LP in the morning and HP in the afternoon. There was no significant effect of treatment on food intake, body weight gain or carcase protein content but carcases were significantly less fat after treatment (B) than treatment (D). 5. It is concluded that growing broiler chicks can compensate well for periods of access to only a low-protein food, by subsequent intake of high-protein food, and vice versa, as long as they are able to gauge the sensory properties of the foods. Offering two complementary foods in alternating half-day periods is an effective method of choice-feeding, allowing birds to select a diet to meet the requirements for growth.

Aging↗