Fowl Plasma Research Articles

Elevated bile acids in the plasma of laying hens fed rapeseed meal

A simple procedure was developed for the estimation of bile acid taurine conjugates in fowl plasma. Laying hens fed a diet containing rapeseed meal (RSM) (400 g kg-1) for 12 weeks had higher bile acid levels (154 mumol litre-1) than hens fed a control soyabean diet (116 mumol litre-1) (P less than 0.01). The incidence of liver haemorrhages was higher (34.8 per cent) in RSM-fed hens than in controls (21 per cent), but the severity of the lesions did not correlate with the bile acid concentration in affected birds. Histological examination of sections from livers of RSM-fed birds did not reveal significant hepatocyte degeneration outside the immediate vicinity of the haemorrhage. Canalicular bile plugs were never seen. The incidence of liver haemorrhages (13 per cent) and plasma bile acids (85 mumol litre-1) were lower in hens fed a diet containing beta-aminopropionitrile (0.5 g kg-1), a known lathyrogen. Acute treatment of hens with the hepatotoxin alpha-naphthyl isothiocyanate over four days induced necrosis of hepatocytes and resulted in elevated bile acid concentrations (262 mumol litre-1) compared with controls (73 mumol litre-1) given arachis oil. It was concluded that laying hens fed high levels of RSM develop cholestasis but the toxic principle is not known.

Radioimmunoassay of fowl angiotensin I

Angiotensin I (ANG I) from various nonmammalian vertebrates does not bind the antibody to human ANG I, and thus radioimmunoassay (RIA) is not applicable for measurement of renin activity. It is largely due to this difficulty that biochemistry and physiology of the renin-angiotensin system of nonmammalian vertebrates have not been studied. The present work was therefore undertaken to develop a system to determine plasma renin activity by measuring the ANG I generation rate with RIA in fowl plasma. Antibody to [Asp 1,Val 5,Ser 9]-angiotensin I (fowl ANG I) was produced by immunizing goats and rabbits with a conjugate of human immunoglobulin and fowl ANG I. An antiserum, with a maximum binding of 86.3%, was obtained from the goat after six intradermal injections of the conjugate (each contained 0.25 mg fowl ANG I). This antiserum (at a final dilution of 1:200,000), and 125I-labeled fowl ANG I, purified by DEAE-cellulose and Sephadex G-25 column chromatography, were used for the RIA of natural fowl ANG I. Fowl ANG I generated by incubating plasma and kidney extract, was measured by both RIA and rat vasopressor assay. The values obtained from these two methods correlated well. The formation of angiotensin increased when the duration of incubation was prolonged, and was optimum at pH 5.0–5.5. Plasma renin activity (rate of fowl ANG I generation) in 50 μl plasma from anesthetized female white leghorn chickens (24 weeks, n = 6) was 15.04 ± 2.26 ng/ml·hr, and that in plasma from chronically cannulated conscious chickens (30 weeks, n = 5) was 4.58 ± 0.61 ng/ml·hr. The plasma angiotensinogen level was 500–600 ng/ml in both groups. Thus this sensitive RIA method may provide a useful tool to study the functions and regulation of the renin-angiotensin system in fowl.

Studies on the Phylogenetic Relationships of the Japanese Native Fowl Breeds

Blood samples were taken from about 600 fowls of 24 Japanese native breeds to study the phylogenetic relationships of these breeds. Genetic polymorphisms of albumins, esterases and alkaline phosphatases of fowl plasma were detected using horizontal starch gel electrophoresis.Plasma albumins of the fowl are controlled by three autosomal alleles, AlbA, AlbB and AlbC (STRATIL, 1968). In most of Japanese native breeds, the gene frequency of AlbB was high. AlbA was observed only Jitokko. AlbC was present in Shamo (Japanese Games), and Minohiki and Satsumadori, both of which were phylogenetically related to Shamo breed, and the breeds of Tosajidori, Uzurao and Ohiki.Plasma esterases are controlled by three autosomal codominant alleles (GRUNDER, 1971). Gene frequency of Es-1B was higher than those of Es-1A and Es-1C in most of Japanese native breeds, Es-1C being the lowest, whereas Es-1A was predominant in Mediterranean breeds. A higher frequency of Es-1C was observed in Ohiki and Koshamo (Japanese Game Bantam), small breeds.Plasma alkaline phosphatase of the fowl was observed in two electophoretic regions. One is controlled by the dominant (Akp) and recessive (akp) alleles (WILCOX, 1966), and the other is controlled by the dominant (Akp-2o) and recessive (Akp-2a) alleles GODA et al., 1971). Gene frequency of Akp was higher in Onagadori, Totenko and Uzurao, which were charactaristic to have white ear lobes, than the other breeds. Low frequency of Akp was observed in Shokoku having red ear lobes. It is postulated that Totenko was a more possible ancestor of Onagadori (Long Tailed Japanese fowl) than Shokoku.

The use of osmolality as an aid to establishing consistent fixation quality: Studies on the kidney of the domestic fowl

1. Using the proximal convoluted tubule of the fowl kidney as indicator tissue the immersion application of fixatives of differing composition and osmolality was studied. 2. The best results were obtained with Dalton's buffered osmium tetroxide and with sodium cacodylate buffered glutaraldehyde followed by osmium post-fixation. 3. The most satisfactory component concentrations in both cases were those which most reproduced a total osmolality close to that of fowl plasma. With both types of fixation the ultrastructural image of the kidney tubule was sensitive to changes in fixative osmolality and differences were clearly identified over a 30 to 50 mOsm range. 4. With glutaraldehyde/osmium double fixation it was found that it was the total osmolality of the glutaraldehyde buffer combination that was to be considered in a study of osmolotic effects, not the osolality of the buffer vehicle alone. 5. The 30 to 50 mOsm difference in fixative osmolality that was able to be detected by studying the image of the kidney tubule was of the order of magnitude commonly experienced as experimental error in the laboratory as a result of changes in reagent batches or error in making up the solutions. The measurement of osmolality therefore provides an ideal quality control to the laboratory make-up of fixative solutions.

Changes in Fowl Plasma α-Lipoproteins Caused by Starvation

Splitting and retardation of the a-lipoprotein bands on starch gel electrophoresis was observed in the plasmas of 15-week old birds starved for as little as 15 hours and in birds given reduced feed levels (10 gm./bird) for as little as four days. Feeding restored the normal condition within 3 hours. The changes were essentially the same as those produced by the addition of turpentines, acids or cationic soaps to plasma in vitro. These changes may provide a sensitive method for evaluating the extent of anorexia caused by disease or experimental conditions.

Morphogenesis and functional differentiation of the rat parotid gland in vivo and in vitro

ABSTRACT In comparison with the submandibular and the sublingual glands, the parotid develops slowly in the rat. The foetal rudiment appears a day later than that of the submandibular gland and the formation of adenomeres is slower, leading to a more diffusely branched structure. Cytodifferentiation, in the form of traces of mucopolysaccharide in the tubules and terminal buds, begins at, or just before, birth. There is a transitory increase in mucopolysaccharide production for a few days after birth until the presumptive acinar cells become pyramidal in shape with basal nucleus and granular cytoplasm. Amylase activity of the gland begins to rise between the second and third day after birth and reaches the adult level at weaning. That of the submandibular gland remains at the foetal level. Parotid rudiments were cultivated on a film of agar over a medium of fowl plasma and chick embryo extract. The oxygen in the gas phase of air and 5 % CO2 was increased to 50 % after the first 9 days in vitro. Under these conditions the mass of the rudiments increased tenfold during 18 days cultivation and the initially unbranched rudiment formed adenomeres in which the cytodifferentiation followed the same course as in vivo. The rise in amylase activity of the explants was only slightly delayed compared with that in vivo, suggesting that systemic or environmental factors are not obligatory in the early postnatal development of the rat parotid.

Kinetics of Hydrolysis of p-Nitrophenyl Phosphate by Fowl Plasma Alkaline Phosphatase

THE rapid hydrolysis of p-nitrophenyl phosphate (King and Delory, 1939) and the high extinction coefficients of the p-nitrophenol ion, one of the products of hydrolysis, were utilized by Bessey et al. (1946) in the development of a simple procedure for measuring alkaline phosphatase activity in minute quantities in human serum. Our interest in p-nitrophenyl phosphate was the possibility of its use in measuring quantitatively changes in plasma alkaline phosphatase in the same birds over long periods during the growth of chicks and egg production by hens.Although p-nitrophenyl phosphate has been used as the substrate for the determination of phosphatase of tissues of a number of species, including fowl (Bell, 1960, 1964; Bell and Sillers, 1962; Hurwitz and Griminger, 1961; Tanabe and Wilcox, 1961), very little has been published on the kinetics of hydrolysis of this compound by alkaline phosphatase of chicken plasma. The present paper describes studies directed toward…

Properties of turpentine relative to the effects on fowl plasma in vitro. I. The simulation of "leukemic" changes in normal plasma.

The addition of some turpentines to avian plasma in vitro caused α-lipoprotein changes similar to those seen in electropherograms of plasma from birds affected with erythroblastosis. The effective turpentines were old oxidized samples containing large amounts of acids and peroxides. Acids, pure peroxides, and cationic soaps had similar effects on plasma but much more material was required. Neither the acids nor the peroxides could be shown to be the cause of the plasma changes but their presence was indicative of activity of turpentine on plasma lipoproteins. The α-lipoprotein changes which appear to result from many different treatments are discussed in terms of possible common mechanisms.

Properties of turpentine relative to the effects on fowl plasma in vitro. II. The separation of peroxide components of turpentine by thin-layer chromatography.

Thin-layer chromatograms were made of some turpentines, organic peroxides, and epoxides, and sprayed to detect peroxides. Benzoyl peroxide, cumene hydroperoxide, and painters' turpentine gave the most prominent spots. Only one of the three epoxides tested, styrene oxide, showed peroxide. Many more peroxides were present in "aged" turpentine than in freshly prepared samples. As aged turpentine has most effect on fowl plasma α-lipoprotein, chemical differences between samples should be considered when turpentines are used in experimental pathology.

Studies on the evolution of plasma kinins: Reptilian and avian blood

Contact with a foreign surface triggers the release of a plasma kinin in turtle or alligator plasma. The peptide cannot be distinguished from mammalian plasma kinins. The equivalent of 1.4 ug of bradykinin was found in one ml of alligator and 2 ug in one ml of turtle plasma. The plasma of these two reptiles contains a contact factor, possibily Factor XII, a prekallikrein, a kininogen and a kininase. Fowl plasma does not have the contact factor, but this can be transferred from mammalian or reptilian plasma. Thus pigeon or duck plasma can be activated to release a kinin with properties similar to those of bradykinin. (0.1 resp. 0.034 ug per ml). Acid treatment of duck plasma releases an agent or more likely agents that are active in vitro and in vivo , but they are different from bradykinin.

Inheritance of two alkaline phosphatase variants in fowl plasma.

Starch-gel electrophoresis of plasina reveals two mnutually exclusive forms of alkaline phosphatase. Analysis of the plasma of 931 birds from 96 sib families of two inbred lines through two generations shows that the faster-moving isozymne is determined by an autosomal gene Ap(2) which is completely dominant to its allele Ap(4) determining the slower form.

Inheritance of Two Alkaline Phosphatase Variants in Fowl Plasma

Starch-gel electrophoresis of plasma reveals two mutually exclusive forms of alkaline phosphatase. Analysis of the plasma of 931 birds from 96 sib families of two inbred lines through two generations shows that the faster-moving isozyme is determined by an autosomal gene Ap 2 which is completely dominant to its allele Ap 4 determining the slower form.

The Differential Growth-Response of Embryonic Chick Limb-Bone Rudiments to Triiodothyronine in vitro. III. Hormone Concentration

ABSTRACT Limb-bone rudiments from 6-day-old chick embryos were explanted in a medium of fowl plasma and embryo extract and the effect of various concentrations of T3 on the growth in length and wet weight of the rudiments was studied. Treatment with 0·01 to 0·32μg.T3/ml. produced a progressive retardation in the growth in length of the tibia, and a stimulation in the growth of the radius, which was maximal at intermediate concentrations. The growth in length of the ulna was increased by low levels of added hormone and retarded by high levels. The final wet weight of the tibia was slightly increased by low T3 concentrations and reduced by high doses. The final wet weights of the radius and ulna were greater than the controls at all T3 concentrations, but the greatest increase in the size of the ulna was reached at a lower dose level than that needed for the radius. These results suggested that the growth stimulation of the radius and retardation of the leg bones when exposed to the same concentration of T3 reflect a difference in sensitivity of the rudiments which may respond in a fundamentally similar way. The response of the ulna to 0 ·16μg. T3/ml. was a continuous process of growth stimulation followed by retardation. A difference in the threshold of response between the third metatarsus and radius was found at low concentrations of T3: the flattened cell zone of the metatarsus was affected by 0·0005 μg./ml. to which the radius was insensitive.

Comparative starch-gel electrophoretic studies of fowl egg white and plasma.

ELECTROPHORETIC studies of egg white of domestic fowl have been repeatedly carried out by the Tiselius moving boundary method in the past1. Forsythe and Foster2 working with pooled egg albumen samples indicated that small but statistically significant differences could be established between egg white patterns of six different genetic lines of chickens. The high resolving power of starch-gel electrophoresis, with the slightly modified discontinuous buffer system of Poulik3, was used recently by J. E. Lush for the investigations of egg albumen patterns4. He found a genetically conditioned polymorphism in the egg white from 16 closed populations of hens.

Cholinesterases in the domestic fowl and the specificity of some reversible inhibitors

The properties of cholinesterases from several tissues of the domestic fowl have been studied in vitro. Substrate-inhibitor relationships showed that the enzyme corresponding to mammalian acetylcholinesterase resembled the mammalian enzyme in most respects but was considerably less active in its ability to combine with bisquaternary substrates or inhibitors. This suggests some difference between the active sites in the mammalian and the fowl enzymes. The enzyme corresponding to mammalian butyrocholinesterase was found to have properties intermediate between those of mammalian butyrocholinesterase and those of the fowl acetylcholinesterase. “Specific” inhibitors of mammalian butyrocholinesterase were less potent against the corresponding enzyme in the fowl while “specific” inhibitors of mammalian acetylcho-linesterase possessed considerable inhibitory potency against the fowl plasma enzyme. Thus no specific inhibition could be obtained. The enzyme corresponding to butyrocholinesterase, like the fowl acetykholinesterases, was unable to combine with bis-quaternary compounds.

Tissue culture and tooth development

Tissue culture has been used for the study of the morphogenetic field within the tooth and within the jaw. In studies using a culture medium composed of chicken embryo extract and fowl plasma it was found that when isolated tooth rudiments were explanted at an early stage of differentiation, normal cusps formed and histological differentiation of all dental tissues occurred. In pieces of jaws explanted at a very early stage before the formation of tooth germs whole teeth developed, those from the anterior part of the jaw forming incisors and those from the posterior portion molars: these findings illustrate the presence of a morphogenetic field within the jaw and within the tooth germ itself. In tooth germs cut into two before cusps were present, and in which the two halves were grown in tissue culture, regulative changes occurred and each part formed a whole tooth. When tooth germs were grown in a chemically defined medium, cusps formed during the period of cultivation and there was histological differentiation of the component cells and tissues of the tooth germ.

Action de la dicarnitine’ sur la croissance et l’ossification d’ébauches de tibias et de fémurs d’embryons de poulet cultivés in vitro

ABSTRACT Tibias and femurs of 7-day-old chick embryos were cultivated in the absence and in the presence of racemic dicarnitine on a solid medium (fowl plasma + embryo extract, the latter usually diluted 20-to 200-fold). The growth in length of the bones was increased by 1/3 to 2/3 in comparison with the controls, when 2·5 and 5 mg. / I. of dicarnitine was added and when the embryo extract was not diluted more than 100-fold. With embryo extract diluted 200-fold, the shape of the bone was abnormal. Dicarnitine in this case restored the normal shape without increasing the elongation. Dicarnitine in a medium containing embryo extract diluted 200-fold was found to increase the maximal thickness of the periosteal bone in comparison with the controls. This effect was statistically highly significant in the case of dicarnitine concentrations of 2·5 and 5 mg./l. The effect was not found with non-diluted embryo extract. Mitoses were counted in tibias and femurs cultivated in vitro for periods of 1 to 7 days. 2·5 mg./l. dicarnitine was shown to promote mitotic activity. In the periosteum, it maintained for 4 days a mitotic activity equal to 45 per cent, of the mitotic activity in vivo. In vitro controls retained only 6 per cent, of this activity. The usefulness of frequent transfers was demonstrated. The percentage of prophases was higher in the periosteum than in the epiphyses. The relative frequency of metaphases increased with the age of the cultures. The frequency distribution of the phases of mitosis was shown to be very little changed by dicarnitine. After 7 days of culture, the number of cells was more increased in the hypertrophic cartilage than in other tissues. Dicarnitine did not much increase the total number of cells; there was practically no effect on the number of cells of the epiphyses but a large increase in the number of cells of the periosteum and of the hypertrophic cartilage was observed.

Studies on the Development of Wool Follicles in Tissue Culture

Explants of' skin from four sheep foetuses of difIerent ages have beell cultivated for periods of up t,o 38 clays in a medium of fowl plasma lind ehicken embryo extraet. 'I'he development of wool follicles of difforent types has beon st,uilied both in these living ()xplants and in serial sections from them.

Nutrition of animal cells in tissue culture. VI. Low toxicity of barium.

SummaryRelatively high concentrations of barium (1 to 1000 μg per ml) reduce the area of outgrowth of chick fibroblasts propagated in a medium containing fowl plasma and chick embryo extract. But these same concentrations of barium do not shorten the period of survival of chick embryo mesenchyme tissues maintained in a synthetic medium. Chemical analyses showed the barium content of the natural and synthetic media that were used to be less than 5 μg per 10 ml.

Growth and form in vitro of avian femur rudiments on clots in which dried products are substituted for fresh fowl plasma

Growth and form in vitro of avian femur rudiments on clots in which dried products are substituted for fresh fowl plasma