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Pathogenicity by parenteral injection of fowl adenovirus isolated from gizzard erosion and resistance to reinfection in adenoviral gizzard erosion in chickens.

The pathogenicity of a serotype-1 fowl adenovirus (FAV-99ZH), which causes adenoviral gizzard erosion by oral inoculation in chickens, was investigated in specific pathogen-free white leghorn chickens. In trial 1, 14 chickens were inoculated intravenously with the virus at 21 days of age and euthanatized for necropsy within 1-14 days of inoculation. Gizzard erosion was grossly observed from day 7 postinoculation (PI), and histologically, FAV-99ZH antigen-positive, basophilic intranuclear inclusion bodies were seen in the gizzard lesions from day 7 to 11 PI. Necrotizing pancreatitis, and cholecystitis and cholangitis associated with the inclusions were observed from day 3 to 14 PI (pancreatitis) and from day 5 to 9 PI (cholecystitis and cholangitis), respectively. The inclusions were also observed in the epithelial cells of the cecal tonsils from day 3 to 5 PI. The virus was recovered from samples of the lesions. It was revealed that FAV-99ZH causes not only gizzard erosion but also pancreatitis, cholecystitis, and cholangitis by intravenous inoculation in chickens. In trial 2, 10 chickens were inoculated orally with the virus twice, at 13 and 36 days of age, and euthanatized for necropsy within 4-17 days after reinfection. Macroscopically, focal gizzard lesions were observed; however, neither necrosis nor inclusions were observed by microscopy. Moreover, FAV was not recovered from the gizzard or rectum of any of the chickens at necropsy. This suggests that the gizzard lesions occurred as a result of the primary infection, and that the chickens were able to resist reinfection.

Adenoviridae Infections↗

Lack of interaction between avian leukosis virus subgroup J and fowl adenovirus (FAV) in FAV-antibody-positive chickens.

Unfounded field speculation has suggested that avian leukosis virus subgroup J (ALV-J) predisposes young meat-type chickens to inclusion body hepatitis caused by fowl adenovirus (FAV). To address this hypothesis, we infected 1-day-old grandparent meat-type chickens carrying maternal antibodies against FAV with a field isolate of FAV associated with inclusion body hepatitis in broilers, ALV-J, or both FAV and ALV-J. We examined the effects of FAV alone or in combination with ALV-J on the basis of clinical signs, overall mortality, growth rate, and gross and microscopic lesions. With such criteria for evaluating possible interactions, we found no significant differences in the dually infected birds in comparison with chickens that received a monovalent challenge with either FAV or ALV-J.

Adenoviridae Infections↗

Characterization of monoclonal antibodies against fowl adenovirus serotype 1 (FAV1) isolated from gizzard erosion.

Six clones of monoclonal antibodies (Mabs) to fowl adenovirus (FAV) serotype 1 were produced. All Mabs reacted positively by enzyme-linked immunosorbent assay. Three Mabs recognized the putative 100-kD hexon protein and reacted to serotype 1 specifically by western blot analysis but did not react to other FAV serotypes (2, 3, 4, 5, 6, 7, and 8a). These Mabs will be useful for immunodiagnosis of FAV serotype 1 infection in chickens with gizzard erosion and in further research studies involving the genomes and proteins of FAV serotype 1.

Animals↗

Characterisation of fowl adenoviruses from chickens affected with infectious hydropericardium during 1994-1998 in India.

In the present study characterisation has been done for six group I fowl adenoviruses (FAV) isolated from outbreaks of infectious hydropericardium (IHP) of chickens that occurred in different states/regions of India during the years 1994-98. These six viruses were identified as FAV serotype 4 by virus neutralisation and restriction endonuclease analyses. Antigenic analyses of the viruses revealed close relationship (R-values 0.93-0.96). Under the experimental conditions, we have been able to induce IHP using FAV serotype 4 isolate AD: 411 and were also able detect FAV antigens in myocardial tissues by immunofluorescence assay (a new observation), an indication that IHP causing FAV serotype 4 strain replicate in myocardial tissue. Restriction endonuclease analysis of the viral genomes (approximately 46 Kb), using Hind III, Sma I, Xba I, Bam HI, Pst I and Dra I produced identical genetic profiles. Pst I and Bam HI profiles for these six vitus isolates were identical to those published earlier for an IHP causing Pakistani FAV serotype 4 isolate KR31. The identical genetic profiles of viruses, chronology of the outbreaks of IHP in Pakistan during 1989 onward and later in Jammu and Kashmir, India (1994), suggest that FAV serotype 4 isolates involved in outbreaks of IHP in India had probably spread from Pakistan. In order to prevent further spread and economic losses due to IHP in India, based on the antigenic relatedness data in this paper, any one of the six studied FAV serotype 4 isolates can be used as a candidate for mass production of CEH culture based killed vaccine.

Adenoviridae Infections↗

Pulmonary macrophages in birds (barn owl, Tyto tyto alba), domestic fowl (Gallus gallus f. domestica), quail (Coturnix coturnix), and pigeons (Columbia livia).

BACKGROUND: Birds have a limited number of resident macrophages in the normal steady-state respiratory tract. The discovery of phagocytes in lavages of lung from birds contrasts with findings that phagocytes are seldom seen in investigations in situ. An electron microscopic study was performed in the respiratory units, the parabronchi, and air capillaries in particular in several adult bird species to localize the seat of respiratory macrophages. METHODS: Lung tissue of barn owl, domestic fowl, quail, and town and homing pigeons was subjected to standard processing for light and electron microscopy after immersion fixation, intratracheal instillation, and intravascular perfusion. RESULTS: Clusters of macrophages were predominantly housed in the loose connective tissue at the floor of atria at the entrance to the infundibula and gas-exchange tissue proper. Scattered solitary phagocytes were also found in connective tissue of air sacs, interatrial septa, and adventitia of inter- and intraparabronchial arteries and veins and in peribronchial lymphoid tissue. Phagocytized foreign particulate material mostly consists of hard, dense, crystalline formations surrounded by a limiting membrane. The transport of small airborne particles occurs via the squamous atrial epithelium to the underlying macrophages. The macrophages are often accompanied by mast cells. CONCLUSIONS: The present results demonstrate that avian respiratory macrophages are predominantly located in atrial connective tissue compartments and do not seem to migrate to the airway surfaces.

Animals↗

Development of the follicle-associated epithelium and the secretory dendritic cell in the bursa of fabricius of the guinea fowl (Numida meleagris) studied by novel monoclonal antibodies.

Two stromal elements, follicle-associated epithelium and secretory dendritic cells of the bursa of Fabricius were studied by light microscopy and two novel MAbs, that were produced against splenic cell suspensions of guinea fowls. Both antigens recognized by these MAbs, designated GIIF3 and NIC2, are localized in the cytoplasm of the stromal cells, and their molecular weights are 50 and 30 kD, respectively. During embryogenesis the GIIF3 and NIC2 cells emerge in the mesenchyme of the folds before follicle formation. The GIIF3 and the NIC2-positive cells accumulate under the surface epithelium of the plicae and migrate into the epithelium, that precedes the bud-formation. From the bud, the GIIF3-positive cells migrate up to the luminal surface, and they transform to distinct, highly polarized follicle-associated epithelial cells. Single GIIF3-positive cells are also present in the interfollicular epithelium. The NIC2 MAb recognized mesenchymal cells harbor in the lymphoepithelial compartment of the folliculus, and they elaborate cytoplasmic granules. Around Day 20 of embryogenesis large amount of NIC2-positive substance appear extracellularly in the medulla and around it. This period well correlates with the starting up of the bursal functions; clonal expansion of B cells, and generation of immune repertoire. After hatching the NIC2 stainability diminishes, and it is restricted to the medullary bursal secretory dendritic cells. The NIC2-positive, possibly elderly bursal secretory dendritic cells, are capable for migration into the follicle-associated epithelium. In eight-day old birds some cells of the follicle-associated epithelium reveals temporary NIC2 positivity, that may prove the transport of the follicle-associated epithelial cells into luminal direction. By 12 weeks of age the presence of NIC2-positive substance in the intercellular space of the FAE, rather than in the cells of FAE may indicate the termination of the transport of secretory substance. In conclusion, two types of mesenchymal cells enter the surface epithelium of the bursal folds. The GIIF3-positive cells appear on the luminal surface of the follicles and occupy the place of the follicle-associated epithelial cells. The NIC2-positive cells become secretory in nature and differentiate to bursal secretory dendritic cells. The follicle formation possibly, requires the joint presence of both GIIF3 and NIC2 cells in the epithelium.

Animals↗

Steel and c-kit in the development of avian melanocytes: a study of normally pigmented birds and of the hyperpigmented mutant silky fowl.

We describe here the expression of c-kit and Steel (Sl) genes during the development of melanocytes in normally pigmented strains of chick and quail compared to unpigmented (White Leghorn) and hyperpigmented (Silky Fowl) strains of chickens. By using the quail/chick chimera system, we found that the neural crest cells, which migrate dorso-laterally in the subectodermal mesenchyme to give rise to the melanocytes, express c-kit as early as E4, that is about 2 days after they have left the neural primordium. The Sl gene is expressed from E4 onward in the epidermis but not at all in the dermis at any developmental stage. As feather buds develop, Sl mRNA becomes restricted to the apical region of the feather filaments. During formation of the barbs and barbules of the down feather, production of the Steel factor is restricted to the external epidermal cells of the barbules. The cell bodies of the c-kit-positive melanocytes are then located in the internal border of the epidermal ridges and extend their processes toward the source of the Steel factor. We propose that the spatial restriction of Sl gene activity at that stage accounts for the morphology of the melanocytes and their vectorial secretion of melanin to the external barbule cells. As a whole, these results show that during skin development c-kit positive cells are present in the Steel factor-producing areas at the time when melanoblasts proliferate and differentiate. Interestingly, in the mouse, previous studies showed that the Sl gene is activated in the dermis where melanoblasts undergo most of their expansion (Nishikawa et al. [1991] EMBO J. 10:2111-2118). In the unpigmented and hyperpigmented mutants that we studied, expression of the Sl message, as judged quantitatively in Northern blots (for the SF embryos) or spatially by in situ hybridization, is similar to that observed in normal birds. In SF embryos the c-kit expressing melanoblasts migrate initially in the dorso-lateral migration pathway as in normal birds. However their number increases considerably in the dermis from E5 onward. From E7, they invade mesodermally derived organs that do not express the Sl gene. This suggests that another, still unknown, factor(s) is responsible for the survival, the proliferation, and the extensive spreading of melanocytic cells within the mesoderm of this mutant.

Animals↗

Isolation, culture, and preliminary characterization of mucin-producing cells from trachea of the domestic fowl.

Primary cell cultures enriched in mucin-producing cells and basal cells were established from the trachea of the domestic fowl. Epithelial cells were selectively removed from the trachea after incubation in 0.1% pronase/0.1% EDTA in Moscona's saline. The majority of the ciliated cells were removed during the initial 30 minutes of incubation. After 50 minutes of incubation, aggregates of mucin-producing cells and basal cells were removed in large numbers. The cellular aggregates rapidly attached to a collagen-coated substratum and the cells spread out on the culture surface. The mucin-producing cells retained their AB/PAS-reactive secretory granules. The basal cells replicated and as the culture approached confluency, these cells developed a fine dusting of AB/PAS-reactive material; later, larger secretory granules appeared in the cells. These observations suggest that mucin-producing cells are capable of retaining their AB/PAS-reactive secretory products in primary culture and that basal cells are capable of differentiating into mucin-producing cells in vitro.

Animals↗

Density of arteriovenous anastomoses in some skin areas of the domestic fowl (Gallus domesticus).

The vascularity and the density of arteriovenous anastomoses (AVAs) were studied in the skin of the domestic fowl by using vascular injections and histological sections. The density of AVAs and associated blood vessels were low in the wattles and in the thoracic skin (five to 23 AVAs per cm2 and five to 14 AVAs per cm2, respectively), whereas the eyelids were highly vascular and contained many AVAs (170-172 AVAs per cm2). It is suggested that the blood vessels in the eyelids are important in the temperature regulation of the eye and that AVAs in the thoracic skin may contribute to the control of blood flow through the brood patch.

Animals↗

Substance P, somatostatin, and methionine enkephalin immunoreactive elements in the spinal cord of the domestic fowl, Gallus domesticus.

The occurrence and distribution of substance P (SP), somatostatin (SOM), and enkephalin (ENK) immunoreactive elements were examined in the spinal cord of the domestic fowl, Gallus domesticus. SP immunoreactive fibers and their varicosities were densest in laminae I and II, although they were also found within deeper regions of the dorsal horn. In contrast, the intermediate gray area, the area around the central canal, and the ventral horn, contained fewer SP immunoreactive fibers. The distribution of ENK immunoreactivity in the gray matter was similar to that described for SP although immunoreactive fibers were denser around the central canal and in the intermediate zone. Few SOM immunoreactive fibers were present in the dorsal horn, the area around the central canal, and the ventral horn. All three peptidergic immunoreactive elements were found in and around the nucleus of Terni, an autonomic area. Throughout the lumbosacral enlargement SP, SOM, and ENK immunoreactive varicosities were found adjacent to the lumbosacral sinus and in fibers traversing the glycogen body. In addition, at caudal lumbar and rostral sacral levels a plexus of SP and SOM immunoreactive fibers was observed to be in close relationship with presumed motoneurons.

Animals↗

Quail melanoblast migration in two breeds of fowl and in their hybrids: evidence for a dominant genic control of the mesodermal pigment cell pattern through the tissue environment.

In the Silkie fowl large numbers of melanocytes invade most internal tissues and organs. The factors involved in this internal pigment cell pattern were studied by grafting quail neural tube segments into White Leghorn, White Silkie, and F1 hybrids (White Silkie male X White Leghorn female). Sections of quail neural tube five somites long, excised at the level of the last formed somites, were grafted isotopically and ischoronically. Various tissues and organs (mesenteries, muscles, testis, ovary, mesonephros, metanephros, and adrenals) excised from the internal region corresponding to the peripheral transverse strip of quail melanocytes, were studied after staining by the Feulgen-Rossenbeck technique. Despite some variations in pigment cell density, Silkie and hybrid grafted embryos exhibited an extensive quail internal pigmentation similar to the melanocyte distribution in the Silkie breed. In white Leghorn host embryos, the internal pigmentation remained limited. These results show the part played by tissular factors in the expression of the Silkie pigment phenotype and that this genetic tissular character is dominant. On the contrary, White Leghorn embryos, grafted with Silkie neural tube segments, never exhibited any internal pigmentation; the melanocytes deriving from the grafted Silkie neural tube were only localized at the dermoepidermal level. Thus, the migrating and/or differentiating capabilities of the Silkie premelanoblasts are different from those of quail premelanoblasts. The sex-linked inhibitor of the White Leghorn tissue interferes at the level of the pigment cells of chickens but not of quails.

Animals↗

C pigment locus mutants of the fowl produce enzymatically inactive tyrosinase-like molecules.

Three albino mutants of the fowl were tested for tyrosinase activity. Two of these mutants (c and ca) are alleles at the autosomal C locus, while the third mutant (sal) is sex-linked. Both the standard type, E, and sal are tyrosinase positive whereas the two C mutants are tyrosinase negative. Anti-chicken tyrosinase mouse serum was produced and all four genotypes were found to have cross-reacting material to this antiserum. Tyrosinase from the standard type was isolated and its location on denaturing two-dimensional gels determined. A co-migrating series of spots was found within the protein pattern of both the standard type and the tyrosinase positive albino, sal. The same pattern of spots was also observed for c and ca with no apparent change in either the pI or the molecular weight. Transmembrane blots also showed spots that reacted with anti-tyrosinase serum in all four genotypes and that migrated to the same location as that of standard tyrosinase. It is proposed that both c and ca are CRM+ mutants which produce tyrosinase-like molecules that are inactive due to a change that is electrophoretically and antigenically "silent".

Alleles↗

[Electronoptical studies of the effect of 1-[p-(methylnitrosamino)-benzylidenamino]-adamantane on the fowl plague virus (FPV) in cell culture].

The adamantanamine derivative 1-[p-(methylnitrosamino)-benzylidenamino]-adamantane (MBAA) at a concentration of 40 microgram/ml demonstrated no effect on adsorption of fowl plague virus (FPV) on chick embryonal cells. The penetration of the virions took place by means of pinocytosis. In the final stages of penetration the virions became gradually disintegrated. Under the influence of MBAA, after break-down of the membrane of pinocytic vesicles a swollen part of the virus core remained in cytoplasm. The morphologically visible replication stages were completely blocked by MBAA. From these results it was concluded that the antiviral action of MBAA most probably depends on a block of virus replication between the final stages of the penetration process and the beginning of production of virus specific structural antigens.

Adamantane↗

Fertilizing competency of multiple ovulated eggs in the domestic fowl (Gallus domesticus).

Fertilizing competency of multiple ovulated eggs in the domestic fowl was examined by fertilization in vitro and early development in culture. Normal laying hens (White Leghorn) were treated with 75 IU of PMSG for 7 days followed by injection of anterior pituitary extracts from chickens (CAPE). Ovulation began to occur 7.5 h after injection of CAPE. These hens ovulated 1-7 ova but some premature ovulation of GV stage ova were observed. In vitro fertilization of the multiple ovulated ova was examined by inseminating 10(6)-10(7) sperm onto the germinal disks in m-Ringer's solution. The gamete or zygote nuclei were detected by DNA specific fluorescence using DAPI (4',6'-diamidino-2-phenylindole) in the histological section prepared from the germinal disk. Process of fertilization was examined in the eggs incubated for 4 h after insemination in DMEM + liquid albumen at 41 degrees C under the atmosphere of 5% CO2 in air. Fertilization rate of the total multiple ovulated eggs was 55% (11/20), in which 90% (9/10) and 10% (1/10) in the eggs recovered 7.5-8.5 h and 9.0-9.5 h after CAPE injection were obtained, respectively. Normal pronuclei were formed in five eggs of those recovered 7.5-8.5 h after CAPE injection. Early development after fertilization in vitro was also examined by incubation for 12 h in DMEM + liquid albumen at 41 degrees C under the atmosphere of 5% CO2 in air. Although development in vitro was delayed compared to that in utero condition, normal development was observed in naturally and multiple ovulated eggs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Changes of elemental concentrations around and on the surface of fowl sperm membrane during maturation in the male reproductive tract and after in vitro storage.

X-ray microprobe analysis was performed to investigate the changes of elemental concentrations around or on the membrane of the head, midpiece, and principal piece regions of individual fowl spermatozoa during maturation in the male reproductive tract and after storage in vitro at 4 degrees C. The pattern of change of elemental concentrations during maturation and postejaculation was, in general, similar in the three different subcellular regions; i.e., concentrations of sodium, potassium, chlorine, and calcium decreased gradually during sperm passage through the male reproductive tract and after storage. Phosphorus concentration remained almost constant in the male tract and decreased gradually after storage. In contrast, magnesium, zinc, and copper concentrations showed an interesting pattern: concentrations increased significantly during maturation to a maximum at ejaculation and decreased again after storage. The ratios of sodium to potassium in the midpiece region showed patterns similar to those of magnesium, zinc, and copper concentrations.

Animals↗

Crystallization, preliminary X-ray diffraction study, and crystal packing of a complex between anti-hen lysozyme antibody F9.13.7 and guinea-fowl lysozyme.

The complex formed between the Fab fragment of a murine monoclonal antihen egg lysozyme antibody F9.13.7 and the heterologous antigen Guinea-fowl egg lysozyme has been crystallized by the hanging drop technique. The crystals, which diffract X-rays to 3 A resolution, belong to the monoclinic space group P2(1), with a = 83.7 A, b = 195.5 A, c = 50.2 A, beta = 108.5 degrees and have two molecules of the complex in the asymmetric unit. The three-dimensional structure has been determined from a preliminary data set to 4 A using molecular replacement techniques. The lysozyme-Fab complexes are arranged with their long molecular axes approximately parallel to the crystallographic unique axis. Fab F9.13.7 binds an antigenic determinant that partially overlaps the epitope recognized by antilysozyme antibody HyHEL10.

Animals↗

The influence of cryopreservation on parameters of energetic metabolism and motility of fowl spermatozoa.

The objective of this study was to estimate the effect of cryopreservation on the main pathways of energetic metabolism and motility of fowl spermatozoa. Sperm diluted 1:5 with the cryoprotective medium containing ethylene glycol (1.4 M final concentration) was frozen at the rate of 2-3 degrees C/min to -25 degrees C with a pause on the plateau of crystallization and then at an exponentially increasing rate to -196 degrees C. The frozen sperm was thawed in two successive water baths at 0 and at 41 degrees C. After cryopreservation, the rate of radioactive glucose oxidation to 14CO2 slightly decreased, the rate of labeled glutamate oxidation remained unchanged, and the rate of labeled succinate oxidation increased two-fold. After freeze-thawing, the rates of endogenous respiration with and without 2,4-dinitrophenol decreased; the oxidation rate of exogenous succinate in the presence of 2,4-dinitrophenol, rotenone, and digitonin slightly decreased; and the rate of respiration in the presence of ascorbate, N,N,N',N'-tetramethyl-p-phenylenediamine, antimycin A, 2,4-dinitrophenol, and digitonin did not differ from that seen in control. Sperm respiration was highly sensitive to rotenone; antimycin A and cyanide blocked oxygen consumption completely. Succinate, added after 2,4-dinitrophenol and rotenone, stimulated respiration of thawed spermatozoa, which indicated plasma membrane damage. The addition of exogenous malate in the presence of 2,4-dinitrophenol and digitonin restored the respiration rate of thawed spermatozoa to that of unfrozen cells. The rate of respiration of thawed spermatozoa with oligomycin was higher than that of control cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Testosterone and aggression in male red jungle fowl.

We investigated the relationship between aggression, plasma testosterone level (T), and change in T in captive male red jungle fowl (Gallus gallus). T measured on day 1 of our experiment was positively correlated with T on day 8, suggesting that T remains constant in males when the social environment is stable. During aggressive encounters that escalated to include physical combat, males that attacked first (won) had increased T relative to their opponent. Males did not differ in T measured 1 week before the aggressive encounter. Our data suggest that an increase in T during aggression is associated with winning in escalated fights.

Aggression↗