Untreated Chicks Research Articles

Suppression of NK activity of spleen cells by chicken fetal antigen present on Marek's disease lymphoblastoid cell line cells

Chicken fetal antigen (CFA) expressed on the cell surface of Marek's disease (MD) lymphoblastoid cell line (MDCC-MSBI) cells was purified by affinity chromatography using a monoclonal antibody (2H3). A 4-hr 51Cr release assay was performed using spleen cells as effector cells to investigate the role of the CFA in the immune response of chickens, especially in relation to natural killer (NK) activity. Spleen cells of specific-pathogen-free (SPF) chickens showed reduced NK activity in the presence of CFA in vitro, as did those cells treated with MSBI soluble antigen. NK activity of spleen cells from chickens treated with CFA was also suppressed when compared to the cells from untreated or ovalbumin-treated chickens. In addition, MSBI clo. 18 transplantable tumors grew progressively in some of the 14-day-old chickens treated with CFA, whereas the tumors regressed in age-matched chickens with or without treatment by ovalbumin.

Alterations in beta-adrenoceptor number and catecholamine content of chick atria after reversible sympathetic denervation with 6-hydroxydopamine.

1. The characteristics of [3H]-dihydroalprenolol (DHA) binding were determined in atria from untreated chicks. [3H]-DHA binding to atrium homogenates was rapid (k1 = 8.52 X 10(8) l mol-1 min-1), reversible (k-1 = 0.47 min-1), saturable, and of high affinity (KD = 0.6-1.0 nmol/l). Isoprenaline competed for specific [3H]-DHA binding in a stereoselective manner; IC50 values (mumol/l) were: (-)isoprenaline 0.12, (+)isoprenaline 4.7. 2. The number of [3H]-DHA binding sites and catecholamine content of left and right atria were examined after injection of chicks with a single dose of 6-hydroxydopamine hydrobromide (100 mg/kg). There were transient increases in the number of [3H]-DHA binding sites in both the left and the right atrium after 6-OHDA treatment. These increases were quicker in onset and in offset in the right atrium than in the left atrium. [3H]-DHA binding was significantly increased in the left atrium at 5 and 7 days, and in the right atrium at 3 and 5 days after 6-OHDA injection. 3. Saturation binding isotherms indicated that the increase in [3H]-DHA binding was due to an increase in beta-adrenoceptor number with no change in affinity for [3H]-DHA. 4. Twenty four hours after 6-OHDA treatment there was a significant (80%) decrease in noradrenaline content of left and of right atria. 5. The decrease in noradrenaline content was reversible, noradrenaline levels returning to 55% of control in left atrium and to 71% of control in right atrium by 21 days after 6-OHDA treatment. These changes are consistent with reversible sympathetic denervation of the atria.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of the estrogen-sensitive cells expressing progesterone receptor in the bursa of Fabricius

Cells expressing the progesterone receptor (PR) in the bursa of Fabricius (BF) were studied with immunohistochemistry at light-microscopic level, with immunoelectron microscopy (immuno-EM) and with non-specific esterase histochemistry. The antibody (IgG-RB) directed to the B component of the chick oviduct progesterone receptor was shown by immunoblotting to be specific for the PR and to recognize the PR also in the bursa. Two cell types in the BF contain the PR: stromal cells in the interfollicular-subepithelial area and smooth muscle cells lining the BF. The PR was localized in the nuclei of these cells. The bursal epithelium and the cells inside the follicles were not stained for PR. Electron microscopically the immunoreaction precipitate was localized on condensed heterochromatin and on dispersed euchromatin. The cells expressing the PR resembled electron microscopically fibroblasts. Their cytoplasm was rich in rough endoplasmic reticulum indicating active protein synthesis. By non-specific esterase histochemistry we showed that the PR-containing cells were not macrophages, which are morphologically indistinguishable from stromal cells. In the bursae of young untreated chicks the PR was not seen, but was inducible by estradiol treatment and was spontaneously expressed after the onset of sexual maturation. It is concluded that both the stromal fibroblasts and the smooth muscle cells in the BF are estrogen and progesterone sensitive. The expression of PR after the onset of sexual maturation indicates that the BF is directly affected by sexual maturation-associated factors. We suggest that estrogen and progesterone participate in tissue remodelling during bursal involution via the stromal cells and may affect bursal functions via the smooth muscle cells.

Effects of experimental immunosuppression on reovirus‐induced tenosynovitis in light‐hybrid chickens

Groups of specific pathogen-free light-hybrid chickens which had been immunosuppressed either by surgical thymectomy (Tx) or surgical bursectomy (Bx) or cyclophosphamide (Cy) treatment or Tx plus Cy treatment (Tx + Cy), as well as intact (untreated) birds, were inoculated with graded doses of an arthrotropic avian reovirus at 1 day of age and observed up to 5 weeks post-inoculation (p.i.). Cy-treatment with or without Tx considerably increased the mortality, incidence of gross leg lesions and severity of microscopic lesions due to reovirus infection. The Bx group showed only a significant increase in mortality, and the Tx group response was generally similar to the untreated group. Dead birds showed hepatic necrosis, which in Cy-treated groups (Cy, Tx + Cy) was associated with calcification. Surviving Cy-treated birds had acute tenosynovitis characterised grossly by large amounts of serous exudate in leg tendon sheaths, and microscopically by a massive heterophilic but only mild lymphocytic infiltration of tendon sheaths. Tenosynovitis lesions in Bx birds were generally similar to those of the untreated chickens, i.e. grossly small amounts of yellowish brown gelatinous exudate and microscopically moderate chronic inflammatory changes in leg tendon sheaths. In Tx birds gross lesions were rarely seen and the microscopic lesions were often very mild. Reovirus could be recovered from cloacal swabs from untreated and Tx birds for 2 weeks, Bx birds for 3 to 4 weeks, and Cy and Tx + Cy chickens continuously throughout. Reovirus was isolated from tendon tissue of all Cy and Tx + Cy infected birds examined at 5 weeks p.i. and gross tenosynovitis lesions were seen in all birds. The virus was recovered from the tendons of only a proportion of the infected untreated, Tx and Bx groups, and overall more frequently from apparently normal birds. This was especially marked in the infected Tx group. Antibody responses as shown by gel precipitation and virus neutralisation were positive and similar in untreated and Tx birds, were delayed in the Bx group with the precipitation test only, and absent from most of the Cy and Tx + Cy birds. The results of these experiments indicate that recovery from reovirus infection probably involves both B- and T-cell systems but that the B-cell system is predominantly protective.

Synthetic seleno-organic compound with glutathione peroxidase-like activity in the chick

The glutathione peroxidase activity catalyzed by the seleno-organic anti-inflammatory drug Ebselen (registered under the trademark of the Natterman Corp. Cologne, FRG) [PZ51, 2-phenyl-1,2-benzisoselenazol-3(2 H)on], as measured by NADPH oxidation, was inhibited in vitro by the selenium-dependent glutathione peroxidase (SeGSHpx) inhibitors aurothioglucose and D-(−)penicillamine HCl. Vitamin E- and selenium-deficient chicks were given 0, 80 or 320 ppm PZ51 in diets devoid of vitamin E and supplemented with low levels of sodium selenite (0.04 ppm selenium added to the basal diet containing ca. 0.015 ppm selenium) when a small number of chicks ( ca. 13%) had exudative diathesis (ED). By 24 hr, the high PZ51 dose (320 ppm) delayed the onset of ED compared to untreated controls. Similarly, vitamin E-deficient chicks fed diets containing 0, 80, 160, 320, 640 or 1280 ppm PZ51 and supplemented with 0.04 ppm selenium showed ED in inverse proportion to log PZ51 dose. Plasma and liver post-mitochondrial supernatant samples from these chicks also exhibited log-linear relationships between dietary PZ51 level and selenium content or SeGSHpx-like activity. The amount of SeGSHpx-like activity for chicks given PZ51 above that determined for untreated chicks was extractable into ethanol, indicating that those PZ51-associated increases were not due to protein-bound selenium or SeGSHpx. This suggests that selenium from PZ51 was not available to support synthesis of SeGSHpx. Dietary PZ51 (1280 ppm) or selenium (0.1 ppm) alone or in combination decreased the acute lethalities of nitrofurantoin or paraquat in vitamin E-adequate chicks. The results indicate that SeGSHpx-like activity in selenium-deficient chicks is increased by oral administration of PZ51, which appears to mimic the true enzyme by affording protection against clinical signs of selenium deficiency (i.e. ED) and proxidant drug lethality.

Role of the supraoptic decussation in the development of asymmetry of brain function in the chicken

The supraoptic decussation plays an important role in the development of asymmetry of brain function for visual discrimination learning in the chicken. Lesioning the decussation on day 2 has no effect on the asymmetry normally revealed by unilaterally treating either the left or right forebrain hemisphere with cycloheximide on day 4 of life. However, the lesion removes the asymmetry revealed by cycloheximide treatment on day 8 and reverses that revealed by treatment on day 10, albeit to a lesser extent. The asymmetry of learning performance between the left and right eyes present in untreated chickens tested monocularly in the second week of life is also removed by lesioning the supraoptic decussation. Thus, in the first week of life functional asymmetry is present but not dependent on left-to-right side coupling via the supraoptic decussation. The difference between lesioning during the first and second week of life may relate to the relative degree of maturation in the supraoptic decussation.

Acute hematogenous staphylococcal osteomyelitis: the effects of surgical drilling and curettage in an animal model

Surgical drilling and curettage of metaphyseal medullary bone were performed in chickens 4, 8 and 15 d after bacterial inoculation with Staphylococcus aureus producing osteomyelitis. Intramedullary drilling and curettage resulted in extensive vascular damage and necrosis of medullary bone but failed to remove all viable bacteria. An effective continuing drainage channel for the pus was not established due to the formation of blood clot within the drill hole. Surgery performed before the formation of a sequestrum resulted in further spread of bacteria within the medullary cavity and the formation of a larger sequestrum than that commonly observed in the bones of infected, untreated chickens. Surgery performed after the formation of a sequestrum destroyed the abscess wall thus disrupting a natural defence mechanism of the host. Whilst a direct comparison of surgical drilling in human osteomyelitis and in this avian model cannot be made, the results suggest that production of an artificial drill hole across the cortex and curettage of the bone in human osteomyelitis would be destructive and may not be warranted.

Expression of Viral Arthritis-Tenosynovitis Infection in Immunosuppressed Chickens

Previously untreated chicks and those treated neonatally with either cyclophosphamide (CY) or infectious bursal disease virus (IBDV) were subcutaneously inoculated with viral arthritis virus (VAV) to determine whether suppression of humoral immunity would affect VAV pathogenicity. No difference was apparent in the disease process that occurred in the immunosuppressed versus the nonimmunosuppressed chickens. However, in the immunosuppressed birds, VAV was recovered from the heart and tendon tissues earlier in the postinoculation (PI) period. Also, VAV inoculated immunosuppressed birds carried higher levels of serum gamma globulins at certain PI intervals than the VAV inoculated positive controls or the untreated negative control chickens. It is suggested that the suppression of the bursa of Fabricius (BF) by CY or IBDV did not interfere in the antibody production to VAV, but the antibody produced was qualitatively defective.

Stimulation of lipid-linked oligosaccharide assembly during oviduct differentiation.

Regulation of dolichyl phosphate-linked oligosaccharide assembly has been studied during the course of diethylstilbestrol-induced chick oviduct differentiation. Oviduct membranes from treated chicks form 4.6 times as much GlcNAc-P-P-Dol and GlcNAc2-P-P-Dol upon incubation with UDP-[14C]GlcNAc and MgCl2 than do membranes from untreated chicks. Assembly of oligosaccharide-lipid was studied by incubation of membranes with purified exogenous [14C]GlcNAc2-P-P-Dol and GDP-Man. Man transfer required a divalent cation (10 mM Mg2+) and detergent (0.5% Nonidet P-40 is optimal) and occurs in the presence of amphomycin (500 micrograms/ml). The apparent Km for GDP-Man is 1 microM and for [14C]GlcNAc2-P-P-Dol is 0.45 microM. The products are a series of sequentially formed dolichyl pyrophosphate-linked saccharides up to Man5GlcNAc2, the first of which is Man beta 1,4GlcNAc2. The same products are formed either in the presence or absence of amphomycin. Conversion of GlcNAc2-P-P-Dol to higher oligosaccharides is stimulated 3-fold by estrogen treatment of chicks. Similarly, the conversion of partially purified exogenously added Man beta-[14C]GlcNAc2-P-P-Dol is 4.6-fold higher after diethylstilbestrol treatment.

Distribution of IgM-, IgG-, and IgA-positive cells in lymphatic organs of normo- and dysgammaglobulinemic UM-B19 chickens.

Lymphatic organs of normo- and dysgammaglobulinemic chickens were studied by immunohistochemical methods concerning IgM-, IgG-, and IgA-positive cells. IgM- and IgA-positive lymphocytes and plasmacytic cells are similarly distributed in dys- and normogammaglobulinemic chickens. Follicles of the bursa of Fabricius contain cortical lymphocytes weakly and medullar lymphocytes distinctly stained by anti-IgM, as well as some larger irregularly shaped cells with intensely labeled cytoplasm. In spleen, periellipsoid lymphocytes, some red pulp lymphocytes, and varying numbers of plasmacytic cells are also distinctly labeled by anti-IgM. IgA-positive lymphocytes are not present in the bursa of Fabricius, but the bursal medulla is populated by some irregular, mostly longish cells with cytoplasmic IgA reaction. Spleen contains a few IgA-labeled lymphocytes and plasmacytic cells. The only difference between normo- and dysgammaglobulinemic chickens is found in IgG-positive cells, which are absent in permanently dysgammaglobulinemic chickens. The results from dysgammaglobulinemic chickens show that IgA can also be produced in untreated chickens lacking IgG synthesis and suggest that the theory of an IgM leads to IgG leads to IgA sequence is unlikely.

Ultrastructural immunocytochemical localization of carbonic anhydrase in normal and calcitonin-treated chick osteoclasts.

Carbonic anhydrase was localized in osteoclasts by light and electron microscopy using a preembedding peroxidase-antiperoxidase staining method. Osteoclasts on the endosteal surface of the metatarsi of calcitonin-treated and untreated (control) chicks were studied. A positive staining reaction was seen in the cytosol, in the Golgi apparatus, in and on vesicles, on the plasma membrane of the ruffled border, and on the bone surface beneath control osteoclasts. After calcitonin treatment, positive staining reactions were seen at the same sites except that staining was absent on the plasma membrane and endosteal bone surface. The morphology of osteoclasts from calcitonin-treated chicks was indicative of cell inactivity. The carbonic anhydrase which was bound to the plasma membrane of the ruffled border is appropriately arrayed for hydrogen ion secretion and subsequent mineral dissolution. The presence of the enzyme within lysosomelike vesicles and on the endosteal surface beneath active cells suggests that it may be released into the resorbing zone along with the lysosomal hydrolases. The function of the extracellular enzyme is also unknown.

Adherence of Salmonellae and Native Gut Microflora to the Gastrointestinal Mucosa of Chicks

Adherence of salmonellae to the mucosa of the cecum was demonstrated using scanning electron microscopy. In the absence of other microflora, adherence was also shown at all other levels of the alimentary tract tested. Comparative development of native gut microflora in the ceca was also investigated in chicks treated with selected fecal microflora and in untreated chicks using scanning electron microscopy. In the treated chicks, there was early colonization by adherent bacteria interconnected with fibers, forming a mat of microflora. This mat of microflora may be responsible for preventing colonization of salmonellae in the ceca.

Detection of Inhibitors in Chicken Tracheal Washings against Mycoplasma gallisepticum

A chicken tracheal ring tissue-culture system was used to demonstrate that specific and nonspecific substances in chicken tracheal secretions inhibit the cilia-stopping effect (CSE) of Mycoplasma gallisepticum (MG). In chickens less than six weeks old, specific inhibitory substances were found in tracheal washings from chickens inoculated with MG. Nonspecific inhibitory substances were found in tracheal washes from untreated chickens seven weeks or older. Complement may play a role in the inhibition of CSE. Secretory immunoglobulin A was not detected in the tracheal washes.

Reduction of Salmonella Excretion into Drinking Water Following Treatment of Chicks with Nurmi Culture

Day-old chicks (Gallus domesticus) were treated with cultured feces of adult chickens according to the Nurmi concept and were challenged 2 days later with Salmonella typhimurium. Treated chicks were less susceptible to infection than untreated chicks (16% vs. 79% infected). Those treated chicks that did become infected, contaminated their drinking water with fewer Salmonella than the untreated chicks (maximum of 104/ml vs. ⩾ 107/ml). Fecally contaminated water may be a major source for spreading Salmonella infection within a flock.

Prevention of Salmonella Infection in Chicks by Treatment with Fecal Cultures from Mature Chickens (Nurmi Cultures)

Chicks (Gallus domesticus) were treated per os with 24-h-old anaerobic cultures of feces from mature chickens 1 day after hatching, challenged with Salmonella typhimurium in the drinking water 2 days later, and sacrificed on day 11 or 12; then the lower third of the intestinal tract was examined for salmonellae. Cultures of feces inoculated directly into the crop or added to the drinking water, even after holding at −70 C for 21 days, protected chicks against infection by S. typhimurium. Cultures serially subcultured daily up to four times were protective, and dilution to 1:80 in drinking water containing 4 % skim milk powder did not decrease their protective effect. Treated chicks were about 1000-fold more resistant to infection by Salmonella than untreated chicks.

Increased growth of RSV-induced tumors in chickens partially tolerant to MHC alloantigens.

Chickens with B2B2 MHC genotypes were made partically tolerant to B5 MHC cell-surface antigens and the fate of their Rous-sarcoma-virus (RSV)-induced tumors was determined. B2B2 chickens partially tolerant to viable or lysed white blood cells (WBC) or viable red blood cells (RBC) from B5B5 chickens had a significantly higher incidence of tumor progression than untreated, PBS-treated, or B2B2 chickens inoculated with WBC from other B2B2 chickens. The criteria for tolerance were absence of antibody titer to the cell type inoculated and acceptance of allografts from B5B5 donors by B2B2 chickens. Graft-vs-host reactions occurred only in B2B2 chickens inoculated with viable WBC from B5B5 chickens. It appears that B2B2 chickens partially tolerant to B5 antigens failed to mount a successful immune response to RSV-induced tumors partly because of a B5 MHC antigen(s) cross-reacted with a tumor associated antigen(s) thereby severely limiting B2B2 host recognition of the tumor as foreign. Since WBC and RBC cell-surface antigens appear to contribute similarly to the effect, the B-F- region of the MHC may be involved.

In vivo effects of protease inhibitors on chickens with hereditary muscular dystrophy.

Beginning on day 4 ex ovo, and every 3 d thereafter, genetically dystrophic Line 413 chickens were given intraperitoneal injections (4 mg/kg body wt) of a protease inhibitor, leupeptin, pepstatin, or antipain. Experimental chickens received protease inhibitors dissolved in a water:ethanol:dimethyl sulfoxide solution (50:40:10, vol:vol:vol). Control untreated animals received diluent injections. Untreated dystrophic chickens typically reach around day 30 ex ovo a maximum ability to right from the supine position in a standardized functional test for muscle weakness. After day 30 ex ovo, the dystrophic chickens are found to decline progressively in their ability to right, compared with normal, nondystrophic controls, which have an unimpaired ability to right. Concomitantly, dystrophic chickens exhibit characteristically high levels of plasma creatine phosphokinase enzyme activity. In addition, an increased frequency of degenerating, regenerating, and vacuolated myofibers, and inflammatory cells appear in the affected pectoralis major muscles from the dystrophic chicken. Throughout the duration of the trial, there was no major enhancement in the functional righting ability of dystrophic chickens receiving any one of the protease inhibitors tested. However, there was a significant reduction in the abnormally high levels of plasma creatine phosphokinase in the treated chickens. Also, there was an apparent reduction in the mean number of vacuolated fibers in the pectoralis muscle from the protease inhibitor-treated birds. No significant reductions were observed in the relative frequency of degenerating and regenerating myofibers or inflammatory cells. In addition to the plasma creatine phosphokinase decrease, however, therapeutic benefit was seen in 31.0, 30.5, and 14.8% increases in the wet weight (and total noncollagen protein) of pectoralis muscle from dystrophic chickens receiving leupeptin, pepstatin or antipain, respectively.

The in vivo effect of levamisole on phytohaemagglutinin stimulation of lymphocytes in normal and Marek’s disease virus inoculated chickens

Subcutaneous administration of levamisole al 2·5 mg/kg enhanced phytohaemagglulinin (PHA) stimulation of peripheral blood lymphocytes in normal chickens. In a sequential sludy, levamisole initially enhanced PHA stimulation in Marek’s disease virus inoculated chickens compared to untreated virus inoculated chickens. Significant virus induced suppression of PHA stimulation was observed on day 6 after inoculation in levamisole treated chickens compared to days o and 9 alter inoculation in untreated chickens.

Enhanced chick oviduct dolichol kinase activity during estrogen-induced differentiation.

Crude microsomal preparations from hen oviduct catalyze the transfer of [32P]phosphate from [gamma-32P]CTP or [gamma-32P]dCTP to endogenous dolichol, forming dolichyl [32P]monophosphate. The oviduct kinase activity assayed with [gamma-32P]CTP is stimulated by divalent cations and exogenous dolichol. The enzymatic formation of dolichyl [32P]monophosphate is inhibited by dCDP and CDP, but not CMP, ADP, GDP, or UDP. The hen oviduct kinase is inhibited 50% by the addition of 38 microM CDP, but 101 microM dCDP is required for 50% inhibition. The amount of dolichol kinase activity in chick oviduct microsomes increases 3.7-fold within 10 days of estrogen administration. The hormone-induced increase in kinase activity is also observed when membranes from untreated and estrogen-treated chicks are assayed in the presence of saturating levels of exogenous dolichol. The microsomal preparations from oviducts of untreated chicks and fully induced birds both exhibit an apparent Km value of 7.1 microM for CTP. An apparent Km of 14 microM has been determined for dCTP. Thus, the developmental change in dolichol kinase activity does not appear to be the result of a difference in the amount of available endogenous dolichol or an alteration in the reactive site for the nucleoside triphosphate substrate, but is probably due to an increased level of the enzyme.

Studies on the metabolism of glycogen and adenine nucleotides in embryonic chick liver at the end of incubation

1. In embryonic chick liver glycogen mobilization proceeds parallel to the hatching process. The glycogen stores are degraded earlier than normal when the hatching process is induced by L-thyroxine, and at a later stage of incubation when the period of incubation is prolonged by thiourea. 2. In untreated chicks glycogenolysis does not seem to be mediated by cyclic AMP (cAMP), although glycogen metabolism responds to changes of cAMP levels caused by exogenous glucagon. 3. Similar to glycogenolysis, ATP concentrations decrease towards the end of incubation. The fall in ATP is accelerated in L-thyroxine treated embryos. 4. Due to the limited O 2 supply, it seems possible that the decrease ofglycogen and ATP before hatching is released by hypoxia.