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Expression of type 1 pili by Escherichia coli strains of high and low virulence in the intestinal tract of gnotobiotic turkeys.

Highly virulent (strain 1) and weakly virulent (strain 3) Escherichia coli were examined using immunofluorescent and electron microscopic techniques to determine their ability to express type 1 pili in the intestinal tract of 3-week-old gnotobiotic turkeys. Turkeys were necropsied on postinoculation day (PID) 1, 2, 5, 8, and 12. Nonpiliated forms of strains 1 and 3 were more numerous than piliated forms in cecal and colonic contents examined by negative staining electron microscopy. A piliated form of strain 1 was seen in intestinal contents on each PID and was more numerous in cecal contents than in colonic contents. The mucus blanket of the cecum and colon contained large numbers of bacteria, although organisms were rarely intimately associated with the intestinal epithelium. Immunofluorescent staining indicated large numbers of piliated forms of strains 1 and 3 within the mucus blanket of the cecum and colon on PID 2, 5, 8, and 12. Piliated bacteria were infrequently seen in the ileal mucus blanket. Serum antibody titers to type 1 pili increased markedly by PID 5 and persisted in turkeys inoculated with strain 1. In contrast, antibody titers in turkeys exposed to strain 3 increased gradually and varied markedly among birds at each PID. Type 1 pili may not be important for adherence of pathogenic E coli to intestinal epithelium of turkeys.

Animals↗

Isolation and serial propagation of turkey rotaviruses in a fetal rhesus monkey kidney (MA104) cell line.

Turkey rotaviruses from the intestinal contents of poults were isolated and serially propagated in MA104 cell monolayers by a simple procedure. The initial virus isolation was done by low-speed centrifugation of the inoculum onto the monolayers, and subsequent passages were accomplished in roller-tube monolayers using trypsin-treated virus suspensions. Each of the turkey rotavirus isolates possessed the morphologic, antigenic, and genomic attributes characteristic of turkey group A rotaviruses. Attempts to isolate and serially propagate turkey rotavirus-like viruses in MA104 cell monolayers by this procedure were unsuccessful. Turkey reoviruses also did not serially propagate in MA104 cell monolayers by this procedure.

Animals↗

Turkey erythrocyte membranes as a model for regulation of phospholipase C by guanine nucleotides.

Phosphoinositides of human, rabbit, rat, and turkey erythrocytes were radiolabeled by incubation of intact cells with [32P]Pi. Guanosine 5'-O-(thiotriphosphate) (GTP gamma S) and NaF, which are known activators of guanine nucleotide regulatory proteins, caused a large increase in [32P]inositol phosphate release from plasma membranes derived from turkey erythrocytes, but had no effect on inositol phosphate formation by plasma membranes prepared from the mammalian erythrocytes. High performance liquid chromatography analysis indicated that inositol bisphosphate, inositol 1,3,4-trisphosphate, inositol 1,4,5-trisphosphate, and inositol 1,3,4,5-tetrakisphosphate all increased by 20-30-fold during a 10-min incubation of turkey erythrocyte membranes with GTP gamma S. The increase in inositol phosphate formation was accompanied by a similar decrease in radioactivity in phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2). GTP gamma S increased inositol phosphate formation with a K0.5 of 600 nM; guanosine 5'-(beta, gamma-imido)trisphosphate was 50-75% as efficacious as GTP gamma S and expressed a K0.5 of 36 microM. Although GTP alone had little effect on inositol phosphate formation, it blocked GTP gamma S-stimulated inositol phosphate formation, as did guanosine 5'-O-(2-thiodiphosphate). Turkey erythrocytes were also shown to express phosphatidylinositol synthetase activity in that incubation of cells with [3H] inositol resulted in incorporation of radiolabel into phosphatidylinositol, PIP, and PIP2. Incubation of membranes derived from [3H]inositol-labeled erythrocytes with GTP gamma S resulted in large increases in [3H] inositol phosphate formation and corresponding decreases in radiolabel in PIP and PIP2. The data suggest that, in contrast to mammalian erythrocytes, the turkey erythrocyte expresses a guanine nucleotide-binding protein that regulates phospholipase C, and as such, should provide a useful model system for furthering our understanding of hormonal regulation of this enzyme.

Animals↗

Serum resistance and virulence of Escherichia coli isolated from turkeys.

Twenty-five strains of Escherichia coli isolated from turkeys were characterized for their serum resistance and virulence. An in vitro bactericidal assay was used to determine the serum resistance of E coli. Virulence was determined by survival time after IV inoculation of each strain into 3-week-old turkeys. Serum-resistant E coli strains were generally found to be virulent for turkeys, whereas serum-sensitive E coli strains were avirulent. Of the 25 strains, 18 strains were placed in the 2 categories of serum-resistant/virulent and serum-sensitive/avirulent. Five strains were serum-resistant and avirulent, and 2 strains were serum-sensitive and virulent. Serum resistance appears to be an important determinant of virulence for E coli in turkeys; however, the requirement for other virulence factors, in addition to serum resistance, was suggested by the finding that 5 serum-resistant strains were avirulent in turkeys.

Animals↗

Diagnostic summary of 1986 turkey, broiler breeder, and layer necropsy cases at the University of Georgia.

Turkey, broiler breeder, and layer cases submitted for necropsy in 1986 to the University of Georgia were categorized by diagnosis, grouped by clinical features, season, age, and presenting history, and evaluated for statistically significant patterns. The most commonly diagnosed diseases were: fowl cholera in turkeys, 64 cases (34.4% of cases; 26.2% of diagnoses); staphylococcal arthritis/synovitis (SA/S) in broiler breeders, 35 cases (20.7%; 17.7%); and osteomalacia in layers, 17 cases (21.0%; 15.6%). In turkeys, fowl cholera, colibacillosis, and aspergillosis were significantly more common in the fourth, second, and third quarters of the year, respectively; enteritis and viral enteritis were more common in early grow-out, and osteomyelitis, SA/S, and synovitis were greatest in late grow-out. Overall, musculoskeletal problems in turkeys were more common in the third quarter and were less common in the first and fourth quarters. Erysipelas occurred most frequently in turkeys 50 to 112 days of age. In both broiler breeders and layers, SA/S was more common in the second quarter, and a history of musculoskeletal problems was less common in both post-peak-production broiler breeders and layers. Fowl pox was more common in the fourth quarter and in late-production broiler breeders. In layers, osteomalacia and Marek's disease were more common in the first and fourth quarters, respectively.

Age Factors↗

Passive immunization versus adhesion of Bordetella avium to the tracheal mucosa of turkeys.

Three-week-old turkeys were passively immunized with convalescent serum or treated with tracheal washings from turkeys infected with Bordetella avium. Western blot analysis of the convalescent serum and tracheal washings revealed at least two bands of interaction with outer membrane protein preparations of B. avium. Adherence of B. avium in vivo to tracheal mucosa was determined and compared in treated and untreated turkeys. Passive immunization with convalescent serum reduced adherence of B. avium to the tracheal mucosa in a dose- and time-dependent manner. Adherence was significantly inhibited (P less than 0.01) when turkeys were treated intravenously with 1 ml of undiluted serum either 1 or 6 hours previously. Incubation of the bacterial inoculum with convalescent tracheal washings or application of the washings to tracheal segments before adherence determination in vivo resulted in a significant (P less than 0.01) decrease in adherence. These results indicate that adherence of B. avium to tracheal mucosa is inhibited by substances (antibody) present in both serum and tracheal secretions of convalescent turkeys.

Animals↗

Effects of Newcastle disease virus infection on the binding, phagocytic, and bactericidal activities of respiratory macrophages of the turkey.

Effects of Newcastle disease virus (NDV) infection on the binding, phagocytic, and bactericidal activities of turkey respiratory macrophages were studied. Respiratory macrophages of the turkey demonstrated the presence of immunoglobulin (Ig) G and complement receptors but lacked IgM receptors. Respiratory macrophages from NDV-infected turkeys showed little or no depression of binding of sheep erythrocyte-IgG complexes and sheep erythrocyte-IgM-complement complexes to their appropriate membrane receptors. In contrast, respiratory macrophages from NDV-infected turkeys showed significant (P less than or equal to 0.05) depression of phagocytosis of similar complexes. Bacterial killing by respiratory macrophages from NDV-infected turkeys was significantly (P less than or equal to 0.05) inhibited.

Animals↗

Effects of exogenous iron on Escherichia coli septicemia of turkeys.

The effect of inoculation with Escherichia coli on serum iron concentrations of turkeys and the effect of exogenous iron, as ferric ammonium citrate, on E coli septicemia in turkeys were determined. Inoculation of air sacs with E coli produced hypoferremia in 18-day-old turkeys. Administration of iron with E coli significantly (P less than 0.01) increased mortality, frequency and degree of bacteremia, and severity of lesions in inoculated turkeys, compared with those in turkeys given E coli but not given iron. Similar results were seen whether iron was inoculated at the same location as E coli or at a different location.

Animals↗

Clinical outbreak of Bordetella avium infection in two turkey breeder flocks.

An acute upper respiratory disease was observed in two broad-breasted white (BBW) turkey primary breeder flocks. Associated clinical signs included sneezing, depression, and a deep dry cough originating from large conducting airways. Morbidity reached approximately 15-20% of the hens in an affected house. None of the turkeys died, and total feed consumption was not affected. A minimal effect upon egg production was noticed. Sera from an acutely affected flock exhibited a marked rise in titer to Bordetella avium compared with preinfection sera samples. In Case 1, B. avium was isolated in pure culture from affected birds. In Case 2, B. avium was diagnosed by serological results and clinical signs; bacteriological examination was not attempted. The findings presented here are consistent with an acute clinical outbreak of B. avium-induced turkey rhinotracheitis (turkey coryza) in BBW turkey breeder hens.

Animals↗

Association of complement sensitivity with virulence of Pasteurella multocida isolated from turkeys.

Two strains of Pasteurella multocida, both derivatives of strain P1059, were compared for virulence for 14-week-old turkeys and sensitivity to turkey plasma. Strain P1059-1, a nalidixic-acid-resistant mutant of P1059 with an LD50 of approximately 10(3) colony-forming units (CFU), was more resistant to the bactericidal effects of fresh turkey plasma at 37 C than avirulent strain P1059-1A. P1059-1A, with an LD50 of approximately 10(8) CFU, is an acapsular variant of P1059-1 that spontaneously arose after prolonged passage on artificial medium. The bactericidal effect on P1059-1A was removed when turkey plasma was treated with heat or with zymosan, maneuvers that removed hemolytic complement activity from turkey plasma.

Animals↗

Influence of temperature on the growth of Bordetella avium in turkeys and in vitro.

Effects of temperature on growth of three strains of Bordetella avium were determined in young turkeys and in vitro. Colonization of the tracheal mucosa by two virulent strains of B. avium was significantly greater in cold-stressed turkeys than in heat-stressed turkeys. The avirulent vaccine strain, ART-VAX, colonized tracheas of cold-stressed turkeys to a limited extent but failed to colonize heat-stressed turkeys. Growth rates of the three B. avium strains were determined in brain-heart infusion broth at 30, 35, 40, and 45 C. All three strains grew best at 35 C but were killed by 45 C. Compared with virulent strains, ART-VAX grew markedly less at all temperatures, and most cultures of ART-VAX grew at 40 C only after a variable period of declining numbers of viable bacteria. This study indicates that temperature affects growth of B. avium in vivo and in vitro and that growth of the ART-VAX strain is fundamentally different from growth of virulent strains.

Agglutination Tests↗

Prevalence of certain antibodies to selected disease-causing agents in wild turkeys in Texas.

In Texas in 1976 and 1977, Rio Grande turkeys (Meleagris gallopavo intermedia) from seven counties and Eastern turkeys (M. g. silvestris) from one county were tested for antibodies to selected poultry pathogens. Standardized serological tests disclosed reactors to Salmonella pullorum (2.4%), S. typhimurium (2.3%), and Mycoplasma gallisepticum (4.8%). There were no reactors to Newcastle disease virus or Chlamydia psittaci. Prevalence of M. gallisepticum antibody in wild turkeys was significantly higher for counties with commercial turkey operations than for counties lacking domestic turkeys, whereas the incidence of S. pullorum and S. typhimurium did not differ significantly.

Animal Population Groups↗

Neurotoxic and antihypertensive effects of phenytoin in turkeys.

Broad-Breasted White male turkeys were fed a control diet until 4 weeks of age, at which time they were randomized into a control group and two treatment groups which received 0.06% (38.7 mg/kg) and 0.09% (58.2 mg/kg) phenytoin, respectively, until termination of the experiment at 12 weeks of age. Plasma concentrations of phenytoin on the two dosages were 8.0 +/- 0.8 and 15.7 +/- 1.7 micrograms/ml. Systemic arterial blood pressure in the control turkeys was 214 +/- 5/171 +/- mm Hg and was reduced in a dose-related fashion to 185 +/- 8/143 +/- 11 and 156 +/- 5/125 +/- 4 mm Hg in the two treatment groups; likewise, the rate of systolic ejection (dp/dt maximum) was less in the phenytoin-treated turkeys. Heart rate also dropped significantly with drug administration but the difference between the two treatment groups was not significant. Evidence of neurotoxicity developed in 25% of the turkeys on the lower drug schedule; these birds had mean plasma phenytoin levels of 12.8 micrograms/ml as contrasted to the concentration of 8.0 micrograms/ml for the entire group. On 0.09% phenytoin 50% of the birds had abnormal signs and a mean phenytoin concentration of 20.5 micrograms/ml, whereas the mean drug level for the entire group on this drug level was 15.7 micrograms/ml. Early signs of neurotoxicity developed within 2 to 3 days of initiation of phenytoin and consisted of extensor rigidity of the neck and hyperactivity; at the higher drug concentrations, back pedaling and somersaulting appeared. General health and weight gain were not affected. No qualitative or quantitative changes were found in the Purkinje cells in the cerebellum of the affected turkeys.

Animals↗

Effect of lymphoproliferative disease virus and of niridazole on the in vitro blastogenic response of peripheral blood lymphocytes of turkeys.

Turkeys inoculated at 5 weeks of age with lymphoproliferative disease (LPD) virus developed typical lesions in the spleen, thymus, and pancreas. The in vitro blastogenic response of peripheral blood lymphocytes to the mitogens phytohemagglutinin and concanavalin A was drastically (up to 90%) suppressed in the inoculated turkeys 1 to 4 weeks postinoculation compared with uninoculated controls, and even at 11 weeks the response was about 50% inhibited. A lethal (about LD33) dose of antihelminthic drug niridazole, 100 mg/kg given each day for 3 days to 4-week-old turkeys, caused a transient inhibition of the blastogenic response within 32 days of treatment, which was less pronounced than that observed in turkeys inoculated with LPD virus, whether pretreated with niridazole or not. Virus-associated reverse transcriptase activity in the plasma was significantly higher in the turkeys pretreated with niridazole, and LPD lesions developed to the same extent in the untreated and treated groups, as determined 9 weeks post virus inoculation. A sublethal dose of niridazole, 50 mg/kg given each day for 4 days, did not suppress the blastogenic response to mitogens at any time determined (starting 10 days post-treatment) and did not affect the pathogenesis of LPD and the viremia. Body weights were significantly decreased by virus infection and by treatment with lethal doses of niridazole.

Animals↗

Colonization and persistence of Escherichia coli in axenic and monoxenic turkeys.

Sites of colonization and persistence of virulent (O78) and avirulent (O2) Escherichia coli were determined in 3-week-old axenic turkeys. Colonization of O78 in turkeys infected with Streptococcus faecalis was also determined. Colonization and persistence of E coli in the intestinal tract and lung were not correlated with virulence. Both serotypes persisted in high titer in crop, ileum, ceca, colon, and lung. Persistence of E coli in the liver was positively correlated with virulence, although both serotypes were recovered from the liver in the absence of bacteremia, indicating possible intestinal translocation of E coli to the liver. A reduction in intestinal colonization and translocation of O78 was determined in monoxenic turkeys infected with S faecalis. In contrast, S faecalis had no effect on lung titers of O78. Air sacculitis, pericarditis, and pneumonia were observed in axenic and monoxenic turkeys infected with O78. Bacteremia was detected in a turkey exposed to O78.

Animals↗

Hormonal control of protein phosphorylation in turkey erythrocytes. Phosphorylation by cAMP-dependent and Ca2+-dependent protein kinases of distinct sites in goblin, a high molecular weight protein of the plasma membrane.

In previous studies, a correlation was observed between isoproterenol-responsive Na+-K+ co-transport in turkey erythrocytes and increased phosphorylation of goblin, an Mr = 230,000 protein of the turkey erythrocyte plasma membrane. The phosphorylation of specific sites in goblin has now been analyzed by tryptic fingerprinting. Three major phosphopeptides were detected in goblin prepared from intact, 32P-labeled erythrocytes. One of the peptides 1, was maximally phosphorylated in the absence of hormonal agents. Two additional peptides, 2 and 3, were phosphorylated only following exposure of cells to the beta-adrenergic agonist isoproterenol, to cAMP plus isobutylmethylxanthine, or to cholera toxin. In cells stimulated by isoproterenol, phosphorylation of goblin peptides 2 and 3 could be selectively and completely reversed by subsequent addition of the beta-adrenergic antagonist propranolol. Addition of either cAMP or of Ca2+ plus calmodulin to purified turkey erythrocyte plasma membranes increased incorporation of 32P into goblin. Peptides 2 and 3 of goblin were phosphorylated by addition to the membranes of cAMP or of purified cAMP-dependent protein kinase. Two additional goblin peptides, 4 and 5, were phosphorylated in the plasma membrane preparation by addition of purified calmodulin plus Ca2+, whereas peptides 2 and 3 of goblin were not phosphorylated under these conditions. Peptide 1 did not incorporate 32P in the plasma membranes under any condition tested. Both calmodulin and cAMP-dependent protein kinase were identified directly in turkey erythrocytes. The three major phosphopeptides of goblin phosphorylated in intact cells (peptides 1, 2, and 3) contained phosphothreonine and represented distinct phosphorylation sites. In contrast, the two phosphopeptides of goblin phosphorylated in plasma membranes by addition of Ca+/calmodulin (peptides 4 and 5) contained phosphoserine. It is concluded that goblin, a plasma membrane protein possibly involved in the hormonal regulation of Na+-K+ co-transport, contains at least 3 distinct threonine residues and 1 or more serine residues which serve as specific substrates for three or more distinct protein kinases of the turkey erythrocyte, namely a cAMP-dependent enzyme, a Ca2+/calmodulin-dependent enzyme, and a third enzyme with undetermined regulatory control.

Animals↗

In vitro evaluation of B-lymphocyte function in turkeys infected with hemorrhagic enteritis virus.

Studies were conducted on B-lymphocyte function in turkeys infected with hemorrhagic enteritis (HE) virus. Hemolytic plaque-forming technique was used to detect antibody-forming cells in turkeys. The plaque-forming cell responses in HE virus-infected and noninfected controls were compared. Results of this study indicated a decreased capability of HE virus-infected turkeys to produce antibodies to sheep RBC. The greatest inhibition of antibody-forming cell production was seen in the turkeys 19 days after exposure to the virus. However, after this period, the turkeys gradually recovered their immunocompetence to sheep RBC.

Adenoviridae Infections↗

Marek's disease in turkeys: lack of protection by vaccination.

Herpesvirus of turkeys, a highly effective vaccine against Marek's disease (MD) in chickens, was ineffective in protecting turkeys against MD. Another tissue-culture attenuated vaccine virus also protected chickens, but not turkeys, from MD. Intact and immunosuppressed turkey poults inoculated with herpesvirus of turkey developed a persistent viremia, but did not have detectable gross or microscopic lesions.

Animals↗