Evaluation of a dacitic (rhyolitic) tuff breccia use on performance, inflammatory, and antioxidant responses in broilers mildly challenged with Eimeria spp.

The objective of the study was to investigate the effects of a dacitic tuff breccia (DTB) on Eimeria-infected broilers. A total of 600 one-day-old Cobb 500 male chickens were randomly assigned to 5 treatments with 10 replicates of 12 birds. Treatments were: an unchallenged control (UC), a challenged (CC) control (0% DTB), and 3 challenged groups with 0.125, 0.25, or 0.5% DTB. At d 14, birds in the CC and DTB groups were orally gavaged with mixed Eimeria spp., while the UC received water. Growth performance was evaluated during prechallenge, challenge, and postchallenge periods (0-14 d; 14-20 d; and 20-26 d, respectively). Gastrointestinal permeability was measured at 5 days postinfection (dpi). Intestinal histology and nutrient digestibility of dry matter (DM), crude protein (CP), and ileal digestible energy (IDE) were measured at 6 dpi. Liver activity of glutathione peroxidase (GSH-Px) was determined at 6 dpi, and concentrations of reduced (GSH) and oxidized glutathione (GSSG) were analyzed at 6 and 12 dpi. Data were analyzed using a linear mixed model analysis and Tukey's test (P ≤ 0.05). From 0 to 14 d, similar average daily gain (ADG) and average daily feed intake (ADFI, P > 0.05) were observed. Gain:feed ratio (GF) was higher in 0.125, 0.25, and 0.5% of DTB than the CC and UC (P < 0.001). From 14 to 20 d, the UC had the highest ADG, ADFI, and GF (P < 0.001). At 5 dpi, intestinal permeability was higher in the challenged groups than the UC. Additionally, the UC showed the highest apparent ileal digestibility of CP, whereas 0.125% DTB had higher CP digestibility than the CC and 0.5% DTB (P < 0.001). At 6 dpi, 0.125% DTB increased GSH-Px activity compared to the CC, 0.5% DTB, and UC (P < 0.001). At 12 dpi, 0.125% DTB showed increased GSH concentration compared to the CC, 0.25% DTB, and 0.5% DTB (P < 0.01). The mild coccidia infection negatively impacted growth performance, apparent ileal nutrient digestibility, intestinal histology, and gastrointestinal integrity in broilers. The use of 0.125% DTB exhibited potential in improving antioxidant responses, apparent ileal digestibility of CP, and growth performance.

Influence of dietary dacitic tuff breccia on laying hen performance and egg quality parameters and bone structure at 85 weeks of age after a non-anorexic molt program at 73 to 77 weeks.

The objective of the study was to assess the efficacy of AZOMITE (AZM), a dacitic tuff breccia, in laying hens through egg quality and production parameters. A total of ninety six 73-wk-old Hy-Line W-36 commercial laying hens were randomly assigned to 2 dietary treatments, a control diet and the same diet containing 0.25% AZM, with 24 replicates of 2 hens/replication. From 73 to 77 wk, hens went through nonanorexic molt, and, from 77 to 85 wk, the hens were evaluated for egg production, eggshell quality, and bone health. At wk 85, tibiotarsi were collected for ash and mineral composition, ileal contents were collected for calcium, phosphorus, apparent metabolizable energy corrected for N (AMEn), and apparent nitrogen retention (ANR) evaluation. AZM-fed hens tended to have higher body weight (P = 0.07) from 82 to 83 and 84 to 85 wk, and higher hen day egg production than control (90.54 vs. 79.51%, P = 0.005) from 84 to 85 wk. In general, no differences were reported in feed intake, eggshell color, egg weight, albumen height, Haugh units, or eggshell thickness (P > 0.05). However, shell strength and elasticity were improved (P < 0.02) and yolk color was decreased (P = 0.03) in AZM-fed hens than control. Moreover, the digestibility of Ca, AMEn, and ANR was increased with 0.25% AZM compared to control (P < 0.01). Tibiotarsi P and Ca percentage were lower in AZM-fed birds than control (P < 0.01), without affecting bone strength and mineral density (P > 0.36). Therefore, the use of 0.25% AZM showed a potential in improving egg production and eggshell strength, while maintaining bone quality in post-molt laying hens.

Dietary inclusion of AZOMITE improves feed efficiency in broilers and egg production in laying and broiler breeder hens.

The dietary inclusion of aluminosilicates has been reported to enhance pellet quality, improve feed mill throughput, bind toxins, improve feed efficiency, and promote immunological function across a variety of production systems. AZOMITE is a product marketed as a hydrated sodium calcium aluminosilicate containing macro and trace minerals, and rare earth elements and the potential benefits of its dietary inclusion in broiler, layer, and broiler breeder diets was investigated. In a battery study, broilers were fed diets containing 0, 0.125, 0.250, or 0.500% AZOMITE from 0 to 21 d of age. Laying hens were fed a control diet or this diet supplemented with 0.25% AZOMITE from 54 through 98 wk of age, with the hens fed a standard molting diet or this diet supplemented with 0.25% AZOMITE from 71 to 72 wk of age. Broiler breeder hens were fed a control diet or this diet supplemented with 0.25% AZOMITE from the onset of photostimulation at 21 wk of age through 65 wk of age. All 3 dietary inclusion rates of AZOMITE improved (P < 0.05) the feed to body weight gain ratio in broilers fed these diets relative to broilers fed the control diet. In laying hens total marketable eggs, and in broiler breeder hens total settable eggs were increased (P < 0.05) with the dietary inclusion of AZOMITE by 8 eggs per hen. The inclusion of dietary AZOMITE also improved apparent Ca and P digestibility in broilers and tibia ash content in laying hens. The results indicate the dietary inclusion of AZOMITE in poultry diets improves bird performance.

Comparison of the inhibitory activities of human tissue factor pathway inhibitor (TFPI)α and TFPIß.

BACKGROUND: Tissue factor pathway inhibitor (TFPI) is an alternatively spliced protein with two isoforms, TFPIα and TFPIß, which differ in their C-terminal structure and cellular localization. Detailed characterization of their inhibitory activity is needed to define potentially unique inhibitory roles in tissue factor (TF)-mediated thrombotic and inflammatory disease, and to understand how pharmaceuticals targeted to different structural regions of the TFPI isoforms alter hemostasis in hemophilia patients. METHODS: The TF inhibitory activity of TFPIß localized to the surface of CHO cells was compared with that of soluble TFPIα by the use of in vitro and in vivo assays. RESULTS: In TF-factor VIIa-mediated FXa generation assays, TFPIß was a slightly better inhibitor than TFPIα, which was approximately three-fold better than TFPI-160, a soluble, altered form of TFPI similar to TFPIß. In direct FXa inhibitory assays, TFPIß had an IC50 2.5-fold lower than that of TFPIα and 56-fold lower than that of TFPI-160. TFPIß inhibited TF-mediated CHO cell migration though Matrigel, whereas TFPIα and TFPI-160 were poor inhibitors, demonstrating that TFPIß effectively blocks TF-initiated signaling events during cellular migration through matrices that are not permeable to soluble forms of TFPI. Furthermore, TFPIß inhibited TF-dependent CHO cell infiltration into lung tissue following tail vein injection into SCID mice, and blocked the development of consumptive coagulopathy. CONCLUSIONS: TFPIß is a slightly better inhibitor of TF procoagulant activity than TFPIα. As a surface-associated protein, TFPIß is a much better inhibitor of TF-mediated cellular migration than soluble TFPIα, and may specifically act in the inhibition of TF-mediated signaling events on inflamed endothelium and/or monocytes.

Temporal expression of alternatively spliced forms of tissue factor pathway inhibitor in mice.

BACKGROUND: Mouse tissue factor pathway inhibitor (TFPI) is produced in three alternatively spliced isoforms that differ in domain structure and mechanism for cell surface binding. Tissue expression of TFPI isoforms in mice was characterized as an initial step for identification of their physiological functions. METHODS AND RESULTS: Sequence homology demonstrates that TFPIalpha existed over 430 Ma while TFPIbeta and TFPIgamma evolved more recently. In situ hybridization studies of heart and lung did not reveal any cells exclusively expressing a single isoform. Although our previous studies have demonstrated that TFPIalpha mRNA is more prevalent than TFPIbeta or TFPIgamma mRNA in mouse tissues, western blot studies demonstrated that TFPIbeta is the primary protein isoform produced in adult tissues, while TFPIalpha is expressed during embryonic development and in placenta. Consistent with TFPIbeta as the primary isoform produced within adult vascular beds, the TFPI isoform in mouse plasma migrates like TFPIbeta in SDS-PAGE and mice have a much smaller heparin-releasable pool of plasma TFPIalpha than humans. CONCLUSIONS: The data demonstrate that alternatively spliced isoforms of TFPI are temporally expressed in mouse tissues at the level of protein production. TFPIalpha and TFPIbeta are produced in embryonic tissues and in placenta while adult tissues produce almost exclusively TFPIbeta.

Tissue factor pathway inhibitor-gamma is an active alternatively spliced form of tissue factor pathway inhibitor present in mice but not in humans.

BACKGROUND: Tissue factor pathway inhibitor (TFPI) is a potent inhibitor of tissue factor procoagulant activity produced as two alternatively spliced isoforms, TFPIalpha and TFPIbeta, which differ in domain structure and mechanism for cell surface association. 3' Rapid amplification of cDNA ends was used to search for new TFPI isoforms. TFPIgamma, a new alternatively spliced form of TFPI, was identified and characterized. METHODS: The tissue expression, cell surface association and anticoagulant activity of TFPIgamma were characterized and compared to those of TFPIalpha and TFPIbeta through studies of mouse and human tissues and expression of recombinant proteins in Chinese hamster ovary (CHO) cells. RESULTS: TFPIgamma is produced by alternative splicing using the same 5'-splice donor site as TFPIbeta and a 3'-splice acceptor site 276 nucleotides beyond the stop codon of TFPIbeta in exon 8. The resulting protein has the first two Kunitz domains connected to an 18 amino acid C-terminal region specific to TFPIgamma. TFPIgamma mRNA is differentially produced in mouse tissues but is not encoded within the human TFPI gene. When expressed in CHO cells, TFPIgamma is secreted into conditioned media and effectively inhibits tissue factor procoagulant activity. CONCLUSIONS: TFPIgamma is a third alternatively spliced form of TFPI that is widely expressed in mouse tissues but not made by human tissues. It contains the first two Kunitz domains and is a secreted, rather than a cell surface-associated, protein. It is a functional anticoagulant and may partially explain the resistance of mice to coagulopathy in tissue factor-mediated models of disease.

A GPI-anchored co-receptor for tissue factor pathway inhibitor controls its intracellular trafficking and cell surface expression.

BACKGROUND: Tissue factor pathway inhibitor (TFPI) lacks a membrane attachment signal but it remains associated with the endothelial surface via its association with an, as yet, unidentified glycosyl phosphatidylinositol (GPI)-anchored co-receptor. OBJECTIVES/METHODS: Cellular trafficking of TFPI within aerolysin-resistant ECV304 and EA.hy926 cells, which do not express GPI-anchored proteins on their surface, was compared with their wild-type counterparts. RESULTS AND CONCLUSIONS: Although aerolysin-resistant cells produce normal amounts of TFPI mRNA, TFPI is not expressed on the cell surface and total cellular TFPI is greatly decreased compared with wild-type cells. Additionally, normal, not increased, amounts of TFPI are secreted into conditioned media indicating that TFPI is degraded within the aerolysin-resistant cells. Confocal microscopy and studies using metabolic inhibitors demonstrate that aerolysin-resistant cells produce TFPI and transport it into the Golgi with subsequent degradation in lysosomes. The experimental results provide no evidence that cell surface TFPI originates from secreted TFPI that binds back to a GPI-anchored protein. Instead, the data suggest that TFPI tightly, but reversibly, binds to a GPI anchored co-receptor in the ER/Golgi. The co-receptor then acts as a molecular chaperone for TFPI by trafficking it to the cell surface of wild-type cells or to lysosomes of aerolysin-resistant cells. TFPI that escapes co-receptor binding is secreted through the same pathway in both wild-type and aerolysin-resistant cells. The data provide a framework for understanding how TFPI is expressed on endothelium.

Resistance plasmids of Pseudomonas aeruginosa: change from conjugative to nonconjugative in a hospital population.

Properties of a population of carbenicillin- and gentamicin-resistant, tobramycin-susceptible Pseudomonas aeruginosa at Veteran's Administration Hospital, Cincinnati, Ohio, have been followed during a 16-month period. As originally described, the strains were isolated from patients with urinary tract colonizations and were predominantly Parke-Davis immunotype 7. For the majority of these organisms, antibiotic resistance was correlated with the presence of a self-conjugative plasmid of incompatibility group P-2. The source and relative incidence of multiply resistant isolates have remained constant during the current study, but the immunotype has shifted form type 7 to type 2. Concomitantly, the population has lost the property of conjugative transfer of resistance, and resistant strains are now compatible with P-2 plasmids. A group P-2 R plasmid, pMG5, will mobilize resistance markers, demonstrating that the multiple resistance of the nonconjugative strains is mediated by R plasmids. Additionally, gentamicin resistance due to either conjugative or nonconjugative plasmids is correlated with the presence of similar gentamicin acetyltransferase activity. pMG5-mobilized plasmids are shown to be incompatible with pMG5. pMG5 is also shown to mobilize resistance markers from nontransferring antibiotic-resistant strains representing populations from Parkland Memorial Hospital, Dallas, Texas, and Cleveland Clinic Foundation, Cleveland, Ohio.

Pseudomonas aeruginosa R factors determining gentamicin plus carbenicillin resistance from patients with urinary tract colonizations.

R factors determining multiple resistance including both gentamicin and carbenicillin have been identified in high incidence among hospital isolates of Pseudomonas aeruginosa. The factors are readily transmitted to other P. aeruginosa but not to Escherichia coli strains K-12 or C, or to Proteus mirabilis. R factor-containing isolates are predominantly immunotype 7 isolated from urinary sources.