Trace mineral source influences ruminal distribution of copper and zinc and their binding strength to ruminal digesta1,2,3.

Eight crossbred steers (BW 719.0 ± 65.0 kg) with ruminal and duodenal cannulae were used to examine the effect of trace mineral (TM) source on digestibility; ruminal and duodenal solubility of Cu, Zn, and Mn; and in vitro release of Cu, Zn, and Mn from the solid fraction of ruminal digesta. Experiment 1 determined the effect of TM source on DM and NDF digestibility in steers fed a corn silage and steam-flaked corn-based diet. Treatments consisted of 10 mg Cu, 20 mg Mn, and 30 mg Zn/kg DM from either sulfate TM (STM) or hydroxy TM (HTM) sources. Following a 14-d adaptation period, total fecal output was collected for 5 d. Dry matter digestibility was not affected by treatment, but NDF digestibility tended (P < 0.09) to be greater in HTM vs. STM supplemented steers. In Exp. 2, steers were fed a diet without supplemental Cu, Zn, or Mn for 19 d. Steers were then administrated a pulse dose of STM or HTM (2× the National Research Council requirements for Cu, Mn, and Zn) via the rumen fistula. Ruminal and duodenal samples were obtained at 2-h intervals starting at -4 and ending at 24 h relative to dosing. Ruminal soluble Cu and Zn concentrations were affected by treatment, time, and treatment × time. Soluble concentrations and percent soluble Cu and Zn in ruminal digesta increased (P < 0.05) above 0-h values for 10 h following dosing with STM, but not HTM. Concentrations of Cu and Zn in ruminal solid digesta were also affected by treatment, time, and treatment × time. Steers dosed with STM had greater (P < 0.05) solid digesta Cu concentrations at 2 and 4 h but lesser (P < 0.05) concentrations from 6 to 20 h post-dosing than those receiving HTM. Ruminal solid digesta Zn concentrations were greater (P < 0.05) in HTM vs. STM-dosed steers from 6 through 24 h post-dosing. Distribution of Mn in ruminal digesta was affected by TM source, but to a lesser extent than Zn and Cu. Duodenal soluble TM concentrations were variable and not affected by treatment. Binding strength of TM to ruminal solid digesta was estimated at 0, 6, and 12 h post-dosing using dialysis against chelating agents. The percentage of Cu and Zn released from ruminal solid digesta by dialysis against Tris-EDTA was greater (P < 0.05) at 12 h post-dosing from steers receiving HTM vs. STM. Results indicate that Cu and Zn from HTM have low solubility in the rumen and appear to be less tightly bound to ruminal solid digesta than Cu and Zn from STM.

The biological effects of five feline IFN-alpha subtypes.

IFN-alpha has been shown to induce both antiviral and antiproliferative activities in animals. This report describes the biological activity of five recently identified feline IFN-alpha subtypes expressed in the Chinese hamster ovary (CHO) cell line (rfeIFN-alpha1[CHO], rfeIFN-alpha2[CHO], rfeIFN-alpha3[CHO], rfeIFN-alpha5[CHO] and rfeIFN-alpha6[CHO]) and the feIFN-alpha6 subtype expressed in and purified from Pichia pastoris (rfeIFN-alpha6[P. pastoris]). The rfeIFN-alpha[CHO] subtypes were tested for antiviral activity against either Vesicular stomatitis virus (VSV) or feline calicivirus (FCV) infected feline embryonic fibroblast cell line (AH927) or Crandell feline kidney cell line (CRFK). Antiviral activity was induced against both VSV and FCV infected AH927 cells and VSV infected CRFK cells by all five of the rfeIFN-alpha[CHO] subtypes and rfeIFN-alpha6[P. pastoris]. In addition, the IFN-alpha inducible Mx gene (associated with antiviral activity) was upregulated in vivo 24 h following treatment with rfeIFN-alpha6[P. pastoris], compared to baseline levels seen prior to treatment. All of the rfeIFN-alpha[CHO] subtypes and rfeIFN-alpha6[P. pastoris] exhibited antiproliferative activity in the FeT-J cell line (an IL-2 independent feline T-cell line). Both necrosis and apoptosis were observed in rfeIFN-alpha6[P. pastoris]-treated FeT-J cells. The rfeIFN-alpha3[CHO] subtype consistently exhibited lower antiviral and antiproliferative activity compared to that observed with the other four rfeIFN-alpha[CHO] subtypes. In summary, this paper demonstrates that five previously described feIFN-alpha subtypes induce both antiviral and antiproliferative activities in vitro and are capable of upregulating the feMx gene in vivo.

Selection of HPRT primers as controls for determination of mRNA expression in dogs by RT-PCR.

Reliable housekeeping gene controls are critical for measuring and comparing gene expression at the transcription level by Northern blot and RT-PCR. In order to develop such controls for studying cytokine mRNA expression in dogs, DNA sequence encoding a full-length canine HPRT protein has been obtained. Numerous primer pairs derived from the canine HPRT sequence have been tested on canine genomic DNA as well as cDNA. The data from the present study suggest that there may be processed HPRT pseudogenes in dogs. Three pairs of canine HPRT primers designed and tested in the present study were able to differentiate between cDNA and genomic DNA under specific PCR conditions. These primers would be useful controls for measurement of mRNA expression by RT-PCR in the dog.

Canine mammary tumor cells transfected with B7-1 or B7-2 stimulate proliferation of peripheral blood mononuclear cells.

Neoplastic cells are believed to evade the immune system due, in part, to their inability to successfully provide a secondary, costimulatory signal for a T lymphocyte proliferative response. This report describes the generation and investigation of genetically engineered canine mammary tumor (CMT) cells that express canine B7-1, canine B7-2, or human B7-2. These transfected cells were used as stimulators in an allogeneic, costimulation assay. CMT cells transfected with canine B7-1 induced the greatest proliferation (7-fold increase), followed by CMT cells transfected with canine B7-2 (5-fold increase). The specificity of the canine B7-2 stimulatory response was demonstrated by a 38% reduction in proliferation caused by an anti-canine B7-2 blocking antibody. These results suggest that canine mammary tumor cells transfected with canine B7-1 or canine B7-2 may be useful for immunotherapeutic purposes.