An improved dry ash procedure for the detection of titanium dioxide in cattle feces

We improved the dry ash procedure for detecting titanium dioxide (TiO₂) in cattle feces containing chromium dioxide (Cr₂O₃). First, the effect of amount of sodium sulfate (Na₂SO₄) on the recovery of TiO₂ from cattle feces that contained Cr₂O₃ was evaluated. Average recovery of TiO₂ at the 2.5 g Na₂SO₄ level was significantly higher (P < 0.0001) than that at 0.75 g Na₂SO₄. Second, the effect of Cr₂O₃ concentration on the recovery of TiO₂ of cattle feces by using two levels of Na₂SO₄ addition was examined. The recovery of TiO₂ decreased with the increase in the amount of Cr₂O₃ at the 0.75 g Na₂SO₄ level but was consistently high at 2.5 g Na₂SO₄. Third, the recovery of Cr₂O₃ from cattle feces was checked. The recoveries of TiO₂ and Cr₂O₃ were high enough at the 2.5 g Na₂SO₄ level. Fourth, the improved dry ash procedure (5 mL of concentrated H₂SO₄ and 2.5 g of Na₂SO₄ were used for sample digestion) was compared to the wet ash procedure. Average recovery of TiO₂ by the improved dry ash procedure was significantly higher (P = 0.0077) than that by the wet ash procedure. Thus, the improved dry ash procedure can be used for TiO₂ analysis in cattle feces containing Cr₂O₃.     

Hypothermic storage of porcine zygotes in serum supplemented with chlorogenic acid

The current study was conducted to investigate the effects of 100% foetal bovine serum (FBS) and 100% porcine follicular fluid (pFF) as a storage medium on the developmental competence of porcine zygotes stored at 25°C for 24 hr. Moreover, we evaluated the additive effects of chlorogenic acid (CGA) in the storage medium. When in vitro‐produced zygotes were stored at 25°C for 24 hr in tubes containing either tissue culture medium (TCM) 199 supplemented with 1 mg/ml bovine serum albumin (BSA), 100% of FBS or 100% of pFF, the rate of blastocyst formation was significantly higher in 100% of FBS than in BSA‐containing TCM 199. When the effects of CGA supplementation in 100% of FBS on the development of zygotes stored at 25°C for 24 hr was evaluated, more zygotes stored with 50 µM CGA developed to blastocysts compared with the other concentrations of CGA. When the formation date and quality of blastocysts derived from zygotes stored in 100% of FBS supplemented with 50 µM CGA were investigated, the highest ratio of blastocysts formation in the storage group appeared 1 day later than in the non‐stored control group. However, a higher proportion of blastocysts with apoptotic nuclei was observed in the stored group as compared to the non‐stored group. In conclusion, 100% of FBS is available for a short storage medium of porcine zygotes. The supplementation of 50 µM CGA into the storage medium improves the rates of blastocyst formation of zygotes after storage, but the quality of embryos from the stored zygotes remains to be improved.     

Follicle formation in the canine ovary after autografting to a peripheral site

This study reports about follicular development on the surface of canine ovarian tissue after autografting under the fascia of the thoracolumbar muscle and about meiotic resumption of follicle-derived oocyte after maturation culture. After ovarian excision from a bitch, each ovary of the pairs was cut approximately into half. The hemi-ovaries were transplanted into the bitch of origin at three different body sites (under the fascia of the quadriceps femoris muscle and the thoracolumbar muscle, and in the deltoid muscle in the scapular region). All grafted ovaries were recovered from the bitch at 35 days post-transplantation. A visible antral follicle was observed on the surface of the ovary grafted under the thoracolumbar fascia. Histological examination revealed viable follicles at different stages of development irrespective of graft site. Most granulosa cells in the follicles at different stages of development expressed proliferating cell nuclear antigen (PCNA). A total of three oocytes were collected from an ovary grafted under the fascia of the thoracolumbar muscle, wherein an oocyte reached metaphase I after maturation culture. This is the first report to demonstrate follicular development and meiotic resumption of oocytes recovered from autografted canine ovarian tissues.     

Urinary transforming growth factor-beta1 in feline chronic renal failure

Transforming growth factor-beta1 (TGF-beta1), an inflammatory cytokine, plays a role in tissue fibrosis, such as glomerular sclerosis and tubulointerstitial fibrosis of the kidneys. In the present study, the urinary TGF-beta1 level of cats diagnosed with chronic renal failure (CRF) was measured to investigate its relationship to the pathogenesis of feline CRF. Urinary TGF-beta1 levels (TGF-beta1/creatinine ratio) were significantly increased compared with healthy controls, whereas serum levels of TGF-beta1 were not. These results indicate that TGF-beta1 is expressed in the kidneys of CRF cats, and that it was reflected in the urinary TGF-beta1 level. Therefore, TGF-beta1 may play a role in feline CRF, and urinary TGF-beta1 could be used as a clinical marker for renal fibrosis.     

Evaluation of RNA interference in developing porcine granulosa cells using fluorescence reporter genes

Gene silencing using small interfering RNA (siRNA) may be useful for functional analyses of unidentified genes expressed during cell differentiation. The present study was performed to evaluate RNA interference (RNAi) in porcine granulosa cells stimulated with bovine FSH, by using two fluorescence reporter genes: a plasmid encoding green fluorescent protein (GFP) and a plasmid encoding red fluorescent protein (RFP). The siRNA targeting GFP mRNA sequence (GFP-siRNA) with both plasmids was introduced into cultured cells by lipofection. GFP- and RFP-expressing cells were observed under fluorescence microscopy and analyzed by flow cytometry. Strong fluorescence was observed after introduction of both plasmids into cells. The intensity of green fluorescence generated by GFP was greatly suppressed by introduction of GFP-siRNA, showing an approximate 70% decrease in the ratio of green to red fluorescence. Consequently, we concluded that gene silencing by siRNA can be used to analyze the functions of genes of interest during differentiation of porcine granulosa cells.     

IgG antibody subclass response against equine herpesvirus type 4 in horses

In this study, IgG subclass responses against equine herpesvirus type 4 (EHV-4) were examined by enzyme-linked immunosorbent assay (ELISA) using a type-specific region of EHV-4 glycoprotein G (gG). ELISA using sera collected from horses experimentally infected with EHV-4 revealed that IgGa and IgGb antibodies were detected at high level, but IgGc and IgG(T) antibody responses were detected at low level or were undetectable. The IgGa antibody response reached its peak on day 10 post-infection, and then dropped. The IgGb antibody response reached its maximum level on day 12 post-infection, and then the level was sustained during at least 28 days after infection. Forty healthy racehorses that had already been infected with EHV-4 possessed antibody against EHV-4. Although IgGa antibodies specific for EHV-4 were not detected in any horses, IgGb antibodies were detected and the levels correlated with total IgG antibodies against EHV-4 gG. The results suggest that EHV-4-specific IgGa and IgGb antibodies are induced in EHV-4-infected horses, and that IgGb antibody, but not IgGa, is long lasting.     

Effect of light irradiation on dynamics of vitamin A compounds in rotifers and Artemia

ABSTRACT:   The dynamics of vitamin A (VA) compounds in live food during enrichment were examined under different light conditions. Rotifers Brachionus plicatilis and Artemia nauplii ( Artemia ) were enriched with or without 10 mg VA palmitate (VAp) in 1 L of culture medium for 24 h under either bright (2000 lx), or dark (<1 lx) conditions. VAp, retinol (ROH), retinal (RAL) and retinoic acid (RA) contents were analyzed at 0 h (before enrichment) and at 3, 6, 12, 18 and 24 h after the onset of enrichment. Retinoid content in rotifers enriched in darkness was always higher than that enriched under light. VAp content showed two peaks at 3 and 18 h in rotifers enriched in darkness, but it showed one peak at 3 h in rotifers enriched under light. ROH and RA contents increased over the 24-h period in rotifers enriched in darkness, whereas they decreased 12 h onward in rotifers enriched under light. In Artemia , VAp and ROH contents were always higher than when enriched under the bright condition, but their dynamics showed a similar pattern in Artemia enriched under dark and bright conditions.     

Effects of wood kraft pulp feeding on feed digestibility and rumen fermentation of Japanese Black steer in the middle fattening stage

We investigated the effects of wood kraft pulp (KP) feeding on feed digestibility and rumen fermentation of Japanese Black (JB) steer in the middle fattening stage. The feeding experiment was carried out a replicated 3 × 3 Latin square design using six JB steers (16.2 ± 0.8 months of age). Steers were fed rice straw and three concentrated feeds: commercial formula feed (control), formula feed containing 7.5% KP on a dry matter (DM) basis (7.5% KP), and formula feed containing 15% KP on a DM basis (15% KP). Feed digestibility and rumen fermentation parameters were recorded for 3 days during the measurement period, after an adaptation phase of 11 days. There was no significant difference in DM intake per day among diets. A slightly higher digestibility of neutral detergent fiber was observed in steers fed 15% KP, compared with those fed other diets (P = 0.059). The butyric acid ratio in rumen fluid was higher in steers fed KP diets than in steers fed control (P = 0.083). The average ruminal pH was unaffected by KP diets. This study suggests that replacing 15% of concentrated feed with KP has little effect on feed digestibility and rumen fermentation in JB fattening steer.