Effects of zeolite a or clinoptilolite in diets of growing swine

Growth, nutrient balance, plasma ammonia levels and urinary p-cresol excretion were evaluated in growing pigs fed diets containing various levels of zeolite A or clinoptilolite. In one growth trial, crossbred pigs averaging 25 kg initial body weight were assigned to diets containing no zeolite, .3% zeolite A or .5% clinoptilolite for a 6-wk growing phase trial. Average daily gain (ADG), average daily feed intake (ADF) and feed/gain (F/G) were unaffected by supplementation of either zeolite in the diet, but metabolizable energy (ME) utilization was improved by feeding diets containing either zeolite. A second growth trial utilized the same crossbred pigs, which averaged 65 kg initial body weight, and were assigned to diets containing no zeolite, 1% zeolite A or 5% clinoptilolite for an 8-wk finishing phase trial. Average daily gain, ADF, and ME utilization were unaffected by feeding either zeolite diet, while F/G was increased in pigs fed the diet containing 5% clinoptilolite. In two nutrient balance trials, 16 crossbred pigs averaging 7.5 kg in initial body weight were fed diets containing 0, 1, 2 or 3% zeolite A in one trial and 16 crossbred pigs averaging 7.0 kg initial body weight were fed diets containing 0, 2.5, 5.0 or 7.5% clinoptilolite in a second trial. In both trials, digestible energy, ME, N-corrected ME and ME corrected for N balance and zeolite levels were linearly reduced as increasing amounts of either zeolite were fed. Daily fecal N increased and apparent digestibility of N was linearly reduced by feeding increasing amounts of zeolite A or clinoptilolite. Biological value of protein was improved linearly as higher levels of zeolite A were fed, indicating that there may be some ammonia binding to zeolite A in the gastrointestinal tract. Net protein utilization was reduced by feeding increasing levels of clinoptilolite in the diet. Calcium, P, Mg, Na, K and Fe retentions were linearly reduced by feeding increasing amounts of zeolite A in the diet, while increasing levels of clinoptilolite caused only P retention to be linearly reduced. Both free and conjugated forms of urinary p-cresol were linearly reduced by feeding increasing levels of clinoptilolite. Plasma ammonia levels were reduced at subsequent bleedings after a meal and by increasing levels of clinoptilolite.     

Ruminal and plasma concentrations of 3-methylindole associated with tryptophan-induced pulmonary edema and emphysema in cattle.

Five Hereford cows were given an intraruminal dose of L-tryptophan (0.35 g/kg of body weight), and 2 cows were used as controls. Of the 5 treated cows, 3 developed clinical signs of interstitial pul monary edema, and emphysema and severe pulmonary lesions were seen at necropsy after 96 hours. Another cow developed moderate clinical signs and pulmonary lesions, and the remaining cow had few clinical signs and mild pulmonary lesions. The severity of clinical signs in each cow was related to the severity of pulmonary lesions at necropsy. The 3-methylindole (3MI) was present in ruminal fluid and plasma within 6 hours after administration of tryptophan, and the concentrations increased to 3.0 and 9.0 mug/ml within 12 to 24 hours. Severity of pulmonary lesions was related to maximal concentration and duration of 3MI in the plasma. At necropsy, gross lesions were characterized by diffuse, pulmonary edema and interstital emphysema; and the lungs were dark red, firm, and heavier than normal. Predominant microscopic changes included accumulation of proteinaceous residue, hypertrophy and hyperplasia of alveolar lining epithelium, thickening of alveolar septums, and emphysematous thickening of interstitial tissues. These changes were similar to previously reported 3MI-induced pulmonary lesions. The presence of 3MI in ruminal fluid and plasma after administration of tryptophan and the relationship between concentration of 3MI and severity of clinical signs indicate that 3MI is the principal metabolite of ruminal fermentation which leads to the development of acute pulmonary edema and emphysema in cattle given tryptophan.     

Molecular characterization of Theileria orientalis piroplasm protein encoded by an open reading frame (To ORF2) in a genomic fragment

In the present study, a novel antigenic protein expressed in the piroplasm stage of Theileria orientalis was characterized. A 4,707 bp genomic fragment amplified by PCR contained two open reading frames (ORFs). The deduced amino acid sequence of the first ORF showed significantly high similarlity to the ubiquitin carboxy terminal hydrolases/proteases while the second ORF (To ORF2) showed homology to several surface antigens of plasmodia. To ORF2 was expressed to determine whether the protein product is expressed by the parasite. In western blot analysis, bovine antiserum from a T. orientalis-infected calf recognized the recombinant protein containing a C-terminal part of the ORF expressed by baculovirus system. Western blot analysis with the anti-To ORF2 mouse serum recognized a 48 kDa protein in T. orientalis piroplasm lysates. Indirect immunofluorescence antibody test by confocal scanning laser microscopic analysis showed that antisera against the recombinant protein recognized T. orientalis piroplasm in the infected erythrocyte. The results from this study indicate that To ORF2 protein is expressed at the piroplasm stage and is immunogenic. This novel antigenic To ORF2 protein could be exploited for vaccine development against bovine piroplasmosis.     

Effect of tissue deterioration on postmortem BSE diagnosis by immunobiochemical detection of an abnormal isoform of prion protein

Surveillance for bovine spongiform encephalopathy (BSE) in fallen stock in Japan is conducted with a commercial enzyme-linked immunosorbent assay (ELISA) for mass screening, with Western blotting (WB) and immunohistochemistry performed for confirmation of the ELISA. All tests are based on immunological detection of an abnormal isoform of the prion protein (PrP(Sc)) in brain tissues, which have sometimes deteriorated by the time samples from fallen stock reach a diagnostic laboratory. To evaluate BSE surveillance procedures for fallen stock, we examined PrP(Sc) detection from artificially deteriorated BSE-affected bovine brain tissues with a commercial ELISA kit and compared the results with those of WB. The optical density (OD) values of the ELISA decreased with advancing deterioration of the tissues, whereas no reduction in the signal for PrP(Sc) was observed in WB, even when performed after 4 days of incubation at 37 degrees C. The progressive decrease in the OD values in the ELISA appear to be caused by a partial loss of the N-terminal moiety of PrP(Sc) due to digestion by endogeneous and/or contaminated microbial enzymes, and by the presence of ELISA inhibitors that are generated in deteriorated tissues. These results suggest that WB is the most reliable test for fallen stock, especially for cattle brains within decaying carcasses.     

Dietary Soy Peptide Enhances Thermotolerance and Survival of Juvenile Japanese Flounder,Paralichthys olivaceus

Soy peptide (SP), a soy protein enzymatic hydrolysate, contains bioactive substances that could be utilized as an immune‐stimulating feed ingredient. The experiment evaluated the efficacy of dietary SP on promoting growth, and enhancing tolerance and survival to heat stress in juvenile Japanese flounder, Paralichthys olivaceus. Four diets were incorporated with different levels of SP (0, 2, 5, and 10%) and a 6‐wk feeding trial ensued. Following the feeding trial, the experimental groups were subjected to heat stress to measure survival rate and heat shock protein 70s (HSP70s) in gill, liver, and skin. Fish fed diets with SP inclusion showed considerable decrease in percent weight gain. Significantly higher lethal time values to 50% mortality (LT₅₀) value were recorded for fish fed 10% SP. Moreover, LT₅₀ values of fish fed 2 and 5% SP were significantly higher compared with fish fed control diet. HSP70s produced in all the tissues were significantly highest in fish fed 10% SP. HSP70s values were significantly higher in fish fed 2 and 5% SP compared with fish fed control diet. A significant reduction in HSP70s among all groups during recovery period was also observed. These results suggest that SP can be used to enhance the immune response and survival of P. olivaceus under heat stress.     

Genetic evidence for paternal inheritance of the chloroplast in four Australian Callitris species (Cupressaceae)

The mode of chloroplast inheritance was investigated in the four Australian conifer species; Callitris columellaris, C. intratropica, C. gracilis and C. verrucosa (Callitroideae, Cupressaceae), as a basis for future use of chloroplast genetic markers for the genus. Open-pollinated seeds and the foliage tissue of the mother trees were collected from one population per species, in which two distinct chloroplast haplotypes at the trnL (UAA) 5′ exon–trnF (GAA) region were co-distributed. In total, the DNA sequences of the chloroplast fragments in 255 samples were determined, and the haplotypes of the germinated seedlings and the mother trees were compared. Among the 16 mother trees examined across all species, 12 trees were shown to have produced at least one offspring with a different haplotype from the mother, indicating the occurrence of paternal inheritance of chloroplast in the fertilization processes. In binomial GLMM analyses, the haplotype of mother trees in C. columellaris, C. gracilis, and C. verrucosa did not significantly affect those of their offspring, and the predicted frequency of seedlings’ haplotypes was compatible with the observed frequency in adult canopy trees. These findings provide a genetic support that paternal inheritance of chloroplast DNA via pollen dispersal would be dominant in the Callitris species.     

Aluminum-26 in deep-sea sediment

The activity of (26)A1 in a North Pacific core was found to be 0.081 +/- 0.046 disintegration per minute per kilogram of dry sediment, which corresponds to a (26)A1/(10)Be ratio of 0.018 +/- 0.011 (or 0.019 +/- 0.012 when this ratio is corrected for decay). This ratio is in good agreement with that measured in Greenland ice, 0.017 +/- 0.008. These ratios are also in agreement with the calculated values for the production of these isotopes by cosmic rays in the atmosphere: 0.013 +/- 0.006. The contribution of cosmic dust bearing (26)Al seems small in comparison with the production of this nuclide in the atmosphere.     

Maintenance of mouse trophoblast stem cells in KSR-based medium allows conventional 3D culture

Mouse trophoblast stem cells (TSCs) can differentiate into trophoblast cells, which constitute the placenta. Under conventional culture conditions, in a medium supplemented with 20% fetal bovine serum (FBS), fibroblast growth factor 4 (FGF4), and heparin and in the presence of mouse embryonic fibroblast cells (MEFs) as feeder cells, TSCs maintain their undifferentiated, proliferative status. MEFs can be replaced by a 70% MEF-conditioned medium (MEF-CM) or by TGF-ß/activin A. To find out if KnockOut^TM Serum Replacement (KSR) can replace FBS for TSC maintenance, we cultured mouse TSCs in KSR-based, FBS-free medium and investigated their proliferation capacity, stemness, and differentiation potential. The results indicated that fibronectin, vitronectin, or laminin coating was necessary for adhesion of TSCs under KSR-based conditions but not for their survival or proliferation. While the presence of FGF4, heparin, and activin A was not sufficient to support the proliferation of TSCs, the addition of a pan-retinoic acid receptor inverse agonist and a ROCK-inhibitor yielded a proliferation rate comparable to that obtained under the conventional FBS-based conditions. TSCs cultured under the KSR-based conditions had a gene expression and DNA methylation profile characteristic of TSCs and exhibited a differentiation potential. Moreover, under KSR-based conditions, we could obtain a suspension culture of TSCs using extracellular matrix (ECM) coating-free dishes. Thus, we have established here, KSR-based culture conditions for the maintenance of TSCs, which should be useful for future studies.     

Polyphenol-induced inhibition of the response of na(+)/glucose cotransporter expressed in Xenopus oocytes

To study the effects of polyphenols on the Na(+)/glucose cotransporter (SGLT1) response, SGLT1 was expressed in Xenopus oocytes by injecting cRNA synthesized from the cloned cDNA of the small intestine cotransporter of rats, and the electrical response elicited by glucose or galactose was measured by a voltage clamping method. Most phenol derivatives had no effect on the response. However, the polyphenols (+)-catechin, (-)-epicatechin gallate (ECg), and (-)-epigallocatechin gallate (EGCg), which are components of green tea, caused an inhibition of the response, which was almost independent of glucose concentration. The inhibition constants were estimated to be 2.3 mM for (+)-catechin and 0.45 mM for both ECg and EGCg, assuming the noncompetitive inhibition mechanism. Saponin prepared from tea seeds also inhibited the response significantly. Tannic acid and aqueous extracts of teas induced nonspecific electrical responses in both cRNA-injected and noninjected oocytes at lower concentrations than those that caused an inhibition of the SGLT1 response when their dose-dependent effects were examined. These results are possibly helpful in the development of a dietary supplement for diabetic patients.