Effects of a high-protein diet versus dietary supplementation with ammonium chloride on struvite crystal formation in urine of clinically normal cats

OBJECTIVE: To evaluate the effects of a high-protein diet versus dietary supplementation with ammonium chloride (NH4Cl) on struvite crystal formation in the urine of clinically normal cats by measuring the urine concentration of hydrochloric acid (HCl)-insoluble sediment, urine pH, struvite activity product (SAP), number of struvite crystals in urine, and urine volume. ANIMALS: 23 healthy adult cats. PROCEDURE: Urine was fractionated by centrifugation with subsequent extraction of the sediment with 1 N HCl (study 1). Diets containing either 29% crude protein or 55% crude protein were fed to cats in a crossover trial of 3 weeks/period (study 2). Diets supplemented with either sodium chloride (NaCl) or NH4Cl were fed, by use of a 3 x 3 Latin-square design with 3 wk/period (study 3). In studies 2 and 3, urine samples were collected for the last 7 days of each period. RESULTS: The HCl-insoluble sediment contained Tamm-Horsfall glycoprotein (THP; study 1). The high-protein diet (study 2) and dietary supplementation with NH4Cl (study 3) resulted in a decrease in urine pH, SAP, and the number of struvite crystals in urine. However, the high-protein diet decreased urine concentrations of HCl-insoluble sediment containing THP (study 2), in contrast to the NH4Cl supplementation that increased urine volume without a significant effect on the urine concentration of the HCl-insoluble sediment (study 3). CONCLUSIONS AND CLINICAL RELEVANCE: Our results indicate that compared with dietary supplementation with NH4Cl, the high-protein diet is preferable as a urine acidifier for the prevention of struvite crystal formation in clinically normal cats.     

Difference in ultraweak photon emissions between sulfonylurea-resistant and sulfonylurea-susceptible biotypes of Scirpus juncoides following the application of a sulfonylurea herbicide

We examined the ultraweak photon emissions from a paddy weed, Scirpus juncoides , to assess the availability of photon emissions for the identification of weed biotypes resistant to sulfonylurea herbicides. The emission intensity from the plant organs increased when treated with a sulfonylurea herbicide in a concentration-dependent manner. The increment in emissions was higher in the sulfonylurea-resistant biotypes than in the sulfonylurea-susceptible biotypes. The difference between the biotypes was greater in the culms than in the roots and remained so through the vegetative growth stage to the flowering stage. This difference was independent of the seed source or mutations in the acetolactate synthase genes of the resistant biotypes. These results suggest that the determination of ultraweak photon emissions can be a useful method for identifying the sulfonylurea-resistant biotypes of S. juncoides.     

A 90-day Feeding Toxicity Study of l-Serine in Male and Female Fischer 344 Rats

A subchronic feeding study of l-serine (l-Ser) was conducted with groups of 10 male and 10 female Fischer 344 rats fed a powder diet containing 0, 0.06, 0.5, 1.5 or 5.0% concentrations of l-Ser for 90 days. There were no toxicologically significant, treatment-related changes with regards to body weight, food intake, water intake or urinalysis data. In several of the hematology, serum biochemistry and organ weight parameters, significant changes were observed between some of the treated groups and the controls. All these changes, however, were subtle and lacked any corresponding pathological findings. In addition, the increased or decreased values remained within the range of the historical control values. In fact, histopathological assessment revealed only sporadic and/or spontaneous lesions. In conclusion, the no-observed-adverse-effect-level (NOAEL) for l-Ser was, therefore, determined to be at least a dietary dose of 5.0% (2765.0 mg/kg body weight/day for males and 2905.1 mg/kg body weight/day for females) under the present experimental conditions.     

Polymorphisms in growth hormone gene and their associations with calf weight in Japanese Black cattle

The objectives of this study were to detect effective genetic polymorphisms of bovine growth hormone (bGH) gene associated with calf weight in Japanese Black cattle. Fifty‐eight sires and 47 breeding cows were used to detect the polymorphisms in exons by single‐strand conformation polymorphism (SSCP). Four homozygous and six heterozygous SSCP genotypes were identified in exon 5. Although each single nucleotide polymorphism (SNP) had been reported, these genotypes were caused by three SNPs at the nucleotide positions 2141, 2277 and 2291. Four haplotypes C‐C‐A, G‐C‐A, C‐C‐C and G‐T‐A were newly identified. It was suggested that other haplotypes not detected in this study may not exist, considering the allele frequencies reported in Bos taurus and Bos indicus, and the migrating process of native Japanese cattle. Thereafter, we examined associations between the detected polymorphic sites in exon 5 by PCR – restriction fragment length polymorphism and calf weight using 53 breeding dams and 135 calves. The birth weights of calves with haplotype G‐C‐A are significantly lighter and calves' weights produced by cows with such haplotype are also lighter at 30 days old, using regression analysis. Although further research is necessary, these results may serve as a useful criterion to select breeding stocks, especially in maternal abilities.     

Molecular changes associated with adaptation of equine influenza H3N8 virus in embryonated chicken eggs

Embryonated chicken eggs (ECEs) are routinely used to isolate equine influenza virus. Propagation of the virus in ECEs results in selection of variants. In the present study, we determined nucleotide sequences of entire coding regions of parent A/equine/Tottori/1/07 (H3N8) and its derivatives that have different passage histories in ECE. After 12 passages, nucleotide sequence analysis predicted 3 amino acid substitutions in hemagglutinin (HA; 2 in HA1 and 1 in HA2). The two amino acid substitutions in HA1 were located in the vicinity of the cell receptor-binding site. Three other amino acid substitutions were predicted in internal proteins, 1 in the M1, 1 in the NP and 1 in the PA. This is the first report showing mutations in the internal protein genes of equine influenza virus associated with adaptation to ECE.     

Treatment of cattle with DNA-encoded Flt3L and GM-CSF prior to immunization with Theileria parva candidate vaccine antigens induces CD4 and CD8 T cell IFN-γ responses but not CTL responses

Theileria parva antigens recognized by cytotoxic T lymphocytes (CTLs) are prime vaccine candidates against East Coast fever in cattle. A strategy for enhancing induction of parasite-specific T cell responses by increasing recruitment and activation of dendritic cells (DCs) at the immunization site by administration of bovine Flt3L and GM-CSF prior to inoculation with DNA vaccine constructs and MVA boost was evaluated. Analysis of immune responses showed induction of significant T. parva-specific proliferation, and IFN-γ-secreting CD4(+) and CD8(+) T cell responses in immunized cattle. However, antigen-specific CTLs were not detected. Following lethal challenge, 5/12 immunized cattle survived by day 21, whereas all the negative controls had to be euthanized due to severe disease, indicating a protective effect of the vaccine (p<0.05). The study demonstrated the potential of this technology to elicit significant MHC class II and class I restricted IFN-γ-secreting CD4(+) and CD8(+) T cells to defined vaccine candidate antigens in a natural host, but also underscores the need to improve strategies for eliciting protective CTL responses.     

Effects of hydroxy-aluminum on flocculation and permeability of silt and clay fractions separated from paddy soils

Flocculation tests and permeability measurements were carried out for silt and clay fractions separated from surface and subsurface horizons of four paddy soils. There was not much difference in the flocculating power between A1(OH)_2.2C1_0.8 and CaSO₄. Their flocculation value, 0.3 to 3 mM, was attained by adding the reagents at the rate of 0.24 to 2.4 mmol/100 g of the silt and clay. The flocculation with A1(OH)_2.2C1_0.8 or CaSO₄ did not affect the sediment volume of the silt and clay. This sediment volume was smaller for the surface than for the subsurface horizons of each soil, suggesting that soil organic matter and repetition of wetting and drying contribute to the stable aggregation of the silt and clay in the surface horizon. The permeability was higher for the silt and clay fractions separated from the surface horizon than that from the subsurface horizon, and the addition of 2.5 to 4.2 mmol of A1(OH)_2.2C1_0.8 or CaSO₄/100g of the silt and clay increased its permeability by 1.5 to 4 times. The effect of A1(OH)_2.2C1_0.8 was more lasting than that of CaSO₄ and was recognized even after 200 ml of water passed through 1 g of the silt and clay. The effect of the flocculating reagent on the permeability was more or less enhanced by air-drying the treated silt and clay.     

Nursery‐Box Total Fertilization Technology (NBTF) Application for Increasing Nitrogen Use Efficiency in Chinese Irrigated Riceland: N–Soil Interactions

High N fertilizer and flooding irrigation applied to rice in anthropogenic‐alluvial soil often result in N leaching and low use efficiency of applied fertilizer N from the rice field in Ningxia irrigation region in the upper reaches of the Yellow River. Sound N management practices need to be established to improve N use efficiency while sustaining high grain yield levels and minimize fertilizer N loss to the environment. We investigated the effects of Nursery Box Total Fertilization technology (NBTF) on N leaching at different rice growing stages, N use efficiency and rice yield in 2010 and 2011. The four fertilizer N treatments were 300 kg N ha⁻¹ (CU, Conventional treatment of urea at 300 kg N ha⁻¹), 120 kg N ha⁻¹ (NBTF120, NBTF treatment of controlled‐release N fertilizer at 120 kg N ha⁻¹), 80 kgN ha⁻¹ (NBTF80, NBTF treatment of controlled‐release N fertilizer at 80 kg N ha⁻¹) and no N fertilizer application treatment (CK). The results showed that the NBTF120 treatment increased N use efficiency, maintained crop yields and substantially reduced N losses to the environment. Under the CU treatment, the rice yield was 9634 and 7098 kg ha⁻¹, the N use efficiency was 31·6% and 34·8% and the leaching losses of TN were 44·51 and 39·89 kg ha⁻¹; NH₄⁺‐N was 5·26 and 5·49 kg ha⁻¹, and NO₃⁻‐N was 27·94 and 26·22 kg ha⁻¹ during the rice whole growing period in 2010 and 2011, respectively. Compared with CU, NBTF120 significantly increased the N use efficiency and decreased the N losses from the paddy field. Under NBTF120, the N use efficiency was 56·3% and 51·4%, which was 24·7% and 16·6% higher than that of CU, and the conventional fertilizer application rate could be reduced by 60% without lowering the rice yield while decreasing the leaching losses of TN by 16·27 and 14·36 kg ha⁻¹, NH₄⁺‐N by 0·90 and 1·84 kg ha⁻¹, NO₃⁻‐N by 110·6 and 10·14 kg ha⁻¹ in 2010 and 2011, respectively. Our results indicate that the CU treatment resulted in relatively high N leaching losses, and that alternative practice of NBTF which synchronized fertilizer application with crop demand substantially reduced these losses. We therefore suggest the NBTF120 be a fertilizer application alternative which leads to high food production but low environmental impact. Copyright © 2014 John Wiley & Sons, Ltd.     

Effects of omega-7 palmitoleic acids on skeletal muscle differentiation in a hyperglycemic condition

Maternal obesity and diabetes are known to be involved in fetal myogenesis, but the later stages of myogenesis are not well understood. In this study, we investigated the influence of a hyperglycemic environment on L6 skeletal myoblast differentiation and the function of omega-7 palmitoleic acids. Exposure to a high concentration of glucose (25 mM) in high-glucose culture medium (HG) increased the expression of myogenic genes (MyoD, Myogenin, MRF4, Myhc2x, and Myhc2a) and the synthesis of myosin. HG also activated the PI3K/AKT pathway revealed muscle cell differentiation. Furthermore, the levels of reactive oxygen species (ROS) and an inflammatory cytokine (Tnfaip3; tumor necrosis factor alpha-induced protein 3), which are crucial for the growth and differentiation of skeletal muscle, were increased by HG. Palmitoleic acids suppressed the expression levels of myogenic regulatory genes and increased the expression level of a cell proliferation-related gene (Pax3). Trans-palmitoleic acid and eicosapentaenoic acid (TPA and EPA) increased the phosphorylation level of MAPK/ERK1/2 and downregulated ROS generation and Tnfaip3 expression. In contrast, cis-palmitoleic acid inactivated MAPK/ERK1/2, leading to increased ROS generation. In conclusion, a hyperglycemic environment mediated by HG induced excessive muscle differentiation. Palmitoleic acids inhibited myoblast differentiation by downregulating muscle-specific genes. Moreover, trans-palmitoleic acids may have beneficial antioxidant and/or anti-inflammatory effects in cells.