Feeding Solanum glaucophyllum to preparturient multiparous cows prevents postparturient hypocalcemia

Solanum glaucophyllum (SG) contains 1,25‐dihydroxyvitamin D₃ (1,25‐(OH)₂D₃) glycosides. We investigated the effect of SG on hypocalcemia in cows. Serum levels of 1,25‐(OH)₂D₃, total calcium and phosphorus dose‐relatedly increased after feeding with SG, while serum magnesium and chloride levels fell (P < 0.05). We also performed an ethylenediaminetetraacetic acid (EDTA) infusion to induce artificial hypocalcemia. Cows that had been fed 4.0 mg/kg body weight of SG daily for 2 weeks had a higher serum concentration of total calcium at the end of EDTA infusion than those not fed SG (P < 0.05). In a field trial, multiparous cows were assigned to one of four groups: (1) no SG, (2) 1.3 g or (3) 2.6 g of SG daily from 14 days before the estimated calving day until 3 days after calving, or (4) a single feed of 35.75 g SG at 3 days before the estimated calving day. The concentrations of serum total calcium after the calving in each treatment group were (1) 7.4, (2) 7.9, (3) 8.0 and (4) 8.9 mg/dL and higher for (4) than for (1) (P < 0.05). The data suggests that feeding a high dose of SG before the calving may maintain higher concentrations of serum calcium after the calving.     

Simultaneous steroids measurement in dogs with hyperadrenocorticism using a column-switching liquid chromatography-tandem mass spectrometry method

We developed an analytical method using an on-line column-switching liquid chromatography with triple quadrupole mass spectrometry (LC/MS/MS) for quantifying multiple steroids in serum. Using the developed method, we evaluated the serum concentration of nine steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 21-deoxycortisol, deoxycorticosterone, progesterone, 17α-OH-progesterone and aldosterone) in dogs with hyperadrenocorticism (HAC). Serum was mixed with stable isotope internal standards and thereafter purified by the automated column-switching system. The limit of detection ranged 2–16 pg/ml for nine steroids. In the baseline samples, five steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, and 17α-OH-progesterone) were detected in all dogs. The concentrations of cortisone, 11-deoxycortisol, and 17α-OH-progesterone in dogs with HAC (n=19) were significantly higher those in dogs without HAC (n=15, P<0.02). After the adrenocorticotropic hormone stimulation test, six steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 17α-OH-progesterone, and deoxycorticosterone) were above the limit of quantification in all dogs. Cortisol, corticosterone, cortisone, and deoxycorticosterone concentrations of dogs with HAC were significantly higher than those of dogs without HAC (P<0.02). In addition, 11-deoxycortisol and 17α-OH-progesterone concentration was higher in dogs with HAC than in dogs without HAC (P=0.044 and P=0.048, respectively). The on-line column-switching LC/MS/MS would be feasible for measuring multiple steroids in dog serum. The results suggest that cortisone, 11-deoxycortisol, and 17α-OH-progesterone would be related to HAC. Further studies are warranted to assess the clinical feasibility of steroid profile in dogs with HAC.     

Rice cultivation and its environmental conditions in the mediterranean countries I. Climate and physiography in relation to rice cultivation

In order to characterize the rice cultivation in the Mediterranean countries, climate and physiography of rice growing areas in Italy, Spain, and Portugal were studied. In northern Italy rice is extensively cultivated on terraces and fluvial plains of the Po and its tributaries, making use of the supply of water from Alpine glaciers in addition to the relatively abundant rainfalls of the area. In the Iberian peninsula summer climate is so dry that rice is cultivable only where the ratio of catchment area to rice land area is sufficiently large, thus rice lands are practically limited to the floodplains and deltas of the big rivers, such as the Sado and the Tejo in Portugal and the Guadalquivir and the Ebro in Spain.

In order to characterize the rice cultivation in the Mediterranean countries, climate and physiography of rice growing areas in Italy, Spain, and Portugal were studied. In northern Italy rice is extensively cultivated on terraces and fluvial plains of the Po and its tributaries, making use of the supply of water from Alpine glaciers in addition to the relatively abundant rainfalls of the area. In the Iberian peninsula summer climate is so dry that rice is cultivable only where the ratio of catchment area to rice land area is sufficiently large, thus rice lands are practically limited to the floodplains and deltas of the big rivers, such as the Sado and the Tejo in Portugal and the Guadalquivir and the Ebro in Spain.     

Antigenic differences between H5N1 human influenza viruses isolated in 1997 and 2003

To assess whether the antigenic properties of H5 hemagglutinin (HA) change over time due to antigenic drift, we produced a panel of monoclonal antibodies (mAbs) against the HA of the index H5N1 human influenza A virus, A/Hong Kong/156/97. By immunizing mice with a plasmid expressing this HA and boosting the initial immunization with cell lysates transfected with the plasmid, a total of six hybridomas producing HA-specific mAbs were established: four to the HA1 subunit with hemadsorption-inhibiting activity and two to the HA2 subunit. None of the mAbs to HA1 could bind to the HA of a recent human isolate, A/Hong Kong/213/2003, indicating that there are substantial antigenic differences between the H5N1 human influenza virus isolated in 1997 and that isolated in 2003.     

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 6-hydroxydopamine on the development of the immune system in chickens

It has been suggested that the sympathetic nervous system communicates with lymphocytes expressing cell surface receptors for neurotransmitters such as norepinephrine (NE), on the basis of the finding that neurotransmitters modify immune responses in mammalian species. We confirmed that chicken lymphocytes in the brusa of Fabricius, thymus and spleen expressed beta-adrenergic receptor (beta-AR) mRNA from embryonic day (E) 10 and that intracellular cAMP level was elevated by NE, suggesting that lymphocytes express functional beta-AR on their surface at an early embryonal stage. To clarify whether the nervous system is involved in the development of the immune system, the effects of 6-hydroxydopamine (6-OHDA), one of sympathectomizing agents, on chicken lymphocytes was investigated. A single injection of 6-OHDA at a dose of 400 microg into a chicken embryo was carried out at E7 or 14 (as referred to E7 group and E14 group, respectively). NE level and the relative proportion of Bu-1a(+), CD4(+) and CD8(+) cells in the spleen of 3-week-old chickens were not altered by 6-OHDA treatment. However, the proliferative responses and expression of IL-2 mRNA in spleen cells cultured with pokeweed mitogen were reduced in E7 group compared with those of control. Furthermore, in CD8(+) spleen cells of E14 group of 3-week-old chickens, the expression of beta-AR mRNA and the relative increase of intracellular cAMP stimulated with NE were significantly decreased. These results suggest that the sympathetic nervous system affects the development of the immune system.     

Detection of mitogen-induced lymphocyte proliferation by bromodeoxyuridine (BrdU) incorporation in the chicken

3H-thymidine (3H-TdR) incorporation assay has been generally used to measure lymphocyte proliferation in the chicken. Disadvantages of this assay are that radioisotope is biological hazard to the person and environment and that it can not measure which subset of lymphocytes proliferates. In this study, bromodeoxyuridine (BrdU) incorporation assay by flow cytometry was compared with 3H-TdR incorporation assay. As a result, BrdU incorporation assay showed a strong correlation with 3H-TdR incorporation assay, and it could be applied simultaneously to detect BrdU incorporation and expression of cell surface marker antigens. These results suggest that the BrdU incorporation assay by flow cytometry is useful to analyze lymphocyte proliferation in detail.     

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.