Virological and serological studies of porcine respiratory coronavirus infection on a Japanese farm

We detected transmissible gastroenteritis virus (TGEV) antibodies in pig farms in Tochigi prefecture, although the farms had no past record of TGEV vaccination or TGE. Among the farms, Farm A showed a high antibody incidence. We could not confirm if either TGEV or porcine respiratory coronavirus (PRCV) induced the antibodies, since conventional tests failed to discriminate PRCV from TGEV. Therefore, we conducted virological and serological examinations of this farm for 4 years to establish the etiology - TGEV or PRCV. Although no TGEV was detected, PRCVs were isolated from the nasal samples of pigs. Using a commercial ELISA kit, it was found that the antibodies detected in pigs of all the raising stages and sows were raised against PRCV but not TGEV. The phylogenetic analysis of the nucleotide sequences of the isolates showed that they were closely related to each other, and formed a separate cluster apart from the U.S.A. and European strains. In Cesarean-derived, colostrums-deprived piglets inoculated with a PRCV isolate, no clinical signs were seen, and the viruses were mainly isolated from the nasal samples. Moreover, viral genes were detected from the nasal sample of the contact pig. The result suggested that PRCV infection was located in the nasal cavity of pigs, and horizontal transmission easily occurs. From these results, PRCVs with different origins from the exotic PRCVs might be prevalent in pig farms in Japan.     

Morphological and histochemical changes in the regenerating vomeronasal epithelium

Receptor cell degeneration and regeneration within the vomeronasal organ (VNO) of the rat was studied using both electron microscopy and histochemical methods. Electron microscopy was employed to examine the morphological changes along the surface of the sensory epithelium, and histochemical markers were used to monitor the changes in the epithelial cell layers. Transection of the vomeronasal nerves induced selective degeneration of the receptor cells, and within six days, a significant decrease in the number of receptor cells was observed. During the subsequent stage of receptor cell regeneration, cilia and bud-like structures characteristic of a developing sensory epithelium were seen. By day 15, thin microvilli covering the surface of the receptor cells reappeared in the sensory epithelium. The neural cell adhesion molecule (NCAM) and two vomeronasal system-specific lectins; 1) Bandeiraea simplicifolia lectin (BSL-I) and 2) Vicia villosa agglutinin (VVA) were used as the histochemical markers. NCAM immunoreactivity on the surface of the epithelium was observed to be decreased significantly six days after nerve transection, and was restored during receptor cell regeneration (day 15). The reactivity of the two lectins, BSL-I and VVA, was decreased slightly during degeneration, but was still detectable at the time of maximum receptor cell degeneration (day 6). Lectin reactivity was restored to control levels by day 15. These findings suggest that (1) NCAM is a useful marker for vomeronasal receptor cells and that the vomeronasal system-specific lectins may bind to both receptor and supporting cells and (2) degeneration of vomeronasal receptor cells occurs during the first week (day 6) following nerve transection and the receptor cell population begins to recover within 15 days. The morphological changes observed during receptor cell regeneration suggest that the stages of VNO receptor cell regeneration are similar to those observed during development.     

Effects of low and high levels of maternal nutrition consumed for the entirety of gestation on the development of muscle,adipose tissue,bone, and the organs of Wagyu cattle fetuses

This study aimed to investigate the effects of high and low levels of energy intake during the entire gestation period on the skeletal muscle development, organ development, and adipose tissue accumulation in fetuses of Wagyu (Japanese Black) cows, a breed with highly marbled beef. Cows were allocated to a high-nutrition (n = 6) group (fed 120% of the nutritional requirement) or low-nutrition (n = 6) group (fed 60% of the nutritional requirement). The cows were artificially inseminated with semen from the same sire, and the fetuses were removed by cesarean section at 260 ± 8.3 days of fetal age and slaughtered. The whole-body, total muscle, adipose, and bone masses of the fetal half-carcasses were significantly higher in the high-nutrition group than the low-nutrition group (p = 0.0018, 0.009, 0.0004, and 0.0362, respectively). Fifteen of 20 individual muscles, five of six fat depots, nine of 17 organs, and seven of 12 bones that were investigated had significantly higher masses in the high-nutrition group than the low-nutrition group. The crude components and amino acid composition of the longissimus muscle significantly differed between the low- and high-nutrition groups. These data indicate that maternal nutrition during gestation has a marked effect on the muscle, bone, and adipose tissue development of Wagyu cattle fetuses.     

Model Intercomparison Study of Long Range Transport and Sulfur Deposition in East Asia (MICS-ASIA)

To help improve the use of models in science & policy analysis in Asia it is necessary to have a better understanding of model performance and uncertainties. Towards this goal an intercomparison exercise has been initiated as a collaborative study of scientists interested in long-range transport in East Asia. An overview of this study is presented in this paper. The study consists of a set of prescribed test calculations with carefully controlled experiments. Models used the same domain, emission inventory, model parameters, meteorological conditions, etc. Two periods (January and May 1993) were selected to reflect long-range transport conditions under two distinct seasons. During these periods measurements of sulfur concentrations and deposition were made throughout the study region using identical sampling and analysis protocols. The intercomparison activity consists of four tasks (Blind Test, Fixed Parameter Test, Source Receptor test, and Tuning Test). All participants were asked to do Task A, and as many of the other tasks as possible. To date seven different models have participated in this study. Results and key findings are presented.     

Identification of six phenylpropanoids from garlic skin as major antioxidants

The extract of garlic skins (peels) showed strong antioxidant activity, and some responsible constituents were isolated and identified. Garlic (Allium sativum L.) has been used as an herbal medicine, but there is no report on the health benefits of the skin or peel. In this study, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of garlic skin extract was evaluated. Using chromatographic techniques, the active constituents were isolated and subsequently identified. Analyses by high-performance liquid chromatography coupled with a photodiode array detector (HPLC-PDA) suggested that these compounds were phenylpropanoids, which had a characteristic absorbance at 300-320 nm. Liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance analyses allowed the chemical structures of the isolated constituents to be postulated. The proposed compounds were subsequently synthesized and compared with the constituents in the extract using HPLC-PDA and LC-MS. N-trans-Coumaroyloctopamine, N-trans-feruloyloctopamine, guaiacylglycerol-beta-ferulic acid ether, and guaiacylglycerol-beta-caffeic acid ether were identified as were trans-coumaric acid and trans-ferulic acid. Also, the antioxidant activities of these compounds were determined.     

Geographical distribution and seasonality of the prevalence of Leucocytozoon lovati in Japanese rock ptarmigans (Lagopus mutus japonicus) found in the alpine regions of Japan

In this study, we investigated the geographical distribution and seasonality of Leucocytozoon lovati infection in the Japanese rock ptarmigan (Lagopus mutus japonicus); this bird is one of the special natural monuments of Japan that inhabits the Japanese alpine regions. We examined blood samples from birds captured in the Kubiki, Hida, and Akaishi mountain ranges for three years from 2002 to 2005. Seventy-three blood samples from 42 males, 19 females, and 12 birds of unknown sex were used for this study. The rate of infection with L. lovati was 78.1% in the 73 birds examined. We demonstrated that the L. lovati infection was distributed across wide ranges of ptarmigan populations from the northern to the southern alpine zones. There was no sex bias in the prevalence ratio. The prevalence of L. lovati and the level of parasitization of the blood cells tended to increase from spring through summer; in contrast, a decrease was observed from summer through autumn. Although L. lovati infection was observed in a number of local populations inhabiting three mountainous regions, no infected birds were found in Mt. Johnen-dake and Mt. Maejohnen-dake. It is necessary to continue surveying the relationship between the population dynamics of the ptarmigan and the density of the arthropod vector from the perspective of in situ conservation of this endangered species.     

Experimental studies of the load reducing effects of nitroglycerin in heart failure

The load-reducing effect of nitroglycerin (NTG), a vasodilator, was studied in dogs with heart failure. The chordae tendineae of the mitral valve were transected to induce acute mitral regurgitation (MR) for hemodynamic evaluation. By such surgical treatment, preload indices such as left ventricular end-diastolic pressure (LVEDP) and left atrial pressure (LAP) increased significantly, and subsequent cardiac dysfunction and heart failure were indicated by another decrease in stroke volume, myocardial contractility, forward flow, and myocardial oxygen consumption. To dogs with artificially established acute MR, 3 micrograms/kg/min of NTG was administered intra-arterially by means of a continuous infusion, that resulted in decrease of LVEDP, LAP and central venous pressure (CVP). Thus, a reduction of preload was determined. Simultaneously, afterload indices such as aortic systolic pressure (Aos), aortic mean pressure (Aom) and total peripheral resistance (TPR) decreased remarkably. Afterload reduction depended on the amount of venous return; therefore, an extra-corporeal circulation system was applied in order to supply a constant venous return before NTG administration. This caused a significant decrease in aortic diastolic pressure (Aod), Aos, Aom, left ventricular systolic pressure (LVSP) and TPR, and an increase in myocardial contractility and cardiac output. This suggested that afterload reduction might be realized by the vasodilatory effect of NTG on the resistance vessels.     

Effects of Intensification of the Amazon River Prawn,Macrobrachium amazonicum,Grow‐out on Effluent Quality

Studies to determine suitable levels of intensification are essential for developing sustainable aquaculture. The objective of this study was to evaluate the quality of effluents discharged from ponds stocked with 10 (D10), 20 (D20), 40 (D40), and 80 (D80) postlarvae of Macrobrachium amazonicum/m². Intake and effluent water samples were taken throughout a 5.5‐mo grow‐out cycle. In that study, twelve 0.01‐ha earthen ponds were stocked postlarvae with 0.01 g. Average water exchange rate was 15%/d; water was discharged from the bottom of the ponds. Prawns were fed a commercial feed with 38% crude protein according to their biomass (3–10%) and the concentration of dissolved oxygen (DO). In our research, temperature, turbidity, total suspended solids, conductivity, DO, pH, biochemical oxygen demand (BOD), chemical oxygen demand (COD), N‐ammonia, N‐nitrite, N‐nitrate, N‐Kjeldahl nitrogen, total phosphorus, and soluble orthophosphate were measured every 15 d throughout the experiment in the early morning (0630 to 0730 h). Turbidity was lower in D10 than in D20 and D40 and total phosphorus was higher in D80 than in D10 and D20. An analysis of principal components comparing treatments and intake water showed three groups: intake, D10 and a cluster of D20, D40, and D80. On the basis of the water characteristics found in our study it appears that the farming of M. amazonicum is likely to have a low environmental impact, at least up to a stocking density of 80 prawns/m².     

Alkaloids from the poisonous plant Ipomoea carnea: effects on intracellular lysosomal glycosidase activities in human lymphoblast cultures

There is natural intoxication of livestock by the ingestion of Ipomoea carnea (Convolvulaceae) in Brazil and other parts of the world. The alkaloidal glycosidase inhibitors swainsonine, 2-epi-lentiginosine, and calystegines B(1), B(2), B(3), and C(1) have been identified as constituents of this plant. Swainsonine is a potent inhibitor of rat lysosomal alpha-mannosidase, with an IC(50) value of 0.02 microM, whereas calystegines B(1), B(2), and C(1) are potent inhibitors of rat lysosomal beta-glucosidase, with IC(50) values of 2.1, 0.75, and 0.84 microM, respectively. The action of swainsonine results in a lysosomal storage disorder that closely mimics alpha-mannosidosis in humans. To determine whether the toxicity of I. carnea to livestock is due to purely swainsonine or due to a combination of effects by swainsonine and calystegines, intracellular lysosomal glycosidase activities in normal human lymphoblasts grown with inhibitors in the medium were examined. Incubation of lymphoblasts with 0.1 microM swainsonine for 3 days resulted in approximately 60% reduction of alpha-mannosidase activity. On the other hand, calystegines B(2) and C(1) showed no inhibition of beta-glucosidase up to 1 mM; instead inclusion of calystegines B(2) and C(1) at 100 microM in the culture medium increased its activity by 1.5- and 1.6-fold, respectively. Calystegines B(2) and C(1) seem to act as chemical chaperones, enhancing correct folding of the enzyme and enabling smooth trafficking to the lysosome. The lysosomal beta-glucosidase inhibitory calystegines seem to have little risk of inducing intoxication of livestock.