Characterization of turkey coronavirus from turkey poults with acute enteritis.

The present study was to characterize turkey coronavirus associated with turkey poult enteritis and mortality. Intestinal contents or intestines from affected turkey poults and inoculated turkey embryos contained coronaviruses as revealed by electron microscopy or were positive for turkey coronavirus by immunofluorescent antibody assay. Sucrose density gradient ultracentrifugation of the virus-containing intestinal homogenate yielded two opalescent bands corresponding to the buoyant densities of 1.14-1.15 and 1.18-1.20 g/ml, respectively. Coronaviral particles from intestinal contents or the sucrose density gradient preparation were mainly spherical in shape and had envelope and central depression. They were surrounded by a fringe of regularly spaced petal-shaped projections attached to the particles by a short stalk. Purified viruses hemagglutinated rabbit erythrocytes with a titer of 16. Major protein bands of purified viruses analyzed by SDS-PAGE were located at 200, 100-110, 50-60, and 30-35 kDa. The patterns of protein bands were consistent with those of Minnesota or Quebec turkey coronavirus isolates. A 568 bp nucleotide fragment of turkey coronavirus spike protein gene was amplified from RNA of inoculated turkey embryo intestine or purified virus. Sequence analysis of the 568 bp PCR product revealed high degree of identity with the corresponding spike protein gene sequence of human and bovine coronaviruses. The results indicated that turkey coronavirus was associated with turkey poults with acute enteritis.     

Mapping of QTLs in tomato line FLA456 associated with resistance to a virus causing tomato yellow leaf curl disease

Tomato (yellow) leaf curl disease (TYLCD) is a serious threat to tomato production in the tropics and subtropics. The genetics of resistance to Tomato yellow leaf curl Thailand virus Taiwan strain (TYLCTHV-[TW]) in a highly resistant tomato line FLA456 was studied through quantitative trait loci (QTL) analysis. Four QTLs named qTy4.1, qTy6.1, qTy10.1 and qTy11.1 were detected on chromosomes 4, 6, 10, and 11, respectively, through evaluation of an F₆ recombinant inbred line (RIL) population derived from a cross between FLA456 (resistant) and CLN1621L (susceptible). Gene action of all QTLs was recessive based on disease reaction of the F₁. The markers SINAC1 and SLM4-34 flanked qTy4.1 on chromosome 4, and SLM11-12 and SLM11-17 defined qTy11.1, which co-located with the previously identified Ty-5 and Ty-2 loci, respectively. qTy6.1 was flanked by the markers SLM6-55 and TES-0014, and qTy10.1 by the markers SLM10-80-SLM10-46 on chromosomes 6 and 10. The LOD values of the putative QTLs ranged from 2.79 to 13.76. The phenotypic variance explained by each QTL ranged from 7.1 to 31.9 %. The four QTLs collectively contributed about 60.5 % of the phenotypic variation in resistance against TYLCTHV-[TW]. Group mean severity scores of those RILs possessing three or four qTy were generally lower than RIL groups with only one or no qTy. Given the diversity of begomoviruses that cause TYLCD across the regions, the new QTLs from FLA456 would be valuable in tomato breeding for developing varieties with durable resistance. Two QTL intervals (qTy4.1 and qTy10.1) contained virus resistance candidate genes such as CTV.22 and eukaryotic translation initiation factor 4E.     

The Combinatorial Effect of Different Equex STM Paste Concentrations,Cryoprotectants and the Straw‐Freezing Methods on the Post‐Thaw Boar Semen Quality

This study was to evaluate the combinatorial effect (14 treatments, A–N) of different Equex STM paste concentrations, cryoprotectants and the straw‐freezing method on the post‐thaw boar semen quality. Two ejaculates were collected from each of nine boars (three boars from each of three breeds). Semen was diluted in extenders with different concentrations of Equex STM paste and different cryoprotectants [glycerol or dimethylacetamide (DMA)] before cryopreserving via liquid nitrogen or dry ice. Motility, viability, percentage of spermatozoa with intense acrosomal staining and with normal morphology of post‐thaw sperm were evaluated. The qualities of thawed semen were best preserved in treatment H (extender with 0.5% Equex STM paste and 5% glycerol and freezing by dry ice) and were worst in treatment B (extender with 0% Equex STM paste and 5% DMA and freezing by dry ice). Significant difference (p < 0.05) was present in post‐thawed sperm motility (63% vs 27%), sperm viability (70% vs 33%) and sperm acrosomal integrity rate (68% vs 29%) between treatments H and B. However, sperm proportion with normal morphology showed no significant difference among treatments (66% vs 66%; p > 0.05). Moreover, statistical analysis suggests that no significant difference was present in semen quality among breed or individual donors (p > 0.05). These findings suggest that Equex STM paste improved the cryosurvival efficiency of boar sperm, and the favourable straw‐freezing method changes between glycerol and DMA.     

High processing tolerances of immunomodulatory proteins in Enoki and Reishi mushrooms

This study investigated the processing tolerances of two mushroom proteins with immunomodulatory activities, including FVE from Enoki ( Flammulia velutipes ) and LZ8 from Reishi ( Ganoderma lucidum ) mushrooms, under food processing treatments such as heating, sterilization, frozen storage, extraction in acid/alkaline conditions, and dehydration. Results showed that the capability of these two proteins to induce IFN-gamma secretion by murine splenocytes remained after 100 degrees C heating for 30 min, 121 degrees C autoclaving for 15 min, and -80 degrees C freezing. The retained activities of both proteins on cell proliferation and IFN-gamma production did not decrease at 0.6 M hydrochloric acid (at pH 2) but strikingly dropped at 5 M sodium hydrate (at pH 13). After vacuum dehydration, FVE and LZ8 retained most of their activities on cell proliferation; nevertheless, the IFN-gamma secretion decreased to about half of the initial values. These findings suggest that these two mushroom proteins have a good thermal/freezing resistance, acid tolerance, and dehydration stability and are candidates for processing in food and nutraceutical utilization.     

Treatment with taurine attenuates hepatic apoptosis in NZB/W F1 mice fed with a high-cholesterol diet

Cholesterol-rich diets are known to cause hepatic apoptosis, which has been associated with the pathogenesis of systemic lupus erythematosus (SLE). However, the mechanisms and treatments for hepatic apoptosis in SLE are poorly understood. To clarify the effects of taurine on hepatic apoptosis in SLE, NZB/W F1 mice received control, cholesterol, and cholesterol/taurine diets. Significant reductions of caspase-3 activity, TUNEL-positive cells, and Fas- and mitochondrial- dependent apoptosis were detected in liver from the cholesterol/taurine group as compared to the cholesterol group. Moreover, significant increases of phosphorylated AKT, NF-kappaB (p65), and ERK1/2 proteins were detected in liver from the cholesterol/taurine group as compared to the cholesterol group. In contrast, a significant reduction of phosphorylated p38 protein was observed in the cholesterol/taurine group. These experimental results demonstrated positive effects of taurine against hepatic apoptosis in NZB/W F1 mice fed a high-cholesterol diet and suggested the therapeutic potential of taurine in SLE.     

A cell-based screening identifies compounds from the stem of Momordica charantia that overcome insulin resistance and activate AMP-activated protein kinase

Treatment of insulin resistance is a critical strategy in the prevention and management of type 2 diabetes. The crude extracts from all parts of Momordica charantia L. have been reported by many studies for the effective treatment of diabetes and related complications. However, the exact ingredients responsible for the hypoglycemic effect and the underlying mechanism of their actions have not been well characterized because of the lack of a proper assay and screening system. A new cell-based, nonradioactive, and nonfluorescent screening method was demonstrated in this study to screen for natural products from the stem of M. charantia, aiming to identify hypoglycemic components that can overcome cellular insulin resistance. The results suggest triterpenoids being potential hypoglycemic components of the plant and the mechanism underlying their action involving AMP-activated protein kinase.     

Characterization of porcine circovirus type 2 (PCV2) infection in swine lymphocytes using mitogen-stimulated peripheral blood lymphocytes from healthy PCV2-carrier pigs

Information regarding the susceptibility of swine lymphocytes to PCV2 is rather limited. To further explore and characterize the PCV2 infection in swine lymphocytes, an in vitro model using concanavalin A (Con A)-stimulated peripheral blood lymphocytes (PBLs) obtained from clinically healthy PCV2-carrier pigs was introduced. It was found that the PCV2 antigen-containing rate was below 2% in PBLs from healthy PCV2-free pigs following treated simultaneously with Con A and PCV2. However, significantly higher PCV2 antigen- and nucleic acid-containing rates could be seen in Con A-stimulated PBLs from clinically healthy PCV2-carrier pigs. Prior to Con A treatment, both of the PCV2 antigen- and nucleic acid-containing rates in PBLs from healthy PCV2-carrier pigs were less than 1%; however, they reached 22.1+/-5.7% by flow cytometry and 27.1+/-6.5% by in situ hybridization, respectively, at 4-day post-incubation with Con A. Phenotyping of PCV2 antigen-containing cells revealed that PCV2-positive cells could be detected in both T and B lymphocyte populations within which IgM-positive B lymphocytes appeared to have a relatively higher positive rate. The Con A-stimulated PBLs also displayed a significantly higher viral load by the measurement of either PCV2 DNA copy number or viral titer when compared with the non-treated PBLs from healthy PCV2-carrier pigs. The results indicate that PBLs, especially IgM-bearing B lymphocytes, are indeed susceptible to PCV2 infection and PCV2 is capable of replicating in dividing lymphocytes. This activation-induced replication may explain in part the pathogenesis of lymphoid depletion in PMWS-affected pigs.     

Immune response of largemouth bass, <Emphasis Type="Italic">Micropterus salmoides</Emphasis>, to whole cells of different <Emphasis Type="Italic">Nocardia seriolae</Emphasis> strains

Largemouth bass Micropterus salmoides were immunized with four different N. seriolae strains—two α-glucosidase-positive (961113, KU040801) and two α-glucosidase-negative (94260, OTTS) strains—along with Freund’s incomplete adjuvant. After primary immunization (week 0), a booster was administered at weeks 4 and 8. Nonspecific immune responses to multiple immunizations with the different N. seriolae strains were determined based on serum lysozyme activity and nitroblue tetrazolium (NBT)-positive cells in peripheral blood. The serum lysozyme activity and NBT-positive cells in peripheral blood were not significantly increased even after the two booster immunizations. Specific antibody responses against N. seriolae cells were investigated by enzyme-linked immunosorbent assay. At 4 weeks after immunization, all groups immunized with N. seriolae antigens showed significant increases in their specific antibody levels. The sera from fish immunized with different N. seriolae strains exhibited reactivity with N. seriolae sonicated antigens of 28, 30, 36 and 84 kDa by western blot analysis. After two boosters, fish were challenged with live N. seriolae to assess the vaccine’s efficacy; however, multiple injections of the N. seriolae strains did not reduce mortality, irrespective of the bacterin.     

Averting the baiji syndrome: conserving habitat for critically endangered dolphins in Eastern Taiwan Strait

• 1. Numbering no more than 100 individuals and facing many threats, the geographically isolated Eastern Taiwan Strait population of Indo‐Pacific humpback dolphins (Sousa chinensis) is in peril. The estuarine and coastal waters of central‐western Taiwan have historically provided prime habitat for these dolphins, but environmental conditions today bear little resemblance to what they were in the past.
• 2. The humpback dolphins must share their habitat with thousands of fishing vessels and numerous factories built upon thousands of hectares of reclaimed land.
• 3. They are exposed to chemicals and sewage released from adjacent terrestrial activities. Noise and disturbance associated with construction, vessel traffic and military activities are features of everyday life for these animals.
• 4. Measures to slow the pace of habitat deterioration and reduce the many risks to the dolphins are urgently needed. As one practical step in this direction, this paper describes the habitat needs of these small cetaceans so that decision makers will be better equipped to define ‘priority habitat’ and implement much needed protection measures under the terms of local legislation.
• 5. The preferred habitat of these dolphins in Taiwan consists of shallow (<30 m), near‐shore marine waters with regular freshwater inputs.
• 6. For such a small, isolated and threatened population, ‘priority habitat’ should not be limited to areas of particularly intensive dolphin use or high dolphin density, but rather it should encompass the entire area where the animals have been observed (their current ‘habitat’), as well as additional coastal areas with similar bio‐physical features (‘suitable habitat’). Such a precautionary approach is warranted because the loss of only a few individuals could have serious population‐level consequences.
• 7. While conventional socio‐economic analysis might suggest that implementing protection measures over an area stretching ～350 km north–south along Taiwan's west coast and ～3 km out to sea would be too ‘costly’, the loss of this charismatic species from Taiwan's waters would send a troubling message regarding our collective ability to reconcile human activities with environmental sustainability. Copyright © 2010 John Wiley & Sons, Ltd.

     

Development and testing of a Bayesian population model for the bigeye thresher shark,Alopias superciliosus,in an area subset of the western North Pacific

A Bayesian population modelling tool integrating separable virtual population analysis, per‐recruit models and age‐structured demographic analysis was developed for the bigeye thresher Alopias superciliosus (Lowe) population in an area subset of the western North Pacific. The mortality rates for years 1989–2016 were estimated, various biological reference points and associated risks of decline were also estimated, and alternative harvest strategies for the stock were evaluated. Estimates of the posterior mean of fishing mortality for bigeye thresher shark suggest fishing pressure has been high in recent years (2011–2016). The estimated population growth rate (λ) (without fishing) obtained from age‐structured demographic model was relatively low (λ = 1.01 per year; 95% confidence intervals of 1.00 and 1.03 per year). Risk analyses revealed that only low levels of fishing pressure (10% of the current fishing pressure) over a wide range of ages could maintain a relatively low risk of population decline for bigeye threshers. Sensitivity testing indicated that the model is robust to prior specification. The developed framework could be used as an assessment tool to evaluate the risk of decline for other widely distributed pelagic shark species where insufficient catch and effort data are available.