Effects of Developmental Stage, Embryonic Interferon-τ Secretion and Recipient Synchrony on Pregnancy Rate after Transfer of in vitro Produced Bovine Blastocysts

Three separate trials of bovine embryo transfers were performed consisting of 32, 41 and 33 transfers, respectively, to examine the effects of (a) the developmental stage of in vitro‐derived blastocysts, (b) the amount of interferon‐τ (IFN‐τ) they secreted during culture and (c) the cyclic stage of the recipient at the time of transfer on the probability of establishment of pregnancy. One blastocyst was transferred into the ipsilateral uterine horn to the CL. At the time of transfer, blastocysts were classified into one of three developmental stages (early blastocyst, blastocyst and expanded blastocyst) and the cyclic stage of each cow was assessed (?12 h, on time, +12 h, +24 h, >24 h). Prior to the second and third trials, blastocysts were individually cultured for 24 h in 50 μl medium droplets and the IFN‐τ concentration in the droplet was determined. Logistic regression analyses revealed that expanded blastocysts had a significantly higher likelihood of establishing pregnancy (p = 0.009), and that there was a significant interaction with the cyclic stage of the recipient in this group with lower rates of pregnancy resulting from decreasing synchrony with the recipient (p = 0.033). IFN‐τ secretion during culture was significantly higher in expanded blastocysts than in the other two groups (p < 0.05). A significant effect of the pre‐transfer level of IFN‐τ secretion was found only in the ‘Blastocyst’ group where transfer of embryos with lower IFN‐τ production prior to transfer resulted in higher pregnancy rates (p = 0.047). These results demonstrate that IFN‐τ secretion may be a useful tool to predict pregnancy outcome, but only within certain developmental stages.     

Antigenic and genotypical characterization of Newcastle disease viruses isolated in Taiwan between 1969 and 1996

Three major epidemics of Newcastle disease (ND) occurred in Taiwan over the past three decades (in 1969, 1984, and 1995). In order to gain a better understanding of the relationships between past ND epizootics in Taiwan, 36 ND viruses (NDVs) isolated between 1969 and 1996 were characterized antigenically and genotypically. The antigenicity of these viruses was analysed by their ability to cause binding of mouse monoclonal antibodies (mAbs) to cell cultures infected with the isolate. Using a panel of 22 mAbs to divide NDVs into subgroups, a total of 18 binding patterns were revealed. The sequences covering the cleavage site of the fusion protein gene of these isolates were also determined. The results of the phylogenetic analysis placed 36 NDVs into I, II, VIb, VIIa, VIII and two novel genotypes (provisionally termed X and VIh). The 1969 velogenic isolates were of genotypes X and VIh; the 1984-1985 velogenic isolates were genotyped VIb, VIh, VIIa, and X; while the 1995-1996 velogenic isolates were genotyped VIIa or VIII. Some 1969 and 1984 velogenic isolates were of the same mAbs binding pattern and genotype, and the mAbs binding patterns of the 1995-1996 isolates have not been seen before. It is concluded that velogenic NDVs of different genotype and antigenic type have co-circulated in Taiwan at least since 1969. Also there were epizootiological links between strains isolated in 1969 and 1984, whereas the 1995-1996 epidemic was caused by new antigenic variants.     

Effects of different forages on the chemical compositions and antiosteoporotic activities of velvet antlers

For this study, we aimed to assess the dose–response antiosteoporotic effects of the middle section of velvet antlers (VAs) from sika deers (Cervus nippon) fed with different types of fodders. VAs prepared from farmed sika deers fed with feed mixtures containing sorghum distillery residue (VA‐SDR) or without SDR (SDR replaced with hay, VA‐Hay) were divided into upper (VAU), middle (VAM) and basal (VAB) sections. The chemical constituents of the middle sections obtained from each VA type were compared, and their antiosteoporotic activities were evaluated using rats with ovaries removed surgically (ovariectomy, OVX). The VA‐Hay exhibited markedly increased iron and cysteine levels, whereas the VA‐SDR exhibited markedly increased level of alcoholic extract and testosterone. Both VA‐Hay‐ and VA‐SDR‐treated rats exhibited increased femur strength compared with the control group. However, VA‐SDR exhibited greater bone‐strengthening effects than did VA‐Hay. The serum osteocalcin and estradiol levels were significantly moderated in the VA‐Hay group alone. These results suggest that VA‐SDR and VA‐Hay prevent the loss of bone strength, and preserve trabecular architecture connectivity in an estrogen‐deficient state. However, differences in the chemical compositions of different forages may be responsible for the varying antiosteoporotic mechanisms observed. Thus, the addition of SDR in deer forage may enhance antiosteoporosis activity in VAs, and confer considerable economic and ecological benefits.     

Expression profiles of immunity and reproductive genes during transition period in Holstein cattle

The transition period is a critical time for dairy cows as the animal is subjected to the physiological stress accompanying parturition. Immunosuppression and health status were examined during this period in 80 Holstein cows. Blood samples were taken from each cow 3, 2 and 1 week before and after calving, and at calving (0 day). RNA was extracted and subjected to real‐time PCR to determine mRNA levels for the immune‐related genes TLR 2, 4, 6, 7 and β‐defensin 5 in addition to the reproduction‐related genes prolactin and IGF‐I. Results showed significant up‐regulation of pro‐inflammatory‐selected genes, TLR 4, 6 7 and β‐defensin 5 at the third‐week post‐calving; however, earlier periods had lower expression of such genes. In contrast, the immunosuppression biomarker TLR2 gene was up‐regulated at calving and 1 week after parturition and then down‐regulated again at second and third week. Prolactin and IGF‐I genes expression levels were significantly and gradually increased mainly post‐partum. This research highlights that the expression patterns of TLRs, BNBD5, PRL and IGF‐I could be biomarkers to follow up immune and reproductive status of dairy cow at peri‐parturient period to predict the most susceptible risk time for disease incidence and to build up management protocol.     

Multiple‐Particle Tracking Study of Microheterogeneity of Nutrim‐10 Suspensions

Nutrim is a newly developed food product containing dietary soluble fiber β‐glucan. The microstructural heterogeneities of Nutrim‐10 suspensions were investigated by monitoring the thermally driven displacements of well‐dispersed microspheres through video fluorescence microscopy. By comparing the distribution of the time‐dependent mean‐square displacement (MSD) of polystyrene microspheres embedded in three concentrations of Nutrim‐10 suspensions, we found that the degree of heterogeneity of suspensions increased dramatically within a narrow range of Nutrim‐10 concentrations. The ensemble‐averaged MSD of 5.5% Nutrim‐10 suspension exhibited a power‐law behavior scaling linearly with time, which was similar to the behavior for a homogeneous aqueous glycerol solution. But the MSD distribution was wider and more asymmetric than for glycerol. Increasing Nutrim‐10 concentration rendered the MSD distribution much more asymmetric and skewed. This study provided a quantitative method to characterize the organization of Nutrim‐10 in suspension.     

Effect of dietary protein on growth, feed conversion, body composition and survival of pike perch fingerlings (Sander lucioperca)

Triplicate groups of pike perch (Sander lucioperca) juveniles were fed six experimental diets containing protein levels varying from 263 to 619 g kg⁻¹ dry matter (d.m.) for 56 days. Dietary protein was supplied by graded amounts of fish meal (with 720 g kg⁻¹ crude protein). Crude lipid and gross energy content of 101–107 g kg⁻¹ and 19.9–20.6 MJ kg⁻¹ remained constant between experimental diets. Pike perch with an initial body weight of 1.05 ± 0.05 g were randomly distributed in 18 tanks of two similar recirculation systems and fed on gradually decreasing feeding rates of 10 to 6% of their body weight per day. Growth performance and feed conversion increased with dietary protein level from 263 to 549 g kg⁻¹ d.m. but did not decline at highest dietary protein level. Protein efficiency ratio declined linearly with increasing dietary protein. Survival ranged between 89.7 and 93.9% and was not affected by dietary composition. Dry matter and crude lipid content of pike perch fingerlings decreased with increasing dietary protein supply and significantly the lowest dry matter and crude lipid levels were observed in fish fed diets containing 619 g kg⁻¹ of crude protein. The dietary protein requirement for pike perch fingerlings calculated by broken‐line and second‐order polynomial regression ranged between 529 and 577 g kg⁻¹, respectively.     

Effect of lipopolysaccharide on intranasal administration of liposomal Newcastle disease virus vaccine to SPF chickens

In order to potentiate the low immunogenicity of the inactivated Newcastle disease virus immunized into chickens by mucosal route, liposomes as a drug delivery system and LPS (lipopolysaccharide) as an immuno-stimulator were evaluated. Here, we report a new nasal delivery system of inactivated Newcastle disease virus (NDV) vaccine. The intranasal vaccine was based on different lipids to form MLV (multi-lamellar vehicles) liposomes. The liposomes had combined carrier and adjuvant activities, which induced strong systemic (serum) and local (lung and nasal) humoral responses in SPF (specific-pathogen-free) chickens, and provided protective immunity. PC-Lip (phosphatidylcholine-liposome) elicited significant mucosal secretary immunoglobulin A (s-IgA) levels (p < 0.05) in tracheal lavage fluid and serum IgG levels (p < 0.05). In response to virulent viral challenge, birds treated with PBS (phosphate buffered saline) as control group died, whereas 80% of chickens which received PC-Lip, PC-Lip-LPS, PS-Lip (phosphatidylserine-liposome), and PS-Lip-LPS survived. HAI titers were 1:2560 in the PS-Lip-LPS group and 1:1280 in the PC-Lip, PC-Lip-LPS, and PS-Lip groups after two vaccinations. The results suggest that PC-Lip or PS-Lip might thus be suitable as a potential adjuvant for mucosal vaccination against NDV in chickens.     

Yeast-expressed classical swine fever virus glycoprotein E2 induces a protective immune response

Classical swine fever (CSF) is an economically important swine disease worldwide. The glycoprotein E2 of classical swine fever virus (CSFV) is a viral antigen that can induce a protective immune response against CSF. A recombinant E2 protein was constructed using the yeast Pichia pastoris expression system and evaluated for its vaccine efficacy. The yeast-expressed E2 (yE2) was shown to have N-linked glycosylation and to form homodimer molecules. Four 6-week-old specified-pathogen-free (SPF) piglets were intramuscularly immunized with yE2 twice at 3-week intervals. All yE2-vaccinated pigs could mount an anamnestic response after booster vaccination with neutralizing antibody titers ranging from 1:96 to 1:768. Neutralizing antibody titers at 10 weeks post booster vaccination ranged from 1:16 to 1:64. At this time, the pigs were subjected to challenge infection with a dose of 1 × 10⁵ TCID₅₀ (50% tissue culture infective dose) virulent CSFV strain. At 1 week post challenge infection, all of the yE2-immunized pigs were alive and without symptoms or signs of CSF. Neutralizing antibody titers at this time ranged from 1:4,800 to 1:12,800 and even to 1:51,200 one week later. In contrast, the control pigs continuously exhibited signs of CSF and had to be euthanized because of severe clinical symptoms at 6 days post challenge infection. All of the yE2-vaccinated pigs were E^rns antibody negative and had seroconverted against E^rns by post challenge day 11, suggesting that yE2 is a potential DIVA (differentiating infected from vaccinated animals) vaccine. The yeast-expressed E2 protein retains correct immunogenicity and is able to induce a protective immune response against CSFV infection.     

Seasonal changes of azimuthal,radial, and tree-to-tree variations in sap flux affect stand transpiration estimates in a <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> forest,central Taiwan

Stand transpiration (E) estimated using the sap flux methods is affected by the azimuthal, radial, and tree-to-tree variations of sap flux. Although several studies have examined the relative importance of the three variations in estimating E, the seasonality of the three variations remains unknown. In the current study, we attempted to clarify whether the relative importance of these three variations could show seasonal changes. Using sap flux data measured in a subtropical cloud forest from August 2010 to July 2011, we calculated the differences resulting from omitting the three variations in estimating E. The effects of the three variations in estimating E showed seasonality. The azimuthal and tree-to-tree variations were more pronounced during winter, whereas the radial variation was more pronounced during summer. However, the effect of tree-to-tree variation was consistently much larger than the other two variations throughout the study period. The tree-to-tree variation is more important in estimating E monthly, seasonally and annually than both the azimuthal and radial variations, although all three variations have shown seasonality. In addition, the sensor allocation for summer would be acceptable for the practical estimation of E if aiming at the long time scale.