Effects of lignin‐modified Populus tremuloides on soil organic carbon

Several genes in the aspen genome have been modified to generate stem wood with lower lignin content and an altered lignin composition. Lower lignin in wood reduces the time and energy required for pulping. Further, this modification can also increase the allocation of photosynthate to cellulose and total biomass production, potentially increasing CO₂‐sequestration capacity. However, widespread planting of trees with altered lignin content and composition could alter soil organic‐C dynamics in complex ways. To further examine the effects of altered lignin biosynthesis on plant growth and accrual of soil organic C (SOC), we conducted a repeated greenhouse study with four lines of transgenic aspen (Populus tremuloides Michx.) and one wild‐type (control) aspen. Accrual of aspen‐derived SOC was quantified by growing aspen trees (C3 plants) in C4 soil and measuring changes in the natural abundance of δ¹³C. We measured plant growth, biomass, and C content and combined these data with SOC measurements to create C budgets for the plant mesocosms. Lignin modifications resulted in differences in the accrual of aspen‐derived SOC and total mesocosm C, primarily due to differences in biomass between genetically modified lines of aspen. One genetic alteration (low lignin, line 23) was able to perform similarly or better than the wild‐type aspen (control, line 271) without altering SOC. Alterations in lignin structure (S : G ratios) had negative effects on biomass production and SOC formation. The addition of new (aspen‐derived) SOC was proportional to the loss of existing SOC, evidence for a priming effect. The pool of new SOC was related to total plant biomass, suggesting that the effects of lignin modification on SOC are driven by changes in plant growth.     

Immunopathological characterization of porcine circovirus type 2 infection-associated follicular changes in inguinal lymph nodes using high-throughput tissue microarray

The immunopathogenesis of porcine circovirus type 2 (PCV2) infection in conventional pigs is complicated by various environmental factors and individual variation and is difficult to be completely reproduced experimentally. In the present field-based study, a tissue microarray (TMA) consisting of a series of lymphoid follicles having different PCV2-loads was constructed using formalin-fixed and paraffin-embedded superficial inguinal lymph nodes (LNs) from 102 pigs. Using the TMA, a wide range of parameters, including co-infected viral pathogens, immune cell subsets, and cell apoptosis/proliferation activity by immunohistochemical (IHC) staining or in situ hybridization (ISH) were measured, characterized, and compared. The signal location and area extent of each parameter were interpreted by pathologists, semi-quantified by automated image analysis software, and analyzed statistically. The results herein demonstrated a significant negative correlation between PCV2 and CD79a (p<0.001) and a significant positive correlation between PCV2 and lysozyme (p<0.001) or TUNEL (p<0.001) using Pearson correlation analysis. The amount of porcine respiratory and reproductive syndrome virus (PRRSV) and porcine parvovirus antigens did not correlate with the tissue loads of PCV2 nucleic acid. Multiple regression analysis further predicted that PCV2 contributed major effects on CD79a, lysozyme, and TUNEL but PRRSV showed relatively less effects on these parameters. In addition, the total signal intensity of Ki67 (index of cell proliferation activity) did not change significantly among cases with different PCV2 loads; however, as the loading of PCV2 nucleic acid increased, the main contribution of Ki67 signal gradually shifted from B cells in the germinal center to T cells and macrophages in the interfollicular regions. In the present study, the use of TMA to establish a mathematical model with a wider range of statistical analysis can bring us a step forward to understand the immunopathogenesis of PCV2 infection-associated follicular changes in LNs.     

Molecular characterization of Cryptosporidium spp. in native breeds of cattle in Kaduna State, Nigeria

Despite numerous molecular epidemiologic studies of cryptosporidiosis in dairy cattle in industrialized countries, there are very few studies on the diversity and public health significance of Cryptosporidium species in native cattle in developing countries. In this study, a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the small-subunit (SSU) rRNA gene was used to detect and identify Cryptosporidium spp. in 194 fecal specimens from 2 to 365 days old calves in 20 White Fulani and Sokoto Gudali herds in Nigeria. Thirty one (16.0%) of the specimens were positive for Cryptosporidium. Restriction digestion of the PCR products showed the presence of Cryptosporidium bovis (7.2%), Cryptosporidium ryanae (4.1%), Cryptosporidium andersoni (2.5%), and concurrent occurrence of C. bovis and C. ryanae (1.5%), and C. bovis and C. andersoni (0.5%). There were no significant differences (p>0.05) in Cryptosporidium infection rates by sex, herd location, management system, breed of calves, or fecal consistency. However, calves 180 days or younger had a higher infection rate of Cryptosporidium than older calves (p=0.034). Likewise, younger calves also had higher occurrence of C. bovis and C. ryanae (p=0.022). The absence of zoonotic Cryptosporidium parvum in the calves studied suggests that native breeds of cattle may not be important in the transmission of human cryptosporidiosis in Kaduna State, Nigeria.     

Evaluation of the immunogenicity of a transgenic tobacco plant expressing the recombinant fusion protein of GP5 of porcine reproductive and respiratory syndrome virus and B subunit of Escherichia coli heat-labile enterotoxin in pigs

Escherichia coli heat-labile enterotoxin B subunit (LTB) can be used as an adjuvant for co-administered antigens. Our previous study showed that the expression of neutralizing epitope GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) in transgenic tobacco plant (GP5-T) could induce PRRSV-specific immune responses in pigs. A transgenic tobacco plant co-expressing LTB and PRRSV GP5 as a fusion protein (LTB-GP5-T) was further constructed and its immunogenicity was evaluated. Pigs were given orally three consecutive doses of equal concentration of recombinant GP5 protein expressed in leaves of LTB-GP5-T or GP5-T at a 2-week interval and challenged with PRRSV at 7 weeks post-initial immunization. Pigs receiving LTB-GP5-T or GP5-T developed PRRSV-specific antibody- and cell-mediated immunity and showed significantly lower viremia and tissue viral load and milder lung lesions than wild type tobacco plant (W-T). The LTB-GP5-T-treated group had relatively higher immune responses than the GP5-T-treated group, although the differences were not statistically significant.     

In vitro predatory activity of nematophagous fungi <Emphasis Type="Italic">Duddingtonia flagrans</Emphasis> on infective larvae of <Emphasis Type="Italic">Oesophagostomum</Emphasis> spp. after passing through gastrointestinal tract of pigs

One isolate of predator fungi Duddingtonia flagrans (AC001) was assessed in vitro regarding the capacity of supporting the passage through pigs' gastrointestinal tract without loss of the ability of preying infective larvae Oesophagostomum spp. Fungal isolates survived the passage and were efficient in preying L₃ since the first 8 h of collection (p < 0.01) in relation to the control group (without fungus). Compared with control, there was a significant decrease (p < 0.01) of 59.6% (8 h), 71.7% (12 h), 76.8% (24 h), 81.0% (36 h), 78.0% (48 h), 76.1% (72 h), and 82.7% (96 h) in means of infective larvae Oesophagostomum spp. recovered from treatments with isolate AC001. Linear regression coefficients of L₃ of recovered Oesophagostomum spp. regarding the collections due to time were −0.621 for control, −1.40 for AC001, and −2.64 for NF34. Fungi D. flagrans (AC001) had demonstrated to be promising for use in the biological control of pig parasite Oesophagostomum spp.     

Porcine circovirus type 2 (PCV2) infection decreases the efficacy of an attenuated classical swine fever virus (CSFV) vaccine

ABSTRACT: The Lapinized Philippines Coronel (LPC) vaccine, an attenuated strain of classical swine fever virus (CSFV), is an important tool for the prevention and control of CSFV infection and is widely and routinely used in most CSF endemic areas, including Taiwan. The aim of this study was to investigate whether PCV2 infection affects the efficacy of the LPC vaccine. Eighteen 6-week-old, cesarean-derived and colostrum-deprived (CDCD), crossbred pigs were randomly assigned to four groups. A total of 105.3 TCID50 of PCV2 was experimentally inoculated into pigs through both intranasal and intramuscular routes at 0 days post-inoculation (dpi) followed by LPC vaccination 12 days later. All the animals were challenged with wild-type CSFV (ALD stain) at 27 dpi and euthanized at 45 dpi. Following CSFV challenge, the LPC-vaccinated pigs pre-inoculated with PCV2 showed transient fever, viremia, and viral shedding in the saliva and feces. The number of IgM+, CD4+CD8-CD25+, CD4+CD8+CD25+, and CD4-CD8+CD25+ lymphocyte subsets and the level of neutralizing antibodies against CSFV were significantly higher in the animals with LPC vaccination alone than in the pigs with PCV2 inoculation/LPC vaccination. In addition, PCV2-derived inhibition of the CSFV-specific cell proliferative response of peripheral blood mononuclear cells (PBMCs) was demonstrated in an ex vivo experiment. These findings indicate that PCV2 infection decreases the efficacy of the LPC vaccine. This PCV2-derived interference may not only allow the invasion of wild-type CSFV in pig farms but also increases the difficulty of CSF prevention and control in CSF endemic areas.     

Activation of goat primordial follicles in vitro: Influence of alginate and ovarian tissue

The present study aimed to evaluate the effect of three culture systems on caprine primordial follicle activation in vitro: follicles cultured either in the isolated form within alginate (Isolated follicles + Alginate treatment), or enclosed in ovarian tissue (in situ), with or without alginate (Fragment + Alginate, and Fragment alone treatments, respectively). After culture, the Isolated follicles + Alginate treatment presented a percentage of morphologically normal follicles (MNF) similar to both the non-cultured control and the Fragment Alone treatments. Nevertheless, Fragment + Alginate treatment showed a significant reduction in the number of MNF when compared to the other treatments. Regarding follicle development, our results showed that regardless of the alginate, the presence of ovarian tissue limited primordial follicle activation during in vitro culture. Remarkably, the Isolated primordial follicle + Alginate treatment was the only one that significantly promoted follicle activation and increased both follicle and oocyte diameters during IVFC, pointing out a higher cell proliferation. In conclusion, the presence of ovarian tissue with or without alginate limited follicle development (activation) after culture. Nevertheless, when primordial follicles were isolated and encapsulated in alginate they presented suitable survival rates, higher rates of follicle activation and continued to grow throughout the culture period.     

Tyrosine phosphorylation is not a relevant mechanism to modulate aquaporin 2 activity in gestational queen endometrium and placenta

Aquaporins have been shown to be regulated by phosphorylation of serine residues, but the possible role of tyrosine residues phosphorylation has not been evaluated. Changes in the localization of aquaporin 2 (AQP2) in the queen endometrium have been related to serum progesterone levels. The aim of this study was to determine whether these AQP2-localization changes are mediated by variations in its tyrosine phosphorylation levels. Twelve queens were included in the study and divided into (a) non-macroscopically pregnant with low levels of progesterone; (b) non-macroscopically pregnant with high levels of progesterone; (c) 30 days of pregnancy; and (d) 60 days of pregnancy. Samples from endometrium and placental transference zone were obtained, immunoprecipitated and analysed by immunoblotting to determine the abundance of AQP2 and its relative levels of tyrosine phosphorylation. No significant differences in the tyrosine phosphorylation levels of immunoprecipated-AQP2 were observed between groups. We can thus conclude that changes in the localization of AQP2 in the queen endometrium are not modulated by tyrosine phosphorylation.     

Automatic adjustment of harrowing intensity in cereals using digital image analysis

Precision farming technologies were implemented into a commercial harrow to increase selectivity of weed harrowing in spring cereals. Digital cameras were mounted before and after the harrow measuring crop cover. Crop soil cover (CSC) was computed out of these two images. Eight field experiments were carried out in spring cereals. Mode of harrowing intensity was changed in four experiments by speed, number of passes and tine angle. Each mode was varied in five intensities. In four experiments, only intensity of harrowing was changed. Weed control efficacy (WCE) and CSC were measured immediately after harrowing. Crop recovery was assessed 14 days after harrowing. Modes of intensity were not significantly different. However, intensity had significant effects on WCE and CSC. Cereals recovered from 10% CSC, and selectivity was in the constant range at 10% CSC. Therefore, 10% CSC was the threshold for the decision algorithm. If the actual CSC was below 10% CSC, intensity was increased. If the actual CSC was higher than 10%, intensity was decreased. Image analysis, decision support system and automatic control of harrowing intensity by hydraulic adjustment of tine angle were installed on a controller mounted on the harrow. The new system was tested in an additional field study. Threshold values for CSC were set at 10%, 30% and 60%. Automatic tine angle adjustment precisely realised the three different CSC values with variations of 1.5% to 3%. This development contributes to selective weed control and supports farmers during harrowing.