Verification by polymerase chain reaction of vertical transmission of Theileria sergenti in cows

To evaluate the transplacental transfer of Theileria sergenti infection in cattle, we used DNA probes to detect T. sergenti in 6 pregnant cows and their calves. All the animals were monitored by parasitologic, serologic, and polymerase chain reaction (PCR) assays for a predicted 875-base-pair (bp) DNA product and a 684-bp amplicon detected by nested PCR in the blood and spleens of aborted fetuses. An open reading frame (ORF) starting at nucleotide 170 and terminating at position 1021 was shown to code for a polypeptide of 283 amino acid residues. All 6 dams and 5 calves were positive for T. sergenti in all tests. One calf was positive only with nested PCR. We conclude that transplacental transmission of T. sergenti is a significant problem. The relevance of the data in the programmed introduction of new (especially pregnant) animals into established clean herds needs serious consideration with regard to control of theileriosis and other tickborne diseases.     

Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Antiviral Effect of Saccharomyces cerevisiaeβ‐glucan to Swine Influenza Virus by Increased Production of Interferon‐γ and Nitric Oxide

The aim of these experiments was to investigate the potential antiviral effect of Saccharomyces cerevisiaeβ‐glucan on the pneumonia induced by swine influenza virus (SIV). Forty colostrum‐deprived 5‐day‐old piglets were randomly divided into four groups of 10. The 20 pigs in groups 1 and 2 were administered Saccharomyces cerevisiaeβ‐glucan orally (50 mg/day/pig; En‐Bio Technology Co., Ltd) for 3 days before SIV infection and those in groups 3 and 4 were given culture medium/diluent alone. Groups 1 and 3 were inoculated intranasally with 3 ml of tissue culture fluid containing 2 × 10⁶ tissue culture infective doses 50% (TCID₅₀)/ml of SIV and those in groups 2 and 4 were exposed in the same manner to uninfected cell culture supernatant. The microscopic lung lesions induced by SIV infection (group 1 pigs) were significantly more severe than those induced by infection in animals pre‐administered β‐glucan (group 3) (P < 0.05). Significantly more SIV nucleic acid was detected in the lungs of pigs experimentally infected with SIV only (group 1) at 5, 7 and 10 days post‐inoculation (dpi) compared with lungs from pigs pre‐administered β‐glucan and infected with SIV (group 3) (P < 0.05). The concentrations of interferon‐γ (IFN‐γ) and nitric oxide (NO) in bronchoalveolar lavage fluid from pigs pre‐administered β‐glucan and infected with SIV (group 3) were significantly higher than for any other group at 7 and 10 dpi for IFN‐γ, and at 5, 7 and 10 dpi for NO (P < 0.05). Saccharomyces cerevisiaeβ‐glucan reduced the pulmonary lesion score and viral replication rate in SIV‐infected pigs. These findings support the potential application of β‐glucan as prophylactic/treatment agent in influenza virus infection.     

Effects of Dietary Supplementation of Barodon on Growth Performance,Innate Immunity and Disease Resistance of Juvenile Olive Flounder,Paralichthys olivaceus,Against Streptococcus iniae

This study was conducted to evaluate the effects of dietary supplementation of Barodon, an anionic alkali mineral complex, on growth, feed utilization, humoral innate immunity and disease resistance of olive flounder. A basal experimental diet was used as a control and supplemented with 0.1, 0.2, 0.3, 0.4, or 0.5% Barodon. Triplicate groups of fish (26.4 ± 0.2 g) were fed one of the diets to apparent satiation twice daily for 10 wk. The growth performance was enhanced (P < 0.05) linearly and quadratically in fish fed diets containing Barodon compared with that in fish fed the control. Feed utilization was significantly improved by Barodon supplementation. Serum lysozyme and antiprotease activities were increased quadratically in Barodon fed groups. Also, significantly higher superoxide dismutase activity was found in Barodon‐fed fish. Dietary supplementation of 0.1–0.3% Barodon resulted in significant enhancement of fish disease resistance against Streptococcus iniae. The findings in this study indicate that dietary supplementation of Barodon can enhance growth, feed utilization, innate immunity, and disease resistance of olive flounder and that the optimum level seems to be 0.1% in diets.     

Comparison of Salt and Drought‐Stress Effects on Maize Growth and Yield Formation with Regard to Acid Invertase Activity in the Kernels

The long‐term effects of salt stress (11 dS m^?1) and drought stress (35 % WHC) were investigated for two maize genotypes, focusing on the relation between metabolic changes around the time of pollination and the impact on yield determinants at maturity. The relatively salt‐resistant hybrid Pioneer 3906 and the relatively drought‐resistant hybrid Fabregas were compared. The experiments were conducted in large plastic containers in a vegetation hall in two consecutive years (2011 and 2012). Plant height and leaf area were significantly reduced under both stress conditions. The transpiration rate was only slightly reduced under drought stress; but under salt stress, a significant reduction occurred 40–53 days after sowing. As a significant increase in sucrose concentrations was observed in the salt‐treated maize kernels 2 days after pollination, the availability of assimilates was not limiting and the plants could afford to save water by reduced stomatal opening. Although under both stress conditions the soluble acid invertase activity was reduced 2 days after pollination, concomitantly, an increase in hexose concentrations was observed. Thus, in these experiments, the delivery of hexoses by acid invertase activity did not limit kernel development. Differences in grain yield at maturity between salt and drought stress were most likely caused by salt‐specific effects (Na⁺ toxicity), Fabregas being more affected than Pioneer 3906.     

Rapidly quantitative detection of Nosema ceranae in honeybees using ultra-rapid real-time quantitative PCR

BackgroundThe microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen.ObjectivesThe present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees.MethodsA procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration.ResultsUR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 10⁴ spores/bee, and the stable detection level was ≥ 2.40 × 10⁵ spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 10⁴ copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min.ConclusionsUR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.     

Effect of Dark,Hard, and Vitreous Kernel Content on Protein Molecular Weight Distribution and on Milling and Breadmaking Quality Characteristics for Hard Spring Wheat Samples from Diverse Growing Regions

Kernel vitreousness is an important grading characteristic for segregation of subclasses of hard red spring (HRS) wheat in the United States. This research investigated the protein molecular weight distribution (MWD) and the flour and baking quality characteristics of different HRS wheat market subclasses. The U.S. regional crop quality survey samples obtained from six regions for three consecutive growing years were used for subclass segregation based on the dark, hard, and vitreous (DHV) kernel percentage. Flour milled from HRS wheat with greater percentages of DHV kernel showed higher water absorption capacity for breadmaking. Protein MWD parameters could be related to the association between DHV kernel level and water absorption. Specifically, flour protein fractions rich in gliadins and high‐molecular‐weight polymeric proteins in the SDS‐unextractable fraction were identified to have significant and positive correlations with both DHV kernels and flour water absorption levels. An example further showed the importance of flour water absorption on potential economic incentives that can be gained with having a greater percentage of vitreous kernels. This information could help the flour milling and baking industry to segregate the different subclasses of HRS wheat with varying DHV content for their intended end‐use applications.     

Relationship Between Solvent Retention Capacity and Protein Molecular Weight Distribution,Quality Characteristics,and Breadmaking Functionality of Hard Red Spring Wheat Flour

This research aims to investigate the relationship between the solvent retention capacity (SRC) test and quality assessment of hard red spring (HRS) wheat flour samples obtained from 10 HRS cultivars grown at six locations in North Dakota. The SRC values were significantly (P < 0.05) correlated with flour chemical components (protein, gluten, starch, and damaged starch contents, except arabinoxylan); with farinograph parameters (stability [FST], water absorption, peak time [FPT], and quality number); and with breadmaking parameters (baking water absorption [BWA], bread loaf volume [BLV], and symmetry). Differences in locations and cultivars contributed significantly to variation in quality parameters and SRC values. Suitability of SRC parameters for discriminatory analysis of HRS wheat flour is greatly influenced by molecular weight distribution (MWD) of SDS‐unextractable proteins. SRC parameters, except for sucrose SRC, showed significant (P < 0.01) and positive correlations with high‐molecular‐weight (HMW) polymeric proteins in SDS‐unextractable fractions, whereas only lactic acid SRC exhibited significant (P < 0.01) correlations with low‐molecular‐weight polymeric proteins. HMW polymeric proteins also exhibited positive associations with FPT, FST, BWA, and BLV. The discrepant variation in association of SRC parameters with respect to MWD of SDS‐unextractable proteins could improve segregation of HRS wheat flour samples for quality.