Individual detection of genetically modified maize varieties in non-identity-preserved maize samples

In many countries, the labeling of grains and feed- and foodstuffs is mandatory if the genetically modified organism (GMO) content exceeds a certain level of approved GM varieties. The GMO content in a maize sample containing the combined-trait (stacked) GM maize as determined by the currently available methodology is likely to be overestimated. However, there has been little information in the literature on the mixing level and varieties of stacked GM maize in real sample grains. For the first time, the GMO content of non-identity-preserved (non-IP) maize samples imported from the United States has been successfully determined by using a previously developed individual kernel detection system coupled to a multiplex qualitative PCR method followed by multichannel capillary gel electrophoresis system analysis. To clarify the GMO content in the maize samples imported from the United States, determine how many stacked GM traits are contained therein, and which GM trait varieties frequently appeared in 2005, the GMO content (percent) on a kernel basis and the varieties of the GM kernels in the non-IP maize samples imported from the United States were investigated using the individual kernel analysis system. The average (+/-standard deviation) of the GMO contents on a kernel basis in five non-IP sample lots was determined to be 51.0+/-21.6%, the percentage of a single GM trait grains was 39%, and the percentage of the stacked GM trait grains was 12%. The MON810 grains and NK603 grains were the most frequent varieties in the single GM traits. The most frequent stacked GM traits were the MON810xNK603 grains. In addition, the present study would provide the answer and impact for the quantification of GM maize content in the GM maize kernels on labeling regulation.     

Effect of using subirrigation and slit tillage to increase soybean (Glycine max) yield in clayey soils in rice paddies converted to uplands

Water stress reduces soybean (Glycine max) yield in fields converted from clayey-soil rice (Oryza sativa) paddies to uplands (upland converted paddies [UCPs]). Subirrigation and/or allowing roots to penetrate to a deeper soil layer were expected to effectively mitigate water stress. Therefore, we examined the effects of subirrigation and slit tillage on mitigating water stress in clayey soils (52% clay) in UCPs. Four treatment experiments (subirrigation, slit tillage, subirrigation control, and slit tillage control) were conducted in each of three growing seasons, with sampling for yield analysis conducted in triplicate. Following conventional tillage, a slit tillage blade was used to cut 15-mm-wide vertical slits in the surface of the soil (cutting depth = 15–20 cm) and seeds were sown 15 mm from the cut lines. Subirrigation then occurred for approximately 21–47 days, from the end of the rainy season, when water table elevation was at 35 cm below ground. We found that subirrigation only resulted in improved soybean yield in 2012, when precipitation after the flowering stage was low, whereas slit tillage significantly increased yield in all three growing seasons, including a particularly wet growing season. Overall, subirrigation and slit tillage increased soybean yield by 2% and 13%, respectively. Slit tillage also resulted in a significantly greater taproot length and root density. Stomatal conductance at midday in about the R5–6 stages was higher in slit-tilled plots than in control plots, but only in the subirrigated field.     

In situ observation of pinewood nematode in wood

In pine wilt disease, xylem dysfunction occurs in relation to nematode migration and proliferation in host tissue, but the detection accuracy of pinewood nematode (PWN), Bursaphelenchus xylophilus, in pine stem tissue remains unclear. This study describes the use of cryo-scanning electron microscopy (cryo-SEM) and fluorescein-conjugated wheat germ agglutinin (F-WGA) staining to detect PWN. After PWN invasion, the frequency of surface fluorescence in PWN increased in pine stems from the day of inoculation to 3 weeks after inoculation. However, the fluorescence frequency decreased significantly during the advanced disease stage after 5 weeks. Thus, detecting PWN based on fluorescent staining of the nematode body surface coat protein can be misleading when used to examine the correlation between the development of disease symptoms and the nematode population. In contrast, all cut body segments were fluorescent, and their fluorescent components were common in pine-stem cross sections, regardless of the timing after inoculation. In addition, PWN were observed under cryo-SEM only in empty resin canals and this distribution was confirmed by F-WGA staining of PWN cut in a cross section. Thus, PWN detection based on fluorescent staining of surface coat proteins by F-WGA was not reliable in longitudinal sections of pine stems because of changes in nematode stainability during disease progression. To detect PWN in infected plants, we concluded that a combination of both methods is most effective.     

A cell preparation of Enterococcus faecalis strain EC-12 stimulates the luminal immunoglobulin A secretion in juvenile calves

The immune system in juvenile calves is immature, so calves are susceptible to several diarrheal and respiratory diseases. Oral administration of lactic acid bacteria (LAB) is known to improve the growth performance and prevent diarrheal and respiratory diseases by stimulating the immune system in juvenile calves. Most of the immunostimulation by LAB is achieved by their cell wall components, and therefore we evaluated the immunostimulation of the cell preparation of Enterococcus faecalis strain EC-12 (EC-12) in juvenile calves in a clinical field. Twenty-nine 1-week old calves were used. Fourteen calves were administered 0.2% (w/w) of an EC-12 preparation that supplemented a milk replacer, and other calves were not supplemented. Feces and serum was collected at day 0, 7 and 49 after the administration to measure the IgA and IgG concentration. The fecal IgA concentration was increased by EC-12 administration at day 49, and the serum IgA concentration was also increased at day 7. These results suggested that oral administration of EC-12 in juvenile calves might have an immunostimulatory effect and provide earlier recovery of IgA levels in mucosal immunity.     

Evaluation of Low Level Laser Therapy on Primary Healing of Experimentally Induced Full Thickness Teat Wounds in Dairy Cattle

Objective —The purpose of this study was to evaluate the effect of low-level laser therapy (LLLT) on sutured wounds of the teat in dairy cattle.
Study Design —By using the Latin square design, the effect of LLLT was evaluated by radiography, measurement of microcirculation flow, histopathology, tensiometry, and hydroxyproline analysis.
Animals or Sample Population —Sixteen teats of four dairy cattle.
Methods —Full thickness wounds were made on the cranial surface of the teats. Teats were distributed into four groups; group A and B wounds were closed with a Gambee pattern, group C and D wounds were closed with three-layers of continuous suture pattern. Group B and D wounds were treated with 3.64 J/cm² of LLLT using a helium-neon system continuous wave (632.8 nm) output of 8.5 mW.
Results —The teat wall in non-LLLT groups was significantly thicker than in LLLT groups on day 7, 14 and 21. The mean blood flow differences between control and sutured sites in LLLT groups were significantly lower than those in non-LLLT groups. The morphology of the epidermis in LLLT groups more closely resembled the normal epidermis than that of non-LLLT groups. Collagen fibers in LLLT groups were denser, thicker, better arranged and more continuous with existing collagen fibers than those in non-LLLT groups. The mean tensile strength was significantly greater in LLLT groups than in non-LLLT groups.
Conclusion —The LLLT affects various aspects of the healing process, including minimizing inflammation, formation of edema, improvement of skin regeneration and enhancement of collagen synthesis.
Clinical Relevance —The LLLT could accelerate healing of sutured wounds of the teat in dairy cattle.     

Decrease in stearic acid proportions in adipose tissues and liver lipids in fatty liver of dairy cows

Samples of liver and perirenal, mesenteric and subcutaneous fat were collected from 16 sick necropsied dairy cows to evaluate the fatty acid profiles in the hepatic and adipose tissues associated with advanced fatty liver or hepatic lipidosis. Hepatic triglyceride and eight fatty acids were measured in the hepatic and adipose tissues. Six cows had more than 3% triglyceride on fresh weight in their livers and were classified as having fatty liver. Stearic and linoleic acid proportions in the liver decreased markedly with increased hepatic triglyceride levels, while the proportion of palmitic and oleic acids increased. The most striking fluctuations in hepatic lipidosis were manifested as decreased stearic acid in the adipose tissues including subcutaneous fat with the trend of decreasing stearic acid. Palmitic acid was elevated in hepatic and perirenal fat in fatty liver cows. In instances of advanced hepatic lipidosis, palmitoleic acid increased in only subcutaneous fat and not in perirenal or mesenteric fat. In addition to the proportions of hepatic fatty acids in fatty liver, this study also clarified the fluctuations observed in the profiles of fatty acids of the adipose tissues in cows with advanced hepatic lipidosis, particularly the decline in the proportions of stearic acid.     

Oral administration of Lactobacillus plantarum Lq80 and Megasphaera elsdenii iNP‐001 induces efficient recovery from mucosal atrophy in the small and the large intestines of weaning piglets

Weaning causes atrophy of intestinal mucosa and a drop of IgA protection in piglets which increases vulnerability to pathogenic infections. Probiotic lactobacilli may support recovery from such weaning stresses. Butyrate‐produce bacteria may support the growth of colonic mucosa. Megasphaera elsdenii, a lactate‐utilizing butyrate producer, may help butyrate production particularly when combined with lactobacilli. Weaned piglets (Experiment 1: 20 days old, Experiment 2: 28 days old) were orally dosed once a day with either (L) 10¹⁰ (cell/dose) L. plantarum Lq80, or (LM) 10¹⁰ (cell/dose) Lq80 with 10⁹ (cell/dose) M. elsdenii iNP‐001. Lq80 was contained in capsules resistant to gastric digestion. M. elsdenii was contained in capsules resistant to gastric and intestinal digestion. An untreated control (C) was also prepared. After 2 weeks of administration, L. plantarum enhanced the recovery from the villous atrophy in both experiments. The rectal and colonic IgA tended to be higher in L and LM than in C in Experiment 1. Colonic butyrate was higher in LM than in the others in Experiment 1. The thickness of the colonic mucosa was greater in LM than in the others in Experiment 1. In early weaned piglets, the effects of L. plantarum and M. elsdenii were clear.     

Microscopic structure of the large intestinal mucosa in piglets during an antibiotic-associated diarrhea

Antibiotic-associated diarrhea (AAD) is caused by the treatments of broad-spectrum antimicrobials that seriously affect the activity and composition of the large intestinal microflora. The pathogenic bacteria or low concentration of short-chain fatty acids (SCFAs) has been repeatedly discussed in relation to AAD. Recently, we reported the detection of a large amount of succinate and lactate in the diarrheal feces in AAD-induced piglets. In this study, we investigated histologically the large intestinal mucosa in AAD-induced piglets, in which succinate and lactate were accumulated. AAD was induced in the piglets by an oral dose of polymyxin B sulfate (PL) or by an intra-muscular injection of enrofloxacin (ERFX). When the piglets were defecating diarrheal feces with a high concentration of succinate and/or lactate, the large intestine was removed and separated into four segments (cecum, gyri centripetales, gyri centrifugales, and rectum). Healthy piglets were used as the control. In the AAD-induced piglets, the lamina propria was edematous in the gyri centripetales. Piglets treated with ERFX were also edematous in gyri centrifugales. These edematous lamina propria contained larger amounts of inflammatory cells than observed in control tissues. ERFX-treated piglets had a more shallow crypt than PL-treated and control piglets. The mucosal tissue of the large intestine was more seriously damaged in the ERFX- than in the PL-treated piglets, which might have been caused by the high succinate and low SCFAs concentration in the digesta.     

A cell wall preparation of Enterococcus faecalis strain EC-12 stimulates β-defensin expression in newly hatched broiler chicks

In our previous study, a cell wall preparation of Enterococcus faecalis strain EC‐12 (EC‐12) prevented the colonization of vancomycin‐resistant enterococci (VRE) in newly hatched broiler chicks. Relatively early prevention against VRE colonization by EC‐12 administration may be related to the enhanced innate immune system. In this study we examined the effect of EC‐12 on the kinetics of immunoglobulin A (IgA) concentration in the cecal digesta of chicks from 1 day to 14 days old to evaluate humoral immunity. We also examined the effect of EC‐12 on β‐defensin mRNA expression and lysozyme production. The IgA concentration in the cecal digesta showed a decline in chicks from 2 to 8 days old and then a recovery up to when they were 14 days old. The concentration tended to be higher in EC‐12‐administered chicks than in control chicks at 14 days old. EC‐12 administration did not stimulate the lysozyme activities in the alimentary tract. However, β‐defensin mRNA expression in the tongue and the bursa of Fabricius was higher in chicks administered EC‐12 than in the control chicks at 5 days old. These results indicate that β‐defensin may be one of the major defense mechanisms inhibiting VRE colonization in young chicks.