Pulsed electric field treatment of seeds altered the endophytic bacterial community and promotes early growth of roots in buckwheat.

BACKGROUND: Endophytic bacteria provide nutrients and stimulate systemic resistance during seed germination and plant growth and development, and their functional properties in combating various stresses make them a powerful tool in green agricultural production. In this paper we explored the function of the endophyte community in buckwheat seeds in order to provide a theoretical basis for the application and scientific research of endophytes in buckwheat cultivation. We used pulsed electric field (PEF) technology to treat buckwheat seeds, monitored the effect of high-voltage pulse treatment on buckwheat seed germination, and analyzed the diversity of endophytic bacteria in buckwheat seeds using the amplicon sequencing method. RESULTS: PEF treatment promoted root development during buckwheat seed germination. A total of 350 Operational taxonomic units (OTUs) that were assigned into 103 genera were obtained from control and treatment groups using 16SrRNA amplicon sequencing technology. Additionally, PEF treatment also caused a significant decrease in the abundance of Actinobacteria, Proteobacteria, and Bacteroidetes. The abundance of 28 genera changed significantly as well: 11 genera were more abundant, and 17 were less abundant. The number of associated network edges was reduced from 980 to 117, the number of positive correlations decreased by 89.1%, and the number of negative correlations decreased by 86.6%. CONCLUSION: PEF treatment promoted early root development in buckwheat and was able to alter the seed endophytic bacterial community. This study thus makes a significant contribution to the field of endophyte research and to the application of PEF technology in plant cultivation.

Colorimetric detection of 2-tert-butyl-1,4-benzoquinone in edible oils based on a chromogenic reaction with commercial chemicals.

2-tert-butyl-1,4-benzoquinone (TBBQ) is the major oxidative product of tert-butylhydroquinone which is a widely used antioxidant in edible oils. The biotoxicity of TBBQ is a risk to human health, that the rapid and accurate monitoring of TBBQ is needed. Herein, a specific chromogenic reaction between TBBQ and polyethyleneimine (PEI) could generate adducts with maximum absorption at 478 nm. Amine groups of PEI are prone to link with TBBQ through Michael addition to form colored adducts. A colorimetric method for detecting TBBQ in edible oils was developed based on the aforesaid chromogenic reaction. The linear range for TBBQ was from 3.0 to 100.0 µg g-1, having a limit of detection of 1.8 µg g-1. Recoveries results ranged from 88.4 % to 93.1 %, which had a good agreement with that of high-performance liquid chromatography. Our study provides a rapid and simple strategy for the sensitive detection of TBBQ using commercial chemicals.

The Impact of Tributyrin on Performance and Intestinal Health of Broiler Chickens Post Coccidiosis Vaccination.

Coccidiosis is a major intestinal disease affecting broiler chickens. Tributyrin (TB) is a valid alternative to butyrate acid, which was associated with the improvement of performance and attenuation of intestinal inflammation in animal production. However, there are few reports on TB as a prophylactic treatment against coccidiosis in broilers. The aim of the study was to investigate the effects of TB supplementation on performance and intestinal health of broiler chickens post coccidiosis vaccination with a mixed-species Eimeria. In the first experiment, 612 broiler chicks were randomly assigned to two treatments with six replicates. Treatments included no TB supplementation and coccidiosis vaccination (CV1), or TB supplementation (400 mg/kg) and coccidiosis vaccination (TBCV1). On day 5, all broilers received a single vaccine dose. Performance, intestinal histopathology, clinical severity, and fecal oocyst counts were evaluated from day 1 to day 63. TB supplementation resulted in a nonsignificant effect on body weight gain (BWG), feed intake (FI), and mortality-corrected feed conversion ratio (FCR), except in increased FI on days 22-42 (P < 0.05). The TBCV1 group had increased (P < 0.05) villi heights in the duodenum and increased (P < 0.05) villi widths in the ileum on day 63 of age and reduced oocyst shedding on days 19-26 compared to CV1(P < 0.05). The route of administration in the second experiment was different from the first experiment in which the seeder birds (half of birds from each pen) received a tenfold dose on day 5. TB supplementation in broilers resulted in increased (P < 0.05) BWG and reduced (P < 0.05) FCR on days 22-42, and increased (P < 0.05) villi heights in the duodenum and increased (P < 0.05) villi widths in the ileum on day 63 of age, as well as a lower frequency (P < 0.05) of intestinal hemorrhage on days 13-62 and reduced (P < 0.001) oocyst shedding on day 5 post-Eimeria challenge. In conclusion, the study demonstrated that TB can be considered as a feed additive for protecting broilers from coccidiosis on days 22-42.

Impacto de la tributirina en el rendimiento y la salud intestinal de pollos de engorde después de la vacunación contra la coccidiosis. La coccidiosis es una enfermedad intestinal importante que afecta al pollo de engorde. La tributirina (TB) es una alternativa viable para el ácido butirato, que se ha asociado con mejora en el rendimiento y atenuación de la inflamación intestinal en la producción animal. Sin embargo, hay pocos informes sobre la tributirina como tratamiento profiláctico contra la coccidiosis en pollos de engorde. El objetivo del estudio fue investigar los efectos de la suplementación de tributirina en el rendimiento y la salud intestinal de pollos de engorde después de la vacunación contra coccidiosis con especies mixtas de Eimeria. En el primer experimento, se asignaron aleatoriamente 612 pollos de engorde en dos tratamientos con seis repeticiones. Los tratamientos incluyeron no suplementación con tributirina y vacunación contra coccidiosis (CV1), o suplementación con tributirina (400 mg/kg) y vacunación contra coccidiosis (TBCV1). En el quinto día de edad, todos los pollos de engorde recibieron una sola dosis de vacuna. El rendimiento, la histopatología intestinal, la severidad clínica y los recuentos de oocistos fecales se evaluaron desde el día primer hasta el día 63. La suplementación con tributirina resultó en un efecto no significativo sobre la ganancia de peso corporal (BWG), en el consume de alimento (FI) y la tasa de conversión alimenticia corregida por la mortalidad (FCR), excepto en un aumento en el consumo de alimento entre los días 22 a 42 (P <0.05). El grupo TBCV1 mostró aumento (P <0.05) de la altura de las vellosidades en el duodeno y aumento (P <0.05) del ancho de las vellosidades en el íleon en el día 63 de edad y reducción de la eliminación de ooquistes entre los días 19-26 en comparación con el tratamiento CV1 (P <0.05). La vía de administración en el segundo experimento fue diferente del primer experimento en el que aves sembradoras (la mitad de las aves de cada corral) recibieron una dosis diez veces mayor en el quinto día. La suplementación de tributirina en pollos de engorde resultó en un aumento (P <0.05) sobre la ganancia de peso corporal y una reducción (P <0.05) en la tasa de conversión alimenticia corregida por la mortalidad entre los días 22-42, aumento (P <0.05) de la altura de las vellosidades en el duodeno y aumento (P <0.05) en el ancho de las vellosidades en el íleon en el día 63 de edad, así como una frecuencia más baja (P <0.05) de hemorragia intestinal entre los días 13 a 62 y reducción (P <0.001) de la eliminación de oocistos en el quinto después de la exposición a Eimeria. En conclusión, el estudio demostró que la tributirina podría considerarse como un aditivo alimentario para proteger a los pollos de engorde de la coccidiosis entre los días 22 a 42.

Comparative effects of pollen limitation, floral traits and pollinators on reproductive success of Hedysarum scoparium Fisch. et Mey. in different habitats.

BACKGROUND: Reproduction in most flowering plants may be limited because of the decreased visitation or activity of pollinators in fragmented habitats. Hedysarum scoparium Fisch. et Mey. is an arid region shrub with ecological importance. We explored the pollen limitation and seed set of Hedysarum scoparium in fragmented and restored environments, and examined whether pollen limitation is a significant limiting factor for seed set. We also compared floral traits and pollinator visitation between both habitats, and we determined the difference of floral traits and pollinators influenced reproductive success in Hedysarum scoparium. RESULTS: Our results indicated that supplementation with pollen significantly increased seed set per flower, which is pollen-limited in this species. Furthermore, there was greater seed set of the hand cross-pollination group in the restored habitat compared to the fragmented environment. More visits by Apis mellifera were recorded in the restored habitats, which may explain the difference in seed production between the fragmented and restored habitats. CONCLUSIONS: In this study, a positive association between pollinator visitation frequency and open flower number was observed. The findings of this study are important for experimentally quantifying the effects of floral traits and pollinators on plant reproductive success in different habitats.

Identification and characterization of miRNAs associated with sterile flower buds in the tea plant based on small RNA sequencing.

BACKGROUND: miRNAs are a type of conserved, small RNA molecule that regulate gene expression and play an important role in the growth and development of plants. miRNAs are involved in seed germination, root development, shoot apical meristem maintenance, leaf development, and flower development by regulating various target genes. However, the role of miRNAs in the mechanism of tea plant flower sterility remains unclear. Therefore, we performed miRNA sequencing on the flowers of fertile male parents, female parents, and sterile offspring. RESULTS: A total of 55 known miRNAs and 90 unknown miRNAs were identified. In the infertile progeny, 37 miRNAs were differentially expressed; 18 were up-regulated and 19 were down-regulated. miR156, miR157, miR164, miR167, miR169, miR2111 and miR396 family members were down-regulated, and miR160, miR172 and miR319 family members were up-regulated. Moreover, we predicted that the 37 differentially expressed miRNAs target a total of 363 genes, which were enriched in 31 biological functions. We predicted that miR156 targets 142 genes, including ATD1A, SPL, ACA1, ACA2, CKB22 and MADS2. CONCLUSION: We detected a large number of differentially expressed miRNAs in the sterile tea plant flowers, and their target genes were involved in complex biological processes. Among these miRNAs, the down-regulation of miR156 may be one of the factor in the formation of sterile floral buds in tea plants.

Innovative health risk assessments of heavy metals based on bioaccessibility due to the consumption of traditional animal medicines.

Few studies reported the extent of heavy metal accumulation in traditional Chinese medicines (TCMs). Currently, oral bioaccessibility of lead (Pb), cadmium (Cd), arsenic (As), mercury (Hg), and copper (Cu) present in traditional animal medicines was investigated with physiologically based extraction test-extracted in vitro model. We are the first to develop a health risk assessment strategy by combinational analysis of bioaccessible heavy metal levels to calculate target hazard quotient (THQ), target hazard index (THI) and cancer risk (CR), which has capacity to evaluate the heavy metal associated heath risk of traditional animal medicines. To precisely acquire a realistic risk assessment, questionnaire data was adopted to measure the frequency and duration of the exposure to traditional animal medicines, and the safety factor was highlighted as well. Our data revealed that the bioaccessibility of Hg was the lowest among the five heavy metals. After the adjustment with the bioaccessibility of each heavy metal to target hazard index (THI) values, excitingly, the results manifested that the consumption of traditional animal medicines might not exert an unacceptable health risk in a broad community. In addition, the CR values of As and Pb indicated that the risk of developing cancers was quite lower than their acceptable levels in the clinic.

Transcriptome profiling of the fertile parent and sterile hybrid in tea plant flower buds.

BACKGROUND: The tea plant is a crucial economic crop. The floral organ development consumes a large amount of nutrients, which affects the leaf yield. To understand the mechanism by which the tea plant produces sterile floral buds, we obtained a sterile tea plant by artificial hybridization. RNA-sequencing based transcriptome analysis was implemented in three samples to determine the differentially expressed genes (DEGs) related to flower development. RESULTS: In this study, a total of 1991 DEGs were identified; 1057 genes were up-regulated and 934 genes were down-regulated in sterile hybrid floral buds. These were mainly distributed in the regulation of biological and metabolic processes. Significantly, auxin biosynthesis genes YUCCA, AUX1 and PIN were dramatically down-regulated, and ARF gene was up-regulated in the sterile hybrid floral buds, and flower development-related genes AP1, AP2 and SPL were changed. A total of 12 energy transfer-related genes were significantly decreased. Furthermore, the expression of 11 transcription factor genes was significantly different. CONCLUSION: The transcriptome analysis suggested that the production of sterile floral buds is a complex bioprocess, and that low auxin-related gene levels result in the formation of sterile floral buds in the tea plant.

Primary biliary cholangitis associated with drug-induced liver injury and alcoholic liver fibrosis: A case report.

RATIONALE: Primary biliary cholangitis (PBC) is a liver autoimmune disease. If this disease is associated with other liver injury factors, both misdiagnosis and missed diagnosis will easily occur. Therefore, detailed disease history collection and related laboratory examination should be performed on patients with liver injury for unidentified causes. When necessary, liver biopsy should be performed to confirm the histopathological diagnosis. PATIENT CONCERNS: The subject patient was a 63-year-old Chinese male with chronic liver injury who had a drinking history of about 30 years and drank 500 g daily on average and began to take health products and dietary supplements (multivitamins) since June 2014. DIAGNOSES: Drug-induced liver injury (DILI) and alcoholic fatty liver disease (AFLD) were initially considered because the patient had a history of using health products (HP) and dietary supplements (DS) and drinking alcohol. However, he was subsequently considered with PBC based on the findings of anti-mitochondrial antibody positivity and elevated immunoglobulin level. Obstructive jaundice and space-occupying lesion in the liver were excluded by imaging examinations. Liver biopsy was performed to confirm the reasons for liver injury. Histopathological examination was conducted, and the patient was diagnosed with PBC associated with DILI and alcoholic liver fibrosis. INTERVENTIONS: Ursodeoxycholic acid, glycyrrhizic acid, and methylprednisolone (small dose) were used to treat the patient. OUTCOMES: After 2 months, the serum levels of ALT, AST, AKP, GGT, and globulin returned to normal. After 4 months, the patient showed liver injury once again (an increase in ALT, AST, AKP, GGT and GLB) caused by repaglinide administration due to hyperglycemia. Ursodeoxycholic acid and methylprednisolone replaced the repaglinide administration. After 3 weeks, the levels of ALT, AST, AKP, GGT, and GLB returned to normal again. LESSONS: The correct knowledge on PBC and early-stage recognition and diagnosis should be emphasized. When other causes of the liver injury cannot be excluded, liver biopsy is suggested. Histopathological change can be used to further clarify the reasons for liver injury and the principal contradiction as well as to guide the theraputic regimen.

Increased pollinator service and reduced pollen limitation in the fixed dune populations of a desert shrub.

Evaluations of restoration success usually focus on the structural aspects of ecosystems. Pollination, as an important functional aspect, is often overlooked. Here, the shifts in pollinator assemblage and pollen limitation in the desert shrub Caragana microphylla were examined along a restoration gradient in Horqin Sand Land, northern China. We identified seven species of bees; however, only four bee species were found to be effective pollinators, with Xanthosaurus remota dominating in the fixed dunes, and with no bee species or only a single species, X. remota, being observed in the semi-fixed and mobile dunes. Flower visitation rate was nearly ten times higher in the fixed dunes than in the mobile and semi-fixed dunes. Experimental floral manipulations revealed that the fixed dune populations experienced less pollen limitation, along with the increase in pollinator availability. Between the mobile and semi-fixed dune populations, pollen limitation was severe and at similar levels. The intensity of pollen limitation was negatively related to pollinator abundance and richness. Overall, the dependence on pollinators for reproduction may be an important constraint that limits persistence in this system. Increased pollinator service during the restoration process may ameliorate pollen limitation, benefiting the restoration of vegetation in this semiarid sandy area.

Effects of sand burial on the survival and growth of two shrubs dominant in different habitats of northern China.

Plants that grow in dune ecosystems always suffer from sand burial. Shrubs play implications on the healthy functioning of dune ecosystems due to control blowing sand. However, the survival and growth responses of shrubs to sand burial remain poorly understood. The survival rate and seedling height of two shrubs (Artemisia halodendron and Lespedeza davurica) along with the soil properties under different burial depths were examined in order to reveal the causing ecophysiological attributes of sand burial on shrubs in the desertified region. It was found that A. halodendron can survive a burial depth of 6 cm greater than its seedling height, which is a dominant shrub in mobile dunes with intense burial, whereas a burial depth equivalent to three fourths of its seedling height is detrimental to L. davurica, which is dominant in fixed dunes with less burial. The reasons for the shrub death under sand burial were associated with the physical barrier to vertical growth and the reduction in photosynthetic area. In conclusion, A. halodendron can facilitate the stabilization of mobile dunes because of their high tolerance to the frequent and intensive sand burial, while L. davurica can be beneficial for the recovery process because of their higher survival rates under shallow burial following restoration of mobile dunes.

Effects of habitat disturbance on the pollination system of Ammopiptanthus mongolicus (Maxim) Cheng f. at the landscape-level in an arid region of Northwest China.

Ammopiptanthus mongolicus is an ecologically important species in the arid region of Northwest China. Habitat disturbance can significantly affect plant mating success and ultimately species viability. Pollen limitation of plant reproduction occurs in many plant species, particularly those under habitat disturbance. However, previous investigations have demonstrated differences in pollen limitation between conserved and disturbed sites. We compared the phenology, pollen limitation, pollinators and breeding system of both sites to determine whether habitat disturbance has generated changes in these plant components. We found that the species differed in four aspects. First, blooming duration and flowering peak were longer in the disturbed site than in the conserved site. Second, A. mongolicus can be pollen-limited and pollen limitation was more intense in the conserved site than in the disturbed site. Third, Anthophora uljanini was found to be a frequent pollinator in the conserved site, while Apis mellifera was the most effective and frequent flower visitor. More pollinator visits were recorded in the disturbed site, which could explain the differences in reproductive success. Finally, seed set was higher in the disturbed site than in the conserved site. We found that outcrossing was dominant in both sites and that agamospermy and self-pollination played complementary roles to ensure reproduction. Differences in flower production influenced by artificial selection and pollinator type explain the different seed set in both sites, whereas habitat disturbance cause changes differences in the pollination process and limits pollen flow. The balance between artificial management and mating success is crucial to analysis of the pollination process and manipulation of A. mongolicus population size.