Time course of the incidence/multiplicity and histopathological features of murine colonic dysplasia,adenoma and adenocarcinoma induced by benzo[a]pyrene and dextran sulfate sodium

Benzo[a]pyrene (BP) is mutagenic but noncarcinogenic in the murine colon. Recently, we reported rapid induction of colonic tumors by treatment of CD2F₁ mice with BP (125 mg/kg for 5 days) followed by a colitis inducer, dextran sulfate sodium (DSS) (4% in drinking water for 1 or 2 weeks). However, there are no reports on detailed time course and histopathological features of colonic proliferative lesions in this model. Here, we show the detailed time course of colonic dysplasia, adenoma and adenocarcinoma induced by treatment with BP, DSS, and a combination of the two (BP/DSS). In the colon of mice exposed to BP/DSS, 14.6 dysplastic foci per mouse were present one week after DSS treatment (week 4). The number of dysplastic foci decreased with time to 3.1 at week 9 and thereafter remained almost constant. At week 4, 1.5 adenocarcinomas were also observed, with a marked increase in numbers with time, reaching 29.3 at week 14. In contrast, the number of dysplastic foci induced by DSS alone showed a time course similar to that following BP/DSS treatment; however, only a few tumors appeared. Neither dysplastic foci nor neoplastic lesions were induced by BP only. In mice exposed to BP/DSS, β-catenin was demonstrated immunohistochemically in the nucleus and/or cytoplasm of the tumor cells, and this translocation from the cell membrane was evident in subsets of dysplastic foci. In dysplastic foci induced by DSS alone, β-catenin was absent in the nucleus/cytoplasm. These finding suggest that aberrant β-catenin accumulation in dysplastic foci is associated with tumor progression in this BP/DSS model.     

Faecal shedding detected earlier than immune responses in goats naturally infected with Mycobacterium avium subsp. paratuberculosis

Paratuberculosis was diagnosed in a goat herd that participated in a sanitation program against Mycobacterium avium subsp. paratuberculosis. The aim of this study was to characterise the development of gamma interferon (IFN-γ) and antibody responses as well as the occurrence of faecal shedding. Faecal culture appeared surprisingly sensitive as about 18% and 40% of the goats were positive at 9 and 15-17 months of age, respectively, and shedding was often seen prior to peripheral immune responses. Peripheral IFN-γ responses were not related to protection as clinical and high shedding goats often had high responses. An IFN-γ response usually preceded a humoral response. However, positive antibody titers could sometimes be seen simultaneously with, and even prior to, IFN-γ responses. In conclusion, faecal culture appeared as sensitive as IFN-γ testing. Furthermore, the antibody ELISA and the IFN-γ assay may perform equally well in an infected herd if surveillance is conducted annually.     

Effects of concentrate‐to‐forage ratios and 2‐methylbutyrate supplementation on ruminal fermentation,bacteria abundance and urinary excretion of purine derivatives in Chinese Simmental steers

This study evaluated the effects of dietary concentrate levels and 2‐methylbutyrate (2MB ) supplementation on performance, ruminal fermentation, bacteria abundance, microbial enzyme activity and urinary excretion of purine derivatives (PD ) in steers. Eight ruminally cannulated Simmental steers (12 months of age; 389 ± 3.7 kg of body weight) were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement. Moderate‐concentrate (400 g/kg diet [MC ]) or high‐concentrate (600 g/kg diet [HC ]) diets were fed with or without 2MB (0 g/day [2MB ?] or 15.0 g/day [2MB +]). Dry matter intake and average daily gain increased, but feed conversion ratio decreased with the HC diet or 2MB supplementation. Ruminal pH decreased, but total volatile fatty acid increased with the HC diet or 2MB supplementation. Molar proportion of acetate and acetate‐to‐propionate ratio decreased with the HC diet, but increased with 2MB supplementation. Propionate molar proportion and ruminal NH ₃‐N content increased with the HC diet, but decreased with 2MB supplementation. Neutral detergent fibre degradability decreased with the HC diet, but increased with 2MB supplementation. Crude protein degradability increased with the HC diet or 2MB supplementation. Abundance of Ruminococcus albus , Ruminococcus flavefaciens , Fibrobacter succinogenes and Bufyrivibrio fibrisolvens as well as activities of carboxymethyl cellulase, cellobiase, xylanase and pectinase decreased with the HC diet, but increased with 2MB supplementation. However, abundance of Prevotella ruminicola and Ruminobacter amylophilus as well as activities of α‐amylase and protease increased with the HC diet or 2MB supplementation. Total PD excretion also increased with the HC diet or 2MB supplementation. The results suggested that growth performance, ruminal fermentation, CP degradability and total PD excretion increased with increasing dietary concentrate level from 40% to 60% or 2MB supplementation. The observed diet × 2MB interaction indicated that supplementation of 2MB was more efficacious for improving growth performance, ruminal fermentation and total PD excretion with promoted ruminal bacteria abundance and enzyme activity in the MC diet than in the HC diet.     

稻渔综合种养的科学范式

探讨不同养殖模式下金背鲤 (Cyprinus carpio var. Jinbei) 肠道菌群和鱼肉风味品质差异,对金背鲤的繁殖优化具有重要意义。采用高通量测序技术对稻田放养和池塘养殖模式下金背鲤的肠道微生物进行测序,利用液相色谱和顶空固相微萃取-气相色谱-离子迁移谱联用 (HS-SPME-GC-IMS) 技术测定其滋味物质、挥发性风味化合物,结合感觉阈值计算滋味活性值和相对气味活度值。结果显示：两种养殖模式的金背鲤肠道细菌群落结构差异显著,稻田放养 (FGF) 组以弧菌 (Vibrio)、拟杆菌 (Bacteroides)、交替单胞菌 (Alteromonadales)、希瓦氏菌 (Shewanella)、嗜冷假单胞菌 (Pseudomonas psychrophile) 和Brevinema属为主,池塘养殖 (FGP) 组以莫拉克斯氏菌 (Moraxella) 和克雷伯菌属 (Klebsiella) 为主;FGF组中鲜味肌苷酸含量及其滋味活性值 (1.676 g·kg⁻¹, 6.705) 远高于FGP组 (0.246 g·kg⁻¹, 0.985),FGF组鲜味氨基酸和甜味氨基酸含量 (0.143和2.052 g·kg⁻¹) 高于FGP组 (0.109和2.001 g·kg⁻¹),而其苦味氨基酸 (3.193 g·kg⁻¹) 却低于FGP组 (3.836 g·kg⁻¹);金背鲤的挥发性化合物组分复杂,其关键气味化合物 (ROAV≥1) 和对整体风味有修饰作用的化合物 (0.1≤ROAV<1) 的种类存在差异。菌属与风味物质的相关性分析显示弧菌属、拟杆菌属、克雷伯菌属和摩根菌属 (Morganella) 与风味物质呈显著相关 (0.01≤P<0.05 & 0.001≤P<0.01 & P<0.001)。研究表明,养殖模式影响了金背鲤的肠道微生物,并间接影响了其风味品质。     

茉莉酸甲酯对葡萄植株不定根发育的影响

利用不同浓度的茉莉酸甲酯（MeJA）对葡萄组培植株进行处理,进而分析其生理及基因表达变化,结果表明MeJA能抑制植株不定根的分化和伸长,诱导根、茎、叶组织中的脱落酸（ABA）含量升高,但降低赤霉素（GA）、生长素（IAA）和细胞分裂素（ZR）的含量,且影响叶片中叶绿素代谢导致褪绿黄化。MeJA能诱导植物体内茉莉酸合成代谢基因表达进而提高内源茉莉酸的含量,诱导ABA合成代谢和信号路径中的基因表达,但是会抑制生长素、赤霉素、细胞分裂素、油菜素内酯合成代谢基因的表达,抑制了与根发育和延伸相关基因VvXET、VvEXPB2、VvEXPA7和VvWUS的表达,进而抑制不定根的发生,导致植物发育缓慢。因此,MeJA通过影响其他激素的合成代谢来调控它们在植物中的含量,导致内源激素不平衡影响植株发育,并调控植株根的发育状态,在不定根发育方面起负向调控作用。     

A three‐dimensional cell culture system as an in vitro canine mammary carcinoma model for the expression of connective tissue modulators

In this study, derived complex carcinoma (CC) and simple carcinoma (SC) cell lines were established and cultured under two‐dimensional (2D) and three‐dimensional (3D) conditions. The 3D was performed in six‐well AlgiMatrix? (LifeTechnologies®, Carlsbad, CA, USA) scaffolds, resulting in spheroids sized 50–125 µm for CC and 175–200 µm for SC. Cell viability was demonstrated up to 14 days for both models. Epidermal growth factor receptor (EGFR) was expressed in CC and SC in both systems. However, higher mRNA and protein levels were observed in SC 2D and 3D systems when compared with CC (P < 0.005). The connective tissue modulators, metalloproteinases‐1, ‐2, ‐9 and ‐13 (MMPs), relaxin receptors 1 and 2 (RXR1 and RXR2) and E‐cadherin (CDH1) were quantitated. All were upregulated similarly when canine mammary tumour (CMT)‐derived cell lines were cultured under 3D AlgiMatrix, except CDH1 that was downregulated (P < 0.005). These results are promising towards the used of 3D system to increase a high throughput in vitro canine tumour model.     

Replant diseases: Bacterial community structure and diversity in peach rhizosphere as determined by metabolic and genetic fingerprinting

Peach tree replant disease, though reported on in the literature for more than two centuries, has yet to have its causes clearly defined. Decline in peach productivity has been attributed to toxic agents, insects, nutritional disturbances, spray residues, fungi and nematodes. Bacteria has also been indicated as a contributing factor.Peach replant disease was reproduced by using two successive cultures on the same soil. Bacterial communities were isolated and characterized from healthy and diseased peach trees. The potential role of cyanide production by rhizobacteria in the replant problem of peaches was studied. Culture-dependent (evaluation of the number of culturable bacteria, metabolic activities, Biolog^® GN2) and independent (ribosomal intergenic spacer analysis, RISA) methods were used, in order to compare bacterial community structure and diversity in healthy and sick soils and to evaluate the possible role of cyanide.Bacterial densities were significantly increased in sick soils. Metabolic activities (Biolog^® GN2) and genetic structure, observed through RISA, were also significantly modified in sick soils. Changes in the composition of individual microbial groups in the rhizosphere of peach trees excavated from healthy or sick soil indicated the involvement of rhizobacteria in the etiology of the replant sickness of peach soil. More than 60% of the strains isolated from healthy soils corresponded to Pseudomonas sp. and 58% of the isolates from sick soils were Bacillus sp. This study determined that Bacillus were able to produce in vitro HCN. It also appeared that in sick soil, there was a shift in the structure of bacterial communities with an increase noted in phytotoxic microorganisms capable of producing HCN compounds.     

A novel screening method for rice allelopathic potential: the inhibitory‐circle method

Screening crop accessions for allelopathic activity is of paramount importance for crop allelopathy research. Previous bioassays often did not use a mixed culture of donor and target plants, did not use soil and were not conducted under natural conditions. In this study, we designed an inhibitory‐circle method in which a rice accession (donor plant) and Echinochloa crus‐galli (target plant) were cultured together in paddy soil under natural conditions. First, we determined that the highest allelopathic activity of allelopathic rice accession PI312777 was at the 5‐leaf stage, and the suitable distance of rice seedlings and E. crus‐galli was 12 cm apart. This method was then validated by a field test. A further 40 rice accessions were evaluated for allelopathic activity to E. crus‐galli using this method. Two rice accessions, PI312777 and Taichung Native 1, had highly allelopathic activity to E. crus‐galli (inhibitory rate > 50%), while another accession, Lemont, had non‐allelopathic activity. These experimental results were in accordance with previous studies using direct field experiments. The inhibitory‐circle method integrated three necessary conditions, that is donor and target plants grown together, with soil as the medium and under natural conditions for reliable results. The ‘inhibitory‐circle method’, which combined donor and target plants, soil medium and field conditions, can give reliable results in one step, compared with laboratory screening methods. Also, the ‘inhibitory‐circle method’ gave results in 30‐35 days, thereby substantially reducing the requirements for time, labour and cost.     

斯氏艾美耳球虫ITS1-5.8S rRNA-ITS2序列的克隆及PCR检测方法的建立

通过对多种鸡球虫和松鼠球虫18SrRNA和28SrRNA进行序列比对分析,在18SrRNA 3′端和28SrRNA 5′端保守区设计艾美耳属通用引物,以斯氏艾美耳球虫洛阳分离株LY卵囊基因组DNA为模板首次成功克隆到斯氏艾美耳球虫完整的ITS1-5.8SrRNA-ITS2序列,其大小为1 178bp,其中ITS1序列长度为423bp,5.8SrRNA为155bp,ITS2为600bp,斯氏艾美耳球虫LY株ITS1/2序列高度变异,与鸡球虫、啮齿动物球虫的序列相似性低于60%。然后在斯氏艾美耳球虫ITS1/2序列超变区设计种特异引物,建立了灵敏、特异的PCR检测方法。本研究结果将为兔球虫强致病种的临床诊断和揭示兔球虫种群遗传特征提供有效的分子工具。