Mycotoxin-Producing Ability and Chemotype Diversity of Aspergillus Section Flavi from Soils of Peanut-Growing Regions in Iran

Invasion of crops with Aspergillus flavus may result in contamination of food and feed with carcinogenic mycotoxins such as aflatoxins (AF) and cyclopiazonic acid (CPA). In the present study, distribution and toxigenicity of Aspergillus flavus and A. parasiticus in soils of five peanut fields located in Guilan province, Northern Iran was investigated. From a total of 30 soil samples, 53 strains were isolated which all of them were finally identified as A. flavus by a combination of colony morphology, microscopic criteria and mycotoxin profiles. Chromatographic analysis of fungal cultures on yeast extract sucrose broth by tip culture method showed that 45 of the 53 A. flavus isolates (84.9 %) were able to produce either CPA or AFB₁, while eight of the isolates (15.1 %) were non-toxigenic. The amounts of CPA and AFB₁ produced by the isolates were reported in the range of 18.2–403.8 μg/g and 53.3–7446.3 μg/g fungal dry weights, respectively. Chemotype classification of A. flavus isolates based on the ability for producing mycotoxins and sclerotia showed that 43.4 % were producers of CPA, AFB₁ and sclerotia (group I), 13.2 % of CPA and AFB₁ (group II), 9.4 % of AFB₁ and sclerotia (group III), 13.2 % of AFB₁ (group IV), 5.7 % of CPA and sclerotia (group V) and 15.1 % were non-toxigenic with no sclerotia (group VI). No strain was found as producer of only CPA or sclerotia. These results indicate different populations of mycotoxigenic A. flavus strains enable to produce hazardous amounts of AFB₁ and CPA are present in peanuts field soils which can be quite important regard to their potential to contaminate peanuts as a main crop consumed in human and animal nutrition.     

The Phylogenetics of Mycotoxin and Sclerotium Production in Aspergillus flavus and Aspergillus oryzae

Aspergillus flavus is a common filamentous fungus that produces aflatoxins and presents a major threat to agriculture and human health. Previous phylogenetic studies of A. flavus have shown that it consists of two subgroups, called groups I and II, and morphological studies indicated that it consists of two morphological groups based on sclerotium size, called “S” and “L.” The industrially important non-aflatoxin-producing fungus A. oryzae is nested within group I. Three different gene regions, including part of a gene involved in aflatoxin biosynthesis (omt12), were sequenced in 33 S and L strains of A. flavus collected from various regions around the world, along with three isolates of A. oryzae and two isolates of A. parasiticus that were used as outgroups. The production of B and G aflatoxins and cyclopiazonic acid was analyzed in the A. flavus isolates, and each isolate was identified as “S” or “L” based on sclerotium size. Phylogenetic analysis of all three genes confirmed the inference that group I and group II represent a deep divergence within A. flavus. Most group I strains produced B aflatoxins to some degree, and none produced G aflatoxins. Four of six group II strains produced both B and G aflatoxins. All group II isolates were of the “S” sclerotium phenotype, whereas group I strains consisted of both “S” and “L” isolates. Based on the omt12 gene region, phylogenetic structure in sclerotium phenotype and aflatoxin production was evident within group I. Some non-aflatoxin-producing isolates of group I had an omt12 allele that was identical to that found in isolates of A. oryzae.     

鹦哥岭自然保护区鹦哥岭树蛙种群分布及数量调查

于2010年7月～2012年1月,采用在不同海拔梯度设置样带和样方的方法,对海南鹦哥岭省级自然保护区内鹦哥岭树蛙(Rhacophorus yinggelingensis)的分布和种群数量进行了调查。调查表明,鹦哥岭树蛙分布范围非常狭窄,栖息的生境类型较为单一,仅分布于海拔1 200 m以上的临时性水潭内,种群数量为(2 972±831)只。由于该物种的数量极其稀少,建议提高鹦哥岭树蛙保护等级以加强保护。     

Taxonomic comparison of three different groups of aflatoxin producers and a new efficient producer of aflatoxin B1, sterigmatocystin and 3-O-methylsterigmatocystin, Aspergillus rambellii sp. nov

Accumulation of the carcinogenic mycotoxin aflatoxin B, has been reported from members of three different groups of Aspergilli (4) Aspergillus flavus, A. flavus var. parvisclerotigenus, A. parasiticus, A. toxicarius, A. nomius, A. pseudotamarii, A. zhaoqingensis, A. bombycis and from the ascomycete genus Petromyces (Aspergillus section Flavi), (2) Emericella astellata and E. venezuelensis from the ascomycete genus Emericella (Aspergillus section Nidulantes) and (3) Aspergillus ochraceoroseus from a new section proposed here: Aspergillus section Ochraceorosei. We here describe a new species, A. rambellii referable to Ochraceorosei, that accumulates very large amounts of sterigmatocystin, 3-O-methylsterigmatocystin and aflatoxin B1, but not any of the other known extrolites produced by members of Aspergillus section Flavi or Nidulantes. G type aflatoxins were only found in some of the species in Aspergillus section Flavi, while the B type aflatoxins are common in all three groups. Based on the cladistic analysis of nucleotide sequences of ITS1 and 2 and 5.8S, it appears that type G aflatoxin producers are paraphyletic and that section Ochraceorosei is a sister group to the sections Flavi, Circumdati and Cervini, with Emericella species being an outgroup to these sister groups. All aflatoxin producing members of section Flavi produce kojic acid and most species, except A. bombycis and A. pseudotamarii, produce aspergillic acid. Species in Flavi, that produce B type aflatoxins, but not G type aflatoxins, often produced cyclopiazonic acid. No strain was found which produce both G type aflatoxins and cyclopiazonic acid. It was confirmed that some strains of A. flavus var. columnaris produce aflatoxin B2, but this extrolite was not detected in the ex type strain of that variety. A. flavus var. parvisclerotigenus is raised to species level based on the specific combination of small sclerotia, profile of extrolites and rDNA sequence differences. A. zhaoqingensis is regarded as a synonym of A. nomius, while A. toxicarius resembles A. parasiticus but differs with at least three base pair differences. At least 10 Aspergillus species can be recognized which are able to biosynthesize aflatoxins, and they are placed in three very different clades.     

云南兰坪云南红豆杉种群年龄结构与空间分布格局分析

利用点格局分析法对滇西北兰坪县4个云南红豆杉(Taxus yunnanensis)种群的年龄结构、不同生长阶段空间分布格局及其关联性进行分析。结果表明:(1)林窗与人为干扰影响4个云南红豆杉种群的龄级结构,小林窗的种群呈稳定型;较大林窗的种群呈进展型;群落结构完整未出现林窗和有较大人为干扰的种群年龄结构均呈衰退型,幼苗、幼树与小树个体缺乏,种群存在一定程度的更新困难,林窗的大小是种群更新的重要因素。(2)人为干扰、自然生境与本身生物学特性影响4个种群空间分布格局,种群1整体及其不同生长阶段个体均呈聚集分布;种群2和种群4在小尺度呈聚集分布,而在大尺度上呈随机分布;种群3随着空间尺度增大分布格局表现为聚集→随机→聚集趋势。(3)种群2中幼苗、小树和中树在小尺度下表现为聚集分布,其他尺度下表现为随机分布;种群3的小树阶段聚集分布和随机分布皆有出现,中树与大树在不同尺度上表现为聚集分布;种群4聚集分布的强度随生长阶段的增加在减弱。(4)4个种群各发育阶段间皆呈或接近显著正相关,种内竞争弱,有利于种群维持。     

贵州特有植物长柱红山茶种群年龄结构及空间分布格局研究

为评估贵州特有植物长柱红山茶(Camellia mairei var.lapiden)的资源现状,采用样方法对其种群年龄结构和空间分布格局进行了研究。结果表明,长柱红山茶种群年龄结构为衰退型,种群密度以中龄树>老年树>幼龄树;不同发育阶段种群的空间分布格局不同,幼龄树和中龄树均呈显著集群分布,老年树接近集群分布或随机分布;种群的3个发育阶段之空间关系不密切,均表现为负相关或不相关。长柱红山茶由于其繁殖生物学特性和受其群落之种内、种间竞争及生境异质性的影响而趋于濒危和灭绝。因此,建立长柱红山茶就地保护区和开展引种栽培,扩大其栽培种植范围以增加种群数量是保护长柱红山茶种质资源的有效措施。     

Effects of neem leaf extract on production of aflatoxins and activities of fatty acid synthetase,isocitrate dehydrogenase and glutathione S-transferase in Aspergillus parasiticus

The relationship between the activities of 3 cytosolic enzymes with aflatoxin biosynthesis in Aspergillus parasiticus cultured under different conditions has been investigated in order to find out the role of each enzyme in aflatoxin biosynthesis. Basically the activity of isocitrate dehydrogenase (IDH) was higher in non-toxigenic strains as compared to its counterpart toxigenic fungi (p < 0.05). In contrast, the activities of fatty acid synthase (FAS) as well as glutathione S-transferase (GST) were higher (P < 0.05) in toxigenic strains than that of the non-toxigenic fungi. Aflatoxin production was inhibited in fungi grown in presence of various concentrations of neem leaf extract. Aflatoxin was at its lowest level (>90% inhibition) when the concentration of neem extract was adjusted to 50% (v/v). No significant changes in FAS and IDH activities were observed when aflatoxin synthesis was under restraints by neem (Azadirachta indica) leaf extract. During a certain period of time of culture growth, when aflatoxin production reached to its maximum level, the activity of FAS was slightly induced in the toxigenic strains fed with a low concentration (1.56% v/v) of the neem leaf extract. At the time (96 h) when aflatoxin concentration reached to its maximum levels, the activity of GST in the toxigenic fungi was significantly higher (i.e., 7–11 folds) than that of non-toxigenic strains. The difference was highest in mycelial samples collected after 120 h. However unlike FAS and IDH, GST was readily inhibited (67%) in mycelia fed with 1.56% v/v of the neem extract. The inhibition reached to maximum of 80% in samples exposed to 6.25–12.5% of the extract. These results further substantiate previous finding that there is a positive correlation between GST activity and aflatoxin production in fungi.This revised version was published online in October 2005 with corrections to the Cover Date.     

基于无人机高光谱影像的薇甘菊分布提取研究 ——以云南德宏州为例

为有效控制薇甘菊入侵,及时掌握其空间分布和动态变化,基于无人机高光谱数据,通过深度学习（DL）、支持向量机（SVM）、随机森林（RF）等方法提取云南省德宏州微甘菊分布情况。结果表明,DL、SVM和RF等3种方法均有效实现了薇甘菊的分布提取,以DL方法的提取效果最佳,制图精度和用户精度分别为96.61%和95.00%;其次为RF方法,制图精度和用户精度分别为94.83%和91.67%;SVM方法的制图精度和用户精度分别为92.45%和81.67%。这3种方法均能很好提取薇甘菊集中分布区域,且DL和RF方法对零散分布薇甘菊的识别效果优于SVM。因此,无人机高光谱影像为薇甘菊的监测、预警和精准防治提供了支撑和依据,对保护当地生态系统安全具有重要意义。     

Patterns in the Distribution of American Beachgrass (<Emphasis Type="Italic">Ammophila breviligulata</Emphasis>) and the Density and Reproduction of Annual Plants on a Coastal Beach

Beach communities of eastern North America are commonly dominated by the native perennial grass Ammophila breviligulata. It typically co-occurs with a variety of annual grasses and herbs in the pioneer zone. To determine the potential significance of this vigorous clonal species to the annual community, density, spatial distribution, seed production and potential seed rain were quantified for three focus species (Triplasis purpurea, Cenchrus tribuloides, and Heterotheca subaxillaris) in a 45×40 m section of coastal beach on Staten Island, New York, USA. During autumn 2000, five 40 m transects (10 m apart) were established perpendicular to shore; ramet density for Ammophila, and densities and per capita seed production for the three species, were estimated in 150 contiguous quadrats (35 cm × 30 cm) per transect. Both Triplasis and Cenchrus were more likely to occur in quadrats without Ammophila. Per capita seed production of Cenchrus and Heterotheca showed a curvilinear decrease with increasing numbers of Ammophila ramets. Mean seed rain per m² was significantly reduced in the presence of Ammophila for all three focus annuals. In this beach community, Ammophila acts as an ecosystem engineer, but has a mostly negative influence on the distribution, density, and reproduction of the co-occurring annuals. Results suggest that continual, deliberate planting of A. breviligulata could be detrimental to the abundance and diversity of native annual herbs on the eastern coasts of North America.     

盐度对哈氏仿对虾肌肉一般营养成分和氨基酸组成及含量的影响

通过调查不同盐度(12～36)环境下养殖的哈氏仿对虾(Parapenaeopsis hardwickii)肌肉一般营养成分和氨基酸组成及含量,研究了盐度对该虾肌肉营养品质的影响。结果显示,肌肉的水分随环境盐度升高出现显著的直线下降,而粗蛋白含量随环境盐度升高而直线升高;各盐度组的粗脂肪含量虽然没有明显差异,但其含量随盐度升高有一定的线性下降趋势;36盐度组的粗灰分含量比12、16、20和28盐度组的明显高,32盐度组的灰分含量比20和24盐度组的也明显高;所检测的肌肉干样16种氨基酸中,只有3种氨基酸的含量随盐度升高而升高,而其他13种氨基酸的含量随盐度升高明显降低,其中有10种氨基酸的含量在盐度12～24条件下的比盐度28～36条件下的明显高,而这些氨基酸含量在盐度12～24之间没有明显差异。各盐度组间氨基酸总量和鲜味氨基酸含量均没有明显差异;必需氨基酸和半必需氨基酸含量在盐度12～24条件下的均比盐度28～36条件下的明显高,而在12～24盐度组之间以及28～36盐度组之间均没有明显差异。必需氨基酸/氨基酸总量的比值和必需氨基酸/非必需氨基酸比值随盐度升高均明显线性降低;盐度12～24组的必需氨基酸指数(66.13～67.42)高于盐度28～36组的(62.56～64.46)。综上所述,盐度12～24环境下养殖的哈氏仿对虾肌肉营养价值相对较高,表现为低盐趋向,考虑到肌肉的水分含量和生长性能,哈氏仿对虾在养殖期间选择16～24盐度范围比较合适,同时也说明哈氏仿对虾适合大多数沿海地区环境的盐度条件。     

黄土高原紫穗槐丛枝菌根真菌与土壤因子和球囊霉素空间分布的关系

在黄土高原的陕西省甘泉、绥德、米脂、榆林4县(市)选取4个样地,研究紫穗槐不同深度土层丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)侵染率、孢子密度、球囊霉素和土壤因子之间的关系。结果表明:(1)AMF侵染率平均值绥德样地最大(100%),米脂样地最小(75.02%);孢子密度平均值米脂样地最大(5.91个/g),榆林样地最小(1.57个/g);4个样地孢子密度最大值均在0～10cm土层,且随土层加深而降低;AMF侵染率与孢子密度在各样地间差异显著,同一样地侵染率与孢子密度变化规律不一致。(2)榆林样地的有机碳、铵态氮、速效磷、脲酶和碱性磷酸酶活性平均值显著高于其他3个样地;除绥德、米脂样地的蔗糖酶和甘泉样地的碱性磷酸酶外,其他样地的土壤酶活性和球囊霉素含量均随土层加深而降低,且各土层之间差异显著,最大值均在0～10cm土层。(3)AMF侵染率与pH、总球囊霉素含量、易提取球囊霉素含量呈极显著正相关,与有机碳、硝态氮、速效磷含量呈显著正相关;孢子密度与速效磷、蔗糖酶、脲酶活性呈显著正相关,与铵态氮含量呈显著负相关。(4)第1、2主成分的累积方差贡献率达到56.4%,第1主成分主要综合了球囊霉素、AMF侵染率和孢子密度,第2主成分综合了pH和部分土壤因子信息。因此,球囊霉素、AMF侵染率、孢子密度、pH、部分土壤因子对决定土壤生态起主要作用。     

Effects of Saccharomyces cerevisiae supplementation on ruminal parameters, nutrient digestibility and growth in sheep: A meta-analysis

Despite the increased use of dry active Saccharomyces cerevisiae yeast supplementation in ruminant feeding, inconsistent results among studies hamper the prediction of its effects on animal performance. A meta-analysis has been conducted to quantify the magnitude of yeast supplementation effects on ruminal parameters, total tract nutrient digestibility, growth and feed conversion across different studies with sheep. Different methodologies and small numbers of studies necessitated the use of the classical effect size method, in which a unitless standardised effect size (Hedges's g) was used to calculate differences obtained in outcomes between supplemented and non-supplemented sheep. Summary statistics across studies were calculated with fixed and random effects models, whereas subgroup-analysis and meta-regression were applied to identify possible interfering factors that could be responsible for between-study variability. Possible publication bias was evaluated with graphical and statistical tests. Effect sizes for ruminal ammonia nitrogen (33 comparisons), pH (42 comparisons) and total volatile fatty acids (38 comparison) did not (P > 0.10) present heterogeneity among studies, and was not affected by yeast supplementation. No effects (P > 0.05) were detected on the stoichiometry of volatile fatty acids or protozoa counts. Effects sizes calculated for digestibility of dry matter, organic matter, crude protein, acid-detergent fibre and neutral-detergent fibre, which included from 17 to 28 comparisons, showed considerable (>50%) between-study variability. This variability could not effectively be explained by the categorical variables (1) mode of yeast application, (2) feed intake (ad lib versus restricted) or (3) faeces collection method, or the continuous independent variables (1) adaptation period, (2) study period, (3) dietary roughage concentration and (4) dietary crude protein concentration. According to random effects models, digestibility of dry matter, organic matter and crude protein were increased by yeast supplementation, with no effects found for digestibility of fibre components. Substantial unexplained between-study variability (50–90%) was found for growth (13 comparisons) and feed intake (9 comparisons). This meta-analysis presented evidence that addition of dry active S. cerevisiae yeast to diets did not have any effect on growth, feed conversion, ruminal parameters or fibre digestibility in sheep.     

Negative feedback on a perennial crop: Fusarium crown and root rot of asparagus is related to changes in soil microbial community structure

The dynamic equilibrium of an ecosystem is driven by mutual feedback interactions between plants and soil microorganisms. Asparagus exerts a particularly strong influence on its soil environment through abundant production of persistent phenolic acids, which impact selectively soil microorganisms and may be involved in Fusarium crown and root rot (FCRR) of asparagus. In a survey of 50 asparagus plantations of the province of Québec, we found that FCRR was associated with a profound cultivar-specific, reorganization of the soil microbial community, as revealed by phospholipid fatty acid (PLFA) profiling. According to PLFA indicators, microbial biodiversity as well as bacterial and fungal abundance dropped sharply with the onset of FCRR in fields planted with the cultivar Guelph Millenium. This drop was followed by a similar drop in the arbuscular mycorrhizal population. Biodiversity and microbial population size then increased to finally reach a new equilibrium. Discriminant analysis of PLFA profiles obtained from soil samples also indicated a shift in soil microbial community structure associated with FCRR development in fields planted with the cultivar Jersey Giant. Different soil biological conditions, as indicated by microbial biomass C and N and soil enzyme activities, were associated with different cultivars. Preceding crop, manure application, geographical location and tillage depth also influenced the structure of soil microbial communities in asparagus plantations, as determined by PLFA profiling. If higher FCRR incidence is a consequence of the soil microbial community reorganization, means to reduce FCRR incidence in asparagus plantations may be found among practices such as soil organic fertilization, soil tillage and intercropping strategies that would dilute the negative influence of asparagus on the soil microbial community. Finally, FCRR outbreaks were generally promoted by a previous crop of maize. It seems that maize and asparagus host a F. proliferatum teleomorph (Gibberella fujikoroi) of the same mating type.     

基于UAV低空遥感的荒漠林大沙鼠洞群覆盖率及分布特征研究——以新疆古尔班通古特沙漠南缘局部为例

草原鼠害是影响草原生态平衡的重要因素,草原鼠害监测是鼠害防治工作的重要环节。无人机低空遥感是一种新的鼠害监测方法,其具备高空间分辨率、高时效、低成本、低损耗等特点,在新疆鼠害监测防治方法中完成了示范工作。于2015年10月与2016年5月对新疆古尔班通古特沙漠南缘的荒漠林大沙鼠典型鼠害区进行两次无人机低空航拍,分别获取分辨率0.02m与0.024m的实验区超高分辨率影像,对两片实验区整幅影像进行目视解译,获得鼠洞分布情况;在鼠洞分布图基础上进行缓冲区分析并添加趋势线,获取鼠洞空间分布趋势,与研究区山体阴影图叠加分析鼠洞分布与地形间的相互关系。通过GIS格网与GIS叠加分析,获取鼠洞洞群覆盖率。得出如下结论:无人机低空遥感可以为大沙鼠鼠害调查提供准确度甚高的解译结果;古尔班通古特沙漠南缘局部研究区的荒漠林大沙鼠鼠害区洞群覆盖率分别为19.4%、18.8%,已为高密度发生区;研究区大沙鼠鼠洞表现出明显的聚集特征和带状分布特征;聚集特征说明鼠洞是以洞群的形式存在;研究区鼠洞带状分布特征与地形有着密不可分的关系;根据其空间分布特征可以科学规划鼠害治理方案;此研究表明无人机低空遥感在鼠害监测防治方面具有广阔的应用前景。     

A C-NMR study on the influxes into the tricarboxylic acid cycle of a renal epithelial cell line, LLC-PK1/Cl4: the metabolism of [2-C]glycine, l-[3-C]alanine and l-[3-C]aspartic acid in renal epithelial cells

Perchloric acid extracts of LLC-PK₁/Cl₄ cells, a renal epithelial cell line, incubated with either [2-¹³C]glycine l-[3-¹³C]alanine, or d,l-[3-¹³C]aspartic acid were investigated by ¹³C-NMR spectroscopy. All amino acids, except labelled glycine, gave rise to glycolytic products and tricarboxylic acid cycle (TCA) intermediates. For the first time we also observed activity of γ-glutamyltransferase activity and glutathione synthetase activity in LLC-PK₁ cells, as is evident from enrichment of reduced glutathione. Time courseS showed that only 6% of the labelled glycine was utilized in 30 min, whereas 31% of l-alanine and 60% of l-aspartic acid was utilized during the same period. ¹³C-NMR was also shown to be a useful tool for the determination of amino acid uptake in LLC-PK₁ cells. These uptake experiments indicated that glycine alanine and aspartic acid are transported into Cl₄ cells via a sodium-dependent process. From the relative enrichment of the glutamate carbons, we calculated the activity of pyruvate dehydrogenase to be about 61% of when labelled l-alanine was the only carbon source for LLC-PK₁/Cl₄ cells. Experiments with labelled d,l-aspartic, however, showed that about 40% of C-3-enriched oxaloacetate (arising from a de-amination of aspartic acid) reached the pyruvate pool.