An overview of short‐term tests for the mutagenic and carcinogenic potential of pesticides

Abstract

In the last few years, marked progress has been made in the development of methods for evaluating the mutagenic and carcinogenic potential of pesticide chemicals. The correlation of genetic and related biological activity in short‐term tests with carcinogenic activity in whole animals allows the utilization of short‐term mutagenicity bioassays to prescreen chemicals for effects related to mutation induction and presumptive carcinogenicity. In addition, bioassays now available can measure directly the chemical transformation of normal cells in culture into cells capable of producing tumors when injected into animals.

This paper will review briefly the major types of relevant short‐term tests and will develop a rationale for a phased approach to the evaluation of the mutagenic and carcinogenic potential of environmental chemicals. This approach involves the sequential application of bioassays which are organized into a three‐level matrix emphasizing first detection, then confirmation, and finally hazard assessment. Chemicals demonstrating positive results in the short‐term detection systems and confirmatory bioassays are pursued in higher level whole animal tests. A core battery of tests is proposed to operationally define a negative result. The phased approach should facilitate a cost effective utilization of limited testing resources and provide protection for human health in proportion to the anticipated hazard.

Results obtained in evaluating a series of thirty‐eight pesticide chemicals according to the phased approach are discussed in detail.     

Organ targeted prenatal gene therapy—how far are we?

Prenatal gene therapy aims to deliver genes to cells and tissues early in prenatal life, allowing correction of a genetic defect, before long-term tissue damage has occurred. In contrast to postnatal gene therapy, prenatal application can target genes to a large population of dividing stem cells, and the smaller fetal size allows a higher vector-to-target cell ratio to be achieved. Early-gestation delivery may allow the development of immune tolerance to the transgenic protein which would facilitate postnatal repeat vector administration if needed. Targeting particular organs will depend on manipulating the vector to achieve selective tropism and on choosing the most appropriate gestational age and injection method for fetal delivery. Intra-amniotic injection reaches the skin, and other organs that are bathed in the fluid however since gene transfer to the lung and gut is usually poor more direct injection methods will be needed. Delivery to the liver and blood can be achieved by systemic delivery via the umbilical vein or peritoneal cavity. Gene transfer to the central nervous system in the fetus is difficult but newer vectors are available that transduce neuronal tissue even after systemic delivery. Copyright © 2011 John Wiley & Sons, Ltd.     

胜利油田外排水中铁细菌主要类群的检测及群落结构分析

根据油田外排水中常见铁细菌的16SrRNA特异性保守序列,设计和合成了4种荧光探针,利用荧光原位杂交方法,分别利用它们对胜利油田孤岛采油厂、胜利采油厂1号和2号综合处理站外排水中存在的铁细菌的种类和数量进行检测,分析了各外排水中铁细菌的群落结构.结果表明,利用基因探针能快速对胜利油田外排水中的铁细菌进行检测:在孤岛采油厂、胜利采油厂1号和2号综合处理站的外排水中,利用4种探针的组合探针检测到的铁细菌细胞数量分别占样品中微生物总细胞数量的11.0%、12.8%和9.0%,其中Leptothrix和Sphaerotilus,以及Leptospirillum fer-riphilum为胜利油田外排水中的3种优势铁细菌,分别占水样中总微生物细胞数的2.94%±0.52%~3.39%±0.52%和2.24%±0.16%~2.63%±0.49%;而Leptospirillum ferrooxidans和Acidithiobacillusspp.的种群则较小.3个污水处理站中铁细菌群落结构差异较大,而多样性相差不大.利用4种探针的各种组合对外排水中铁细菌的检测表明,单探针能够很好地分析外排水中铁细菌的群落结构,而组合探针能快速、较准确地检测外排水中铁细菌的数量,比传统的绝迹稀释法具有快速、准确、直观等优点.图1表5参14     

Preimplantation genetic diagnosis for single gene disorders: experience with five single gene disorders

We report our experience of 14 preimplantation genetic diagnosis (PGD) cycles in eight couples carrying five different single gene disorders, during the last 18 months. Diagnoses were performed for myotonic dystrophy (DM), cystic fibrosis (CF) [ΔF508 and exon 4 (621+1 G>T)], fragile X and CF simultaneously, and two disorders for which PGD had not been previously attempted, namely neurofibromatosis type 2 (NF2) and Crouzon syndrome. Diagnoses for single gene disorders were carried out on ideally two blastomeres biopsied from Day 3 embryos. A highly polymorphic marker was included in each diagnosis to control against contamination. For the dominant disorders, where possible, linked polymorphisms provided an additional means of determining the genotype of the embryo hence reducing the risk of misdiagnosis due to allele dropout (ADO). Multiplex fluorescent polymerase chain reaction (F-PCR) was used in all cases, followed by fragment analysis and/or single-stranded conformation polymorphism (SSCP) for genotyping. Embryo transfer was performed in 13 cycles resulting in one biochemical pregnancy for CF, three normal deliveries (a twin and a singleton) and one early miscarriage for DM and a singleton for Crouzon syndrome. In each case the untransferred embryos were used to confirm the diagnoses performed on the biopsied cells. The results were concordant in all cases. The inclusion of a polymorphic marker allowed the detection of extraneous DNA contamination in two cells from one case. Knowing the genotype of the contaminating DNA allowed its origin to be traced. All five pregnancies were obtained from embryos in which two blastomeres were biopsied for the diagnosis. Our data demonstrate the successful strategy of using multiplex PCR to simultaneously amplify the mutation site and a polymorphic locus, fluorescent PCR technology to achieve greater sensitivity, and two-cell biopsy to increase the efficiency and success of diagnoses. Copyright © 2002 John Wiley & Sons, Ltd.     

Effects of imazethapyr spraying on plant growth and leaf surface microbial communities in Arabidopsis thaliana

Imazethapyr (IM) is an acetolactate synthase (ALS)-inhibiting herbicide that has been widely used in recent years. However, IM spraying can lead to the accumulation of herbicide residues in leaves. Here, we determined the effects of IM spraying on the plant growth and leaf surface microbial communities of Arabidopsis thaliana after 7 and 14?days of exposure. The results suggested that IM spraying inhibited plant growth. Fresh weight decreased to 48% and 26% of the control value after 7 and 14?days, respectively, of 0.035?kg/ha IM exposure. In addition, anthocyanin content increased 9.2-fold and 37.2-fold relative to the control content after 7 and 14?days of treatment, respectively. Furthermore, IM spraying destroyed the cell structures of the leaves, as evidenced by increases in the number of starch granules and the stomatal closure rate. Reductions in photosynthetic efficiency and antioxidant enzyme activity were observed after IM spraying, especially after 14?days of exposure. The diversity and evenness of the leaf microbiota were not affected by IM treatment, but the composition of community structure at the genus level was altered by IM spraying. Imazethapyr application increased the abundance of Pseudomonas, a genus that includes species pathogenic to plants and humans, indicating that IM potentially increased the abundance of pathogenic bacteria on leaves. Our findings increase our understanding of the relationships between herbicide application and the microbial community structures on plant leaves, and they provide a new perspective for studying the ecological safety of herbicide usage.     

Coupled biodegradation of p-nitrophenol and p-aminophenol in bioelectrochemical system: Mechanism and microbial functional diversity

Biodegradation mechanisms and microbial functional diversity during coupled p-nitrophenol (PNP) and p-aminophenol (PAP) degradation were studied in a bioelectrochemical system. PNP in the biocathode and PAP in the bioanode were almost completely removed within 28hr and 68hr respectively. The degradation followed the steps including hydrating hydroxyalkylation, dehydrogenating carbonylation, and hydrolating ring cleavage, etc. Metagemomic analysis based on the KEGG and eggNOG database annotations revealed the microbial composition and functional genes/enzymes related to phenol degradation in the system. The predominant bacteria genera were Lautropia, Pandoraea, Thiobacillus, Ignavibacterium, Truepera and Hyphomicrobium. The recognized biodegradation genes/enzymes related to pollutant degradation were as follows: pmo, hbd, & ppo for phenol degradation, nzba, amie, & badh for aromatic degradation, and CYP & p450 for xenobiotics degradation, etc. The co-occurrence of ARGs (antibiotic resistant genes), such as adeF, MexJ, ErmF, PDC-93 and Escherichia_coli_mdfA, etc., were annotated in CARD database during the biodegradation process. The Proteobacteria & Actinobacteria phylum was the primary host of both the biodegradation genes & ARGs in this system. The microbial functional diversity ensured the effective biodegradation of the phenol pollutants in the bioelectrochemical system.     

聚丙烯微塑料对污泥厌氧消化效能作用影响

以聚丙烯（PP）微塑料为研究对象,考察不同浓度PP微塑料对污泥厌氧消化产CH₄和产酸效能的作用影响,同时采用荧光定量PCR方法定量检测了乙酸激酶（AK）和mcrA基因在不同PP微塑料作用下的丰度变化.结果表明,PP微塑料对污泥厌氧消化产CH₄和产酸效能具有促进影响,CH₄和乙酸累计产量随PP微塑料投加量的增大而升高,当PP微塑料投加量为0.2g/g VSS时,CH₄和乙酸累计产量与空白对照相比分别提高148.2%和15.2%,达227.1mL/g VSS和1291.2mg/L.相应地,mcrA基因丰度随之提高98.2%,表明PP微塑料对产甲烷菌的生长和繁殖具有促进作用,进而强化污泥厌氧消化产CH₄效能.