Hapten and antibody production for a sensitive immunoassay determining a human urinary metabolite of the pyrethroid insecticide permethrin

Permethrin is the most popular synthetic pyrethroid insecticide in agriculture and public health. For the development of the enzyme-linked immunosorbent assay (ELISA) to evaluate human exposure to permethrin, the glycine conjugate (DCCA-glycine) of a major metabolite, cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), of permethrin was established as the target analyte. Four different types of the cis- and trans-isomers of immunizing haptens were synthesized as follows: N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine (hapten 3), N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)-4-amino-l-phenylalanine (hapten 5), N-(N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine)amino-6-(2,4-dinitrophenyl)aminohexanoic acid (hapten 9), and N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine-4-oxobutanoic acid (hapten 24). Sixteen polyclonal antibodies produced against each cis- or trans-hapten-thyroglobulin conjugate as immunogens were screened against numerous hapten-bovine serum albumin conjugates as coating antigens. Six ELISAs with both a heterologous hapten structure and a heterologous hapten configuration (cis/trans or trans/cis) between antibody and coating antigen showed a high sensitivity for the target analyte. The IC50 was 1.3, 2.1, and 2.2 microg/L for the trans-target analyte and 0.4, 2.3, and 2.8 microg/L for the cis-target analyte. The immunizing haptens, except for hapten 5, provided the target specific antibodies. Molecular modeling of the haptens supported the selection of reasonable immunizing haptens that best mimicked the target analyte. Hapten 5 was suitable as a coating antigen rather than as an immunogen since it had a different geometry. Very low cross-reactivities were measured to permethrin, its free metabolite (DCCA), PBA-glycine conjugate, and glycine. The ELISA will be optimized for the detection of total cis/trans-DCCA-glycine in human urine samples.     

Flea market finds and global exports: Four multistate outbreaks of human Salmonella infections linked to small turtles,United States—2015

Zoonotic transmission of Salmonella infections causes an estimated 11% of salmonellosis annually in the United States. This report describes the epidemiologic, traceback and laboratory investigations conducted in the United States as part of four multistate outbreaks of Salmonella infections linked to small turtles. Salmonella isolates indistinguishable from the outbreak strains were isolated from a total of 143 ill people in the United States, pet turtles, and pond water samples collected from turtle farm A, as well as ill people from Chile and Luxembourg. Almost half (45%) of infections occurred in children aged <5 years, underscoring the importance of the Centers for Disease Control and Prevention recommendation to keep pet turtles and other reptiles out of homes and childcare settings with young children. Although only 43% of the ill people who reported turtle exposure provided purchase information, most small turtles were purchased from flea markets or street vendors, which made it difficult to locate the vendor, trace the turtles to a farm of origin, provide education and enforce the United States federal ban on the sale and distribution of small turtles. These outbreaks highlight the importance of improving public awareness and education about the risk of Salmonella from small turtles not only in the United States but also worldwide.     

耕作方式与施肥对旱农区粮饲兼用玉米耗水特性和干物质积累的影响

采用大田试验,在全膜双垄沟播技术的基础上,比较研究了4种耕作方式(T1:传统翻耕,T2:旋耕,T3:深松耕,T4:免耕)搭配2种施氮水平(N2:施纯氮200 kg·hm~(-2),N3:施纯氮300 kg·hm~(-2))下对玉米耗水特性和干物质积累的影响。结果表明:与施氮量相比,耕作方式对耗水量和土壤贮水消耗量影响显著;深松耕降低了玉米播种至拔节期的阶段耗水量,提高了拔节至灌浆期的耗水量及其占总耗水量的比例;播种至拔节期深松耕较传统翻耕0~30 cm土层的贮水消耗量减少了35.3%,拔节至开花期和开花至灌浆期,30~110 cm土层贮水消耗量增加了54.2%和20.5%;玉米不同生育时期各器官的干物质积累量都表现为深松耕+施纯氮300 kg·hm~(-2)最高,是生物产量和籽粒产量最高的处理,深松耕+施纯氮300 kg·hm~(-2)和深松耕+施纯氮200 kg·hm~(-2)处理间差异不显著;免耕+施纯氮200 kg·hm~(-2)处理的水分利用效率最高,与深松耕+施纯氮300 kg·hm~(-2)处理相比差异不显著。在综合考虑高产以及对水分和肥料的高效利用的条件下,基于全膜双垄沟播技术,深松耕+施纯氮200 kg·hm~(-2)是陇中旱农区适宜的生产方式。     

Influence of genotype and yield and composition of milk on interval to first postpartum ovulation in milked beef and dairy cows.

Our objectives were to investigate the effects of genotype, yield and composition of milk, and changes in BW and body condition on the initiation of cyclic ovarian activity in nonsuckled beef and dairy cows milked twice daily. Nulliparous heifers were selected from three breeds, 12 Angus, 13 Simmentals, and 26 Holsteins, based on their EBV for milk yield. Cows were machine-milked twice daily, and daily milk yield was recorded; composite samples of milk were collected twice weekly for analyses of milk components. Blood was collected from all cows thrice weekly for 75 d postpartum, and concentrations of progesterone in serum measured by RIA were used to estimate day of first postpartum ovulation. Holstein cows produced more (P less than .05) milk (unadjusted or 3.5% fat-corrected) than Angus or Simmental cows during the first 30 d of lactation. Holstein cows had higher (P less than .01) peak yield of milk than Simmental and Angus cows. Days to peak milk yield were similar for Simmental and Holstein cows, and both were more (P less than .05) than those for Angus cows. Percentages of fat, protein, and total solids in milk were highest (P less than .05) for Simmental cows, whereas milk of Holstein cows had the highest (P less than .05) percentage of lactose and lowest (P less than .05) concentration of somatic cells. Average BW at ovulation differed (P less than .05) among breeds. Estimated daily changes in BW from calving to first ovulation were different (P less than .05) for Holstein and Simmental cows but were similar to those of Angus cows.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cellulose synthase-like CslF genes mediate the synthesis of cell wall (1,3;1,4)-beta-D-glucans

A characteristic feature of grasses and commercially important cereals is the presence of (1,3;1,4)-beta-d-glucans in their cell walls. We have used comparative genomics to link a major quantitative trait locus for (1,3;1,4)-beta-d-glucan content in barley grain to a cluster of cellulose synthase-like CslF genes in rice. After insertion of rice CslF genes into Arabidopsis, we detected (1,3;1,4)-beta-d-glucan in walls of transgenic plants using specific monoclonal antibodies and enzymatic analysis. Because wild-type Arabidopsis does not contain CslF genes or have (1,3;1,4)-beta-d-glucans in its walls, these experiments provide direct, gain-of-function evidence for the participation of rice CslF genes in (1,3;1,4)-beta-d-glucan biosynthesis.     

"肉鸡球虫病及其综合防治措施"专栏(七)成功控制禽球虫病的挑战

艾美尔球虫以宿主特异性的方式感染家畜,是引起球虫病的病原体.艾美尔球虫的控制是必要的.目前主要的控制方法是化学疗法,而用由活野毒株或弱毒虫株制成的疫苗进行免疫预防则呈流行趋势.新一代的亚单位疫苗、活栽体疫苗和DNA疫苗正在研究之中,而合适抗原的鉴定是仍需要解决的难题.本文简述了过去和当前控制球虫病的方法,及一种鉴定作为候选疫苗免疫保护性抗原的新方法.     

Isolation of plant transcription factors using a modified yeast one-hybrid system

Background  

The preparation of expressional cDNA libraries for use in the yeast two-hybrid system is quick and efficient when using the dedicated Clontech™ product, the MATCHMAKER Library Construction and Screening Kit 3. This kit employs SMART technology for the amplification of full-length cDNAs, in combination with cloning using homologous recombination.     

Use of immunochemical techniques for the analysis of pesticides

Immunochemical assays for small molecules such as pesticides are rapidly gaining acceptance among analytical chemists. These techniques are rapid, sensitive, cost effective and can easily cope with large sample loads. This review lists the advantages and disadvantages of the technique and describes the steps in assay development using examples from this laboratory, particularly the thiocarbamate herbicide molinate and the triazine herbicides. The focus is primarily on hapten synthesis strategies, assay format considerations, sample preparation and assay validation.