Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.     

Tank-mix adjuvants and pesticide residues: some regulatory and quantitative aspects

Although it is well known that judicious use of adjuvants can increase the performance of foliage-applied sprays of many agrochemicals, little information is available in the public domain about their ultimate effects on pesticide residues in treated crops. In the present work, the influence of Agral (polyoxyethylene nonylphenols), Toil (methyl esters of rapeseed fatty acids) and Bond (styrene-butadiene copolymers) on surface and crop residues of diclofop-methyl/diclofop and propiconazole in wheat and field beans was investigated using a model system simulating field practice. Pesticides were applied as commercial formulations, diclofop-methyl 378 g litre(-1) EC (Hoegrass) and propiconazole 250 g litre(-1) EC (Tilt), at their maximum approved rates, 1135 g AI ha(-1) and 125 g AI ha(-1), respectively, both in the presence or absence of the maximum rate recommended for each candidate adjuvant. No detectable residues of diclofop-methyl or propiconazole were found in wheat 35 days after any of the four applications. However, residues of diclofop were present in this crop, and those from applications containing Agral (0.07 mg kg(-1) fresh weight (FW)) or Bond (0.08 mg kg(-1) FW) were significantly lower than those with no adjuvant (0.14 mg kg(-1) FW) or Toil (0.16 mg kg(-1) FW). Unlike wheat, residues of both diclofop and propiconazole were detected in field beans after harvest. Significantly higher residues of the former were recorded from the applications with Agral or Bond (ca 0.32 mg kg(-1) FW) than with those with no adjuvant or Toil (ca 0.15mg kg(-1) FW). All the propiconazole applications containing adjuvants showed a similar significant increase in residues (0.10-0.16 mg AI kg(-1) FW) over the no-adjuvant treatment (0.05 mg kg(-1) FW) in this crop. There appeared to be little agreement between the apparent amounts of uptake, as indicated by the rates of decline of surface residues up to 5 days after application, and final residues in either target species. On wheat, surface residues of diclofop-methyl decreased from initially ca 20 to as little as 0.02 mg kg(-1) FW using adjuvants; the corresponding values for propiconazole were ca 2 to ca 0.03 mg kg(-1) FW. Recoveries of diclofop-methyl from the surfaces of field beans were much higher than those from wheat, declining from ca 30 to only ca 6 mg kg(-1) FW during the course of 5 days; the corresponding residues for propiconazole were ca 2 mg to 0.15 mg kg(-1) FW. These findings are discussed in relation to uptake results obtained with radiolabelled pesticides and adjuvants in the laboratory, and to the mandatory requirements for pesticide residue data for the authorised use of adjuvants in the UK.     

滇黄芩毛状根的诱导及其黄芩苷含量测定

本文利用发根农杆菌A grobacterizum rhizogenes1.2556感染滇黄芩再生苗的茎段和叶片,建立了毛状根培养及其植株再生体系。毛状根可直接从受伤的茎、叶外植体表面产生,在无外源激素的MS固体和液体培养基上自主生长,表现出典型的发根特征。毛状根茎段的诱导率较叶片高,最高可达到14.44%;经rolB基因PCR分析和甘露碱纸电泳检测,证明Ri质粒T-DNA已整合到滇黄芩基因组中并表达;毛状根在附加6-BA2mg/L和NAA0.2mg/L的MS固体培养基上直接诱导不定芽,并在MS培养基上生根,形成再生植株。获得的毛状根系经MS液体培养基培养30d后通过HPLC都能检测到黄芩苷,其中1个转化系黄芩苷含量为2.59%,是药材黄芩的0.20倍,而从3年单位时间黄芩苷生成量计算,毛状根是药材黄芩的7.18倍。本研究建立的毛状根培养体系,将对滇黄芩转基因技术的完善和利用毛状根生产黄芩苷的生物转化提供了实验基础。     

Aspartic Proteinase Members Secreted by the Ruminant Placenta: Specificity of Three Radioimmunoassay Systems for the Measurement of Pregnancy-associated Glycoproteins

Pregnancy‐associated glycoproteins (PAGs) isolated from the placenta of various ruminant species are enzymatically inactive members of the aspartic proteinase family. The measurement of these proteins in the maternal blood can be a good indicator of the presence of a live embryo. As certain aspartic proteinases are present in biological fluids in physiological and pathological conditions at various concentrations, it was necessary to determine the specificity of three radioimmunoassay (RIA) systems currently used for the detection of PAG molecules. Commercially available members of the aspartic proteinase family like pepsinogen, pepsin, chymosin, rennet, cathepsin D and renin were tested in a wide concentration range (10 ng/ml – 1 mg/ml). Pepsinogen cross‐reacted in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 50 μg/ml and 500 μg/ml concentrations, respectively. In the presence of pepsin, cross‐reaction was observed in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 500 μg/ml and 1 mg/ml concentrations, respectively. Chymosin and rennet could cross‐react in RIA 2 and RIA 3, while renin and cathepsin D did not decrease the binding of the tracer to antisera more, than that of the minimal detection limit. As the plasma/serum concentrations of the examined aspartic proteinases reported in the literature were outside the concentration range where cross‐reaction was observed, it can be concluded that these RIA systems were specific for the detection of PAGs in biological fluids.     

Factors Affecting Plasma Pregnancy-associated Glycoprotein 1 Concentrations Throughout Gestation in High-producing Dairy Cows

This study was designed to establish the factors, if any, which could affect plasma pregnancy-associated glycoprotein-1 (PAG-1) expression in a study population of 87 pregnant, high-producing dairy cows. The factors examined were: semen providing breed (Holstein-Friesian vs Limousin), outcome of gestation (male vs female newborn, and singleton vs twin pregnancies), lactation number, milk production at pregnancy diagnosis, plasma progesterone concentration, season of gestation (warm period, March–November vs cool period, December–February), and day of gestation (40, 90, 120, 150, 180 and 210). Pregnancy was diagnosed by transrectal ultrasound on day 40 post-insemination and by palpation per rectum on days 90, 120, 150, 180 and 210. Blood samples were collected from each animal immediately before each pregnancy diagnosis. The relative contributions of the different factors on PAG-1 concentrations were evaluated by GLM repeated measures analysis of variance. No significant effects of the herd, foetal sex, milk production, lactation number and plasma progesterone concentrations were observed. In contrast, twin pregnancy, the use of Limousin semen and conception during the cool period were correlated with significantly increased plasma PAG-1 concentrations throughout gestation. Our data indicate that both cow well-being during early placental development, determined in our conditions by reduced heat stress when conception occurred in the cool season, and crossbreed pregnancies lead to improved PAG-1 production throughout the gestation period.     

Drift reduction and droplet-size in sprays containing adjuvant oil emulsions

The droplet-sizes and drift produced by sprays containing dilute oil-in-water emulsions of various vegetable and mineral oil adjuvants were measured in wind-tunnel experiments. When compared with water or a surfactant solution, both types of oil adjuvant reduced the proportion of small driftable droplets and, hence, reduced measured spray drift. Vegetable oils were more effective at drift reduction than the mineral oil. Spray drift tended to increase with increasing oil concentration and decreasing emulsifier content.