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Veterinary antimicrobials are introduced to wider environments by manure application to agricultural fields or through leaching or runoff from manure storage areas (feedlots, stockpiles, windrows, lagoons). Detected in manure, manure-treated soils, and surface and ground water near intensive cattle feeding operations, there is a concern that environmental contamination by these chemicals may promote the development of antimicrobial resistance in bacteria. Surface runoff and leaching appear to be major transport pathways by which veterinary antimicrobials eventually contaminate surface and ground water, respectively. A study was conducted to investigate the transport of three veterinary antimicrobials (chlortetracycline, sulfamethazine, tylosin), commonly used in beef cattle production, in simulated rainfall runoff from feedlot pens. Mean concentrations of veterinary antimicrobials were 1.4 to 3.5 times higher in surface material from bedding vs. non-bedding pen areas. Runoff rates and volumetric runoff coefficients were similar across all treatments but both were significantly higher from non-bedding (0.53Lmin(-1); 0.27) than bedding areas (0.40Lmin(-1); 0.19). In keeping with concentrations in pen surface material, mean concentrations of veterinary antimicrobials were 1.4 to 2.5 times higher in runoff generated from bedding vs. non-bedding pen areas. Water solubility and sorption coefficient of antimicrobials played a role in their transport in runoff. Estimated amounts of chlortetracycline, sulfamethazine, and tylosin that could potentially be transported to the feedlot catch basin during a one in 100-year precipitation event were 1.3 to 3.6ghead(-1), 1.9ghead(-1), and 0.2ghead(-1), respectively. This study demonstrates the magnitude of veterinary antimicrobial transport in feedlot pen runoff and supports the necessity of catch basins for runoff containment within feedlots., (Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.)

Melanoplussanguinipes oviposition stimulating protein (MsOSP) was characterized and its role in stimulating oviposition in virgin females was examined. A 967nt MsOSP mRNA sequence with homology to previously characterized N-terminal amino acid sequence data for MsOSP was identified in a RNAseq library generated from an mRNA pool from the long hyaline tubule (LHT) of the male accessory gland complex. This transcript contained a predicted 729nt open reading frame encoding the 242aa putative MsOSP protein and had the second highest read abundance in the library. The MsOSP transcript was detected exclusively in the LHT tissue of adult males and its abundance increased with time until 7 days post-eclosion. Western blot analysis using an anti-MsOSP antibody showed high levels of MsOSP protein in the LHT luminal secretions of virgin males and to a lesser degree was associated with the aedeagus and ejaculatory duct. MsOSP was shown to be a major protein component of the spermatophore packet transferred from the male to female during copulation. However, only minor amounts of MsOSP could be detected in the female bursa, spermatheca and oviduct. Intrahemocoelic injection of LHT luminal protein into mature virgin females stimulated oviposition in ∼ 65% of females. A similar but non-significant trend was observed upon injection of purified recombinant MsOSP protein, and immunoprecipitation of LHT protein with anti-MsOSP antibody led to abrogation of oviposition stimulation upon injection of mature virgin females. Despite the demonstration of stimulation of oviposition upon intrahemocoelic injection of LHT-derived-MsOSP into mature virgin females, the potential mode of action of MsOSP in this process remains to be determined. MsOSP cannot be detected in the tissues other than the bursa, spermatheca and oviduct of female grasshoppers and relatively large quantities of MsOSP are required to stimulate oviposition upon injection., (Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.)

A precise, accurate, nonambiguous and high-throughput method is required to assess nuclear maturation of mammalian oocytes. The objectives of this study were to compare the efficiency and ease of use of a simplified fluorescence imaging (anti-lamin A/C and 4',6-diamidino-2-phenylindole [DAPI]) technique to the existing technique (aceto-orcein staining) for the evaluation of nuclear maturation of bovine oocytes, and to determine the kinetics of bovine oocyte maturation using an anti-lamin A/C-DAPI technique. In Experiment 1, oocytes were matured in vitro and stained with aceto-orcein and anti-lamin A/C-DAPI staining techniques. The proportions of oocytes lost during procedures and those that could not be classified (because of ambiguous morphology) during evaluation were lower (P < 0.0001) in oocytes stained with anti-lamin A/C-DAPI (9% and 2%) than those stained with aceto-orcein (31% and 13%), respectively. Anti-lamin A/C-DAPI was a quick procedure which could be completed within 7 h after completion of the maturation (compared with > 24 h for the aceto-orcein method). Furthermore, > 200 oocytes could be stained in one batch with anti-lamin A/C-DAPI technique. In Experiment 2, nuclear maturation kinetics of bovine oocytes at various time intervals (0, 6, 12, and 22 h) during in vitro maturation (IVM) was evaluated using the anti-lamin A/C-DAPI technique. Germinal vesicle, germinal vesicle breakdown, metaphase I, and metaphase II oocytes were predominant at 0, 6, 12, and 22 h of IVM, respectively. We concluded that the anti-lamin A/C-DAPI was an efficient and simple technique for nonambiguous evaluation of nuclear maturation status of large numbers of oocytes in a short interval., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The occupational exposure resulting from the application of crop protection agents continues to be of great interest for the purposes of identifying hazards or determining safer chemical handling methods. The purpose of the present study was to identify the potential respirator exposure of a mixer/loader to chlorothalonil, with the mixing and loading operation as the only source of aerosols (particles <13 microm diameter). Three worst-case mixing/loading scenarios were simulated in the lab: (1) a spill of undiluted chlorothalonil formulation onto a dry, horizontal metal surface; (2) a spill of undiluted chlorothalonil formulation onto a rapidly rotating shaft; and (3) pouring undiluted chlorothalonil formulation into a container of water. Aerosol generation from these scenarios was compared to that resulting from atomizing dilute chlorothalonil through hydraulic nozzles. Aerosols were captured with a cascade impactor, and quantified by gas chromatography. Results indicated that simulated spill scenarios generated aerosol concentrations between 2.1 and 5.3 ng/l, which were in the same order of magnitude as, and only marginally higher than, the detection threshold (1.7 ng/l) and background levels (2.2 ng/l). In comparison, atomization of dilute chlorothalonil through a hollow cone and flat fan nozzles resulted in airborne concentrations of 354 and 96 ng/l, respectively, related to the atomization characteristics of these nozzles. Measurement of the dimensions of the aerosol cloud indicated that aerosols resulting from a spill amounted to approximately 10(-5)% of the spilled chlorothalonil. It was estimated that a male worker respiring 29 l/min would inhale approximately 0.32-0.78 ng of chlorothalonil during a typical 30 s spill, assuming a 1% transfer efficiency between the spill site and the mixer/loader. These estimates were between 10000 and 480000 times less than literature data for respiratory exposure of chlorothalonil by applicators and harvesters, suggesting that inhalation of aerosols from mixing and loading represents a minor component of overall exposure.