Pesticide residues in honey bee (Apis mellifera) pollen collected in two ornamental plant nurseries in Connecticut: Implications for bee health and risk assessment.

Honey bees (Apis mellifera L.) are one of the most important managed pollinators of agricultural crops. While potential effects of agricultural pesticides on honey bee health have been investigated in some settings, risks to honey bees associated with exposures occurring in the plant nursery setting have received little attention. We sought to identify and quantify pesticide levels present in honey bee-collected pollen harvested in two ornamental plant nurseries (i.e., Nursery A and Nursery B) in Connecticut. From June to September 2018, pollen was collected weekly from 8 colonies using bottom-mounted pollen traps. Fifty-five unique pesticides (including related metabolites) were detected: 24 insecticides, 20 fungicides, and 11 herbicides. Some of the pesticide contaminants detected in the pollen had not been applied by the nurseries, indicating that the honey bee colonies did not exclusively forage on pollen at their respective nursery. The average number of pesticides per sample was similar at both nurseries (i.e., 12.9 at Nursery A and 14.2 at Nursery B). To estimate the potential risk posed to honey bees from these samples, we utilized the USEPA's BeeREX tool to calculate risk quotients (RQs) for each pesticide within each sample. The median aggregate RQ for nurse bees was 0.003 at both nurseries, well below the acute risk level of concern (LOC) of ≥0.4. We also calculated RQs for larvae due to their increased sensitivity to certain pesticides. In total, 6 samples had larval RQs above the LOC (0.45-2.51), resulting from the organophosphate insecticide diazinon. Since 2015, the frequency and amount of diazinon detected in pollen increased at one of our study locations, potentially due to pressure to reduce the use of neonicotinoid insecticides. Overall, these data highlight the importance of considering all life stages when estimating potential risk to honey bee colonies from pesticide exposure.

Impact of Diflubenzuron on Bombus impatiens (Hymenoptera: Apidae) Microcolony Development.

Reliance on the honey bee as a surrogate organism for risk assessment performed on other bees is widely challenged due to differences in phenology, life history, and sensitivity to pesticides between bee species. Consequently, there is a need to develop validated methods for assessing toxicity in non-Apis bees including bumble bees. The usefulness of small-scale, queenless colonies, termed microcolonies, has not been fully investigated for hazard assessment. Using the insect growth regulator diflubenzuron as a reference toxicant, we monitored microcolony development from egg laying to drone emergence using the Eastern bumble bee Bombus impatiens (C.), a non-Apis species native to North America. Microcolonies were monitored following dietary exposure to diflubenzuron (nominal concentrations: 0.1, 1, 10, 100, and 1,000 µg/liter). Microcolony syrup and pollen consumption was significantly reduced by diflubenzuron exposure. Pupal cell production was also significantly decreased at the highest diflubenzuron concentration assessed. Ultimately, diflubenzuron inhibited drone production in a concentration-dependent manner and a 42-d 50% inhibitory concentration (IC50) was determined. None of the dietary concentrations of diflubenzuron tested affected adult worker survival, or average drone weight. These data strengthen the foundation for use of this methodology, and provide valuable information for B. impatiens; however, more work is required to better understand the utility of the bumble bee microcolony model for pesticide hazard assessment.

A cost-effective colourimetric assay for quantifying hydrogen peroxide in honey.

Honey is a natural product with many beneficial properties including antimicrobial action. Production of hydrogen peroxide (H2O2) in diluted honey is central to this action. Here, we describe an optimized method for measuring levels of H2O2 in honey. This method is based on established methods, with the level of dilution, the time between dilution and reading the assay, and aeration of the samples during the assay identified as critical points for ensuring reliability and reproducibility. The method is cost-effective and easy to perform using common laboratory equipment. Using this method, we quantified the hydrogen peroxide content of five different, unprocessed polyfloral honeys collected in NC, USA. Our results show that H2O2 production by these honeys varies greatly, with some samples producing negligible levels of H2O2. We assessed the effect of colour on the assay by measuring the recovery of spiked H2O2 from light and dark honey and from serially diluted dark corn syrup, and found the amount of H2O2 that could be detected was lower in dark corn syrup and darker honey samples.

Development and utilization of a unique in vitro antigen presentation co-culture model for detection of immunomodulating substances.

Current regulatory immunotoxicity studies require the use of animal models. However, evolving regulatory requirements, the need to evaluate large numbers of chemicals efficiently and societal pressures are driving the development and utilization of alternative in vitro methods for identifying potential immunotoxicants. In line with these efforts, we developed a novel in vitro cell-based assay to evaluate effects on antigen presentation - a key step in successful immunization. In this assay, Ch27 B cells acquire and present hen egg lysozyme peptides to antigen-restricted 3A9 T cells, causing them to produce and secrete IL-2. IL-2 levels in the culture medium may be monitored to identify effects of immunotoxicant exposure on antigen uptake, processing or presentation by the Ch27 cells and on antigen recognition and IL-2 production and secretion by the 3A9 cells. IL-2 production was reduced in response to treatment with well-known immunotoxicants cyclosporin A (CYA), dexamethasone (DEX), azathioprine (AZPR), methotrexate (MOT) and benzo(a)pyrene (BAP) but was not affected by treatment with cyclophosphamide (CYPH). A negative control compound mannitol (MANN) altered neither cell viability nor IL-2 levels whereas the lysosomotrophic compound ammonium chloride (AMCL) reduced IL-2 production. This novel in vitro assay of immune function may be suitable for integration into a tiered testing battery for screening and prioritization of potential immunosuppressants.

Lung function changes in mice sensitized to ammonium hexachloroplatinate.

Occupational exposure to halogenated platinum salts can trigger the development of asthma. The risk to the general population that may result from the use of platinum in catalytic converters and its emerging use as a diesel fuel additive is unclear. To investigate pulmonary responses to platinum, we developed a mouse model of platinum hypersensitivity. Mice were sensitized through application of ammonium hexachloroplatinate (AHCP) to the shaved back on days 0, 5 and 19, and to each ear on days 10, 11 and 12. On days 24 and 29, mice were challenged by oropharyngeal aspiration with AHCP in saline. Before and immediately after challenge, pulmonary responses were assessed using whole body plethysmography (WBP). A dose-dependent increase in immediate responses was observed in AHCP-sensitized and challenged mice. On days 26 and 31, changes in ventilatory responses to methacholine (Mch) aerosol were assessed by WBP; dose-dependent increases in Mch responsiveness occurred in sensitized mice. Lymph node cell counts indicate a proliferative response in lymph nodes draining the sites of application. Bronchoalveolar lavage fluid harvested from sensitized mice contained an average of 5% eosinophils compared to less than 0.5% in non-sensitized mice (p < 0.05); significant increases in total serum immunoglobulin E were observed for all sensitized mice. Although a second airway challenge on day 29 affected some results, only one airway challenge was needed to observe changes in lung function.

Development and utilization of an ex vivo bromodeoxyuridine local lymph node assay protocol for assessing potential chemical sensitizers.

The murine local lymph node assay (LLNA) is widely used to identify chemicals that may cause allergic contact dermatitis. Exposure to a dermal sensitizer results in proliferation of local lymph node T cells, which has traditionally been measured by in vivo incorporation of [(3) H]methyl thymidine. A more recent non-isotopic variation of the assay utilizes bromodeoxyuridine (BrdU) incorporation in vivo. To further improve the utility of this assay, we developed an ex vivo BrdU labeling procedure eliminating the need for in vivo injections. The results of this assay correctly identified a strong sensitizer (i.e., trimellitic anhydride) as well as weak/moderate sensitizers (i.e., eugenol, cinnamaldehyde and hexylcinnaminic aldehyde). As anticipated, neither non-sensitizers isopropanol and lactic acid nor the false negative chemical nickel II sulfate hexahydrate induced a positive threshold response in the assay. The results of this assay are in close agreement with those of the in vivo LLNA:BrdU-enzyme-linked immunosorbent assay labeling procedure. We also used the ex vivo BrdU LLNA procedure to evaluate ammonium hexachloroplatinate, ammonium tetrachloroplatinate and cis-diamminedichloroplatinum(II) and the assay correctly identified them as sensitizers based on the calculation of EC2 values. We conclude that this ex vivo BrdU labeling method offers predictive capacity comparable to previously established LLNA protocols while eliminating animal injections and the use of radioisotope. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

Small molecule disruption of G protein beta gamma subunit signaling inhibits neutrophil chemotaxis and inflammation.

G protein betagamma subunit-dependent signaling is important for chemoattractant-dependent leukocyte chemotaxis. Selective small molecule targeting of phosphoinositide 3-kinase (PI3-kinase) gamma catalytic activity is a target of interest for anti-inflammatory pharmaceutical development. In this study, we examined whether small-molecule inhibition of Gbetagamma-dependent signaling, including Gbetagamma-dependent activation of PI3-kinase gamma and Rac1, could inhibit chemoattractant-dependent neutrophil migration in vitro and inflammation in vivo. Small-molecule Gbetagamma inhibitors suppressed fMLP-stimulated Rac activation, superoxide production, and PI3-kinase activation in differentiated HL60 cells. These compounds also blocked fMLP-dependent chemotaxis in HL60 cells and primary human neutrophils. Systemic administration inhibited paw edema and neutrophil infiltration in a mouse carrageenan-induced paw edema model. Overall, the data demonstrate that targeting Gbetagamma-regulation may be an effective anti-inflammation strategy.

Integrated enterprise management: a look at the functions, the enterprise, and the environment--can you see the difference?

The performance of an organization is paced by its use of resources, including its ability to acquire, access, and use knowledge. A high-performance organization, more than likely, has structured its resources around process linkages and is characterized by a horizontal organization chart, teams and teamwork, empowerment, and operational excellence. Organizational researchers hypothesize that performance improves with fuzzy internal boundaries, cross-functional participation, and goals anchored in the interests of customers and other external stakeholders. This article looks at the competitive need for more integration of resources and greater sharing of knowledge, the integrated nature of work within emerging types of organizations, how expanded views can improve the marketplace centering of processes and individuals, and how combining a model of integration and individual work challenges thinking and actions in the new environment.