Spatially resolved quantification of agrochemicals on plant surfaces using energy dispersive X‐ray microanalysis

BACKGROUND: In the present study the principle of energy dispersive X‐ray microanalysis (EDX), i.e. the detection of elements based on their characteristic X‐rays, was used to localise and quantify organic and inorganic pesticides on enzymatically isolated fruit cuticles. Pesticides could be discriminated from the plant surface because of their distinctive elemental composition. RESULTS: Findings confirm the close relation between net intensity (NI) and area covered by the active ingredient (AI area). Using wide and narrow concentration ranges of glyphosate and glufosinate, respectively, results showed that quantification of AI requires the selection of appropriate regression equations while considering NI, peak‐to‐background (P/B) ratio, and AI area. The use of selected internal standards (ISs) such as Ca(NO₃)₂ improved the accuracy of the quantification slightly but led to the formation of particular, non‐typical microstructured deposits. The suitability of SEM‐EDX as a general technique to quantify pesticides was evaluated additionally on 14 agrochemicals applied at diluted or regular concentration. CONCLUSION: Among the pesticides tested, spatial localisation and quantification of AI amount could be done for inorganic copper and sulfur as well for the organic agrochemicals glyphosate, glufosinate, bromoxynil and mancozeb. Copyright © 2009 Society of Chemical Industry     

Hygroscopic salts support the stomatal penetration of glyphosate and influence its biological efficacy

The objective of this study was to better understand the role of chaotropic and kosmotropic salts in stomatal uptake, a process that occurs in parallel with cuticular penetration and affects the bioefficacy of glyphosate. In Trial 1, salt solutions of kosmotropic (NH₄)₂SO₄ and chaotropic NaClO₃, with or without the organosilicone surfactant, Break‐Thru® S233 (BT), and with or without glyphosate, were prepared. In Trial 2, sodium salts with a kosmotropic‐to‐chaotropic nature (Na₂SO₄, NaCl, NaNO₃ or NaClO₃) and BT were prepared with or without glyphosate. At very high concentrations, the salts of a kosmotropic nature induced a slight decrease in the surface tension, whereas those of a chaotropic nature triggered a significant decrease in the surface tension. Solution droplets were deposited onto the leaves of Viola arvensis and Chenopodium album. The dried deposits were analyzed with an environmental scanning electron microscope. The salts formed structures linking the exterior to the interior of the stomata in the crystalline ([NH₄]₂SO₄) or amorphous (NaClO₃) form. However, the low surface tension alone did not appear to be the unique driving force for stomatal uptake. With glyphosate, amorphous globular structures of glyphosate salt were formed in the Na₂SO₄ (kosmotropic) solution. Differently, the salts of a rather chaotropic nature (NaNO₃ or NaClO₃) formed a colloidal matrix. In general, salt residues were observed over the periclinal cell walls and inside the stomatal chamber. Neither (NH₄)₂SO₄ nor Na₂SO₄ had a significant impact on glyphosate bioefficacy. However, the chaotropic salt, NaNO₃, enhanced the bioefficacy of glyphosate the most, with a performance that was comparable to that of the solution glyphosate + organosilicone surfactant.     

Einfluß von Mg- und Ca-Mangel auf Mg- und Ca-Gehalte,Chlorophyllgehalt und Chlorophyllfluoreszenz von Fichtennadeln sowie auf die Mikromorphologie und Benetzbarkeit ihrer epicuticulären Wachse

The influence of Mg- and Ca-deficiency on the micromorphology of epicuticular waxes and on the wettability of spruce needles The influence of Mg- and Ca-deficiency in nutrient solution on epicuticular waxes of needles of Norway spruce (Picea abies [L.] Karst.) was investigated in greenhouse experiments. Spruce clones were cultivated in sand and supplied with nutrient solution. For two vegetation periods, plants of the treatment groups -Mg, -Ca and -Mg/-Ca were provided with only 15 % of the respective mineral element of the control. At the end of the second vegetation period, the nutrient deficiency was documented by reduced Mg- and/or Ca-content of the needles and changes in chlorophyll fluorescence. No effect on micromorphology of epicuticular waxes and wettability of needles of any age was observed following deficient Mg and/or Ca supply.     

Host response to the chondracanthid copepod Chondracanthus goldsmidi,a gill parasite of the striped trumpeter,Latris lineata (Forster), in Tasmania

The chondracanthid copepod, Chondracanthus goldsmidi is an ectoparasite of gills, inner opercula and nasal cavities of cultured striped trumpeter, Latris lineata (Forster). Whilst often present in high numbers (up to 60 parasites per host), little is known about its effect on striped trumpeter. In this study C. goldsmidi was associated with extensive epithelial hyperplasia and necrosis. Pathological changes were most pronounced near the parasite’s attachment site, with papilloma‐like growths surrounding the entire parasite resulting in deformation of the filament. The number of mucous cells increased near the parasite attachment sites on both the opercula and gills. Mast cells were absent in healthy gills; in contrast numerous mast cells were identified in the papilloma‐like growths. Immunostaining identified piscidin‐positive mast cells in the papilloma‐like growths, presenting the first evidence of piscidin in the family Latridae.     

Histopathology of culture‐associated skin erosions and papillary hyperplasia of Atlantic halibut,Hippoglossus hippoglossus (L.)

Atlantic halibut, Hippoglossus hippoglossus (L.), that were cultured in tanks with a smooth bottom (gel‐coated fibreglass) substrate developed papillary hyperplasia and skin erosions on the blind (ventral) side. No viruses, bacteria or parasites were observed in any sections of affected skin. Comparison of microscopic pathology with that of skin from normal, wild‐caught halibut showed severe epidermal proliferation with foci of severe mucous cell hyperplasia. Both epidermal thickness and mucous cell density were significantly greater in fish held on a smooth substrate compared to skin of healthy fish. Spongiosis was present, especially at the base of the papillary, hyperplastic epidermis, and there was a chronic inflammatory infiltrate in the scale pockets composed of lymphocytes, histiocytes and erythrocytes. Skin erosions had various degrees of epidermal loss, in some cases to the basement membrane. A predisposing factor for the epidermal sloughing may have been related to the spongiosis, and the vacuolated, degenerated basal cells. When affected halibut were cultured for an additional 28, 62 or 97 days on a sand substrate, which is conducive to skin lesion healing, there was no apparent change in epidermal thickness over time. However, mucous cell density significantly increased from day 0 to 97 during the healing process.     

Antiparasitic activity of the antimicrobial peptide HbβP‐1, a member of the β‐haemoglobin peptide family

A family of antimicrobial peptides (AMPs) derived from the β‐subunit of haemoglobin was recently isolated from channel catfish, Ictalurus punctatus, infected with Ichthyophthirius multifiliis (ich), an important freshwater fish parasite that causes ichthyophthiriosis. We previously discovered that one of these AMPs, HbβP‐1, had strong cidal activity against ich as well as another ectoparasite, Tetrahymena pyriformis. HbβP‐1 toxicity was specific, primarily affecting the trophozoite (trophont) stage of ich. Here, we show that HbβP‐1 acts more rapidly to kill smaller (presumably less mature) trophonts of ich, taking almost twice as long to kill larger trophonts (P < 0.0001). It acts more rapidly than an unrelated AMP, piscidin 1, which is haemolytic and also lethal to ich trophonts. HbβP‐1 is potently and selectively lethal to the trophont stage of the dinoflagellate ectoparasite, Amyloodinium ocellatum, one of the most important pathogens of warmwater marine fish. HbβP‐1 has no effect on the fish gill cell line feeder layer (G1B cells) used to propagate Amyloodinium, further suggesting a highly selective action. These findings suggest that HbβP‐1 or related AMPs might function in protecting marine as well as freshwater fish and that HbβP‐1 has highly selective activity against specific life stages of important fish ectoparasites.     

Activity of the antimicrobial polypeptide piscidin 2 against fish ectoparasites

Abstract The antiparasitic effects of piscidin 2, an antimicrobial polypeptide (AMPP) first isolated from mast cells of hybrid striped bass, were tested against three protistan ectoparasites of marine fish (the ciliates Cryptocaryon irritans and Trichodina sp., and the dinoflagellate Amyloodinium ocellatum) and one ciliate ectoparasite of freshwater fish (Ichthyophthirius multifiliis). I. multifiliis was the most susceptible parasite, with all theronts killed at 6.3 microg mL(-1) piscidin 2. The most resistant parasite was Trichodina, where a few cells were killed at 12.5 microg mL(-1), but several were still alive even at 100 microg mL(-1). C. irritans was of intermediate sensitivity, with some theronts killed at 12.5 microg mL(-1) and all killed at 25 microg mL(-1). High parasite density apparently exhausted the piscidin 2 before it could attain its maximal effect, but surviving parasites were often visibly damaged. The lower efficacy of piscidin 2 against marine parasites compared with the freshwater ciliate might be related to the inhibitory effects of high sea water cation levels. The tissue concentration of piscidins estimated in healthy hybrid striped bass gill (40 microg mL(-1)) suggests that piscidin 2 is lethal to the parasites tested at physiological concentrations and is thus an important component of innate defence in fish expressing this type of AMPP.