Testing MACRO (version 5.1) for pesticide leaching in a Dutch clay soil

Testing of pesticide leaching models against comprehensive field-scale measurements is necessary to increase confidence in their predictive ability when used as regulatory tools. Version 5.1 of the MACRO model was tested against measurements of water flow and the behaviour of bromide, bentazone [3-isopropyl-1H-2,1,3-benzothiadiazin-4(3H)-one-2,2-dioxide] and imidacloprid [1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine] in a cracked clay soil. In keeping with EU (FOCUS) procedures, the model was first calibrated against the measured moisture profiles and bromide concentrations in soil and in drain water. Uncalibrated pesticide simulations based on laboratory measurements of sorption and degradation were then compared with field data on the leaching of bentazone and imidacloprid. Calibrated parameter values indicated that a high degree of physical non-equilibrium (i.e. strong macropore flow) was necessary to describe solute transport in this soil. Comparison of measured and simulated bentazone concentration profiles revealed that the bulk of the bentazone movement in this soil was underestimated by MACRO. Nevertheless, the model simulated the dynamics of the bentazone breakthrough in drain water rather well and, in particular, accurately simulated the timing and the concentration level of the early bentazone breakthrough in drain water. The imidacloprid concentration profiles and its persistence in soil were simulated well. Moreover, the timing of the early imidacloprid breakthrough in the drain water was simulated well, although the simulated concentrations were about 2-3 times larger than measured. Deep groundwater concentrations for all substances were underestimated by MACRO, although it simulated concentrations in the shallow groundwater reasonably well. It is concluded that, in the context of ecotoxicological risk assessments for surface water, MACRO can give reasonably good simulations of pesticide concentrations in water draining from cracking clay soils, but that prior calibration against hydrologic and tracer data is desirable to reduce uncertainty and improve accuracy.     

Characterization of transport Na+-ATPases in gills of freshwater tilapia

Branchial plasma membranes from the freshwater cichlid teleostOreochromis mossambicus (tilapia) contain two Na⁺-dependent ATPases: Na⁺/K⁺ ATPase, and an amiloride-sensitive ATPase which is postulated to operate as a Na⁺/H⁺ (–NH₄ ⁺) ATPase. It is suggested that both enzyme activities are located in the basolateral membrane system of the chloride cells. K⁺ has opposing effects on the two enzymes: it stimulates Na⁺/K⁺ ATPase and inhibits Na⁺/H⁺ (–NH₄ ⁺) ATPase activity. Na⁺/H⁺ ATPase appears more sensitive to NH₄ ⁺ at low concentrations than Na⁺/K⁺ ATPase and the stimulatory effect by NH₄ ⁺ ions on the first enzyme could be important in facilitating NH₄ ⁺ excretion by tilapia gills under physiological conditions.In vitro maximum stimulation by NH₄ ⁺ is similar for the two enzymes (200%). In contrast to Na⁺/K⁺ ATPase, Na⁺/H⁺ ATPase activity is inhibited by supra-physiological (>20 mM) concentrations of NH₄ ⁺.     

Soil fertility distributions in long-term no-till, chisel/disk and moldboard plow/disk systems

In permanent no-till (NT), soil nutrients are no longer mixed into the topsoil as with moldboard plow/disking (MD), whereas chisel/disking (CD) does limited mixing. Surface broadcast and/or banded nutrient applications may result in high and low fertility zones in permanent NT, with possible implications for soil sampling and nutrient placement. We investigated effects of 25 years of continuous NT, CD and MD with corn planted in the same row locations on organic matter (SOM), pH-H₂O and Mehlich-3 extractable phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg). Vertical distribution at 0–5, 5–10 and 10–15 cm depths was measured as well as horizontal distributions across corn rows. We observed higher SOM and P in NT and CD than in MD in the 0–15 cm layer. SOM content was greatest in the top 5 cm in NT, but declined sharply with depth. SOM content in CD was not as high at the surface as in NT, but did not decline as fast as in NT. SOM was uniform but low throughout the 0–15 cm depth of MD. In all tillage systems, SOM did not vary across rows. Soil pH was higher in the 0–5 cm layer of NT than the deeper layers but the reverse was true in the CD or MD treatments. Concentrations of P, K and Ca were higher in the surface 0–5 cm than 10–15 cm depth of all tillage systems, but most strikingly in NT and CD. Starter fertilizer injection resulted in higher P and lower pH in the injection zone of all tillage treatments, but most notably in NT. The pH was depressed under the band of side-dressed nitrogen with all tillage systems. Potassium accumulated in the rows of the previous crop, probably because it leached from crop residue that accumulated there. Tillage did not affect Mg distribution. Optimal nutrient management in NT should take account of horizontal and vertical nutrient and pH distributions. Samples in long-term NT could potentially be taken to a shallower depth if calibration curves are available. To avoid underestimating P and K availability or overestimate lime needs, high P or decreased pH bands should be avoided, as well as crop rows. Possibilities to reduce P and K applications with banding need more investigation. Results show the importance of regular liming in NT to maintain surface pH in the optimum range, but also show that lime does not have to be incorporated.     

Branchial calcium uptake: possible mechanisms of control by stanniocalcin

The branchial Ca²⁺ uptake by teleost fish is under inhibitory control by the hormone stanniocalcin (STC) which is generated by the corpuscles of Stannius (CS). Removal of the CS in North American eel, Anguilla rostrata LeSueur, induced a rapid rise in blood calcium levels. Branchial Ca²⁺ influx following the extirpation of the CS (stanniectomy, STX) increased during the first four days and stayed elevated thereafter (in agreement with previous studies). The transepithelial potential (TEP) across the gills did not change after STX and this means that the electrochemical gradient for Ca²⁺ is less favourable for passive influx of Ca²⁺ in STX eel. Therefore, the Ca²⁺ influx in STX eels is a transcellular flux, with Ca²⁺ crossing the apical and basolateral membrane barrier. The kinetics of ATP-driven Ca²⁺-transport across basolateral plasma membranes from eel gills did not change after STX. Thus, the increased Ca²⁺-influx after STX is not correlated with changes in ATP-dependent Ca²⁺-extrusion across the basolateral membrane, suggesting a regulation at the apical membrane. Moreover, STC did not affect ATP-driven Ca²⁺-transport in isolated basolateral membranes (in vitro). STC (0.1 nM) reduced cAMP levels in dispersed eel gill cells. It had no significant effect on the IP₃ levels in these cells. We postulate that STC controls the permeability to Ca²⁺ of the apical membranes of the Ca²⁺ transporting cells of fish gills by controlling second messenger operated Ca²⁺ channels in the apical membrane.

---

Résumé L'entrée de calcium au niveau des branchies est sous le controle inhibiteur de la stanniocalcine (STC) qui est synthétisée au niveau des corpuscules de Stannius (CS). L'ablation des CS chez l'anguille d'Amérique du Nord, Anguilla rostrata LeSueur, induit une augmentation rapide des niveaux de calcium dans le sang. Le flux entrant branchial de calcium consécutif à l'ablation des CS (stanniectomie, STX) augmente pendant les 4 premiers jours et reste élevé au-delà (en accord avec des études antérieures). Le potentiel transépithélial (TEP) à travers les branchies ne change pas après STX, ceci indiquant que le gradient électrochimique du Ca²⁺ est moins favorable pour le flux entrant passif du Ca²⁺ chez l'anguille STX. En conséquence, le flux entrant de Ca²⁺ chez l'anguille STX est un flux transcellulaire, avec le Ca²⁺ traversant la barrière membranaire apicale et basolatérale. La cinétique du transport de Ca²⁺ conduit par l'ATP à travers les membranes plasmatiques basolatérales de branches d'anguille n'est pas modifiée après STX. Ainsi, l'augmentation du flux entrant de Ca²⁺ après STX n'est pas corrlée avec des modifications de l'excrétion de Ca²⁺ conduit par l'ATP à travers la membrane basolatérale, suggérant donc une régulation au niveau de la membrane apicale. De plus, la STC ne modifie pas le transport de Ca²⁺ conduit par l'ATP dans des membranes basolatérales isolées (in vitro). La STC (0.1 nM) réduit les niveaux d'AMPc dans des cellules dispersées de branchies d'anguille. Cette hormone n'a pas d'effet significatif sur les niveaux d'IP₃ dans ces cellules. Nous suggérons que la STC régule la perméabilité au Ca²⁺ des membranes apicales des cellules branchiales transporteuses de Ca²⁺ en controlant un second messager agissant sur les canaux calciques de la membrane apicale.

     

Cloning and sequence analysis of hypothalamus cDNA encoding tilapia melanin-concentrating hormone

Melanin-concentrating hormone (MCH) is a neuroendocrine peptide involved in the regulation of skin pigmentation in teleosts. We isolated and sequenced a 543 bp hypothalamic cDNA encoding the MCH-preprohormone of tilapia (Oreochromis mossambicus). Initially, polymerase chain reaction (PCR) experiments were performed on hypothalamic RNA with a synthetic oligonucleotide primer corresponding to a conserved region of salmon and mammalian MCH peptide and an oligo dT primer. A 0.2 kb PCR fragment was obtained and found to have low but significant nucleotide sequence similarity with the 3′ ends of known MCH-mRNAs. Subsequently, the PCR fragment was used to screen λZAP cDNA libraries constructed from tilapia hypothalamic poly(A⁺) RNA. The cloned tilapia MCH preprohormone cDNA encodes a 133-amino acid protein of which 17 amino acids belong to the signal peptide. The MCH peptide sequence is located at the carboxy terminus of the preprohormone structure and is preceded by a pair of arginine residues which can serve as a proteolytic cleavage site. 23 to 25 amino acids further upstream in the prohormone structure three consecutive basic residues are present. Cleavage at this site would yield a 22-amino acid MCH gene-related peptide (Mgrp), which is much larger than (12− to 13-amino acid) salmon and mammalian Mgrp. A comparative structural analysis between tilapia preproMCH and its salmon and mammalian counterparts revealed that the MCH peptide sequence is very well conserved (100% identity with salmon and 75% identity with both rat and human MCH). In contrast, the remaining parts of the preproMCH structures have diverged considerably. Northern blot analysis revealed the presence of tilapia preproMCH mRNA in the hypothalamus and not in other brain regions nor in several peripheral tissues.

---

Résumé La MCH (melanin-concentrating hormone) est un peptide neuroendocrinien impliqué dans la régulation de la pigmentation de la peau chez les Téléostéen. Nous avons isolé et séquencé un ADNC hypothalamique de 543 pairs de bases codant pour la préprohormone de la MCH de tilapia (Oreochromis mossambicus). Ce travail a débuté par des expériences de PCR (Polymerase Chain Reaction) qui ont été réalisées sur des ARNm hypothalamiques avec comme amorces un oligonucléotide synthétique correspondant à une région conservée du peptide MCH de saumon et de mammifère et un oligo dT. Un fragment de 0.2 kb a ainsi été obtenu par PCR et l'analyse de sa séquence nucléotidique montre une similitude faible mais significative avec les extrémités 3′ des ARNms de MCH connus. Ensuite ce fragment a été utilisé pour cribler des banques de cDNA construites dans λZAP à partir d'ARN poly (A⁺) d'hypothalamus de tilapia. L'ADNc de la préprohormone de MCH de tilapia code pour une proteine de 133 acides aminés dont 17 constituent le peptide signal. Le peptide de la MCH est localisé dans l'extrémité carboxy de la préprohormone et se trouve précédé par une pair de résidus arginine qui servent de site de coupure protéolitique. De plus, 3 résidus basiques consécutifs sont présents à 23–25 acides aminés en amont de la structure de la prohormone. La coupure à ce site permettrait d'obtenir un peptide associé au gène de la MCH (Mgrp) de 22 acides aminés, ce qui est beaucoup plus grand (de 12 à 13 acides aminés) que les Mgrp de saumon ou de mammifères. Une analyse structurale comparée entre la preproMCH de tilapia et la preproMCH de saumon et de mammifères montre que la séquence du peptide MCH est très conservée (100% d'identité avec le saumon et 75% d'identité avec la MCH humaine ou de rat). Par contre, les autres parties de la structure de la preproMCH ont divergé considérablement. L'analyse par “Northern Blot” met en évidence la présence d'ARNm de preproMCH dans l'hypothalamus de tilapia mais pas dans les autres régions du cerveau ni dans plusieurs tissus périphériques.

     

A dog with alveolar echinococcosis: the larval stage of the fox tapeworm

In this report we describe the first case of alveolar echinococcosis and leukemic malignant T-cell lymphoma in a dog presented to a general practice in the Netherlands. The owner and the dog moved from Switzerland to The Netherlands a few weeks before. The clinical aspects, diagnostics, pathology, therapy and public health aspects of this severe parasitic zoonosis are described.     

Some physicochemical and environmental factors affecting transformation rates and sorption of the herbicide metamitron in soil

In addition to the molecular structure of a pesticide, environmental conditions may influence its persistence through their effect on the growth and activity of pesticide-degrading micro-organisms. As a result, transformation rates may decrease rapidly when a compound is leached into subsoil. Metamitron sorption isotherms were determined and incubation series were set up for a sandy loam soil, simulating single and combination effects that occur during transport of metamitron into subsoils. K_OC values increased with increasing depth from 185 to 700 litre kg⁻¹. A combination of conditions that are unfavourable for microbial activity, such as low temperature (5°C), low concentrations (0·5 mg kg⁻¹) and a large sorbed fraction (K_OC = 700) resulted in half-lives of over one year. Oxygen inhibition decreased the transformation rate of metamitron from 0·058 to 0·019 day⁻¹. In order of significance, the transformation of metamitron appears to be a function of temperature, oxygen availability and sorption to organic carbon. Increasing doses did not change transformation rates significantly, although different transformation pathways were observed.     

Uptake,distribution and excretion of magnesium inOreochromis mossambicus: dependence on magnesium in diet and water

The euryhaline Mozambique tilapia (Oreochromis mossambicus) shows a more marked ability to adapt to low magnesium levels in food and water than many other fish species. Nonetheless, the internal distribution of magnesium is altered under low-magnesium conditions. The amount of magnesium in scales, and to a lesser extent the vertebral bone, is reduced and hence serve as magnesium reservoirs. The magnesium concentration of muscle is only marginally reduced by low external magnesium, suggesting that magnesium is partitioned to ensure normal muscle functioning. The water magnesium is of vital importance, as exposure to low-magnesium water markedly deminished the ability of tilapia to adapt to low-magnesium feeding. However, magnesium intake from the water, eithervia the integument or drinking, does not increase in lowmagnesium fed fish, despite an increased opercular chloride cell density. The growth related magnesium accumulation of tilapia under low-magnesium conditions approximates the total intake of the element (from the food and from the water), indicating that magnesium losses are minimized and that the magnesium absorption from the gastrointestinal tract may be highly efficient and very important.