Residues of diflubenzuron on horse chestnut (Aesculus hippocastanum) leaves and their efficacy against the horse chestnut leafminer, Cameraria ohridella

Residues of the insect growth regulator diflubenzuron were quantified on horse chestnut (Aesculus hippocastanum L.) leaves treated with a diflubenzuron 480 g litre^?1 SC, Dimilin. To analyse the samples, an analytical procedure was developed involving a simple extraction step followed by high‐performance liquid chromatography on an octadecyl‐modified silica column with methanol + 0.01 M ammonium acetate mobile phase. The results showed diflubenzuron to be highly stable on horse chestnut leaves; more than 4 months (127 days) after application, 38% (on average) of the insecticide still remained on/in the leaves. The data confirmed biological observations showing diflubenzuron's long‐term efficacy against the horse chestnut leafminer, Cameraria ohridella Deschka & Dimi?, which is the most important pest of the horse chestnut in Europe. The hypothesis of possible penetration of diflubenzuron into the leaf mass is explored and discussed. Copyright © 2006 Society of Chemical Industry     

Ultrastructure of the secondary osmoregulatory canals in the scolex and neck region of Silurotaenia siluri (Batsch, 1786) (Cestoda: Proteocephalidae)

The secondary osmoregulatory canals in the scolex and neck region of Silurotaenia siluri, a parasite of the catfish Silurus glanis (L.), terminate below the tegument basal plasma membrane. The basal plasma membrane of the osmoregulatory canal syncytium is in tight contact with the tegument basal plasma membrane.     

A rapid real-time PCR assay for detecting columnar growth in apples (Malus × domestica)

Columnar growth in apples can have considerable positive consequences for orchard maintenance, but the lack of columnar cultivars producing fruits with desired market quality prevents their broader utilization in orchards. New crossings focused on their breeding are therefore conducted throughout the world every year. Unfortunately, the reliable discrimination between columnar growth and normal phenotype is difficult until seedlings are 2–3 years of age. However, molecular marker–assisted selection (MAS) can be used in seedlings several weeks old providing immense help in breeding programmes focused on columnar apple production. Here, we describe a rapid and reliable method for the detection of both wild-type (WT) and columnar (Co) alleles originating from ‘McIntosh Wijcik’ in a single reaction using a real-time PCR allelic discrimination approach. Our assay was tested in 130 genotypes (80 normal and 50 columnar habit) from various crossings and their parents, and in all cases, genotype corresponded to phenotype. The assay is thus suitable for laboratories interested in high-throughput MAS for columnar tree habit breeding.     

Absorption of proinflammatory cytokines from inflamed porcine uterus into the uterine venous blood--preliminary data

The aim of the present study was to estimate the absorption of 125I-labeled proinflammatory cytokines--interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) from inflamed porcine uterus into the uterine venous blood. Moreover, in order to test the hypothesis that the above cytokines penetrate directly into ovaries and oviduct via local destination transfer in the area of the ovarian vascular pedicle and bypassing the systemic circulation, the concentration of IL-1beta, IL-6 and TNF-alpha in ovarian and oviductal tissues was also studied. These cytokine concentrations were also estimated in the ovarian venous blood. IL-1beta, IL-6 and TNF-alpha from both control and inflamed uteri were absorbed into the uterine venous blood, but it was higher (P < 0.05-0.001) from the pathologically changed uteri. The uterine tissues, particularly the endometrium, of both control and inflamed uteri retained all studied cytokines, but to a higher degree (P < 0.001) in the inflamed uteri. Injections of IL-1beta, IL-6 and TNF-alpha into the control and inflammatory changed uteri produced the presence of these proteins in the ovary and oviduct. However, the concentrations of IL-1beta and IL-6 in the ovarian and oviductal tissues was low after injections of control and inflamed uteri with these cytokines. In turn, administration of TNF-alpha into the inflammatory changed uteri lead to an enhancement in the concentration of this cytokine in the ovarian parenchyma (P < 0.05) and oviduct (P < 0.001). All studied cytokines were found in the ovarian venous blood after their injection into both control and inflamed uteri, which indicated its local destination transfer to the ovary. However, the concentration of cytokines increased (P <0.05-0.001) in the gilts with pathologically changed uteri as compared to controls. The study showed that both control and inflamed porcine uteri absorbed IL-1beta, IL-6 and TNF-alpha into the uterine venous blood, but the values of absorbed cytokines from inflamed uteri were higher. Moreover, the quantity and the manner of the studied cytokineS absorption into the uterine venous blood differed.     

Comparison of methods to reduce dioxin and polychlorinated biphenyls contents in fishmeal: extraction and enzymatic treatments

Methods to remove dioxins and polychlorinated biphenyls (PCBs) from fishmeal were investigated and compared. The tested methods include (i) lowering the fat content and simultaneously the level of toxic contaminants using either organic solvents or (ii) lowering the fat content using protease and (iii) removal of dioxins and PCBs using either oil extraction or (iv) breakdown of dioxin and PCBs using oxidoreductase. The results showed that the organic solvents tested (ethanol, isopropanol, and isohexane) were efficiently lowering the oil content of the fishmeal by 80%. However, the treated fishmeal has a low fat content and may contain traces of solvent. The protease alcalase was not as efficient as the solvent extraction and only removed approximately 30% of the oil but presented the advantage of being a mild process. Other proteases, alone or in combination with other enzymes, might give better yield if the reaction conditions are optimized. In contrast, extraction of dioxin and PCBs using olive oil or fish oil was very effective and resulted in 60-75% extraction of dioxin and PCBs, respectively, after a single extraction step. No preference for the oil type was observed. This method is very simple and quick and does not require an important investment for the fishmeal producer. It is expected that with optimization this method could be implemented at an industrial scale without too many difficulties. In contrast, the oxidoreductases tested did not result in a major degradation of dioxins and PCBs with only 10-15% degradation achieved. However, with the recent advancement in biotechnology, it is possible that future research will result in the development of enzymes that effectively degrade recalcitrant contaminants.     

Detection and quantification of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis> in irrigation waters

A quantitative RT real-time PCR method was developed for the detection and quantification of Tomato mosaic virus (ToMV) in irrigation waters. These have rarely been monitored for the presence of plant pathogenic viruses, mostly due to the lack of efficient and sensitive detection methods. The newly developed method presented here offers a novel approach in monitoring the health status of environmental waters. ToMV was reliably detected at as low as 12 viral particles per real-time PCR reaction, which corresponds to the initial concentration of approximately 4.2 × 10^?10 mg (6,300 viral particles) of ToMV per ml of sample. The sensitivity of the method was further improved by including the Convective Interaction Media^® (CIM) monolithic chromatographic columns for quick and efficient concentration of original water samples. Seven out of nine water sources from different locations in Slovenia tested positive for ToMV, after concentrating the sample. Four samples tested ToMV-positive without the concentrating procedure. The presence and integrity of infective ToMV particles in the original sample, as well as in the chromatographic fraction, was confirmed using different methods from test plants, DAS ELISA to electron microscopy and real-time PCR. In this study, we propose a unique and simple diagnostic scheme for rapid, efficient, and sensitive monitoring of irrigation waters that could also be adopted for other plant, human or animal viruses.     

Atypical BSE in Germany--proof of transmissibility and biochemical characterization

Intensive active surveillance has uncovered two atypical German BSE cases in older cattle which resemble the two different atypical BSE phenotypes that have recently been described in France (designated H-type) and Italy (designated L-type or BASE). The H-type is characterized by a significantly higher molecular size, but a conventional glycopattern of the proteinase K treated abnormal prion protein (PrP(Sc)), while the L-type PrP(Sc) has only a slightly lower molecular size and a distinctly different glycopattern. In this paper we describe the successful transmission of both German atypical BSE cases to transgenic mice overexpressing bovine PrP(C). Upon challenge with the L-type, these mice developed BSE after a substantially shorter incubation period than any classical BSE transmission using these mice to date. In contrast, the incubation period was distinctly prolonged when these mice were challenged with the H-type. PrP(Sc) accumulated in the brains of these mice were of the same atypical BSE type that had been used for the transmission. These atypical cases suggest the possible existence of sporadic BSE cases in bovines. It is thus feasible that the BSE epidemic in the UK could have also been initiated by an intraspecies transmission from a sporadic BSE case.