Highly efficient preparation of lipophilic hydroxycinnamates by solvent-free lipase-catalyzed transesterification

Various medium- or long-chain alkyl cinnamates and hydroxycinnamates, including oleyl p-coumarate as well as palmityl and oleyl ferulates, were prepared in high yield by lipase-catalyzed transesterification of an equimolar mixture of a short-chain alkyl cinnamate and a fatty alcohol such as lauryl, palmityl, and oleyl alcohol under partial vacuum at moderate temperature in the absence of solvents and drying agents in direct contact with the reaction mixture. Immobilized lipase B from Candida antarctica was the most effective biocatalyst for the various transesterification reactions. Transesterification activity of this enzyme was up to 56-fold higher than esterification activity for the preparation of medium- and long-chain alkyl ferulates. The relative transesterification activities found for C. antarctica lipase were of the following order: hydrocinnamate > cinnamate > 4-hydroxyhydrocinnamate > 3-methoxycinnamate > 2-methoxycinnamate approximately 4-methoxycinnamate approximately 3-hydroxycinnamate > hydrocaffeate approximately 4-hydroxycinnamate > ferulate > 2-hydroxycinnamate > caffeate approximately sinapate. With respect to the position of the hydroxy substituents at the phenyl moiety, the transesterification activity of C. antarctica lipase B increased in the order meta > para > ortho. The immobilized lipases from Rhizomucor miehei and Thermomyces lanuginosus demonstrated moderate and low transesterification activity, respectively. Compounds with inverse chemical structure, that is, 3-phenylpropyl alkanoates such as 3-(4-hydroxyphenyl)propyl oleate and 3-(3,4-dimethoxyphenyl)propyl oleate, were obtained by C. antarctica lipase-catalyzed transesterification of fatty acid methyl esters with the corresponding 3-phenylpropan-1-ols in high yield, as well.     

Primary infundibular stenosis and pedigree analysis in three Golden Retriever littermates

Three eight-week-old Golden Retriever puppy littermates were evaluated because of left basilar systolic murmurs and were diagnosed with primary infundibular stenosis. Pedigree analysis in this line was also performed to identify a mode of inheritance. All dogs were asymptomatic at the time of diagnosis; two of the three had congenital lesions in addition to primary infundibular stenosis. Two additional affected dogs were identified in the line, and pedigree analysis suggested an autosomal recessive mode of inheritance. Another, unrelated golden retriever was also identified with isolated infundibular stenosis in the record database. Primary infundibular stenosis should be considered in the differential diagnoses for golden retriever dogs with a left basilar systolic murmur, and is often associated with complex congenital cardiac disease. Primary infundibular stenosis may worsen in severity with time, and in this line of dogs an autosomal recessive pattern of inheritance is likely.     

Evaluation of sodium percarbonate as a bath treatment for amoebic gill disease in Atlantic salmon

Amoebic gill disease (AGD), caused by Neoparamoeba perurans, is a major health challenge for Atlantic salmon aquaculture globally. While freshwater bathing for 2 hr is effective in reducing infection severity, there is need for more rapid and lower cost alternatives. To this end, a combination of sodium percarbonate (SPC) in freshwater was examined for its treatment efficacy. Initial in vitro studies showed a reduction in amoeba viability when exposed for 30 min to freshwater containing >500 mg/L SPC. Subsequently, AGD‐affected salmon were bathed for 30 min in 16°C freshwater containing 100, 500 or 1,000 mg/L SPC, or for 2 hr in 16°C freshwater to mimic industry practice. Treatment at the highest SPC concentration caused extensive gill damage and substantial mortality. Neither occurred to a significant extent at lower SPC concentrations. Gill pathology of surviving fish 10 days post‐treatment (dpt) was comparable to or more severe than pre‐treatment, and significantly (p < .001) more severe than in 2 hr freshwater bathed fish. N. perurans DNA was confirmed by qPCR in all treatment groups at 10 dpt. The data indicate that a 30‐min exposure to SPC in freshwater is not a suitable alternative to existing freshwater treatment of AGD.     

Gamma/delta T-cell lymphoma in a dog

A 15-month-old dachshund was presented for examination because of a cough. Thoracic radiographs revealed the presence of a mass in the upper mediastinum. A diagnosis of γδ T-cell lymphoma was made by biopsy and flow cytometry analysis. The dog was treated with chemotherapy and remains asymptomatic after 24 months.     

The effects of metribuzin on early life stages of common carp (Cyprinus carpio)

Metribuzin belongs to the triazine group of herbicides, which are frequently used in agriculture. The aim of this study was to assess the impact of metribuzin on growth and development of early life stages of fish. Subchronic toxic effects of metribuzin at concentrations of 0.9, 4, 14, and 32 mg/L on embryos and larvae of common carp (Cyprinus carpio) were investigated during a 30 day toxicity test under experimental conditions. Exposure to metribuzin at 32 mg/L was associated with increased mortality. Negative effects on total body length, weight, and inhibition of specific growth rate were induced by all experimental concentrations. Length and weight parameters were the most sensitive. The negative impact of metribuzin was observed in the highest tested concentration beginning on day 6 of exposure. Retardation of early ontogeny was associated with concentrations ?4 mg/L. Histological examination revealed changes in liver and caudal kidney after 30 days exposure to 32 mg/L. Based on growth parameters, development, and histological examination, the Lowest Observed Effect Concentration (LOEC) value was 0.9 mg/L.     

Paratuberculosis in a miniature donkey (Equus asinus f. asinus)

Paratuberculosis is mainly an infectious disease of ruminants with worldwide distribution. Infection occurs in early stages of life. Other animal species beyond ruminants are rarely affected, however, experimental and natural infections are possible. A case of paratuberculosis in a miniature donkey (Equus asinus f. asinus) with typical clinical and pathomorphological changes is reported here. Lesions were mainly observed in the intestine. Causative for the profuse diarrhoea with emaciation was massive diffuse granulomatous enteritis involving large quantities of acid-fast organism mainly in macrophages. Granulomatous inflammation with acid-fast bacilli again in macrophages to a lesser degree could be detected in the liver. Mycobacterium avium subsp. paratuberculosis (MAP) was isolated from intestinal contents after an incubation period of four weeks. MAP-specific DNA (IS900 and f57) was detected by polymerase chain reaction in culture material. Additionally MAP-isolates were characterized by multi-target genotyping (MIRU-VNTR- and MLSSR-typing). Isolates belonged to the Type II group and exhibited a unique genotype different from other MAP strains in Germany. The donkey originated from a donkey breeding farm in France with intensive free ranging cattle in the neighbourhood and could have been infected there. Donkeys should be considered as paratuberculosis-susceptible animals in exceptional cases and as possible reservoirs or disseminators of infection.     

Besteht das Risiko der Verschleppung der Bakteriellen Ringfäule der Kartoffel durch den Krautschläger?

In 2001 and 2002, two field trials were performed by the Plant Protection Service (PPS) of the Landwirtschaftskammer Weser-Ems in Oldenburg and the Institute for Plant Protection of the Bavarian State Research Center for Agriculture (LfL) in Freising respectively to examine the transmission of the bacterial ring rot pathogen of potato (Clavibacter michiganensis ssp. sepedonicus) by haulm cutting. In the experiment conducted by the PPS in Oldenburg a lawn-trimmer was used to simulate haulm cutting, before a bacterial suspension was sprayed on the ends of the cut haulms. Two cultivars (early and late) were tested and also the effect of an additional chemical measure for haulm destruction on the transmission of the pathogen was determined. In the trial performed at the LfL in Freising plots with infected plants for the contamination of the haulm cutter were treated immediately prior to plots with healthy plants. Infections of stems (re-growth) and tubers of the plants were determined by polymerase chain reaction (PCR) and indirect immunofluorescence antibody staining (IFAS). In the trials conducted with the haulm cutter, ring rot could be detected in the re-growth of 60% of the plants derived from healthy tubers 4 weeks after haulm cutting. It was confirmed in a bioassay, that the isolated pathogens were viable and virulent. Although the ring rot pathogen was transmitted to a great extent to the leaf material, no infection of progeny tubers occurred in a total of 450 tested plants. However in general a migration of the transmitted pathogen to the tubers cannot be excluded. Therefore further on it is advisable to clean and disinfect the haulm cutter before each use.     

Untersuchungen zur Übertragung von Clavibacter michiganensis ssp. sepedonicus, dem Erreger der Bakteriellen Ringfäule, in das Kraut von Kartoffelpflanzen

From 1999 to 2001, field trials were carried out to find out weather there is a risk of the causal agent of Bacterial Ring Rot of potato (Clavibacter michiganensis ssp. sepedonicus) to be transmitted to the leaves of healthy potato plants via contaminated machinery and equipment during the vegetation period and if so weather the bacteria are transported into the newly formed tubers. Two methods were chosen. Firstly, a sharp-edged metal bar contaminated with a bacterial suspension was drawn through the leaves of potato plants up to 6 times during the vegetation period. Secondly, a suspension of the pathogen was directly injected into the stems of potato plants. The harvested tubers were tested in all trials, leaves were additionally tested in the trial with the contaminated metal bar. The pathogen was not found in any trial. Therefore the risk for the transmission of the Ringrot agent to the leaves of potato plants with contaminated machinery might be very low.     

Die neue Datenbank ALPS-BBA — der komfortable, schnelle Weg zu Informationen über Alternativen zum chemischen Pflanzenschutz für Erzeuger und Berater

With the database ALPS-BBA (http://ALPS.BBA.de), the Federal Biological Research Centre for Agriculture and Forestry (BBA) contributes to promoting integrated pest management and to the necessary extent of applying plant protection products in crop farms. ALPS-BBA offers a comfortable way to search for combinations of crops and pests, possible specific measures, and for key words. Running free text search is also possible. The free text search, in particular, has a great potential for random queries. Web front end and layout allow intuitive navigation, and one can say that search is barrier-free. The database provides information for users who are able to determine crops and pests. Therefore diagnostic aids are not included. Specialised scientists will permanently improve and update the database.     

Mycobacterium gordonae as potential cause of granulomatous lesions of the toe tips in the South African clawed frog (Xenopus laevis)

During regular health status monitoring of the colony of amphibian, Mycobacterium (M.) gordonae were isolated from granulomatous lesions of the tiptoes from the South African clawed frog (Xenopus laevis) maintained at the Tierforschungszentrum of the University of Ulm. During a period of three years a total of 21 animals of the colony, consisting of 350-400 frogs, showed granuloma of the tip of the toes and a loss of the claws. The general condition and the behavior of the frogs appeared to be unchanged. Using a selective medium one isolate was cultured and identified by sequencing of the 16S rRNA gene. To apply a rapid diagnostic method for detecting mycobacteria, in particular M. gordonae in the health monitoring programme of the Xenopus laevis colony, we established the rpoB gene PCR followed by HaeIII restriction analysis of the PCR product. We identified M. gordonae from granuloma of the tiptoes and from unaltered tissue samples of the lungs and skin by PCR restriction analysis. Since mycobacterial species apparently are widespread in granulomatous lesions of the tiptoes of Xenopus laevis, we hypothesize a pathogenic potential. This view is supported by an increasing number of reports in the literature on infections with nontuberculous, "non-pathogenic" mycobacteria in Xenopus laevis.