The Welfare of Laboratory Animals

Veterinary Research Communications -     

Molecular Identification of a New Member of the Clover Proliferation Phytoplasma Group (16SrVI) Associated with Centaurea Solstitialis Virescence in Italy

In the United States, yellow starthistle (Centaurea solstitialis) is an annual invasive weed with Mediterranean origins. Malformed plants displaying witches' broom, fasciations, abortion of buds and flower virescence symptoms were observed in central Italy. Attempts to transmit the causal agent from the natural yellow starthistle host to periwinkle by grafting, resulted in typical symptoms of a phytoplasma, i.e. yellowing and shortening of internodes. The detection of phytoplasmas was obtained from both symptomatic yellow starthistle and periwinkle by the specific amplification of their 16S-23S rRNA genes. PCR amplification of extracted DNA from symptomatic plant samples gave a product of expected size. Asymptomatic plants did not give positive results. An amplicon obtained by direct PCR with universal primers P1/P7 was cloned and sequenced. The homology search using CLUSTALW program showed more than 99% similarity with Illinois elm yellows (ILEY) phytoplasma from Illinois (United States) and 97% with Brinjal little leaf (BLL) phytoplasma from India. Digestion of the nested-PCR products with restriction enzymes led to restriction fragment length polymorphism patterns referable to those described for phytoplasmas belonging to the clover proliferation (16S-VI) group. Since this is a previously undescribed disease, the name Centaurea solstitialis virescence has been tentatively assigned to it. This is a new phytoplasma with closest relationships to ILEY and BLL, but distinguishable from them on the basis of 16S rDNA homology, the different associated plant hosts and their geographical origin.     

La Sota vaccination may not protect against virus shedding and the lesions of velogenic Newcastle disease in commercial turkeys

The aim of this project is to study the clinical signs and lesion of velogenic Newcastle disease (vND) in commercial turkeys, and also to find out if La Sota vaccination offered protection against these signs and lesions. The cockerels were included as positive controls. One hundred and twenty turkey poults and cockerels were divided into eight groups as follows: unvaccinated unchallenged turkeys (UUT), unvaccinated challenged turkeys (UCT), vaccinated unchallenged turkeys (VUT), vaccinated challenged turkeys (VCT), and along the same lines, the cockerel groups were UUC, UCC, VUC and vaccinated challenged cockerels (VCC). Vaccination was at 3 weeks of age while challenge was at 6 weeks of age. The unvaccinated turkeys and cockerels (UCT and UCC) showed different degrees of depression, diarrhoea and later paralysis at challenge. Total mortality was 100% in cockerels within 6 days, but 60% in turkeys. Similar but milder clinical signs were found in the VCC with a total mortality of 13.3%. The VCT showed mild drop in feed and water consumption, and no mortality. All the challenged groups had significant (p < 0.05) loss of weight when compared with their controls. Necropsy showed that while the UCC had severe proventricular haemorrhages, intestinal and caecal tonsil ulcers, the UCT had no digestive tract lesion. There was severe atrophy of the lymphoid organs in all the challenged groups. Histopathological sections of the bursa, spleen and thymus in all the challenged groups with special emphasis on the vaccinated and unvaccinated turkeys with mortalities of 0 and 60%, respectively, had very severe necrosis and depletion of the lymphoid tissue. Virus was isolated from the cloacal swabs. The haemagglutination inhibition antibodies were significantly higher (p < 0.05) in the challenged groups than the unchallenged. The above observations in the intestinal tracts of UCT are of diagnostic significance while the gross and microscopic lesions in the UCT and VCT show that La Sota vaccination may not protect turkeys against the destruction of the lymphoid organs by vND as earlier reported in chickens. This may lead to immunosuppression and production problems in areas where vND is enzootic.     

Changes in enzymic activity and distribution of acid-soluble,amino acid-nitrogen in soil during nitrogen immobilization and mineralization

During a period of immobilization of nitrate-¹⁵N and mineralization of organic N in a sandy-loam, changes were recorded in: (a) the concentration of an added carbon source, glucose-¹⁴C: (b) evolution of ¹⁴CO₂: (c) bacterial populations; (d) distribution and concentration of newly-synthesized, acid-soluble, amino acid-¹⁵N; and (e) distribution and activities of several oxidative and hydrolytic enzyme systems.Added glucose-¹⁴C was rapidly metabolized by the soil microflora. After 1.5 day's incubation, when only 3.6 per cent of the added glucose was present, 68 per cent of the ¹⁴C remained in the soil-microbial system. During this period there was a marked increase in viable bacterial numbers and an almost complete immobilization of nitrate-¹⁵N. On continued incubation, microbial metabolites were oxidized at decreasing rates, the more rapid phase corresponding to a period of net decline in the viable bacterial population.Soil was fractionated by a relatively mild procedure into components containing: (a) extractable proteins; (b) extractable amino acids and peptides; (c) particulate material containing microbial cells, cell debris and material bound to larger soil particles; and (d) microbial metabolites mainly bound to soil colloids. Although the total, acid-soluble, amino acid-¹⁵N remained relatively constant for about 50 days, there were marked changes in their concentration in different fractions, especially in the extracts and in the fraction containing fine colloidal material. However, the relatively large decline in labelled, acid-soluble, amino acid-¹⁵N occurred during the active phase of oxidation of microbial metabolites when little net mineralization of labelled compounds occurred.Increases in enzymic activities generally coincided with increased viable bacterial populations although there were some exceptions, notably casein and benzoyl arginine amide-hydrolysing enzymes. The stabilities of the newly-formed enzymes varied markedly. The greatest relative changes in activity occurred with the casein-hydrolysing enzymes. Their activity reached a maximal value after the main flush of bacterial growth, was short-lived and was to a large extent extractable. The formation and disappearance of this extracellular proteolytic activity coincided approximately with that of a secondary peak of extractable, newly-synthesized, protein-¹⁵N. In general however, changes in enzymic activity could not be identified with changes of protein-¹⁵N concentrations of the different fractions.     

A GIS-based plant prediction model for wetland ecosystems

An existing non-spatial model for the prediction of response of wetland plant species on ecological factors has been transformed into a GIS-based prediction model which produces spatial output at the landscape scale. The input, spatial patterns of the ecological factors, were constructed with geostatistical spatial interpolation (kriging). With this GIS-based model the spatial patterns of presence and absence of 78 wetland plant species are predicted for an area with wetlands in the Netherlands of approximately 500 square kilometers. The GIS-based model has been validated, and the estimated uncertainty of the input has been propagated through the model. At the species level the output shows spatially coherent and non-random patterns. The validation is affected by the propagation of input errors through the model. The number of valid predictions declines approximately 10–20% when 95% confidence intervals are used in the validation. This study shows that it is feasible to use a geostatistical interpolation method to construct spatial patterns of ecological factors on a landscape scale and to use these patterns as input for a GIS-based prediction model. The added uncertainty on the input values however, affects the number of valid predictions of the model.     

Zur Chemie der Verunreinigungen des Hexachlorcyclohexans

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