Assessment of an ELISA method to support surveillance of bovine tuberculosis in Albania

Background

Bovine tuberculosis (bTB) is an important bacterial infectious disease in Albania of concern to animal and human health; its prevalence is poorly documented.

Methods

In this longitudinal study, we tested by ELISA 2661 serum samples, from 154 herds, with the aim of establishing the suitability of this approach to screen the bovine population for bTB. In a follow-on survey of 87 animals in three villages, we assessed the usefulness of the Mycobacterium bovis IDEXX ELISA (IDEXX M. bovis Antibody (Ab) Test. IDEXX Europe B.V P.O. Box 1334, 2130 EK Hoofddorp, The Netherlands) assay by comparing IDEXX results with the results of the single intradermal cervical skin test. Skin tests were performed either after or at the time of collection of blood samples, and therefore cattle were not sensitized by tuberculin before serological testing.

Results

The proportion of herds in which serologically positive cattle were found was 18.2 % (95 % CI, 1.9–25.8 %) and the prevalence of seropositive cattle was 1.4 % (95 % CI, 0.8–2.1 %). In the follow-up study, two of the 87 animals reacted positively to the skin test and two produced inconclusive reactions. No overlap was found between the four animals with positive IDEXX ELISA results and the four animals with non-negative skin test results.

Conclusion

The lack of agreement between the results of the two tests may reflect different elements of the immune response (humoral and cell-mediated immunity). In future, cattle should be sensitized by the intradermal injection of tuberculin 14 days prior to the collection of blood samples, which would then be tested by the Mycobacterium bovis IDEXX ELISA Test in order to determine more accurately the prevalence of infection.

     

Study of allozyme diversity in guar [Cyamopsis tetragonoloba (L.) Taub.] germplasm

Guar [Cyamopsis tetragonoloba (L.) Taub.] is an important commercial crop of India, mainly cultivated in the northwestern part and neighboring areas of Pakistan. High morphological diversity is reported in guar. The present study attempts to analyze the genetic diversity in cultivated guar using allozyme polymorphism and compare it with reported morphological diversity. Accessions for the study were selected from a set of guar germplasm conserved at the National Bureau of Plant Genetic Resources, New Delhi. The morphological data of 108 selected accessions was subjected to UPGMA clustering. Fifty-five accessions were then picked randomly from the clusters generated. Tris-borate system (pH 8.3) was found to be most effective for detection of allozyme polymorphism in guar. Ten enzymes were selected for the study out of 18 systems tried, namely, malate dehydrogenase, malic enzyme, phosphogluco isomerase, glutamate oxaloacetate transaminase, esterase, peroxidase, alcohol dehydrogenase, glutamate dehydrogenase, glucose 6-phosphate dehydrogenase and shikimate dehydrogenase. A total of 20 loci were detected with 34 alleles. The number of polymorphic loci in different accessions was observed to be highly variable (0–9). An average of 1.27 allozyme per polymorphic locus was observed. The allele frequency data of the 55 accessions was used to determine genetic diversity parameters. The dendrogram generated on genetic distance showed three clusters in the distance range of 0–0.28. Most of the accessions were in the first clusters in a narrow distance range of 0–0.08. The second cluster consisted of improved varieties and accession from Pakistan. The third cluster of accessions was genetically more distant but there was no definite segregation of accessions according to the place of collection. The accessions from south west Rajasthan were scattered in all the three clusters showing higher variability in this region, also supported by higher morphological variation. The genetic distance was low among accessions of guar, indicating low diversity in India.     

Acyl-homoserine lactones from Erwinia psidii R. IBSBF 435T, a guava phytopathogen (Psidium guajava L.)

The phytopathogen Erwinia psidii R. IBSBF 435T causes rot in branches, flowers, and fruits of guava (Psidium guajava L.), being responsible for crop losses, and has no effective control. It was demonstrated that this strain produces two compounds [S-(-)-N-hexanoyl and N-heptanoyl-homoserine lactone], both belonging to the class of quorum-sensing signaling substances. A protocol using gas chromatography-flame ionization detection with chiral stationary phase is described for the absolute configuration determination of a natural acyl-homoserine lactone. Biological assays with specific reporter and synthesis of identified substances are also described. This is the first report on the N-heptanoyl-homoserine lactone occurrence in the Erwinia genus.     

Effect of ageing on quality of jojoba oil from Indian locations

Jojoba oil is a source of specialty chemicals and its uses for industrial application are gaining in adaptability and importance. For prolong use of jojoba oil, the details of its storage behaviour are required. This information is scarce for the products obtained from Indian locations. In this study, the ageing effect on the quality of oils was studied for oils from five locations of Rajasthan (India). The oils were extracted/expelled from the seeds that contained 2.5–3.9 wt.% moisture and 42–50 wt.% oil.Physico-chemical properties were determined by standard analytical test methods. Effect of storage time on quality of solvent extracted oil samples with respect to acid value, iodine value, saponification value and peroxide value were determined. The samples were stored for 18 months in the laboratory under climatic condition prevailing in Dehradun (India) during study period. The analysis of samples was done at an interval of 3 months. The properties of jojoba oils compare well with the International Jojoba Expert Council (IJEC) specifications. During storage, acid values, iodine values and peroxide values of the oil increase with time while saponification values remain almost unchanged.     

Recovery in soil cover and vegetation structure after ancient landslide in mountain fens under Caltho-Alnetum community and response of soil microarthropods (Hexapoda: Collembola) to natural restoration process

Purpose

The objective of this study was to determine the long-term environmental changes induced by ancient landslide in the mountain fen. Attempts were made to demonstrate the progress of the soil cover restoration process about 200 years after the landslides and associated with changes in vegetation and soil microarthropod biodiversity and occurrence pattern associated with the restoration of soil cover.

Material and methods

The study covered mountain fen of the Caltho-Alnetum in the Babiogórski National Park in Outer Flysch Carpathians, Poland, where ancient landslide deposits are causing disturbance in fen hydrologic regime and over time related to various rate of fen area natural restoration processes. The drill test has been used to assess changes in layering and thickness of the fen soils. The following soil parameters, such as pH and total organic carbon content, were determined at each distinguish layer. The diversity and distribution pattern of soil microarthropods, represented by Collembola, was examined in two parts of the fen: restored and not-restored. The soil parameters, such as pH, electrical conductivity, total exchangeable base, total organic carbon, and nitrogen content, were determined in the soil samples simultaneously. The assessment of the vegetation structure recovery within studied mountain fen after landslide was referred to average parameters of the community completed for other natural mountain fens under the Caltho-Alnetum community. The research results were statistically verified.

Results and discussion

The test drillings showed that landslides change soil layering, site-specific soil properties of mountain fen such as chemistry (the statistically significant differences were noted only in the case of total organic carbon content), vegetation structure, and soil microarthropod communities. By changing site-specific conditions, landslides influence on the biodiversity and peatland ecosystems functioning.

Conclusion

Our results clearly demonstrate that the landslide itself initiated a complex and lengthy process of changes in biological aspects of peatlands including the biodiversity of the underground fauna and plant community. Soil microarthropod communities represented by Collembola can be used as a good indicator of mountain fen restoration process.

     

Landscape relatedness: detecting contemporary fine-scale spatial structure in wild populations

Context

Methods for detecting contemporary, fine-scale population genetic structure in continuous populations are scarce. Yet such methods are vital for ecological and conservation studies, particularly under a changing landscape.

Objectives

Here we present a novel, spatially explicit method that we call landscape relatedness (LandRel). With this method, we aim to detect contemporary, fine-scale population structure that is sensitive to spatial and temporal changes in the landscape.

Methods

We interpolate spatially determined relatedness values based on SNP genotypes across the landscape. Interpolations are calculated using the Bayesian inference approach integrated nested Laplace approximation. We empirically tested this method on a continuous population of brown bears (Ursus arctos) spanning two counties in Sweden.

Results

Two areas were identified as differentiated from the remaining population. Further analysis suggests that inbreeding has occurred in at least one of these areas.

Conclusions

LandRel enabled us to identify previously unknown fine-scale structuring in the population. These results will help direct future research efforts, conservation action and aid in the management of the Scandinavian brown bear population. LandRel thus offers an approach for detecting subtle population structure with a focus on contemporary, fine-scale analysis of continuous populations.

     

Antifungal properties of wheat histones (H1-H4) and purified wheat histone H1

Wheat ( Triticum spp.) histones H1, H2, H3, and H4 were extracted, and H1 was further purified. The effect of these histones on specific fungi that may or may not be pathogenic to wheat was determined. These fungi included Aspergillus flavus , Aspergillus fumigatus , Aspergillus niger , Fusarium oxysporum , Fusarium verticillioides , Fusarium solani , Fusarium graminearum , Penicillium digitatum , Penicillium italicum , and Greeneria uvicola . Non-germinated and germinating conidia of these fungi were bioassayed separately. The non-germinated and germinating conidia of all Fusarium species were highly susceptible to the mixture (H1-H4) as well as pure H1, with viability losses of 99-100% found to be significant (p < 0.001) at ≤10 μM or less for the histone mixture and pure H1. F. graminearum was the most sensitive to histone activity. The histones were inactive against all of the non-germinated Penicillium spp. conidia. However, they significantly reduced the viability of the germinating conidia of the Penicillium spp. conidia, with 95% loss at 2.5 μM. Non-germinated and germinating conidia viability of the Aspergillus spp. and G. uvicola were unaffected when exposed to histones up to 10 μM. Results indicate that Fusarium spp. pathogenic to wheat are susceptible to wheat histones, indicating that these proteins may be a resistance mechanism in wheat against fungal infection.     

1H NMR, a rapid method to monitor organic acids during cupuassu (Theobroma grandiflorum Spreng) processing

The development of an analytical method using 1H nuclear magnetic resonance (1H NMR) spectrometry to monitor cupuassu (Theobroma grandiflorum Spreng) bean fermentation, drying, and roasting processes is reported. The analysis of organic acids and alcohols of crude water extracts of cupuassu ground kernels were monitored by HPLC and 1H NMR spectroscopy. The residual protein signals caused deleterious effects on acid and alcohol quantifications. Therefore, the analytical procedures were optimized by sample cleanup and water suppression pulse sequences in order to obtain compatible data using HPLC and 1H NMR. The quantification of lactic acid, acetic acid, and 2,3-butanediol by NMR is 5- to 10-fold faster than by HPLC, with the advantage of providing the identification of several chemical species in a single experiment. Application of these analytical conditions to some cupuassu samples revealed that this methodology can be applied to the quality profiles of fermentation and roasting processes.     

Isolation and biochemical diagnosis of cell lines of groundnut (Arachis hypogaea L) selected on glyphosate†

Three cell lines of groundnut (Arachis hypogaea L), an important oilseed legume, were selected on glyphosate using in-vitro culture techniques. The cell lines isolated through single as well as stepwise selection procedures showed c 20-fold increase in glyphosate tolerance as compared to the unselected control cell line. Studies on the biochemical mechanism of glyphosate tolerance in these cell lines showed a significant increase in the total extractable activity of the target enzyme, 5-enolpyruvyl shikimate-3-phosphate (EPSP) synthase (EC 2.5.1.19), which was further confirmed with immunological data. The over-expressed EPSP synthase activity was, however, subject to inhibition by glyphosate in vitro. Two other key regulated enzymes of the shikimic acid pathway, 3-deoxy-D -arabino heptulosonate 7-phosphate (DAHP) synthase (EC 4.1.2.15) and chorismate mutase (CM) (EC 5.4.99.5) did not show any change in specific activity in the selected cell lines. The enhanced activity of EPSP synthase in the tolerant cell lines was found to be stably inherited in the absence of selection pressure. © 1999 Society of Chemical Industry