Molecular characterisation of 'strangles' outbreaks in the UK: the use of M-protein typing of Streptococcus equi ssp. equi

Reasons for performing study: Strangles is the most commonly diagnosed and important infectious disease of horses worldwide. Very little is known about the temporo‐spatial and molecular epidemiology of strangles. The disease is not notifiable in the UK and there are few published data on the geographical locations of outbreaks. Objective: To investigate whether typing of a surface protein (SeM) of Streptococcus equi ssp. equi (S. equi), the causative agent of strangles, is a useful epidemiological tool. Methods: The variable region of the SeM gene was amplified from 145 isolates of S. equi by PCR and sequenced. Different SeM gene alleles were assigned based on the SeM database, grouped into phylogenetic clusters using split decomposition analysis and plotted against the submitting veterinary practices. Results: In this study 21 S. equi SeM alleles were found, including 9 previously unidentified alleles and representing 4 phylogenetic groups. S. equi containing SeM alleles 9 and 7 were the most commonly isolated and there was a high number of low frequency alleles. The occurrence of an outbreak cluster in the north‐west of the UK is also reported. Conclusions: Strangles outbreaks can be differentiated on the basis of their SeM allele sequences. The data provide further evidence of SeM mutation leading to the emergence of novel, but related SeM alleles that are geographically linked. Sequencing of the SeM gene is a useful tool for the elucidation of strangles epidemiology at a regional and a national level. Potential relevance: This technique may allow differentiation or linkage of strangles outbreaks and as such may be an effective tool for local as well as national and international disease surveillance.     

Nobody farms here anymore: Livelihood diversification in the Amazonian community of Carvão,a historical perspective

Over the past 15 years income sources in the Amazonian community of Carvão have diversified to include government salaries, retirement and welfare benefits, and wages from an evolving informal service sector. These non-farm incomes are now more important to household incomes than the sale of agricultural products. Out of 80 households only three families were found to depend almost entirely on the sale of agricultural goods for cash income. Agriculture is still a part of most families’ livelihoods; however, production today is mainly a subsistence activity. Recent changes in Carvão are consistent with trends of livelihood diversification observed in rural societies across the globe. However, current research reveals that Carvão is different from other case studies in a number of ways. A history of livelihoods illustrates that residents in Carvão have always engaged in a range of activities, including farming, extractive activities, and wage labor. New incomes are the result of new jobs in the public sector and social policies benefiting the rural poor. In contrast to the literature on livelihood diversification, the decentralization of the federal government in Brazil has resulted in greater opportunities for rural income and employment. Consistent with recorded trends, research shows that small farmers in Carvão have down-sized agricultural production. Farmers cite low market prices (the result of vertical integration of local markets) as one cause of this decline. Residents, especially small farmers, interested in diversifying agricultural production are limited by inadequate extension services and credit, and younger residents seek public sector employment. Income diversification has increased livelihoods security; however, future livelihoods will depend on new economic growth. Given the stagnating public sector and a weak industrial sector, production geared toward growing urban markets is a viable means for further income generation in Carvão.     

Quantitation of silibinin, a putative cancer chemopreventive agent derived from milk thistle (Silybum marianum), in human plasma by high-performance liquid chromatography and identification of possible metabolites

Silibinin has recently received attention as a potential cancer chemopreventive agent because of its antiproliferative and anticarcinogenic effects. A simple and specific reversed-phase high-performance liquid chromatography method was developed and validated for the quantitation of silibinin in human plasma. Sample preparation involved simple protein precipitation, and separation was achieved on a Waters Atlantis C18 column with flow rate of 1.0 mL/min at 40 degrees C and UV detection at 290 nm. Silibinin was detected as two peaks corresponding to trans-diastereoisomers. The peak area was linear over the investigated concentration range (0-5000 ng/mL). The limits of detection were 2 and 1 ng/mL for the two diastereoisomers (d1 and d2), with a recovery of 53-58%. This method was utilized to detect silibinin in plasma of colorectal patients after 7 days of treatment with silipide (silibinin formulated with phosphatidyl choline).     

Development of analyses by high-performance liquid chromatography and liquid chromatography/tandem mass spectrometry of bilberry (Vaccinium myrtilus) anthocyanins in human plasma and urine

Anthocyanins are potent antioxidants that may possess chronic disease preventive properties. Here, rapid, reliable, and reproducible solid-phase extraction, high-performance liquid chromatography (HPLC), and mass spectrometry techniques are described for the isolation, separation, and identification of anthocyanins in human plasma and urine. Recoveries of cyanidin-3-glucoside (C3G) were 91% from water, 71% from plasma, and 81% from urine. Intra- and interday variations for C3G extraction were 9 and 9.1% in plasma and 7.1 and 9.1% in urine and were less than 15% for all anthocyanins from a standardized bilberry extract (mirtoselect). Analysis of mirtoselect by HPLC with UV detection produced spectra with 15 peaks compatible with anthocyanin components found in mirtoselect within a total run time of 15 min. Chromatographic analysis of human urine obtained after an oral dose of mirtoselect yielded 19 anthocyanin peaks. Mass spectrometric analysis employing multiple reaction monitoring suggests the presence of unchanged anthocyanins and anthocyanidin glucuronide metabolites.     

Human beta-globin gene sequences injected into mouse eggs, retained in adults, and transmitted to progeny

Foreign gene sequences were retained in two adult mice (out of 62 analyzed) from fertilized eggs injected with a recombinant plasmid containing the human beta-globin genomic region and the herpes simplex viral thymidine kinase gene. The intact human and viral genes were found in DNA of one of the animals and, in the other, at least part of the human globin gene was present. The latter individual transmitted these sequences to its progeny in a Mendelian ration. Thus, human DNA may be incorporated into the germ line of mice for in vivo studies of regulation of gene expression in development, genetic diseases, and malignancy.     

Dorsal, an embryonic polarity gene in Drosophila, is homologous to the vertebrate proto-oncogene, c-rel

The Drosophila gene, dorsal, is a maternal effect locus that is essential for the establishment of dorsal-ventral polarity in the developing embryo. The dorsal protein was predicted from the complementary DNA sequence; it is almost 50 percent identical, over an extensive region, to the protein encoded by the avian oncogene v-rel, its cellular homolog, c-rel, and a human c-rel fragment. The oncogene v-rel is highly oncogenic in avian lymphoid, spleen, and bone marrow cells.     

90 years of forest cover change in an urbanizing watershed: spatial and temporal dynamics

Landscape structure in the Eastern US experienced great changes in the last century with the expansion of forest cover into abandoned agricultural land and the clearing of secondary forest cover for urban development. In this paper, the spatial and temporal patterns of forest cover from 1914 to 2004 in the Gwynns Falls watershed in Baltimore, Maryland were quantified from historic maps and aerial photographs. Using a database of forest patches from six times—1914, 1938, 1957, 1971, 1999, and 2004—we found that forest cover changed, both temporally and spatially. While total forest area remained essentially constant, turnover in forest cover was very substantial. Less than 20% of initial forest cover remained unchanged. Forest cover became increasingly fragmented as the number, size, shape, and spatial distribution of forest patches within the watershed changed greatly. Forest patch change was also analyzed within 3-km distance bands extending from the urban core to the more suburban end of the watershed. This analysis showed that, over time, the location of high rates of forest cover change shifted from urban to suburban bands which coincides with the spatial shift of urbanization. Forest cover tended to be more stable in and near the urban center, whereas forest cover changed more in areas where urbanization was still in process. These results may have critical implications for the ecological functioning of forest patches and underscore the need to integrate multi-temporal data layers to investigate the spatial pattern of forest cover and the temporal variations of that spatial pattern.     

OPTIMIZING A PROTOCOL FOR 1H‐MAGNETIC RESONANCE SPECTROSCOPY OF THE CANINE BRAIN AT 3T

Intracranial diseases are common in dogs and improved noninvasive diagnostic tests are needed. Magnetic resonance (MR) spectroscopy is a technique used in conjunction with conventional MR imaging to characterize focal and diffuse pathology, especially in the brain. As with conventional MR imaging, there are numerous technical factors that must be considered to optimize image quality. This study was performed to develop an MR spectroscopy protocol for routine use in dogs undergoing MR imaging of the brain. Fifteen canine cadavers were used for protocol development. Technical factors evaluated included use of single‐voxel or multivoxel acquisitions, manual placement of saturation bands, echo time (TE), phase‐ and frequency‐encoding matrix size, radiofrequency coil, and placement of the volume of interest relative to the calvaria. Spectrum quality was found to be best when utilizing a multivoxel acquisition with the volume of interest placed entirely within the brain parenchyma without use of manually placed saturation bands, TE = 144 ms, and a quadrature extremity radiofrequency coil. An 18 × 18 phase‐ and frequency‐encoding matrix size also proved optimal for image quality, specificity of voxel placement, and imaging time.