Molecular characterization of Brazilian strains of Xanthomonas campestris pv. viticola by rep-PCR fingerprinting

Bacterial canker of grapevine (Vitis vinifera), caused by Xanthomonas campestris pv. viticola was first detected in Brazil in 1998, affecting grapevines in the São Francisco river basin, state of Pernambuco. The disease was also reported in Juazeiro, Bahia and later in Piauí and Ceará. Due to its limited geographical distribution and relatively recent detection in Brazil, very little is known about the pathogen's biology and diversity. Repetitive DNA based-PCR (rep-PCR) profiles were generated from purified bacterial DNA of 40 field strains of X. campestris pv. viticola, collected between 1998 and 2001 in the states of Pernambuco, Bahia and Piauí. Combined analysis of the PCR patterns obtained with primers REP, ERIC and BOX, showed a high degree of similarity among Brazilian strains and the Indian type strain NCPPB 2475. Similar genomic patterns with several diagnostic bands, present in all strains, could be detected. Fingerprints were distinct from those of strains representing other pathovars and from a yellow non-pathogenic isolate from grape leaves. The polymorphism observed among the Brazilian strains allowed their separation into five subgroups, although with no correlation with cultivar of origin, geographic location or year collected.

Growth through internationalization of Russian SMEs in Finland

The study examines Russian SMEs motives, needs, prospects and constrains to enter Finland through Lappeenranta. Russian entrepreneurship is a young phenomenon in Finland. The present research becomes a first guide in understanding of Russian SMEs’ capabilities for establishing their subsidiaries in Finland. The extensive model of different theoretical approaches was created for evaluating Russian SMEs growth and internationalization opportunities. The findings of the research indicated that Russian growing companies chose the internationalization to continue their development. Many Finland’ positive conditions were evaluated as important for the Russian entrepreneurship. Finland was the first European country of the expansion to the European Union, for the Russian SMEs established the subsidiaries in Lappeenranta. Their motives for entering Finland were usually based on market and resource views. Generally, the Russian SMEs establish a new company in Finland only when they have sufficient capabilities and resources for internationalization. Entering Finland the Russian firms make better the allocation of their natural, financial and human resources.

Serum neutralization with different types and subtypes of bovine herpesvirus 1 and 5

The serum neutralization (SN) test is the gold standard method to measure neutralizing antibodies to bovine herpesviruses. However, in view of the further subdivisions of bovine herpesviruses in types/subtypes, defining which virus to use at challenge in SN tests may be difficult. In view of that, this study was carried out to re-evaluate (SN) sensitivity with different types/subtypes of bovine herpesviruses types 1 (BoHV-1) and 5 (BoHV-5) as challenge viruses. Bovine sera (n=810) were collected from two distinct geographic regions and tested by SN with three type 1 viruses (BoHV-1.1 strains "Los Angeles" and "EVI123/98"; BoHV-1.2a strain "SV265/96") and three type 5 viruses (BoHV-5a strain "EVI88/95"; BoHV-5b strain "A663" and BoHV-5c "ISO97/95"). SN tests were performed with a 1 hour incubation of the serum-virus mixtures at 37ºC against 100 TCID50 of each of the viruses. SN sensitivity varied greatly depending on the challenge virus used in the test. The highest sensitivity (327 positive/810 total sera tested; 40.37%) was attained when the positive results to the six viruses were added together. No association could be found between any particular type or subtype of virus and the sensitivity of the test. When positive results to each single strain were considered, SN sensitivity varied from 41.7% to 81.7%, depending on the virus and the geographic region of origin of the sera. Variation was detected even when challenge viruses belonged to the same subtype, where disagreement between positive results reached 41%. These results indicate that one hour incubation SN tests against single viruses, as performed here, may display a significantly low sensitivity (p=0.05); performing SN tests against a number of different viruses may increase considerably SN sensitivity. Furthermore, the choice of virus used for challenge is critical in SN tests. In addition, sera from different geographic regions may give rise to disagreeing results with different strains of BoHV-1 and BoHV-5. This might be particularly relevant for control programs and in international trade, were maximum sensitivity should be targeted.

Oestrus ovis infection of grazing sheep during summer in southern Chile

Abstract: Oestrus ovis is a botfly whose larvae cause nasal myiasis, an environmental-dependent disease in small ruminants, generating acute and chronic injuries in the cranial cavities of sheep. Chile is a country of the southernmost worldwide distribution of this parasite, and there is few information about. Whence, the objective of this study was to approximate the epidemiological situation of O. ovis infection in Chilean sheep. From December 2009 to March 2010, a total of 87 samples were obtained by necropsy for skull inspection. The larvae were collected, and microscopically identified. The prevalence in the sampled sheep was 60.9%. From those that were infected, 85.7% (18/21) of sheep were 1 to 3 years old, constituting the stratum with the highest prevalence. The difference of infection in females and males was not significant. The high risk of infection seems to be dependent upon the environmental conditions of this southern region, especially during summer when the first larval stage (L1) could be found as the evidence. Therefore, the disease should be considered as a significant problem for this kind of livestock production.

Biochemical characterization of the GM2 gangliosidosis B1 variant

The deficiency of the A isoenzyme of ß-hexosaminidase (Hex) produced by different mutations of the gene that codes for the alpha subunit (Tay-Sachs disease) has two variants with enzymological differences: the B variant consists of the absence of Hex A isoenzyme and the B1 variant produces an inactive Hex A isoenzyme for the hydrolysis of the GM2 ganglioside and synthetic substrates with negative charge. In contrast to the early childhood form of the B variant, the B1 variant appears at a later clinical stage (3 to 7 years of age) with neurodegenerative symptoms leading to the death of the patient in the second decade of life. The most frequent mutation responsible for the GM2 gangliosidosis B1 variant is R178H, which has a widespread geographic and ethnic distribution. The highest incidence has been described in Portugal, which has been suggested as the point of origin of this mutation. Biochemical characterization of this lysosomal disease is carried out using negatively charged synthetic alpha subunit-specific sulfated substrates, since Hex A isoenzyme heat-inactivation assays are not applicable. However, the determination of the apparent activation energy of Hex using the neutral substrate 3,3'-dichlorophenolsulfonphthaleinyl N-acetyl-ß-D-glucosaminide, may offer a valid alternative. The presence of an alpha subunit in the alphaß heterodimer Hex A means that its activation energy (41.8 kJ/mol) is significantly lower than that of the ßß homodimer Hex B (75.1 kJ/mol); however, as mutation inactivates the alpha subunit, the Hex A of the B1 variant presents an activation energy that is similar to that of the Hex B isoenzyme.

Geographic distribution of hepatitis C virus genotypes in Brazil

Brazil is a country of continental dimension with a population of different ethnic backgrounds. Thus, a wide variation in the frequencies of hepatitis C virus (HCV) genotypes is expected to occur. To address this point, 1,688 sequential samples from chronic HCV patients were analyzed. HCV-RNA was amplified by the RT-PCR from blood samples collected from 1995 to 2000 at different laboratories located in different cities from all Brazilian States. Samples were collected in tubes containing a gel separator, centrifuged in the site of collection and sent by express mail in a refrigerated container to Laboratório Bioquímico Jardim Paulista, São Paulo, SP, Brazil. HCV- RNA was extracted from serum and submitted to RT and nested PCR using standard procedures. Nested PCR products were submitted to cycle sequencing reactions without prior purification. Sequences were analyzed for genotype determination and the following frequencies were found: 64.9% (1,095) for genotype 1, 4.6% (78) for genotype 2, 30.2% (510) for genotype 3, 0.2% (3) for genotype 4, and 0.1% (2) for genotype 5. The frequencies of HCV genotypes were statistically different among Brazilian regions (P = 0.00017). In all regions, genotype 1 was the most frequent (51.7 to 74.1%), reaching the highest value in the North; genotype 2 was more prevalent in the Center-West region (11.4%), especially in Mato Grosso State (25.8%), while genotype 3 was more common in the South (43.2%). Genotypes 4 and 5 were rarely found and only in the Southeast, in São Paulo State. The present data indicate the need for careful epidemiological surveys throughout Brazil since knowing the frequency and distribution of the genotypes would provide key information for understanding the spread of HCV.

Detection of enterotoxins produced by B. cereus through PCR analysis of ground and roasted coffee samples in Rio de Janeiro, Brazil

Coffee is one of the most appreciated drinks in the world. Coffee ground is obtained from the fruit of a small plant that belongs to the genus Coffea. Coffea arabica and Coffea canephora robusta are the two most commercially important species. They are more commonly known as arabica and robusta, respectively. Two-thirds of Coffea arabica plants are grown in South and Central America, and Eastern Africa - the place of origin for this coffee species. Contamination by microorganisms has been a major matter affecting coffee quality in Brazil, mainly due to the harvesting method adopted. Brazilian harvests are based on fruits collected from the ground mixed with those that fall on collection cloths. As the Bacillus cereus bacterium frequently uses the soil as its environmental reservoir, it is easily capable of becoming a contaminant. This study aimed to evaluate the contamination and potential of B. cereus enterotoxin genes encoding the HBL and NHE complexes, which were observed in strains of ground and roasted coffee samples sold in Rio de Janeiro. The PCR (Polymerase Chain Reaction) results revealed high potential of enterotoxin production in the samples. The method described by Speck (1984) was used for the isolation of contaminants. The investigation of the potential production of enterotoxins through isolates of the microorganism was performed using the B. cereus enterotoxin Reverse Passive Latex Agglutination test-kit (BCET-RPLA, Oxoid), according to the manufacturer's instructions. The potential of enterotoxin production was investigated using polymerase chain reaction (PCR) methods for hblA, hblD and hblC genes (encoding hemolysin HBL) and for nheA, nheB and nheC genes (encoding non-hemolytic enterotoxin - NHE). Of all the 17 strains, 100% were positive for at least 1 enterotoxin gene; 52.9% (9/17) were positive for the 3 genes encoding the HBL complex; 35.3% (6/17) were positive for the three NHE encoding genes; and 29.4% (5/17) were positive for all enterotoxic genes.

Technological properties of precooked flour containing coffee powder and rice by thermoplastic extrusion

Although Brazil is a country of tradition in both the production and consumption of coffee, the most of the coffee is consumed as a beverage, which reduces greatly the competitiveness on international market, for reducing the chances of supplying the product under other forms of consumption. Owing to that, the aim of this study was developing a precooked mixed flour containing coffee powder and rice for use in coffee flavored products. Mixtures of rice and coffee in the proportions of 900:100, 850:150 and 800:200 g, respectively, were processed in a single screw extruder (Brabender DS-20, Duisburg, German) and the effect of the extrusion process on the variables moisture content (16%, 18% and 20%) and temperature in the third extruding zone (140 °C, 160 °C and 180 °C) was studied. The results for expansion index ranged from 2.91 to 11.11 mm in diameter; the water absorption index from 4.59 to 6.33 g gel/g sample and the water solubility index varied from 4.05% to 8.57%. These results showed that, despite coffee powder influenced the variables studied, the expanded product after milling resulted in a extruded mixture with good absorption and water solubility indices, which favors the use of the precooked mixed flour for human consumption.

Salted lamb meat blanket of Petrolina-Pernambuco, Brazil: process and quality

Salted lamb meat blanket, originated from boning, salting, and drying of whole lamb carcass, was studied aiming at obtaining information that support the search for guarantees of origin for this typical regional product from the city of Petrolina-Pernambuco-Brazil. Data from three processing units were obtained, where it was observed the use of a traditional local technology that uses salting, an ancient preservation method; however, with a peculiar boning technique, resulting in a meat product with great potential for exploitation in the form of meat blanket. Based on the values of pH (6.22 ± 0.22), water activity (0.97 ± 0.02), and moisture (69.86 ± 2.26) lamb meat blanket is considered a perishable product, and consequently it requires the use of other preservation methods combined with salt, which along with the results of the microbiological analyses (absence of Salmonella sp, score <10 MPN/g of halophilic bacteria, total coliforms between 6.7 × 10³ and 5.2 × 10(6) FUC/g, and Staphylococcus from 8.1 × 10³ CFU/g at uncountable) reinforce the need of hygienic practices to ensure product safety. These results, together with the product notoriety and the organization of the sector are important factors in achieving Geographical Indication of the Salted lamb Meat blanket of Petrolina.

Optimization of the HS-SPME-GC/MS technique for the analysis of volatile compounds in caprine Coalho cheese using response surface methodology

Abstract Caprine Coalho cheese presents great potential for a typical protected designation of origin, considering that this traditional Brazilian cheese presents a slightly salty and acid flavor, combined with a unique texture. This study optimized the HS-SPME-GC-MS methodology for volatile analysis of Coalho cheese, which can be used as a tool to help in the identification of the distinctive aroma profile of this cheese. The conditions of equilibrium time, extraction temperature and time were optimized using the statistical tool factorial experimental design 23, and applying the desirability function. After the evaluation, it was concluded that the optimum extraction conditions comprised equilibrium and extraction time of 20 and 40 minutes, respectively; and ideal extraction temperature of 45 °C. The optimum extraction of volatile compounds in goat Coalho cheese captured 32 volatile compounds: 5 alcohols, 5 esters, 3 ketones, 6 acids, 3 aldehydes, 3 terpenes, and 7 hydrocarbons.

Migrations and gradations : reappraising the health profile of immigrants to Canada

New immigrants to Canada typically have a more favourable health profile than the non-immigrant population. This phenomenon, known as the 'healthy immigrant effect', has been attributed to both the socioeconomic advantage (ie. educational attainment, occupational opportunity) of non-refugee immigrants and existing screening protocols that admit only the healthiest of persons to Canada. It has been suggested that this health advantage diminishes as the time of residence in Canada increases, due in part to the adoption of health-risk behaviours such as alcohol and cigarette use, an increase in excess body weight, and declining rates of physical activity. However, the majority of health research concerning immigrants to Canada has been limited to cross-sectional studies (Dunn & Dyck, 2000; Newbold & Danforth, 2003), which may mask an immigrant-specific cohort effect. Furthermore, the practice of aggregating foreign-bom persons by geographical regions or treating all immigrants as a homogeneous group may also obfuscate intra-immigrant differences in health. Accordingly, this study uses the Canadian National Population Health Surveys (NPHS) and data from the United Nations Development Program (UNDP) to prospectively evaluate factors that predict health status among immigrants to Canada. Each immigrant in the NPHS was linked to the UNDP Human Development Index of their country of birth, which uses a combined measure of health, education, and per capita income of the populace. The six-year change in health function, psychological distress, and self-rated health were considered from a population health perspective (Evans, 1994), using generalized-estimating equations (GEE) to examine the compounding effect of past and recent predictors of health. Demographic

Agrarian revolution in Central America : a comparison of Nicaragua and Honduras using Jeffery Paige's theory of Agrarian revolution

ABSTRACT In 1979 Nicaragua, under the Sandinistas, experienced a genuine, socialist, full scale, agrarian revolution. This thesis examines whether Jeffery Paige's theory of agrarian revolutions would have been successful in predicting this revolution and ln predicting non-revolution in the neighboring country of Honduras. The thesis begins by setting Paige's theory in the tradition of radical theories of revolution. It then derives four propositions from Paige's theory which suggest the patterns of export crops, land tenure changes and class configurations which are necessary for an agrarian and socialist revolution. These propositions are tested against evidence from the twentieth century histories of economic, social and political change in Nicaragua and Honduras. The thesis concludes that Paige's theory does help to explain the occurrence of agrarian revolution in Nicaragua and non-revolution in Honduras. A fifth proposition derived from Paige's theory proved less useful in explaining the specific areas within Nicaragua that were most receptive to Sandinista revolutionary activity.

Vitamin D status and latent tuberculosis infection : a preliminary study in a group of healthy Mexican agricultural workers

Vitamin D metabolites are important in the regulation of bone and calcium homeostasis, but also have a more ubiquitous role in the regulation of cell differentiation and immune function. Severely low circulating 25-dihydroxyvitamin D [25(OH)D] concentrations have been associated with the onset of active tuberculosis (TB) in immigrant populations, although the association with latent TB infection (LTBI) has not received much attention. A previous study identified the prevalence of LTBI among a sample of Mexican migrant workers enrolled in Canada's Seasonal Agricultural Workers Program (SA WP) in the Niagara Region of Ontario. The aim of the present study was to determine the vitamin D status of the same sample, and identify if a relationship existed with LTBI. Studies of vitamin D deficiency and active TB are most commonly carried out among immigrant populations to non-endemic regions, in which reactivation of LTBI has occurred. Currently, there is limited knowledge of the association between vitamin D deficiency and LTBI. Entry into Canada ensured that these individuals did not have active TB, and L TBI status was established previously by an interferon-gamma release assay (IGRA) (QuantiFERON-TB Gold In-Tube®, Cellestis Ltd., Australia). Awareness of vitamin D status may enable individuals at risk of deficiency to improve their nutritional health, and those with LTBI to be aware of this risk factor for disease. Prevalence of vitamin D insufficiency among the Mexican migrant workers was determined from serum samples collected in the summer of 2007 as part of the cross sectional LTBI study. Samples were measured for concentrations of the main circulating vitamin D metabolite, 25(OH)D, with a widely used 1251 250HD RIA (DiaSorin Inc.®, Stillwater, MN), and were categorized as deficient «37.5 nmoI/L), insufficient (>37.5 nmollL, < 80 nmol/L) or sufficient (2::80 nmoI/L). Fisher's exact tests and t tests were used to determine if vitamin D status (sufficiency or insufficiency) or 25(OH)D concentrations significantly differed by sex or age categories. Predictors of vitamin D insufficiency and 25(OH)D concentrations were taken from questionnaires carried out during the previous study, and analyzed in the present study using multiple regression prediction models. Fisher's exact test and t test was used to determine if vitamin D status or 25(OH)D concentration differed by LTBI status. Strength of the relationship between interferongamma (IFN-y) concentration (released by peripheral T cells in response to TB antigens) and 25(OH)D concentration was analyzed using a Spearman correlation. Out of 87 participants included in the study (78% male; mean age 38 years), 14 were identified as LTBI positive but none had any signs or symptoms of TB reactivation. Only 30% of the participants were vitamin D sufficient, whereas 68% were insufficient and 2% were deficient. Significant independent predictors of lower 25(OH)D concentrations were sex, number of years enrolled in the SA WP and length of stay in Canada. No significant differences were found between 25(OH)D concentrations and LTBI status. There was a significant moderate correlation between IFN-y and 25(OH)D concentrations ofLTBI-positive individuals. The majority of participants presented with Vitamin D insufficiency but none were severely deficient, indicating that 25(OH)D concentrations do not decrease dramatically in populations who temporarily reside in Canada but go back to their countries of origin during the Canadian winter. This study did not find a statistical relationship between low levels of vitamin D and LTBI which suggests that in the presence of overall good health, lower than ideal levels of 2S(OH)D, may still be exerting a protective immunological effect against LTBI reactivation. The challenge remains to determine a critical 2S(OH)D concentration at which reactivation is more likely to occur.

Integrating vitrinite reflectance, rock-eval pyrolysis, flourescence microscopy and palynology of the Athabasca oil sands, Kearl Lake area, northeastern Alberta

Three cores from the Kearl Lake Oil Sands area within the Athabasca deposit of northeastern Alberta have been analyzed to understand the thermal history of the McMurray and Clearwater formations of the Lower Cretaceous Mannville Group. The approach involves the integration of vitrinite reflectance (VR), Rock-Eval pyrolysis, fluorescence microscopy, and palynology. Mean VR varies between 0.21 and 0.43% Ro and indicates thermally immature levels equivalent to the rank of lignite to sub-bituminous coal. Although differing lithologies have influenced VR to some extent (i.e., coals and bitumen-rich zones), groundwater influence and oxidation seem not to have measurably altered YR. Rock-Eval analysis points to Type III/IV kerogen, and samples rich in amorphous organic matter (ADM) show little to no fluorescence characteristics, implying a terrestrial source of origin. Palynology reveals the presence of some delicate macerals but lack of fluorescence and abundant ADM suggests some degradation and partial oxidation of the samples.

Identification of renin progenitors in the mouse bone marrow that give rise to B-cell leukaemia

cell of origin and triggering events for leukaemia are mostly unknown. Here we show that the bone marrow contains a progenitor that expresses renin throughout development and possesses a B-lymphocyte pedigree. This cell requires RBP-J to differentiate. Deletion of RBP-J in these renin-expressing progenitors enriches the precursor B-cell gene programme and constrains lymphocyte differentiation, facilitated by H3K4me3 activating marks in genes that control the pre-B stage. Mutant cells undergo neoplastic transformation, and mice develop a highly penetrant B-cell leukaemia with multi-organ infiltration and early death. These reninexpressing cells appear uniquely vulnerable as other conditional models of RBP-J deletion do not result in leukaemia. The discovery of these unique renin progenitors in the bone marrow and the model of leukaemia described herein may enhance our understanding of normal and neoplastic haematopoiesis.