CARACTERÍSTICAS DE CULTURAS LÁCTICAS PROBIÓTICAS PARA USO EM PRODUTOS CÁRNEOS FERMENTADOS: SENSIBILIDADE AOS SAIS DE CURA E USO DE ANTIBIÓTICOS PARA CONTAGEM SELETIVA

Os objetivos deste trabalho foram avaliar a resistência de espécies probióticas de Lactobacillus na presença de sais de cura e testar sua sensibilidade frente a diferentes antimicrobianos para o desenvolvimento de meio de cultura seletivo. As culturas Lactobacillus casei, Lactobacillus paracasei e Lactobacillus rhamnosus foram semeadas em ágar MRS contendo concentrações de 0% a 3% de cloreto de sódio e 0 a 200 ppm de nitrito de sódio. O efeito do uso concomitante dos sais de cura foi verificado utilizando-se 3% de cloreto de sódio e 200 ppm de nitrito de sódio. As bactérias probióticas e a cultura starter Pediococcus pentosaceus foram testadas frente a 20 discos de antimicrobianos pela técnica de disco-difusão. O crescimento dos probióticos não apresentou alteração nas diferentes concentrações de cloreto de sódio, assim como nas concentrações de até 200 ppm de nitrito de sódio. Verificou-se resistência ao uso simultâneo de cloreto e nitrito de sódio. Entre os antimicrobianos testados, a tetraciclina apresentou resultados satisfatórios para a inibição dos probióticos permitindo o crescimento isolado da cultura starter. Os probióticos apresentaram desenvolvimento satisfatório na presença dos sais de cura, demonstrando potencial para sua utilização em produtos cárneos fermentados com ação probiótica. CHARACTERISTICS OF PROBIOTIC CULTURES FOR THE USE IN FERMENTED MEAT PRODUCTS - SENSIBILITY TO CURING SALTS AND ANTIBIOTIC USE FOR THE SELECTIVE ENUMERATION Abstract The objective of this study was to evaluate the resistance of probiotic species of Lactobacillus in the presence of curing salts and to test their sensibility in the presence of antibiotics for the development of a selective culture media. The probiotic cultures, Lactobacillus casei, Lactobacillus paracasei spp paracasei and Lactobacillus casei spp rhamnosus, were plated in MRS agar with concentration of 0 to 3% of sodium chloride and 0 to 200 ppm of sodium nitrite. The inhibitory effect of 3% sodium chloride and 200 ppm sodium nitrite in simultaneous use was evaluated for the probiotic strains. The sensibility of the starter culture, Pediococcus pentosaceus and the probiotic cultures in the presence of antibiotic was carried out using 20 different antibiotic discs by the disc-diffusion technique. The growth of the probiotic cultures wasnt affected at different concentrations of sodium chloride, and even to concentrations of 200 ppm of sodium nitrate. Resistance was verified by simultaneous use of sodium chloride and nitrate. Among the tested antibiotics, tetracycline showed a satisfactory inhibition effect for the probiotic strains, since the growth of starter Pediococcus pentosaceus was not affected. The probiotics showed satisfactory growth in the presence of curin

Justifications of national gambling policies in France and Finland

AIMS - The principles of free trade and free circulation of services within the European Union have created pressures to make the strictly controlled European gambling markets more open. According to the Court of Justice of the European Union, restrictions on gambling are only allowed if they are justified in admissible terms of consumer protection, prevention of criminal activity and protection of public order. This study compares the gambling laws of two European societies, France and Finland, to analyse how their legal frames of gambling have been adjusted to these principles. DESIGN - The data consists of up-to-date legislation on gambling in Finland and France. A qualitative analysis was conducted to study whether new ways of justifying have been included in legislative texts and if these are substantiated by measures related to consumer protection or crime prevention. RESULTS - France has mainly justified its restrictive policies on gambling in terms of preventing criminal activities while the Finnish legislation highlights the charitable causes funded by gambling proceeds, a claim not accepted by the Court of Justice of the European Union. Consumer protection is increasingly stressed in both countries, and the range of rationales has also grown notably since 2007. CONCLUSION - While the vocabularies of justification accepted by the CJEU have expanded since 2007, these have not been substantiated by many new legislative measures. This is not attributed to political ill will but rather to the difficulty of changing existing legislative traditions.Peer reviewe

Identification and Differentiation of the Twenty Six Bluetongue Virus Serotypes by RT–PCR Amplification of the Serotype-Specific Genome Segment 2

Bluetongue (BT) is an arthropod-borne viral disease, which primarily affects ruminants in tropical and temperate regions of the world. Twenty six bluetongue virus (BTV) serotypes have been recognised worldwide, including nine from Europe and fifteen in the United States. Identification of BTV serotype is important for vaccination programmes and for BTV epidemiology studies. Traditional typing methods (virus isolation and serum or virus neutralisation tests (SNT or VNT)) are slow (taking weeks, depend on availability of reference virus-strains or antisera) and can be inconclusive. Nucleotide sequence analyses and phylogenetic comparisons of genome segment 2 (Seg-2) encoding BTV outer-capsid protein VP2 (the primary determinant of virus serotype) were completed for reference strains of BTV-1 to 26, as well as multiple additional isolates from different geographic and temporal origins. The resulting Seg-2 database has been used to develop rapid (within 24 h) and reliable RT–PCR-based typing assays for each BTV type. Multiple primer-pairs (at least three designed for each serotype) were widely tested, providing an initial identification of serotype by amplification of a cDNA product of the expected size. Serotype was confirmed by sequencing of the cDNA amplicons and phylogenetic comparisons to previously characterised reference strains. The results from RT-PCR and sequencing were in perfect agreement with VNT for reference strains of all 26 BTV serotypes, as well as the field isolates tested. The serotype-specific primers showed no cross-amplification with reference strains of the remaining 25 serotypes, or multiple other isolates of the more closely related heterologous BTV types. The primers and RT–PCR assays developed in this study provide a rapid, sensitive and reliable method for the identification and differentiation of the twenty-six BTV serotypes, and will be updated periodically to maintain their relevance to current BTV distribution and epidemiology (http://www.reoviridae.org/dsRNA_virus_proteins/ReoID/rt-pcr-primers.htm)

Complete Genome Characterisation of a Novel 26th Bluetongue Virus Serotype from Kuwait

Bluetongue virus is the “type” species of the genus Orbivirus, family Reoviridae. Twenty four distinct bluetongue virus (BTV) serotypes have been recognized for decades, any of which is thought to be capable of causing “bluetongue” (BT), an insect-borne disease of ruminants. However, two further BTV serotypes, BTV-25 (Toggenburg orbivirus, from Switzerland) and BTV-26 (from Kuwait) have recently been identified in goats and sheep, respectively. The BTV genome is composed of ten segments of linear dsRNA, encoding 7 virus-structural proteins (VP1 to VP7) and four distinct non-structural (NS) proteins (NS1 to NS4). We report the entire BTV-26 genome sequence (isolate KUW2010/02) and comparisons to other orbiviruses. Highest identity levels were consistently detected with other BTV strains, identifying KUW2010/02 as BTV. The outer-core protein and major BTV serogroup-specific antigen “VP7” showed 98% aa sequence identity with BTV-25, indicating a common ancestry. However, higher level of variation in the nucleotide sequence of Seg-7 (81.2% identity) suggests strong conservation pressures on the protein of these two strains, and that they diverged a long time ago. Comparisons of Seg-2, encoding major outer-capsid component and cell-attachment protein “VP2” identified KUW2010/02 as 26th BTV, within a 12th Seg-2 nucleotype [nucleotype L]. Comparisons of Seg-6, encoding the smaller outer capsid protein VP5, also showed levels of nt/aa variation consistent with identification of KUW2010/02 as BTV-26 (within a 9th Seg-6 nucleotype - nucleotype I). Sequence data for Seg-2 of KUW2010/02 were used to design four sets of oligonucleotide primers for use in BTV-26, type-specific RT-PCR assays. Analyses of other more conserved genome segments placed KUW2010/02 and BTV-25/SWI2008/01 closer to each other than to other “eastern” or “western” BTV strains, but as representatives of two novel and distinct geographic groups (topotypes). Our analyses indicate that all of the BTV genome segments have evolved under strong purifying selection

Variable fixation promotes callus formation: an experimental study on transverse tibial osteotomies stabilized with locking plates

Background A new locking screw technology, named variable fixation, has been developed aiming at promoting bone callus formation providing initial rigid fixation followed by progressive fracture gap dynamisation. In this study, we compared bone callus formation in osteotomies stabilized with standard locking fixation against that of osteotomies stabilized with variable fixation in an established tibia ovine model. Methods A 3 mm tibial transverse osteotomy gap was stabilized in three groups of six female sheep each with a locking plate and either 1) standard fixation in both segments (group LS) or 2) variable fixation in the proximal and standard fixation in the distal bone segment (group VFLS3) or 3) variable fixation in both segments (group VFLS6). The implantation site and fracture healing were compared between groups by means of radiologic, micro tomographic, biomechanical, and histological investigations. Results Compared to LS callus, VFLS3 callus was 40% larger and about 3% denser, while VFLS6 callus was 93% larger and its density about 7.2% lower. VFLS3 showed 65% and VFLS6 163% larger amount of callus at the cis-cortex. There wasn’t a significant difference in the amount of callus at the cis and trans-cortex in groups featuring variable fixation only. Investigated biomechanical variables were not significantly different among groups and histology showed comparable good healing in all groups. Tissues adjacent to the implants did not show any alteration of the normal structure in all groups. Conclusions Variable fixation promoted the formation of a larger amount of bone callus, equally distributed at the cis and trans cortices. The histological and biomechanical properties of the variable fixation callus were equivalent to those of the standard fixation callus. The magnitude of variable fixation had a biological effect on the formation of bone callus. At the implantation site, the usage of variable fixation did not raise additional concerns with respect to standard fixation. The formation of a larger amount of mature callus suggests that fractures treated with variable fixation might have a higher probability to bridge the fracture gap. The conditions where its usage can be most beneficial for patients needs to be clinically defined

In vivo and in vitro propagation and transmission of Toggenburg orbivirus

The Toggenburg orbivirus (TOV), a recently discovered virus related to bluetongue virus (BTV), has been identified in goats in Switzerland, Italy and Germany. Isolation of TOV in vitro has not yet been achieved and the transmission mechanisms are still unknown. In the experimental infection of pregnant goats described here, TOV could not be detected in secretion/excretion samples or fetal blood. Material from the goat experiment was used as inoculum for propagating the virus in vitro. To enhance the infectivity of TOV several modified protocols, e.g. pretreatment of the virus with trypsin, polyethylene glycol-mediated infection and lipofection were applied. Isolation of TOV, attempts to infect Culicoides nubeculosus by feeding TOV-positive blood and intracerebral inoculation of newborn mice were unsuccessful. The results of these studies suggest that TOV requires specific but different factors than other BTVs for infection and replication outside of its natural caprine host

Variable fixation promotes callus formation: an experimental study on transverse tibial osteotomies stabilized with locking plates

Background A new locking screw technology, named variable fixation, has been developed aiming at promoting bone callus formation providing initial rigid fixation followed by progressive fracture gap dynamisation. In this study, we compared bone callus formation in osteotomies stabilized with standard locking fixation against that of osteotomies stabilized with variable fixation in an established tibia ovine model. Methods A 3 mm tibial transverse osteotomy gap was stabilized in three groups of six female sheep each with a locking plate and either 1) standard fixation in both segments (group LS) or 2) variable fixation in the proximal and standard fixation in the distal bone segment (group VFLS3) or 3) variable fixation in both segments (group VFLS6). The implantation site and fracture healing were compared between groups by means of radiologic, micro tomographic, biomechanical, and histological investigations. Results Compared to LS callus, VFLS3 callus was 40% larger and about 3% denser, while VFLS6 callus was 93% larger and its density about 7.2% lower. VFLS3 showed 65% and VFLS6 163% larger amount of callus at the cis-cortex. There wasn’t a significant difference in the amount of callus at the cis and trans-cortex in groups featuring variable fixation only. Investigated biomechanical variables were not significantly different among groups and histology showed comparable good healing in all groups. Tissues adjacent to the implants did not show any alteration of the normal structure in all groups. Conclusions Variable fixation promoted the formation of a larger amount of bone callus, equally distributed at the cis and trans cortices. The histological and biomechanical properties of the variable fixation callus were equivalent to those of the standard fixation callus. The magnitude of variable fixation had a biological effect on the formation of bone callus. At the implantation site, the usage of variable fixation did not raise additional concerns with respect to standard fixation. The formation of a larger amount of mature callus suggests that fractures treated with variable fixation might have a higher probability to bridge the fracture gap. The conditions where its usage can be most beneficial for patients needs to be clinically defined.ISSN:1471-247