Characterisation of Neisseria meningitidis cc11/ET-15 variant by whole genome sequencing

BACKGROUND Neisseria meningitidis strains belonging to clonal complex 11 is the cause of numerous outbreaks and epidemics in the United States, Canada and Europe, accounting for 49.5% of cases of meningococcal disease caused by serogroup C worldwide. In Brazil, it is the second most frequent clonal complex within this serogroup. The genetic characterisation of cc11/ET-15 variants is important for the epidemiological monitoring of meningococcal disease, through the identification of circulating epidemic clones, to support specific actions of Health Surveillance aiming outbreaks control. OBJECTIVES The objective of this study was to identify features in the genome of cc11/ET-15 clones through whole-genome sequencing (WGS), that differ from cc11/non-ET-15 strains that could explain their virulence. METHODS The whole genome of three cc11/ET-15 representative strains were sequenced with a minimum coverage of 100X with the MiSeq System and compared to the genome of cc11/non-ET-15 strains. RESULTS Genome analysis of cc11/ET-15 variants showed the presence of resistance factors, mobile genetic elements and virulence factors not found in cc11/non-ET-15 strains. MAIN CONCLUSIONS Our results show that these strains carry virulence factors not identified in cc11/non-ET-15 strains, which could explain the high lethality rates attributed to this clone worldwide.

Childhood aplastic anaemia with paroxysmal nocturnal haemoglobinuria clones: A retrospective single-centre study in South Africa

Background: Paroxysmal nocturnal haemoglobinuria (PNH) clones in children are rare but commonly associated with aplastic anaemia (AA) and myelodysplasia.Objective: This study aimed to determine the prevalence of PNH clones in paediatric patients with idiopathic AA, identify differences in clinical and laboratory features and outcomes, and determine the impact of clone size on clinical presentation.Methods: Patients with confirmed idiopathic AA who were tested for PNH between September 2013 and January 2018 at the Inkosi Albert Luthuli Central Hospital, Durban, KwaZulu-Natal, South Africa, were included. PNH clones were detected in neutrophils and monocytes by flow cytometry using fluorescent aerolysin, CD24, CD66b and CD14. Results: Twenty-nine children with AA were identified and 11 were excluded. Ten patients (10/18, 55.6%) had PNH clones ranging from 0.11% to 24%. Compared to the PNH-negative group, these children were older (median: 10 years vs 4 years, p= 0.02) and had significantly lower total white cell counts (median 1.7 × 109/L vs 3.2 × 109/L; p= 0.04). There was no difference in median absolute neutrophil count or haemoglobin concentration. Four patients in each group received immunosuppressive therapy (IST). At six months, all four patients with PNH clones had responded, compared to one in the PNH-negative group. Conclusion: More than half of children with AA had a PNH clone. The size of the clone did not impact clinical severity; however, IST use may positively impact prognosis. We recommend early initiation of IST in patients with AA to avoid delays associated with human leukocyte antigen typing.

Use and response to phosphorus by potato clones in two cropping systems / Utilização e resposta ao fósforo por clones de batata em dois sistemas de cultivo

The selection of potato plants (Solanum tuberosum L.) that are efficient in the use of phosphorus (P) plays an important role in increasing crop productivity, reducing the cost of production due to the high price of phosphate fertilizers, as well as reducing the pollution of the environment due to the better use of the applied fertilizers. The objective of this work was to compare the method of selection of potato clones for the efficiency of use and response to P between in vitro and off - soil systems with the use of sand as substrate. To that end, potato clones SMIC 148-A, Dakota Rose, SMINIA 793101-3, SMIB 106-7, SMIF 212-3, SMIJ 319-1 and P 150 were cultivated at low and high levels of P in the culture systems in vitro (1,935 and 19,346 mg P L-1) and off-soil (2.32 and 23.2 mg P L-1). The selection of potato clones using only as a criterion the accumulation of P under low nutrient level is not adequate, both in off-soil and in vitro cultivation. Clones selected as being more efficient in the use of P in in vitro cultivation do not prove to be necessarily more efficient in off-soil cultivation. No clone remains in the same classification group regarding the efficiency of use and response to P, based on the production of dry mass, in the two cropping systems.

A seleção de plantas de batata (Solanum tuberosum L.) que são eficientes no uso de fósforo (P) desempenha um papel importante no aumento da produtividade das culturas. reduzindo o custo de produção devido ao alto preço dos fertilizantes fosfatados. além de reduzir a poluição do meio ambiente devido ao melhor uso dos fertilizantes aplicados. O objetivo deste trabalho foi comparar o método de seleção de clones de batata quanto à eficiência de uso e resposta ao P entre sistemas in vitro e fora do solo com o uso de areia como substrato. Para o efeito. os clones de batata SMIC 148-A. Dakota Rose. SMINIA 793101-3. SMIB 106-7. SMIF 212-3. SMIJ 319-1 e P 150 foram cultivados a baixos e altos níveis de P nos sistemas de cultivo in vitro (1.935 e 19.346 mg P L-1) e fora do solo (2.32 e 23.2 mg P L-1). A seleção de clones de batata utilizando apenas como critério o acúmulo de P em baixo nível de nutrientes não é adequada. tanto no cultivo fora do solo quanto no in vitro. Clones selecionados como mais eficientes no uso de P em cultivo in vitro não se mostraram necessariamente mais eficientes no cultivo fora do solo. Nenhum clone permanece no mesmo grupo de classificação quanto à eficiência de uso e resposta ao P. baseado na produção de massa seca nos dois sistemas de cultivo.

Tracking methicillin-resistant Staphylococcus aureus clones in a hospital in Southern Mexico / Seguimiento de clonas de Staphylococcus aureus resistentes a meticilina en un hospital en el sur de México

Abstract: Objective: To describe the clinical and molecular characteristics of methicillin-resistantStaphylococcus aureus(MRSA) strains that were collected in theHospital Regional de Alta Especialidad de Veracruz(HRV). Materials and methods: A total of 107 MRSA strains from individual patients were examined. The strains were collected between September 2009 and September 2010. The clinical and demographic characteristics of patients were analyzed; molecular typing by pulsed-field gel electrophoresis (PFGE), staphylococcal chromosomal cassette mec (SCCmec) typing and multilocus sequence typing were used to characterize the isolates. Results: Two PFGE patterns (NY/J and IB) were identified with 4 and 3 subtypes respectively. The isolates analyzed showed two SCCmec types (I and II) and two sequence types (ST): ST247 and ST5 related with the Iberian and New York/Japan clones respectively. Conclusion: This study establishes the presence of two very important clonal lineages of MRSA: New York/Japan and Iberian clone in the hospital environment.

Resumen: Objetivo: Describir las características clínicas y moleculares de cepas deStaphylococcus aureusresistentes a meticilina (SARM) que fueron recolectadas en el Hospital Regional de Alta Especialidad de Veracruz (HRV). Material y métodos: Un total de 107 cepas de SARM fueron analizadas en el presente estudio. Las cepas estudiadas fueron recolectadas de septiembre de 2009 a septiembre de 2010. Las características clínicas y demográficas de los pacientes fueron analizadas; la tipificación molecular de las cepas se hizo por electroforesis en campos pulsados, casete cromosomal estafilococócico (SCCmec, en inglés) y secuenciación de múltiples locus. Resultados: Dos patrones electroforéticos (NY/J y IB) fueron identificados con 4 y 3 subtipos respectivamente. Los aislamientos analizados mostraron dos tipos de SCCmec (I y II) y dos secuencias tipo (ST): ST247 y ST5 relacionados con las clonas Ibérica y Nueva York/Japón respectivamente. Conclusión: Este estudio estableció la presencia en el medio hospitalario de dos linajes clonales de SARM importantes: Nueva York/Japón e Ibérico.

Application of pulsed field gel electrophoresis for molecular characterization of vancomycin resistant Staphylococcus aureus isolates from hospitals

Aim: The present study was conducted to characterize VRSA isolates on the basis of pulsed field gel electrophoresis (PFGE) and the presence of spa gene, recovered from different Doon Valley Hospitals.Methodology: Six VRSA isolates were analyzed using PFGE and spa typing. spa gene coded Protein A was used as a genetic marker for the characterization of Staphylococcus aureus isolates. Dendrogram were constructed on the basis of unweighted pair group method with arithmetic means (UPGMA method) for clusters analyses.Results: Dendrogram finally showed two major banding patterns at about 85% similarity designated as PFGE type A and PFGE type B exhibiting differences of 4-6 bands. The length of spa gene varied from 1200 to 1500 bp, showing variation in length. The most prevalent length was 1200bp.

Japanese encephalitis virus: Biological clones from a clinical isolate quasispecies show differing neurovirulence in vitro and in a mouse model

@#The Japanese encephalitis virus (JEV), a leading cause of encephalitis, exists as quasispecies in clinical isolates. Using a limiting dilution method combined with immunohistochemistry to detect viral antigens, 10 biological clones were isolated and purified from a clinical JEV isolate (CNS138/9) derived from an autopsy brain. These biological clones were tested for neurovirulence in SK-N-MC and NIE-115 neuronal cells, and a 2-week-old, footpad-infected, JE mouse model. Nine clones were found to be neurovirulent; one clone neuroattenuated. Although further studies are needed to determine genotypic differences, if any, in these clones, the limiting dilution purification and neurovirulence testing methods described herein should be useful for phenotypic studies of quasispecies of neurotropic viruses in general, and JEV and other flaviviruses in particular.

Effect of down-regulation of sex determining region Y-box 9 on epithelial mesenchymal transition and cloning of oral squamous carcinoma cells / 华西口腔医学杂志

OBJECTIVE@#To investigate the effect of sex determining region Y-box 9 (SOX9) on epithelial mesenchymal transition (EMT) and cloning of oral squamous cell carcinoma (OSCC).@*METHODS@#siRNA control, SOX9 siRNA were transfected into BcaCD885 cells in OSCC. Simultaneously, cells that did not undergo transfection were used as the control. Quantitative real time polymerase chain reaction (qRT-PCR) and Western blot were used to select SOX9 siRNA1 with enhanced interference effect. A cell cloning assay was used to determine the cell's clone formation ability. E-cadherin and Vimentin expressions were detected by immunofluorescence. The expressions of E-cadherin, matrix metalloprotease 2 (MMP-2), Vimentin and matrix metalloprotease 9 (MMP-9) were detected by Western blot. Cell invasion and migration were detected in the Transwell compartment.@*RESULTS@#The levels of SOX9 mRNA and protein in SOX9 siRNA cells were significantly lower than those of the control (P<0.05). An increase in the number of SOX9 siRNA1 cell clonesled to the considerable decrease of the number of cell invasion and migration. In addition, levels of MMP-2 and MMP-9 proteins in cells decreased significantly compared with the control (P<0.05). The level of Vimentin expression in SOX9 siRNA1 cells decreased, and expression level of E-cadherin was elevated. Cell EMT was inhibited compared with the control, and the difference was statistically significant (P<0.05).@*CONCLUSIONS@#Down-regulation of SOX9 inhibited EMT, clonogenic formation, cell invasion and OSCC migration.

PLZFposc-KITpos-delineated A1-A4-differentiating spermatogonia by subset and stage detection upon Bouin fixation / 亚洲男科学杂志(英文版)

While hallmarks of rodent spermatogonia stem cell biomarkers' heterogeneity have recently been identified, their stage and subset distributions remain unclear. Furthermore, it is currently difficult to accurately identify subset-specific SSC marker distributions due to the poor nuclear morphological characteristics associated with fixation in 4% paraformaldehyde. In the present study, testicular cross-sections and whole-mount samples were Bouin fixed to optimize nuclear resolution and visualized by immunohistochemistry (IHC) and immunofluorescence (IF). The results identified an expression pattern of PLZFhighc-KITpos in A1 spermatogonia, while A2-A4-differentiating spermatogonia were PLZFlowc-KITpos. Additionally, this procedure was used to examine asymmetrically expressing GFRA1 and PLZF clones, asymmetric Apr and false clones were distinguished based on the presence or absence of TEX14, a molecular maker of intercellular bridges, despite having identical nuclear morphology and intercellular distances that were <25 μm. In conclusion, this optimized Bouin fixation procedure facilitates the accurate identification of spermatogonium subsets based on their molecular profiles and is capable of distinguishing asymmetric and false clones. Therefore, the findings presented herein will facilitate further morphological and functional analysis studies and provide further insight into spermatogonium subtypes.

Dominance of international high-risk clones in carbapenemase-producing Pseudomonas aeruginosa: Multicentric molecular epidemiology report from India

Background: Pseudomonas aeruginosa is one of the most common opportunistic pathogens that cause severe infections in humans. The burden of carbapenem resistance is particularly high and is on the rise. Very little information is available on the molecular mechanisms and its clonal types of carbapenem-resistant P. aeruginosa seen in Indian hospitals. This study was undertaken to monitor the ?-lactamase profile and to investigate the genetic relatedness of the carbapenemase-producing (CP) P. aeruginosa collected across different hospitals from India. Materials and Methods: A total of 507 non-duplicate, carbapenem-resistant P. aeruginosa isolated from various clinical specimens collected during 2014–2017 across seven Indian hospitals were included. Conventional multiplex polymerase chain reaction for the genes encoding beta-lactamases such as extended-spectrum beta-lactamase (ESBL) and carbapenemase were screened. A subset of isolates (n = 133) of CP P. aeruginosa were genotyped by multilocus sequence typing (MLST) scheme. Results: Of the total 507 isolates, 15%, 40% and 20% were positive for genes encoding ESBLs, carbapenemases and ESBLs + carbapenemases, respectively, whilst 25% were negative for the ?-lactamases screened. Amongst the ESBL genes, blaVEB is the most predominant, followed by blaPER and blaTEM, whilst blaVIM and blaNDM were the most predominant carbapenemases seen. However, regional differences were noted in the ?-lactamases profile across the study sites. Genotyping by MLST revealed 54 different sequence types (STs). The most common are ST357, ST235, ST233 and ST244. Six clonal complexes were found (CC357, CC235, CC244, CC1047, CC664 and CC308). About 24% of total STs are of novel types and these were found to emerge from the high-risk clones. Conclusion: This is the first large study from India to report the baseline data on the molecular resistance mechanisms and its association with genetic relatedness of CP P. aeruginosa circulating in Indian hospitals. blaVIM- and blaNDM-producing P. aeruginosa is the most prevalent carbapenemase seen in India. Majority of the isolates belongs to the high-risk international clones ST235, ST357 and ST664 which is a concern.

Initial development and sample dimensioning of rubber tree clones / Desenvolvimento inicial e dimensionamento amostral em clones de seringueira

The current success of rubber cultivation is related to the use of clones adapted to different edaphoclimatic regions. Hence, it is important to evaluate variables that are correlated with yield. However, a common problem is choosing the plot dimensions where these variables will be measured. In view of the above, the objective of this work was to evaluate the initial development of two rubber tree clones and to determine the ideal sample unit size to characterize trunk circumference, tree height and bark thickness. The variables circumference at breast height (CBH), total height (Ht) and bark thickness (BT) were measured in seven plots of 680.4 m², in addition to determining the plot size to satisfactorily sample each of the variables measured in each clone, 52 months after planting. At 52 months, clone RRIM 937 showed better development than RRIM600 in relation to the analyzed variables. The ideal sample unit size is different for the variables in the following order: trunk circumference> total height> bark thickness. The measurement of plots with 15 trees is adequate to represent the variability of the analyzed variables, considering the acceptable error of 10%.

O sucesso atual do cultivo de seringueira está relacionado ao uso de clones adaptados a diferentes regiões edafoclimáticas. Portanto, é importante avaliar as variáveis que estão correlacionadas com o rendimento. No entanto, um problema comum é escolher as dimensões da área onde essas variáveis serão medidas. Este trabalho foi conduzido com objetivo de avaliar o desenvolvimento inicial de dois clones de seringueira e determinar o tamanho da amostra ideal para caracterizar a circunferência do tronco, altura da árvore e espessura da casca. As variáveis circunferência na altura do peito (CBH), altura total (Ht) e espessura da casca (BT) foram medidas em sete parcelas de 680,4 m² e foi estabelecido o tamanho da parcela para amostrar satisfatoriamente cada uma das variáveis medidas em cada clone, 52 meses após o plantio. Aos 52 meses, o clone RRIM 937 apresentou melhor desenvolvimento do que RRIM600 em relação às variáveis analisadas. O tamanho ideal da unidade de amostra é diferente para as variáveis na seguinte ordem: circunferência do tronco > altura total > espessura da casca. A medida de parcelas com 15 árvores é adequada para representar a variabilidade das variáveis analisadas, considerando o erro aceitável de 10%.

Consortium of eucalyptus with forage sorghum in semiarid of Minas Gerais State / Clones de eucalipto em consórcio com o sorgo forrageiro no semiárido de Minas Gerais

ABSTRACT: The objective was to estimate the wood yield and essential oil content in three clones of eucalyptus that were planted in four contrasting arrangements and intercropped with sorghum. Eucalyptus clones MA2001 (Eucalyptus camaldulensis x E. tereticornis), A144 (Eucalyptus urophylla x E. grandis), and GG100 (Eucalyptus urophylla x E. grandis), were planted in single rows (10x2m), double rows (2x3+15m and 2x3+20m); and, triple rows (2x3x2+10m) in a randomized, complete block design experiment with four replicates. Our results demonstrated that planting spacing did not influence the essential oil yield or diameter at breast height in the clones. However, higher density plantings were shown to result in higher fresh weight of branches and leaves per plant. MA2001 grew taller, produced higher quantity of fresh biomass of branches and leaves per plant and volume of wood per hectare, and yielded more essential oil yield than the other clones. We concluded that MA2001 is the most suitable of the clones tested here for cultivation in water deficit conditions.

RESUMO: O objetivo do trabalho foi estimar o rendimento de madeira e o teor de óleo essencial de clones de eucalipto em diferentes arranjos, consorciado com sorgo. Foram utilizados 3 clones de eucalipto: Clone 1 (MA2001-Eucalyptus camaldulensis x E. tereticornis); Clone 2 (A144-Eucalyptus urophylla x E. grandis); Clone 3 (GG100 - Eucalyptus urophylla x E. grandis). Os clones de eucalipto foram plantados nos diferentes arranjos: em linha dupla, sendo 2x3+15m e 2x3+20m; linhas simples 10x2m e em linha tripla, sendo 2x3x2+10m. O delineamento experimental adotado foi o de blocos casualizados, com quatro repetições, sendo os tratamentos referentes a 3 clones de eucalipto e 4 arranjos de plantas diferentes. Os resultados mostraram que menores ou maiores espaçamentos não influenciaram no rendimento de óleo essencial e no diâmetro da altura do peito. Porém, menores espaçamentos podem proporcionar maior matéria fresca de ramos e folhas por planta. O clone MA2001 (Camaldulensis x Tereticornis) por ter apresentado maior altura de plantas, maior matéria fresca de galho e folhas por planta, maior volume de madeira por hectare e maior produtividade de óleo essencial pode ser o mais indicado para cultivo em condições com maior déficit hídrico.

Effects of cell-to-cell communication and histone acetyltransferase on the change of osteogenic differentiation ability among single-cell clones from healthy periodontium with heterogeneity of osteogenic differentiation abilities / 中华口腔医学杂志

Objective@#To investigate the effect of cell-to-cell communication amongst single-cell clones from healthy periodontium with different osteogenic differentiation potentials on change of osteogenic differentiation capabilities and the role histone acetyltransferase partaken in this process.@*Methods@#In order to research the change of osteogenic differentiation ability via cell-to-cell communication, indirect co-culture method was used by placing two single-cell clones with different osteogenesis potentials in each of the 6-well plates. Blank control, weak and strong osteogenic groups were set up, corresponding to Transwell chambers with blank, cells of weak osteogenesis ability and cells of strong osteogenesis ability, respectively. Each group was made in triplicate. After co-culture for four days, Transwell chamber was removed. Quantitative real-time PCR (qPCR) and alizarin red staining were employed to detect the change of osteogenic differentiation ability. The acetylation level of H3 was measured by using Western blotting. Histone acetyltransferases were detected by qPCR.@*Results@#Single-cell clones were ensured from mesenchymal stem cells by flow cytometer, the positive expression of CD29, CD90, CD105, CD146 was (99.80±0.02)%, (99.36±0.18)%, (99.41±0.05)% and (95.10±2.11)%, respectively. And CD31 and CD34 expression were (0.29±0.11)% and (0.22±0.13)%, respectively. Alizarin red and oil red O staining confirmed that single-cell clones had the abilities of adipogenesis and osteogenesis. Alkaline phosphatase (ALP) and alizarin red staining indicated that different single-cell clones were heterogeneity in osteogenesis differentiation. Indirect co-culture indicated that the mRNA expression of osteocalcin (OCN) were 14.24±5.60 and 4.78±2.90, respectively and Runt-related transcription factor 2 (RUNX2) were 2.75±1.44 and 1.61±0.44, respectively, in strong and weak osteogenic groups. They were significantly higher compared to the blank group (the mRNA expression of OCN and RUNX2 were 1.00±0.47 and 1.00±0.39, respectively). The expression of OCN and RUNX2 were also higher in strong osteogenic group than that in weak osteogenic group (P<0.05). The mean gray level of the acetylation of H3 in strong osteogenic group (0.76±0.09) and weak osteogenic group (0.54±0.12) were also higher than that in the blank group (0.30±0.04)(P<0.05). qPCR results showed that KAT6A in strong osteogenic group exhibiting higher expression (P<0.05) compared to weak osteogenic group and the blank group, which were corresponding to the changes of acetylation levels.@*Conclusions@#Single-cell clones from healthy periodontium showed heterogeneity in osteogenic differentiation abilities. Single-cell clones with strong osteogenesis abilities had an advantage over others by promoting others' osteogenesis differentiation and this change mediated by cell-to-cell communication might be caused by modulating KAT6A to affect the acetylation level of histone.

Local circulating clones of Staphylococcus aureus in Ecuador

ABSTRACT The spread of pandemic Staphylococcus aureus clones, mainly methicillin-resistant S. aureus (MRSA), must be kept under surveillance to assemble an accurate, local epidemiological analysis. In Ecuador, the prevalence of the USA300 Latin American variant clone (USA300-LV) is well known; however, there is little information about other circulating clones. The aim of this work was to identify the sequence types (ST) using a Multiple-Locus Variable number tandem repeat Analysis 14-locus genotyping approach. We analyzed 132 S. aureus strains that were recovered from 2005 to 2013 and isolated in several clinical settings in Quito, Ecuador. MRSA isolates composed 46.97% (62/132) of the study population. Within MRSA, 37 isolates were related to the USA300-LV clone (ST8-MRSA-IV, Panton-Valentine Leukocidin [PVL] +) and 10 were related to the Brazilian clone (ST239-MRSA-III, PVL−). Additionally, two isolates (ST5-MRSA-II, PVL−) were related to the New York/Japan clone. One isolate was related to the Pediatric clone (ST5-MRSA-IV, PVL−), one isolate (ST45-MRSA-II, PVL−) was related to the USA600 clone, and one (ST22-MRSA-IV, PVL−) was related to the epidemic UK-EMRSA-15 clone. Moreover, the most prevalent MSSA sequence types were ST8 (11 isolates), ST45 (8 isolates), ST30 (8 isolates), ST5 (7 isolates) and ST22 (6 isolates). Additionally, we found one isolate that was related to the livestock associated S. aureus clone ST398. We conclude that in addition to the high prevalence of clone LV-ST8-MRSA-IV, other epidemic clones are circulating in Quito, such as the Brazilian, Pediatric and New York/Japan clones. The USA600 and UK-EMRSA-15 clones, which were not previously described in Ecuador, were also found. Moreover, we found evidence of the presence of the livestock associated clone ST398 in a hospital environment.

Genetic relationship between Escherichia coli strains isolated from dairy mastitis and from the stable fly Stomoxys calcitrans / Caracterização genotípica de Escherichia coli isolados de leite com mastite e da mosca dos estábulos Stomoxys calcitrans

The stable fly Stomoxys calcitrans (Linnaeus, 1758) has been described as a potential spreader of infectious agents to cattle herds. Among the agents transmitted by this fly, Escherichia coli has attracted attention due to its potential to cause gastrointestinal disorders as well as environmental mastitis in dairy cows. Therefore, the aim of this study was to isolate and to assess the genetic diversity and the clonal relatedness among E. coli isolates from the milk of dairy mastitis and from stable flies anatomical sites by the Random Amplification of Polymorphic DNA (RAPD-PCR) technique. The molecular typing revealed a high degree of genetic polymorphism suggesting that these microorganisms have a non-clonal origin. Identical electrophoretic profiles were observed between E. coli isolates from different flies, different mammary quarters of the same cow and from cows on a single farm. These results reveal the circulation of the same bacterial lineages and suggest the role of the stable fly in bacterial dispersion. Considering the high pathogenic potential of this bacterial species, our findings alert to a more effective health surveillance.(AU)

A mosca dos estábulos Stomoxys calcitrans é descrita como um importante dispersor de agentes infecciosos aos bovinos. Dentre os agentes veiculados por esta mosca a bactéria Escherichia coli ganha relevância devido ao seu potencial em desenvolver alterações gastroentéricas, bem como mastite bovina ambiental. Desta forma, objetiva-se com este estudo isolar e acessar a diversidade genética e relação de clonalidade entre isolados de E. coli provenientes de casos de mastite e de moscas dos estábulos utilizando a técnica da Amplificação Randômica do DNA Polimórfico (RAPD). A tipagem molecular revelou elevado polimorfismo genético sugerindo que esses microrganismos têm origem não clonal. Perfis eletroforéticos idênticos entre si foram observados entre amostras isoladas de diferentes moscas, quartos mamários de uma mesma vaca, bem como de diferentes vacas dentro de uma mesma propriedade. Esses resultados revelam a circulação de uma mesma linhagem bacteriana e sugerem o papel da Stomoxys calcitrans na dispersão bacteriana. Considerando o elevado potencial patogênico dessa espécie bacteriana, nossos achados alertam para uma vigilância sanitária mais efetiva.(AU)

Prevalence of Major Methicillin-Resistant Staphylococcus aureus Clones in Korea Between 2001 and 2008

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) are important pathogens causing nosocomial infections in Korean hospitals. This study aimed to investigate the epidemiological and genetic diversity of clinical S. aureus isolates in healthcare settings from 2001 to 2008. METHODS: Samples and data were obtained from 986 individuals as part of the National Antimicrobial Surveillance Project, involving 10 regions nationwide. Molecular typing studies, including multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing were performed, and a representative clone of Korean MRSA was classified by combinational grouping using a DiversiLab (DL; bioMérieux, France) repetitive element polymerase chain reaction (rep-PCR) system. RESULTS: Nine Korean MRSA clones (KMRSA-1 to -9) were identified by analysis of genetic backgrounds and molecular characteristics. KMRSA-1 to -3, expressing clonal complex (CC) 5 (carrying SCCmec II), CC8 (carrying SCCmec III), and CC72 (carrying SCCmec IV) were spread nationwide. In contrast, KMRSA-6 was highly prevalent in Gyeongsangnam-do, and KMRSA-4 was highly prevalent in Jeollanam-do and Jeollabuk-do. CONCLUSIONS: Epidemic KMRSA clones were genetically similar to major clones identified from the USA, with the exception of KMRSA-2, which had the SCCmec III type. Our results provide important insights into the distribution and molecular genetics of MRSA strains in Korea and may aid in the monitoring of MRSA spread throughout the country.

Diversity among genotypes of conilon coffee selected in Espírito Santo state / Diversidade entre genótipos de café conilon selecionados no estado do Espírito Santo

The use of multivariate techniques for factor analysis is an efficient alternative for coffee breeding programs. This study aimed to evaluate the genetic diversity of 60 genotypes of conilon coffee based on agronomic performance in the northern state of Espírito Santo and to estimate the relative contribution of different agronomic characteristics towards the diversity of the species. The data were collected in an experiment conducted on the Experimental Farm of Bananal do Norte (Instituto Capixaba de Pesquisa, Assistência Técnica e Extenção Rural ­ INCAPER) in the southern state of Espírito Santo, and 12 agronomic characteristics were evaluated over four sequential harvests (4 years). Significant differences between the genotypes were observed for all of the characteristics, indicating the possibility of exploiting the high genetic variability to classify the genotypes into different groups based on their similarities. Of the agronomic characteristics, the duration of the ripening cycle was the variable that contributed the most to the variability among the 60 genotypes, with a relative contribution of 70.02%.

A utilização de técnicas multivariadas de análise de fatores é uma alternativa eficiente utilizada no melhoramento genético do cafeeiro. O presente trabalho objetivou avaliar a divergência genética de 60 clones de café conilon, selecionados pelo seu desempenho no norte do Estado do Espírito Santo, e estimar a contribuição relativa de diferentes características agronômicas para a diversidade da espécie. Os dados foram coletados em experimento conduzido na Fazenda Experimental Bananal do Norte (INCAPER), considerando 12 características agronômicas, avaliadas através de médias de quatro safras. Diferenças significativas entre os genótipos foram observadas para todas as características avaliadas, indicando a possibilidade de exploração da alta variabilidade genética para a classificação dos genótipos em diferentes grupos homogêneos, baseado em suas similaridades. Dentre as características agronômicas, a duração do ciclo de maturação foi a variável que mais contribuiu para a variabilidade entre os 60 genótipos, com contribuição relativa de 70,02%.

Esterase isozymes patterns of grape vine (Vitis vinifera L) are altered in response to fungicide exposure / Padrões de isozimas Esterases de videira (Vitis vinifera L) alterados em resposta à exposição a fungicidas

Current analysis characterizes the effect of different fungicides often applied for pest control on α−and ß-esterase patterns of four economically important table-wine grape cultivars (Italia, Rubi, Benitaka and Brasil) of Vitis vinifera. The α- and ß-esterase patterns in bud leaves of the cultivars were assessed by native PAGE analysis. Cabrio Top® compound inhibited EST-2, EST-5, EST-6, EST-7, EST-8, EST-9 and EST-10 carboxylesterases, whereas EST-4, EST-11, EST-12, EST-13, EST-14 acetylesterases and EST-16 carboxylesterase were detected as weakly stained bands. Carboxylesterases and acetylesterases were also detected as weakly stained bands when exposed to fungicides Orthocide 500®, Positron Duo® and Folicur PM®. No changes in α- and ß-esterase patterns were reported when the vines were exposed to the fungicides Rovral SC®, Kumulus DF®, Curzate M®, Score® or Cuprogarb 500®. The evidence of functional changes in carboxylesterase and acetylesterase levels in current study is a warning to grape producers on the dangers inherent in the indiscriminate use of potent and modern fungicides extensively used in agriculture. The inhibition effect of fungicides on esterase isozyme molecules seems to be independent of the fungicide chemical.

O presente estudo caracterizou o efeito de diferentes fungicidas comumente aplicados como medidas de controle de pragas sobre padrões de α- e ß-esterases de quatro importantes cultivares de uva de mesa (Itália, Rubi, Benitaka e Brasil) de Vitis vinifera. Os padrões de α- e ß-esterases de brotos foliares das cultivares foram avaliados por PAGE. O composto Cabrio Top® inibiu as carboxilesterases EST-2, EST-5, EST-6, EST-7, EST-8, EST-9 e EST-10, enquanto as acetilesterases EST-4, EST-11, EST-12, EST-13, EST-14 e a carboxilesterase EST-16 foram detectadas como bandas fracamente coradas. As carboxilesterases e acetilesterases também foram detectadas como bandas fracamente coradas quando expostas aos fungicidas Orthocide 500®, Positron Duo® e Folicur PM®. Não foram observadas alterações nos padrões de α- e ß-esterases quando as videiras foram expostas aos fungicidas Rovral SC®, Kumulus DF®, Curzate M®, Score® ou Cuprogarb 500®. A evidência de alterações em nível funcional em carboxilesterases e acetilesterases, apresentada neste estudo, pode servir como um alerta aos produtores de uva dos perigos inerentes ao uso indiscriminado de fungicidas potentes e modernos amplamente utilizados hoje na agricultura. O efeito dos fungicidas sobre as enzimas esterases parece ser independente do grupo químico ao qual pertence o fungicida.

Adaptabilidade e estabilidade da produção de café beneficiado em Coffea canephora / Adaptability and stability of Coffea canephora coffee bean yield

O objetivo deste trabalho foi comparar a capacidade discriminatória de métodos para seleção e recomendação de clones de cafeeiros da espécie Coffea canephora, considerando a interação genótipos x anos na distribuição da produtividade de café beneficiado, nos períodos de 2000 a 2004 e de 2007 a 2011. O efeito significativo da interação genótipos x anos indicou que existem clones que apresentaram desempenho não consistente ao longo do tempo. Os métodos de LIN & BINNS (1988) e ANNICCHIARICO (1992) apresentaram resultados similares, de modo que um deles pode ser utilizado em substituição ao outro. O método de EBERHART & RUSSEL (1966) identificou parte dos clones de adaptação específica, enquanto que os métodos, MHPRVG e centroide permitiram caracterizar os clones de maior adaptabilidade e estabilidade, apresentando resultados complementares. A seleção de plantas baseada na distribuição da produtividade de café beneficiado ao longo do tempo permitiu selecionar clones de desempenho superior sem acentuar a bienalidade.

This research aimed to compare methods of selecting Coffea canephoraclones, considering the genotypes x years interaction, based on data of processed coffee yield obtained from 2000 to 2004 and 2007 to 2011. The significant clones x seasons (CS) interaction, indicates that the genotypes showed no consistent performance over the years. The LIN & BINNS (1988) and ANNICCHIARICO (1992) methods showed similar results indicating that one method may be used to replace the other. The EBERHART & RUSSEL (1966) identified some of the specific adaptability clones. The MHPRVG and centroid methods allowed to select clones of higher adaptability and stability. The plant selection based on the distribution of coffee productivity over time allowed the selection of superior clones without accentuate biannuality.

Resurgence of pertussis at the age of vaccination: clinical, epidemiological, and molecular aspects / Ressurgimento da coqueluche na era vacinal: aspectos clínicos, epidemiológicos e moleculares

OBJECTIVE: Report the incidence, epidemiology, clinical features, death, and vaccination status of patients with whooping cough and perform genotypic characterization of isolates of B. pertussis identified in the state of Paraná, during January 2007 to December 2013.METHODS: Cross-sectional study including 1,209 patients with pertussis. Data were obtained through the Notifiable Diseases Information System (Sistema de Informação de Agravos de Notificação - SINAN) and molecular epidemiology was performed by repetitive sequence-based polymerase chain reaction (rep-PCR; DiversiLab(r), bioMerieux, France).RESULTS: The incidence of pertussis in the state of Paraná increased sharply from 0.15-0.76 per 100,000 habitants between 2007-2010 to 1.7-4.28 per 100,000 between 2011-2013. Patients with less than 1 year of age were more stricken (67.5%). Fifty-nine children (5%) developed pertussis even after receiving three doses and two diphtheria-tetanus-pertussis (DTP) boosters vaccine. The most common complications were pneumonia (14.5%), otitis (0.9%), and encephalopathy (0.7%). Isolates of B. pertussis were grouped into two groups (G1 and G2) and eight distinct patterns (G1: P1-P5 and G2: P6-P8).CONCLUSION: The resurgence of pertussis should stimulate new research to develop vaccines with greater capacity of protection against current clones and also encourage implementation of new strategies for vaccination in order to reduce the risk of disease in infants.

OBJETIVO: Relatar a incidência, os aspectos epidemiológicos, clínicos, a morte e a vacinação de pacientes com coqueluche e fazer a caracterização genotípica de isolados de Bordetella pertussisidentificados no Estado do Paraná, de janeiro de 2007 a dezembro de 2013.MÉTODOS: Estudo transversal, incluindo 1.209 pacientes com coqueluche. Os dados foram obtidos no Sistema de Informação de Agravos de Notificação (Sinan) e a epidemiologia molecular foi feita por PCR baseada em sequências repetitivas (rep-PCR; DiversiLab(r), bioMerieux, France).RESULTADOS: A incidência de coqueluche no Estado do Paraná aumentou acentuadamente de 0,15-0,76 por 100.000 habitantes entre 2007-2010 para 1,7-4,28 por 100.000 habitantes entre 2011-2013. Os pacientes com menos de um ano foram os mais afetados (67,5%); 59 crianças (5%) desenvolveram coqueluche mesmo depois de receber três doses da vacina e dois reforços com a vacina tríplice DTP. As complicações mais comuns foram pneumonia (14,5%), otite (0,9%) e encefalopatia (0,7%). Isolados de B. pertussis foram agrupados em dois grupos (G1 e G2) e oito padrões distintos (G1: P1-P5 e G2: P6-P8).CONCLUSÃO: O ressurgimento da coqueluche vem para sugerir novas pesquisas com o objetivo se desenvolverem vacinas com maior capacidade de proteção contra os clones atuais e também implantar novas estratégias de vacinação, a fim de reduzir o risco de doenças em lactentes.

Immunological response to re-infections with clones of the Colombian strain of Trypanosoma cruzi with different degrees of virulence: influence on pathological features during chronic infection in mice

Re-infections with Trypanosoma cruzi are an aggravating factor for Chagas disease morbidity. The Colombian strain of T. cruzi represents multiclonal populations formed by clonally propagating organisms with different tropisms and degrees of virulence. In the present study, the influence of successive inoculations with clones of the Colombian strain, exhibiting different degrees of virulence, on chronic myocarditis and the humoral and cellular immune responses (Col-C1 high virulence, Col-C8 medium virulence and Col-C5 low virulence) were demonstrated. Mice from three groups with a single infection were evaluated during the acute (14th-30th day) and chronic phases for 175 days. An immunofluorescence assay, ELISA and delayed type hypersensitivity (DTH) cutaneous test were also performed. Mice with a triple infection were studied on the 115th-175th days following first inoculation. The levels of IgM and IgG2a were higher in the animals with a triple infection. DTH showed a higher intensity in the inflammatory infiltrate based on the morphometric analysis during a 48 h period of the triple infection and at 24 h with a single infection. The histopathology of the heart demonstrated significant exacerbation of cardiac inflammatory lesions confirmed by the morphometric test. The humoral responses indicate a reaction to the triple infection, even with clones of the same strain.