Inoculation methods and agressiveness of Macrophomina phaseolina isolates in cowpea / Métodos de inoculação e agressividade de isolados de Macrophomina phaseolina em feijão-caupi

The fungi Macrophomina phaseolina is the charcoal rot causal agent, one of the most important cowpea crop disease in semiarid regions can causes 100% yield losses. The search for resistant genotypes requires efficient phenotyping. In addition, there is the problem of great variation in aggressiveness between isolates. This study aimed to 1) test three methods of inoculation in semiarid conditions, and 2) to evaluate the aggressiveness of isolates of M. phaseolina. In the first experiment carried out in greenhouse, the inoculations methods were evaluated, using two cowpea lines, three inoculation methods and three pathogen isolates. On the second experiment, fifteen M. phaseolina isolates were inoculated in one cultivar to evaluate their aggressiveness. By assessing the length of the lesions and the severity of the disease using an index, we identified the toothpick inoculation method as the most efficient. Toothpick method allowed to discriminate the genotypes and the aggressiveness of the pathogen.

O fungo Macrophomina phaseolina é o agente etiológico da podridão cinzenta do caule, uma das mais importantes doenças na cultura do feijão caupi em regiões semiáridas, podendo ocasionar perdas de 100%. A busca por genótipos resistentes exige uma fenotipagem eficiente. Além disso tem o problema da grande variação na agressividade entre isolados. Este trabalho teve como objetivos 1) testar três metodologias de inoculação em condições semiáridas, e 2) avaliar a agressividade de isolados de M. phaseolina. No primeiro experimento, conduzido em ambiente protegido, avaliou-se metodologias de inoculação em duas linhagens de caupi, por três métodos de inoculação e três isolados do patógeno. No segundo experimento, 15 isolados de M. phaseolina foram inoculados em uma cultivar de caupi para avaliar a agressividade do patógeno. Pela avaliação do comprimento das lesões e da severidade da doença por meio de um índice, identificamos o método de inoculação por palito de dente mais eficiente pois permitiu discriminar os genótipos estudados e a agressividade do patógeno.

Characterization of wheat genotypes for drought tolerance and water use efficiency

The selection of wheat genotypes according to their drought tolerance is essential to off-season cultivation. The objective of this work was to characterize wheat genotypes through yield and components, morphological characteristics under water stress, and irrigated water use efficiency in the Cerrado region in Brazil. Genotypes were planted during the winters of 2016 and 2017 since there is no precipitation during this season and water levels can be measured. They were then submitted to four water regimes: WR1, WR2, WR3, and WR4, representing 100 %, 83 %, 50 %, and 30 % of evapotranspiration replacement. The following variables were evaluated: peduncle length (PL), number of ears m-² (NE m-²), hectoliter weight (HW), thousand grain weight (TGW), drought resistance index (DRI), irrigated water use efficiency (IWUE) and yield. Most variables showed correlation with yield and can be a useful tool for breeding programs. PL and HW were best correlated with yield. BRS 264 (irrigated biotype) was productive in treatments receiving the greatest number of irrigation treatments. Given that WR1 registered the highest water level, it was not expected that the rainfed biotype (BR18) would show a higher yield than an irrigated biotype (BRS254). BRS404 (rainfed biotype) was the most productive under moderate stress treatment (WR3). Aliança (rainfed biotype) showed a higher yield under severe stress. Rainfed biotypes presented a higher DRI than the irrigated ones. These genotypes can be used as a reference in breeding programs under each water regime in which their performance was outstanding. None of the variables studied contributed to the selection of the most efficient wheat genotypes in the IWUE.

Characterization of wheat genotypes for drought tolerance and water use efficiency

The selection of wheat genotypes according to their drought tolerance is essential to off-season cultivation. The objective of this work was to characterize wheat genotypes through yield and components, morphological characteristics under water stress, and irrigated water use efficiency in the Cerrado region in Brazil. Genotypes were planted during the winters of 2016 and 2017 since there is no precipitation during this season and water levels can be measured. They were then submitted to four water regimes: WR1, WR2, WR3, and WR4, representing 100 %, 83 %, 50 %, and 30 % of evapotranspiration replacement. The following variables were evaluated: peduncle length (PL), number of ears m-² (NE m-²), hectoliter weight (HW), thousand grain weight (TGW), drought resistance index (DRI), irrigated water use efficiency (IWUE) and yield. Most variables showed correlation with yield and can be a useful tool for breeding programs. PL and HW were best correlated with yield. BRS 264 (irrigated biotype) was productive in treatments receiving the greatest number of irrigation treatments. Given that WR1 registered the highest water level, it was not expected that the rainfed biotype (BR18) would show a higher yield than an irrigated biotype (BRS254). BRS404 (rainfed biotype) was the most productive under moderate stress treatment (WR3). Aliança (rainfed biotype) showed a higher yield under severe stress. Rainfed biotypes presented a higher DRI than the irrigated ones. These genotypes can be used as a reference in breeding programs under each water regime in which their performance was outstanding. None of the variables studied contributed to the selection of the most efficient wheat genotypes in the IWUE.(AU)

Erythrocyte phenotyping for feline AB system in domestic cats from the Ilhéus-Itabuna microregion, Bahia, Brazil

ABSTRACT: This study aimed to determine the erythrocyte phenotypes of the feline AB system and to check the presence of antigens other than those present in the feline AB system in domestic cats from Ilhéus-Itabuna microregion, Bahia, Brazil. Three-hundred feline blood samples were collected at the Veterinary Hospital of the Universidade Estadual de Santa Cruz (UESC) and in home visits to perform blood phenotyping using the tube-method testing. The reverse phenotyping was made between cats that tested phenotype B with blood samples of cats that tested phenotype A to confirm the blood phenotype B. The cross-tested among cats with phenotype A was made in order to verify the presence of different antigens of AB system in this blood phenotype. The results underwent macroscopic and microscopic analyses. Among the 300 animals tested, regarding breed, 290 were mixed-breed cats and among the remaining ten, five were Persians, four Siamese, and one Angora. 297 (99%) presented with phenotype A (including all the breeding cats) and three (1%) with phenotype B, and all this cats were mixed-breed cats. None (0%) of the cats showed the phenotype AB. All phenotype B bloods reacted to reverse phenotyping with phenotype A, confirming the phenotype B of these cats. All phenotype A bloods were compatible among each other, so no further erythrocyte antigens were detected through this test. The mother of one of the phenotype B cats was identified and had phenotype A, demonstrating phenotype A parents with phenotype B offspring. This finding indicates heterozygosis in the studied population. This data enable to conclude that the studied population presented different erythrocyte phenotypes, subsequently highlighting the importance of conducting phenotype analyses in these animals before performing blood transfusion to avoid serious hemolytic complications associated with incompatibility.

RESUMO: O objetivo deste estudo foi determinar a frequência dos fenótipos eritrocitários do sistema AB felino e verificar a presença de outros antígenos, não pertencentes ao sistema AB felino, em gatos domésticos das cidades de Ilhéus e Itabuna, Bahia, Brasil. Amostras de sangue de 300 gatos foram coletadas no Hospital Veterinário da Universidade Estadual de Santa Cruz (UESC) e em visitas domiciliares para realizar a fenotipagem sanguínea usando o método de tubo. A fenotipagem reversa foi realizada em gatos que testaram o fenótipo B com amostras que testaram o fenótipo A, para confirmação do fenótipo sanguíneo. O teste cruzado foi realizado entre gatos do fenótipo A, para pesquisar a presença de diferentes antígenos do sistema AB dentro desse fenótipo sanguíneo. Os resultados foram submetidos a análises macroscópicas e microscópicas. Dos 300 animais testados, 110 eram machos e 190 fêmeas, e suas idades variaram de cinco meses à 15 anos. Sobre as raças, 290 eram gatos sem raça definida e dos 10 restantes, cinco eram Persas, quatro eram Siameses e um Angorá. 297 (99%) apresentaram fenótipo A (incluindo todos os gatos de raça) e três (1%) tiveram fenótipo B, sendo todos esses gatos sem raça definida. Nenhum (0%) dos gatos apresentou fenótipo AB. Todos os sangues com fenótipo B reagiram na fenotipagem reversa com o fenótipo A, confirmando o fenótipo B desses gatos. Todos os sangues com fenótipo A foram compatíveis entre si, portanto nenhum antígeno eritrocitário adicional foi detectado através deste teste. A genitora de um dos gatos com fenótipo B, foi encontrada e a mesma possuía fenótipo A, demonstrando pais com fenótipo A e cria com fenótipo B. Esse achado indica heterozigose na população estudada. Esses dados levam à conclusão de que diferentes fenótipos eritrocitários estão presentes na população estudada e destacam a importância da realização de testes fenotípicos nesses animais antes dos procedimentos de transfusão, a fim de evitar complicações hemolíticas graves decorrentes do envolvimento de animais incompatíveis.

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus)

ABSTRACT: This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.

RESUMO: Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.

Erythrocyte phenotyping for feline AB system in domestic cats from the Ilhéus-Itabuna microregion, Bahia, Brazil / Fenotipagem eritrocitária para sistema AB felino em gatos domésticos da microrregão Ilhéus-Itabuna, Bahia, Brasil

This study aimed to determine the erythrocyte phenotypes of the feline AB system and to check the presence of antigens other than those present in the feline AB system in domestic cats from Ilhéus-Itabuna microregion, Bahia, Brazil. Three-hundred feline blood samples were collected at the Veterinary Hospital of the "Universidade Estadual de Santa Cruz" (UESC) and in home visits to perform blood phenotyping using the tube-method testing. The reverse phenotyping was made between cats that tested phenotype B with blood samples of cats that tested phenotype A to confirm the blood phenotype B. The cross-tested among cats with phenotype A was made in order to verify the presence of different antigens of AB system in this blood phenotype. The results underwent macroscopic and microscopic analyses. Among the 300 animals tested, regarding breed, 290 were mixed-breed cats and among the remaining ten, five were Persians, four Siamese, and one Angora. 297 (99%) presented with phenotype A (including all the breeding cats) and three (1%) with phenotype B, and all this cats were mixed-breed cats. None (0%) of the cats showed the phenotype AB. All phenotype B bloods reacted to reverse phenotyping with phenotype A, confirming the phenotype B of these cats. All phenotype A bloods were compatible among each other, so no further erythrocyte antigens were detected through this test. The mother of one of the phenotype B cats was identified and had phenotype A, demonstrating phenotype A parents with phenotype B offspring. This finding indicates heterozygosis in the studied population. This data enable to conclude that the studied population presented different erythrocyte phenotypes, subsequently highlighting the importance of conducting phenotype analyses in these animals before performing blood transfusion to avoid serious hemolytic complications associated with incompatibility.(AU)

O objetivo deste estudo foi determinar a frequência dos fenótipos eritrocitários do sistema AB felino e verificar a presença de outros antígenos, não pertencentes ao sistema AB felino, em gatos domésticos das cidades de Ilhéus e Itabuna, Bahia, Brasil. Amostras de sangue de 300 gatos foram coletadas no Hospital Veterinário da Universidade Estadual de Santa Cruz (UESC) e em visitas domiciliares para realizar a fenotipagem sanguínea usando o método de tubo. A fenotipagem reversa foi realizada em gatos que testaram o fenótipo B com amostras que testaram o fenótipo A, para confirmação do fenótipo sanguíneo. O teste cruzado foi realizado entre gatos do fenótipo A, para pesquisar a presença de diferentes antígenos do sistema AB dentro desse fenótipo sanguíneo. Os resultados foram submetidos a análises macroscópicas e microscópicas. Dos 300 animais testados, 110 eram machos e 190 fêmeas, e suas idades variaram de cinco meses à 15 anos. Sobre as raças, 290 eram gatos sem raça definida e dos 10 restantes, cinco eram Persas, quatro eram Siameses e um Angorá. 297 (99%) apresentaram fenótipo A (incluindo todos os gatos de raça) e três (1%) tiveram fenótipo B, sendo todos esses gatos sem raça definida. Nenhum (0%) dos gatos apresentou fenótipo AB. Todos os sangues com fenótipo B reagiram na fenotipagem reversa com o fenótipo A, confirmando o fenótipo B desses gatos. Todos os sangues com fenótipo A foram compatíveis entre si, portanto nenhum antígeno eritrocitário adicional foi detectado através deste teste. A genitora de um dos gatos com fenótipo B, foi encontrada e a mesma possuía fenótipo A, demonstrando pais com fenótipo A e cria com fenótipo B. Esse achado indica heterozigose na população estudada. Esses dados levam à conclusão de que diferentes fenótipos eritrocitários estão presentes na população estudada e destacam a importância da realização de testes fenotípicos nesses animais antes dos procedimentos de transfusão, a fim de evitar complicações hemolíticas graves decorrentes do envolvimento de animais incompatíveis.(AU)

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus) / Fenotipagem citométrica do sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus)

This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.(AU)

Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.(AU)

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus) / Fenotipagem citométrica do sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus)

This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.(AU)

Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.(AU)

Erythrocyte phenotyping for feline AB system in domestic cats from the Ilhéus-Itabuna microregion, Bahia, Brazil / Fenotipagem eritrocitária para sistema AB felino em gatos domésticos da microrregão Ilhéus-Itabuna, Bahia, Brasil

This study aimed to determine the erythrocyte phenotypes of the feline AB system and to check the presence of antigens other than those present in the feline AB system in domestic cats from Ilhéus-Itabuna microregion, Bahia, Brazil. Three-hundred feline blood samples were collected at the Veterinary Hospital of the "Universidade Estadual de Santa Cruz" (UESC) and in home visits to perform blood phenotyping using the tube-method testing. The reverse phenotyping was made between cats that tested phenotype B with blood samples of cats that tested phenotype A to confirm the blood phenotype B. The cross-tested among cats with phenotype A was made in order to verify the presence of different antigens of AB system in this blood phenotype. The results underwent macroscopic and microscopic analyses. Among the 300 animals tested, regarding breed, 290 were mixed-breed cats and among the remaining ten, five were Persians, four Siamese, and one Angora. 297 (99%) presented with phenotype A (including all the breeding cats) and three (1%) with phenotype B, and all this cats were mixed-breed cats. None (0%) of the cats showed the phenotype AB. All phenotype B bloods reacted to reverse phenotyping with phenotype A, confirming the phenotype B of these cats. All phenotype A bloods were compatible among each other, so no further erythrocyte antigens were detected through this test. The mother of one of the phenotype B cats was identified and had phenotype A, demonstrating phenotype A parents with phenotype B offspring. This finding indicates heterozygosis in the studied population. This data enable to conclude that the studied population presented different erythrocyte phenotypes, subsequently highlighting the importance of conducting phenotype analyses in these animals before performing blood transfusion to avoid serious hemolytic complications associated with incompatibility.(AU)

O objetivo deste estudo foi determinar a frequência dos fenótipos eritrocitários do sistema AB felino e verificar a presença de outros antígenos, não pertencentes ao sistema AB felino, em gatos domésticos das cidades de Ilhéus e Itabuna, Bahia, Brasil. Amostras de sangue de 300 gatos foram coletadas no Hospital Veterinário da Universidade Estadual de Santa Cruz (UESC) e em visitas domiciliares para realizar a fenotipagem sanguínea usando o método de tubo. A fenotipagem reversa foi realizada em gatos que testaram o fenótipo B com amostras que testaram o fenótipo A, para confirmação do fenótipo sanguíneo. O teste cruzado foi realizado entre gatos do fenótipo A, para pesquisar a presença de diferentes antígenos do sistema AB dentro desse fenótipo sanguíneo. Os resultados foram submetidos a análises macroscópicas e microscópicas. Dos 300 animais testados, 110 eram machos e 190 fêmeas, e suas idades variaram de cinco meses à 15 anos. Sobre as raças, 290 eram gatos sem raça definida e dos 10 restantes, cinco eram Persas, quatro eram Siameses e um Angorá. 297 (99%) apresentaram fenótipo A (incluindo todos os gatos de raça) e três (1%) tiveram fenótipo B, sendo todos esses gatos sem raça definida. Nenhum (0%) dos gatos apresentou fenótipo AB. Todos os sangues com fenótipo B reagiram na fenotipagem reversa com o fenótipo A, confirmando o fenótipo B desses gatos. Todos os sangues com fenótipo A foram compatíveis entre si, portanto nenhum antígeno eritrocitário adicional foi detectado através deste teste. A genitora de um dos gatos com fenótipo B, foi encontrada e a mesma possuía fenótipo A, demonstrando pais com fenótipo A e cria com fenótipo B. Esse achado indica heterozigose na população estudada. Esses dados levam à conclusão de que diferentes fenótipos eritrocitários estão presentes na população estudada e destacam a importância da realização de testes fenotípicos nesses animais antes dos procedimentos de transfusão, a fim de evitar complicações hemolíticas graves decorrentes do envolvimento de animais incompatíveis.(AU)

Papaya recombinant inbred lines selection by image-based phenotyping

The selection of superior Carica papaya (L) genotypes depends on the availability of genetic variability and on the favorable and simultaneous response of the genotypes to those traits of most interest. However, manual phenotyping (MP) demands intensive labor, is time-consuming and expensive. The aim of the current study is to access the efficiency of image-based phenotyping (IBP) in estimating genetic parameters and in selecting F4 recombinant inbred lines. The genetic parameters and values were estimated in accordance with the REML/BLUB procedure and combined selection using the selection index based on standardized genetic values. The majority of traits accessed through IBP showed experimental coefficients of variation similar to those found through MP. Both methodologies showed genetic parameters of similar magnitude, indicating expressive genetic variability between lines in the traits accessed in this study. The same superior lines were indicated in both methodologies and expressive genetic gains obtained through the lines were selected for all traits. IBP performance was similar to that of MP with respect to the estimates of breeding-relevant traits such as commercial fruits and yield. Thus, IBP showed efficient phenotypic assessment, as well as selective accuracy in accessing genetic variability and genetic gains, when it was compared to MP. Since IBP is far less dependent on labor, it is expected to be incorporated into the routine of papaya breeding programs as a way of increasing the number of accessed lines and, consequently, increasing genetic gains.

Phenotypic and molecular characterization of apple (Malus × domestica Borkh) genetic resources in Greece

ABSTRACT: The phenotypic and genetic analysis of apple (Malus × domestica Borkh) genotypes is essential for breeding species. Information on the morphology and genetic structure of apple offers significant help for germplasm maintenance and selection of suitable material to breed superior cultivars. This study shows the results of an investigation on the morphology and the genetic diversity for 19 apple cultivars, which are preserved in an ex situ collection in Naoussa, Central Macedonia, Greece. Information was recorded over a 5-year period for 47 traits describing plant morphology and phenotype, as well as leaf and fruit quality. Data were analyzed using the principal component analysis (PCA) method. The Euclidean distance metric and the Wards agglomeration method were used in an unsupervised hierarchical cluster analysis of all cultivars. The cultivars were grouped into four main clusters, suggesting that the characterized apple collection has a high potential for specific breeding goals. Furthermore, the cultivars were genotyped using seven microsatellite primers. Moderate levels of polymorphism were detected, and 38 distinctive alleles (5.4 alleles per primer pair) were identified. Both multivariate clustering approach (phenotypic data) and the genetic distance clustering approach (genetic data) grouped the apple cultivars according to their type. Hence, these data could be used for protection or patenting processes of existing or new apple cultivars carried out by the EU-Community Plant Variety Office.

Phenotypic and molecular characterization of apple (Malus × domestica Borkh) genetic resources in Greece

The phenotypic and genetic analysis of apple (Malus × domestica Borkh) genotypes is essential for breeding species. Information on the morphology and genetic structure of apple offers significant help for germplasm maintenance and selection of suitable material to breed superior cultivars. This study shows the results of an investigation on the morphology and the genetic diversity for 19 apple cultivars, which are preserved in an ex situ collection in Naoussa, Central Macedonia, Greece. Information was recorded over a 5-year period for 47 traits describing plant morphology and phenotype, as well as leaf and fruit quality. Data were analyzed using the principal component analysis (PCA) method. The Euclidean distance metric and the Wards agglomeration method were used in an unsupervised hierarchical cluster analysis of all cultivars. The cultivars were grouped into four main clusters, suggesting that the characterized apple collection has a high potential for specific breeding goals. Furthermore, the cultivars were genotyped using seven microsatellite primers. Moderate levels of polymorphism were detected, and 38 distinctive alleles (5.4 alleles per primer pair) were identified. Both multivariate clustering approach (phenotypic data) and the genetic distance clustering approach (genetic data) grouped the apple cultivars according to their type. Hence, these data could be used for protection or patenting processes of existing or new apple cultivars carried out by the EU-Community Plant Variety Office.(AU)

Phenotypic and molecular characterization of apple (Malus × domestica Borkh) genetic resources in Greece

ABSTRACT: The phenotypic and genetic analysis of apple (Malus × domestica Borkh) genotypes is essential for breeding species. Information on the morphology and genetic structure of apple offers significant help for germplasm maintenance and selection of suitable material to breed superior cultivars. This study shows the results of an investigation on the morphology and the genetic diversity for 19 apple cultivars, which are preserved in an ex situ collection in Naoussa, Central Macedonia, Greece. Information was recorded over a 5-year period for 47 traits describing plant morphology and phenotype, as well as leaf and fruit quality. Data were analyzed using the principal component analysis (PCA) method. The Euclidean distance metric and the Wards agglomeration method were used in an unsupervised hierarchical cluster analysis of all cultivars. The cultivars were grouped into four main clusters, suggesting that the characterized apple collection has a high potential for specific breeding goals. Furthermore, the cultivars were genotyped using seven microsatellite primers. Moderate levels of polymorphism were detected, and 38 distinctive alleles (5.4 alleles per primer pair) were identified. Both multivariate clustering approach (phenotypic data) and the genetic distance clustering approach (genetic data) grouped the apple cultivars according to their type. Hence, these data could be used for protection or patenting processes of existing or new apple cultivars carried out by the EU-Community Plant Variety Office.

Papaya recombinant inbred lines selection by image-based phenotyping

The selection of superior Carica papaya (L) genotypes depends on the availability of genetic variability and on the favorable and simultaneous response of the genotypes to those traits of most interest. However, manual phenotyping (MP) demands intensive labor, is time-consuming and expensive. The aim of the current study is to access the efficiency of image-based phenotyping (IBP) in estimating genetic parameters and in selecting F4 recombinant inbred lines. The genetic parameters and values were estimated in accordance with the REML/BLUB procedure and combined selection using the selection index based on standardized genetic values. The majority of traits accessed through IBP showed experimental coefficients of variation similar to those found through MP. Both methodologies showed genetic parameters of similar magnitude, indicating expressive genetic variability between lines in the traits accessed in this study. The same superior lines were indicated in both methodologies and expressive genetic gains obtained through the lines were selected for all traits. IBP performance was similar to that of MP with respect to the estimates of breeding-relevant traits such as commercial fruits and yield. Thus, IBP showed efficient phenotypic assessment, as well as selective accuracy in accessing genetic variability and genetic gains, when it was compared to MP. Since IBP is far less dependent on labor, it is expected to be incorporated into the routine of papaya breeding programs as a way of increasing the number of accessed lines and, consequently, increasing genetic gains.(AU)

PREDIÇÃO DA ÁREA DO MÚSCULO Longissimus dorsi A PARTIR DE IMAGENS ULTRASSONOGRÁFICAS EM OVINOS UTILIZANDO VISÃO COMPUTACIONAL

A ovinocultura de corte é um setor que se encontra em constante expansão. Este setor também está ganhando espaço no agronegócio brasileiro e na região Nordeste. Entretanto, a produtividade é baixa. Portanto, este setor exige alternativas tecnológicas dentro do melhoramento genético dos rebanhos, como fenotipagem de precisão, através das coletas de informações fenotípicas com precisão para permitir melhorar o rendimento de carcaça dos animais, a fim de atender às demandas e a avaliação de carcaça com resultados mais precisos, a qual tem sido realizada a partir da análise de imagens ultrassonográficas por profissionais especializados. Sendo assim, o objetivo com pesquisa foi o desenvolvimento de uma abordagem com o uso de inteligência artificial para avaliação de carcaça ovina de forma acurada por meio do reconhecimento de imagens ultrassonográficas do músculo Longissimus dorsi. A metodologia proposta para este trabalho foi dividida em 4 etapas. Na etapa de coleta de dados, 121 imagens ultrassonográficas de fêmeas ovinas foram coletadas com auxílio de aparelho de ultrassom, Durante a segmentação das regiões de interesse, utilizou-se apenas um método de segmentação automatizado com base em algoritmo de redes neurais (U-Net). Para a avaliação de segmentações automáticas, utilizou-se as métricas de coeficiente de dados Dice e a métrica de intersecção sobre união (IoU). Na extração de recursos, objetivou-se encontrar características importantes para a previsão de AOL (Área de Olho de Lombo). Na última etapa, foi realizada uma análise de regressão, sendo a variável independente os valores dos atributos obtidos com os descritores utilizados e a variável dependente a AOL previsto para a imagem de ultrassom do animal, aonde utilizou-se as métricas de quadrado médio do resíduo (QMR) e erro médio absoluto (EMA). Dois algoritmos de regressão foram utilizados, AdaBoost Regressor(ABR)e Random Forest Regressor (RFR). Realizou-se uma análise de variância e o teste t de student para comparar as médias de AOL observada e predita. Os valores obtidos pela métrica Dice foi de 0,94 e a da IoU foi de 0,89 o que demonstra alta similaridade entre o real e o previsto. Os valores de QMR, EMA e R² para o ABR é de, respectivamente, 2,61, 1,22 e 0,51 e para o RFR é de 2,15, 1,12 e 0,61, o que demonstra uma correlação positiva entre os valores preditos e os valores reais. Observou que não houve diferenças significativas entre as médias de AOL observada e predita. Portanto, a medição automatizada da AOL a partir de imagens ultrassonográficas é promissora e possibilitará maior eficiência na realização desta medida em grandes quantidades de imagens com alta precisão, pois dispensa a intervenção humana na delimitação da área do músculo Longissimus dorsi em ovinos de corte de carcaça.

Beef sheep is an expanding sector. This sector is also gaining space in Brazilian agribusiness and in the Northeast region. However, productivity is low. Therefore, this sector requires technological alternatives within the genetic improvement of herds, such as precision phenotyping, through the collection of phenotypic information needed to improve the carcass yield of the animals, to meet the demands and the carcass evaluation with more accurate results. , one that has been carried out from the analysis of ultrasound images by specialized professionals. Therefore, the objective with research was the development of an approach with the use of artificial intelligence to evaluate sheep carcass accurately through the recognition of ultrasound images of the Longissimus dorsi muscle. The methodology proposed for this work was divided into 4 stages. In the data collection stage, 121 ultrasound images of sheep from collections were collected with the aid of an ultrasound device. During the selection of the regions of interest, using only an automated supply method based on a neural network algorithm (U-Net) For the evaluation of automatic segmentations, the Dice data coefficient metrics and the intersection over union (IoU) metrics were used. In the extraction of resources, the objective was to find important characteristics for a forecast of REA (Rib Eye Area). In the last step, a regression analysis was performed, with the independent variable being the values of the resources provided with the descriptors used and the REA-dependent variable predicted for the animal's ultrasound image, where it was used as mean square error (MSE) and mean absolute error (MAE). Two regression algorithms were used, AdaBoost Regressor (ABR) and Random Forest Regressor (RFR). An analysis of variance and the student's t test were performed to compare how REA averages were compared and predicted. The values obtained by the metric Dice was 0.94 and that of the IoU was 0.89, which demonstrates a high similarity between the real and the predicted. The values of MSE, MAE and R² for ABR are 2.61, 1.22 and 0.51, respectively, and for RFR it is 2.15, 1.12 and 0.61, which demonstrates a correlation between predicted and actual values. He observed that there were no significant differences between the observed and predicted REA averages. Therefore, an automated measurement of REA from ultrasound images is promising and will enable greater efficiency in carrying out this measurement in large quantities of images with high precision, since it does not require human intervention in the delimitation of the Longissimus dorsi muscle area in carcass sheep. .

Estomatite vesicular: cinética de multiplicação in vitro e predição de epítopos imunodominantes do Alagoas vesiculovirus e validação de uma RT-qPCR para o Cocal vesiculovirus

A estomatite vesicular (EV) no Brasil é causada principalmente pelo Alagoas vesiculovirus (VSAV),mas surtos nos anos de 1996 e 1998 foram caudados por uma segunda espécie, Cocal vesiculovirus (COCV). Apesar de apresentar uma baixa mortalidade, a EV é a principal doença vesicular que acomete os animais domésticos no país e clinicamente indistinguível da febre aftosa. Estudos filogenéticos mostraram que as amostras de VSAV isoladas no Brasil podem ser divididas em três grupos genéticos, entretanto não se sabe se há uma diferença fenotípica entre estes grupos. O objetivo dessa tese foi analisar a cinética do crescimento do VSAV em cultivo celular; predizer epítopos imunodominantes da glicoproteína do VSAV e validar uma RT-qPCR para o COCV. Técnicas moleculares como droplet digital RT-PCR e RT-qPCR, cultivo viral em linhagens celulares, spot synthesis e bioinformática foram utilizados durante os estudos. Observou-se que os isolados do VSAV brasileiros apresentam diferenças no crescimento in vitro. A partir dos estudos da glicoproteína foi possível predizer peptídeos imunodominantes no domínio de trimerização e que a maior variação está na porção da transmembrana da proteína. A detecção do COCV pode ser realizada a partir da RT-qPCR diretamente do tecido de epitélio com 97% de precisão, com uma sensibilidade de 3950 unidades formadoras de placas.

Vesicular stomatitis (VS) in Brazil is mainly caused by Alagoas vesiculovirus(VSAV), but outbreaks in 1996 and 1998 were caused by a second species, the Cocal vesiculovirus (COCV). Despite presenting low mortality, VS is the main vesicular disease that affects domestic animals in the country and is clinically indistinguishable from foot-and-mouth disease. Phylogenetic studies have shown that VSAV samples isolated in Brazil can be divided into three genetic groups, however, it is not known if there is a phenotypic difference between these groups. The aim of this thesis was to analyze the growth kinetics of VSAV in cell culture; predict immunodominant epitopes of the VSAV glycoprotein and validate an RT-qPCR for COCV. During the studies, molecular techniques such as digital droplet RT-PCR and RT-qPCR, viral culture in cell lines, spot synthesis and bioinformatics were used. The Brazilian VSAV isolates were observed to show differences in in vitro growth. From glycoprotein studies, it was possible to predict immunodynamic peptides in the trimerization domain and that the greatest variation is in transmembrane portion of the protein. COCV detection can be performed from RT-qPCR from the epithelial tissue with 97% accuracy, with a sensitivity of 3950 plaque-forming units.

DIVERSIDADE GENÉTICA E CARACTERIZAÇÃO DA VIRULÊNCIA EM Staphylococcus aureus ISOLADOS DE MASTITE EM REBANHOS BOVINOS BRASILEIROS

A mastite bovina é a doença de maior impacto econômico para a bovinocultura leiteira em todos os continentes e Staphylococcus aureus é um de seus principais agentes etiológicos. Este patógeno possui uma variedade de fatores de virulência que levam a mastite clínica ou subclínica e infecções intramamárias crônicas de difícil tratamento. Além da mastite, este agente é responsável por outras enfermidades nos animais e no homem, tais como, endocardites, osteomielites e septicemia, sendo bastante importante para a saúde pública. Os objetivos desse trabalho foram realizar a caracterização fenotípica, usando como modelo experimental larvas de Galleria mellonella, e genotípica, pelas técnicas de Pulsed-field Gel Electrophoresis (PFGE), Multiple locus VNTR fingerprinting (MLVF) e prospecção de genes de virulência codificadores de adesinas e toxinas em duas populações de S. aureus provenientes de mastite bovina, isolados entre os anos de 1994 e 2014, sendo uma restrita ao Estado de Minas Gerais, contendo isolados de origem clínica e subclínica, e outra contendo isolados de mastite subclínica de rebanhos de sete estados brasileiros. Os resultados das análises fenotípicas mostraram que larvas de G. mellonella constituem um modelo prático e eficiente para avaliar fenótipos de virulência em isolados de S. aureus provenientes de mastite bovina, permitindo a distinção de isolados mais e menos virulentos. A infecção experimental em G. mellonella demonstrou que os isolados provenientes de mastite subclínica foram, em geral, mais virulentos. As técnicas de genotipagem utilizadas se mostraram efetivas para caracterização de S. aureus e apontaram a existência de tipos genéticos predominantes amplamente disseminados entre diferentes rebanhos. Embora a PFGE tenha sido ligeiramente mais discriminatória, a MLVF também se confirmou como uma boa técnica de tipagem, permitindo um agrupamento que refletia parcialmente o agrupamento demonstrado pela PFGE. A MLVF, com um nível de similaridade de 80%, apresentou um cluster com predomínio de amostras clínicas e outro com amostras subclínicas em predominância. O conjunto de genes de virulência que codificam os fatores de aglutinação (ClfA e ClfB), a proteína de ligação à fibronectina (FnBPA), as toxinas (Hla, Hlb e Luk-ED), a proteína de ligação a elastina (Ebps) e a proteína de ligação ao colágeno (Cna) foi amplamente presente na população de isolados de mastite subclínica provenientes de sete estados brasileiros sendo que a frequência desses genes variou de 58 a 97,7%. Os resultados demonstraram que esse conjunto de proteínas deve ser melhor avaliado e pode ser promissor como um conjunto de antígenos para constituir novas vacinas para prevenção da mastite causada por S. aureus em rebanhos bovinos brasileiros. Em conclusão, este trabalho contribuiu para o entendimento da epidemiologia da mastite bovina no Brasil, demonstrando a grande diversidade genética dos isolados que causam mastite no país, porém, com predomínio de linhagens especializadas que devem ser mais bem estudadas para o desenvolvimento de novas formas de controle e prevenção desta doença.

The bovine mastitis is the disease of greatest economic impact for dairy herds worldwide and Staphylococcus aureus is one of its main etiological agents. This pathogen has a variety of virulence factors that lead to clinical or subclinical mastitis, and chronic intramammary infections of difficult treatment. In addition to mastitis, this agent is responsible for other diseases in animals and man, such as endocarditis, osteomyelitis and septicemia, being quite important for public health. The objectives of this work were to accomplish the phenotypic characterization, using the experimental model of Galleria mellonella larvae and genotypic using the Pulsed-field Gel Electrophoresis (PFGE), Multiple Locus VNTR fingerprinting (MLVF) and prospect of virulence genes encoding adhesins and toxins in two populations of S. aureus from bovine mastitis, isolated between 1994 and 2014, being one restricted to the State of Minas Gerais, containing isolates of clinical and subclinical origin and one containing subclinical mastitis isolates from herds of seven Brazilian states. The results of the phenotypic analyzes showed that G. mellonella larvae constitute a practical and efficient model to evaluate virulence phenotypes in S. aureus isolates from bovine mastitis, allowing the distinction of more and less virulent isolates. The experimental infection in G. mellonella showed the predominance of more virulent strains among those coming from subclinical mastitis. The genotyping techniques used were effective for the characterization of S. aureus and indicated the existence of predominant genetic types widely disseminated among different herds. Although PFGE was slightly more discriminatory, MLVF also was confirmed as a good typing technique, allowing a clustering that partially reflected the clustering demonstrated by PFGE. The MLVF, with a level of similarity of 80%, showed a cluster with predominance of clinical strains and another with predominance of subclinical strains. The set of virulence genes encoding agglutination factors (ClfA and ClfB), fibronectin binding protein (FnBPA), toxins (Hla, Hlb and Luk-ED), elastin binding protein (Ebps), and the collagen binding protein (Cna) was widely present in the population of subclinical mastitis isolates from seven Brazilian states and the frequency of these genes ranged from 58 to 97.7%. The results show that this set of proteins should be better evaluated and may be promising as a set of antigens to constitute news vaccines for prevention of mastitis caused by S. aureus in Brazilian dairy herds. In conclusion, this work contributed to the understanding of the epidemiology of bovine mastitis in Brazil, showing the great genetic diversity of the isolates that cause mastitis in the country, but with a predominance of specialized strains that should be better studied for the development of new forms of control and prevention of this disease.

Influência do ciclo da muda de penas nas respostas imune e inflamatória de pinguins-de-Magalhães (Spheniscus magellanicus) mantidos em cativeiro

A mudança anual de penas é um componente crítico do ciclo biológico de todas as espécies de aves e pode modificar as respostas imunológicas individuais, sendo que suas consequências são pouco estudadas. Neste contexto, uma vez que são escassos os estudos acerca do perfil clínico e parâmetros imunológicos de pinguinsde- Magalhães em diferentes fases da muda, este estudo propôs avaliar determinados aspectos relacionados a esses perfis, em animais mantidos em ambiente controlado. Foram estudados 21 indivíduos adultos (dez machos e 11 fêmeas) e os parâmetros avaliados foram: teste de hipersensibilidade cutânea tardia à fitohemaglutinina (FHA) com a avaliação histopatológica de biópsias que incluíram a análise qualitativa e quantitativa dos tipos celulares presentes na reação e no controle às 6, 24 e 48 horas; parâmetros clínicos (massa corpórea e hematologia); fagocitose e burst oxidativo; fenotipagem e resposta linfoproliferativa. O teste de FHA demonstrou diferença significativa na espessura da membrana interdigital de ambas as patas inoculadas com FHA e com o controle (PBS) quando comparadas antes da inoculação e antes da biópsia às 24 e 48 horas. A análise histomorfométrica revelou não haver diferenças significativas na frequência relativa de granulócitos (heterófilos/ eosinófilos), linfócitos, macrófagos ou trombócitos entre as patas inoculadas com FHA e PBS em nenhum dos momentos analisados. Este resultado sugere que a diferença na espessura da membrana interdigital das patas inoculadas seja em decorrência do edema e não do infiltrado inflamatório. Para analisar os efeitos do ciclo da muda nos parâmetros clínicos e hematológicos, os animais foram divididos em três grupos: pré-muda (-5 a 0 dias da muda), muda (10º ao 21º dia da muda) e padrão basal (>120 dias da muda). Os resultados mostraram que em relação ao momento basal: houve aumento significativo na massa corpórea dos animais no momento da pré-muda; diminuição significativa de proteínas totais, hematócrito, eritrócitos totais e leucócitos totais na pré-muda e muda. Não houve diferença no número total de monócitos e heterófilos, contudo, houve eosinofilia e linfopenia na pré-muda e muda, respectivamente. Com relação aos parâmetros imunológicos, houve aumento no burst oxidativo gerado pelo estímulo biológico (Staphylococcus aureus) na pré-muda em relação à muda e ao padrão basal; ocorreu aumento do burst oxidativo gerado pelo estímulo químico (éster de forbol) na pré-muda em relação à muda. Em relação à fagocitose, houve diminuição desta capacidade dos leucócitos dos animais na pré-muda e muda em relação ao padrão basal quando o agente foi bacteriano; entretanto, o mesmo não foi observado para a fagocitose de leveduras (Zymosan A); houve diminuição de linfócitos T CD4+ circulantes na pré-muda e muda em relação ao padrão basal e o índice de proliferação de linfócitos aumentou durante o período de muda em relação ao basal. Esses resultados tomados em conjunto sugerem que as mudanças fisiológicas observadas durante o ciclo da muda interferem na modulação de parâmetros imunológicos e são sugestivas de serem consequência do prejuízo energético sofrido, mesmo em animais mantidos em ambientes controlados.

The annual replacement of feathers is a critical component of the biological cycle of all bird species. Molt can modify their individual immune response, and the consequences of this are not well known. Within this context, and the few studies on the clinical profile and immunological parameters of Magellanic penguins at different stages of their molt, this study aimed to evaluate certain aspects related to the clinical profiles of animals kept in a controlled environment. A total of 21 adult individuals (ten males and 11 females) was analyzed and the parameters evaluated in this study were: delayed-type hypersensitivity test to phytohemagglutinin (PHA) based on the histopathological evaluation of foot web biopsies that included qualitative and quantitative analysis of cell types present in the essay and control at 6, 24 and 48 hours; clinical parameters (body mass and hematology); phagocytosis and oxidative burst; phenotyping and lymphoproliferative response. The PHA test showed significant differences in the thickness of the webbing of both feet inoculated with PHA and control (PBS), when compared before inoculation and before biopsy at 24 and 48 hours. Histomorphometric analysis revealed no significant differences in the relative frequencies of granulocytes (heterophils/ eosinophils), lymphocytes, macrophages, thrombocytes, or between the feet inoculated with PHA and with PBS in any of the samples. This result suggests that the difference in thickness of the foot webbing is due to edema and not an inflammatory infiltrate. To analyze the effects of the molting cycle on clinical and hematological parameters, the birds were divided into three groups: pre-molt (from -5 days to the start of the molt, day 0), molt (10th to 21st days of molt) and baseline (>120 days after the completion of the molt). The results showed that in comparison with baseline values: a significant increase in body mass occurred during pre-molt; and a significant decrease in total protein, hematocrit, total erythrocytes and total leukocytes occurred during pre-molt and molt. There was no difference in the total numbers of monocytes and heterophils, however, there were eosinophilia and lymphopenia during pre-molt and molt, respectively. There was an increase in the oxidative burst generated by the biological stimulus (Staphylococcus aureus) in the pre-molt compared to molt and baseline levels; and there was an increased oxidative burst generated by the chemical stimuli (phorbol esters) in the pre-molt in relation to the molt. There was a decreased capacity of phagocytosis for the leukocytes in animals during pre-molt and molt compared to baseline when the agent was bacterial; however, this was not observed for the phagocytosis of yeast (Zymosan A). There was a decrease of circulating CD4+ T lymphocytes in the pre-molt and molt compared to baseline, and the lymphocyte proliferation index increased during the molt compared to baseline. The results suggest that the physiological changes observed during the molt cycle does interfere with the modulation of immune parameters, and are suggestive of resulting as a consequence of from the high energy demands of molt, even for birds kept in controlled environments

DIAGNÓSTICO, CLASSIFICAÇÃO E ESTADIAMENTO DE LINFOMA MULTICÊNTRICO EM CÃES ATRAVÉS DA REAÇÃO EM CADEIA PELA POLIMERASE PARA REARRANJOS DE RECEPTORES DE ANTÍGENOS (PRRA)

Em medicina veterinária, o linfoma, proliferação neoplásica de linfócitos, é uma das neoplasias mais frequentes e de maior importância para cães. Por ser um grupo muito heterogêneo de neoplasias, é necessário obter o máximo de informações para avaliação de prognóstico e monitoramento do paciente com neoplasia linfoide, para isso sendo necessário o aprimoramento das técnicas diagnósticas disponíveis. Diagnóstico molecular baseado na reação em cadeia pela polimerase (PCR), como a PCR para rearranjos de receptores dos antígenos (PRRA), fornece uma série de informações de forma relativamente rápida e mais acessível que outras técnicas. Dentre suas utilidades, destaca-se a capacidade de classificar o linfoma em fenótipos e a detecção precoce de infiltrado neoplásico em sangue e medula óssea em comparação com a microscopia óptica. O presente estudo tem por objetivos: implementar um ensaio de PRRA para aplicação em amostras de linfonodo, sangue e medula óssea de cães com linfoma multicêntrico; classificar os casos de linfoma multicêntrico em fenótipos; pesquisar infiltrado neoplásico em sangue e medula óssea; e coletar dados epidemiológicos sobre a população acometida por linfoma. A população de estudo foi composta por 13 cães para coleta de linfonodo, sangue e medula óssea, além de 47 lâminas de citopatologia de linfonodo positivas para linfoma. O DNA dessas amostras foi extraído, amplificado usando iniciadores e condições adaptadas com base no descrito na literatura. Produtos de PCR foram submetidos à eletroforese em gel de poliacrilamida a 12%. Dados epidemiológicos demonstraram predomínio em cães de raça pura, adultos a idosos, e frequência pouco maior em machos do que fêmeas. Dados laboratoriais revelaram ocorrência de anemia, leucocitose, trombocitopenia, aumento da atividade enzimática de fosfatase alcalina e gama-glutamil transferase e hipoalbuminemia. A PRRA amplificou com sucesso DNA linfoide em 54/60 (90%) das amostras de linfonodo. Destas, 26/54 (48,1%) foram classificadas como tipo B, 11/54 (20,3%) como tipo T, 1/54 (1,9%) como apresentando dupla clonalidade e 16/54 (29,6%) como indeterminadas. Não houve diferença significativa entre os resultados da microscopia e da PRRA na pesquisa de células neoplásicas no sangue e medula óssea, provavelmente devido à amostragem. Conclui-se que a técnica de PRRA foi desenvolvida com êxito para uso em amostras aspirados de linfonodo, sangue, medula óssea e lâminas secas ao ar e coradas, apresentando sensibilidade de 70,1%.

In veterinary medicine, lymphoma, a neoplastic proliferation of lymphocytes, is one of the most frequent and important neoplasia among dogs. Being a heterogeneous group of neoplasia, one must obtain maximum information in order to evaluate prognosis and monitor patients with lymphoid malignancy, thus diagnostic tool development being required. Molecular diagnosis based on polymerase chain reaction (PCR), such as PCR for antigen receptor rearrangement (PARR), gives a series of information relatively fast and in a more accessible way than other tools. Among its utilities, it can detect the lymphoma cell phenotype and detect neoplastic infiltrate in blood and bone marrow earlier than microscopic evaluation. This study aimed into: implementing a PARR-assay and applying it to lymph node, blood and marrow samples from dogs with multicentric lymphoma; using it to detect multicentric lymphoma phenotype; detecting neoplastic infiltrate in blood and bone marrow; and collecting epidemiological data about the population of the study. There were 13 dogs in the study from which lymph node, blood and bone marrow samples were collected, along 47 lymph nodes cytopathology air-dried slides diagnosed as lymphoma. DNA was extracted from the samples and amplified using iniciadores and PCR-conditions adapted from other studies. PCR products were analysed by polyacrylamide 12% electrophoresis. Epidemiological data show prevalence of the disease in pure-breed, adult to old aged dogs, with a few more male than female dogs. Exam results show anaemia, leukocytosis, thrombocytopenia, increased alkaline phosphatasis and gammaglutamyl transferase and hypoalbuminemia. The PARR-assay successfully amplified lymphoid DNA from 54/60 (90%) of the lymph node samples, from which 26/54 (48,1%) were diagnosed as B-cell type, 11/54 (20,3%) as T-cell type, 1/54 (1,9%) as dual cell type and 16/54 (20,3%) as indeterminate. There were no statistical difference between PARR-assay and optics microscopy in neoplastic cell detection, probably due to sampling. In conclusion, a PARR-assay technique was developed successfully for usage with lymph node, blood, bone marrow and cytopathology slide samples, presenting sensitivity of 70,1%.

Phenotypic aspects of oral strains of Candida albicans in children with down's syndrome

The aim of this article is to characterize the biological aspects of oral strains of C. albicans in children with Down's syndrome. These yeasts were analyzed as to their macromorphological and enzymatic aspects and were tested as to their in vitro susceptibility to antifungal drugs using broth microdilution to determine the minimum inhibitory concentration (MIC). The morphotyping revealed that all oral C. albicans isolates from children with Down's syndrome promoted the formation of fringes regardless of size, while the control group presented smaller fringes. All oral C. albicans strains produced proteinase, but those with phospholipolytic activity showed greater enzyme capacity in the test group. In vitro susceptibility showed that all oral C. albicans isolates were sensitive to the drugs used.

O objetivo deste artigo foi caracterizar os aspectos biológicos de cepas de C. albicans orais em crianças com síndrome de Down. Estas leveduras foram analisadas quanto aos seus aspectos macromorfológicos e enzimáticos e foram testadas quanto a sua suscetibilidade in vitro a drogas antifúngicas, usando a microdiluição em caldo para a determinação da concentração inibitória mínima (CIM). A morfotipagem revelou que todos os isolados de C. albicans orais de crianças com síndrome de Down induziram à formação de franjas independente do tamanho, enquanto o grupo controle teve franjas menores. Todas as cepas de C. albicans orais produziram proteinase, mas aquelas com atividade fosfolipidolítica mostraram maior capacidade enzimática no grupo teste. A suscetibilidade in vitro mostrou que todos os isolados de C. albicans orais foram sensíveis a drogas empregadas.