Potassium fertilization as salt stress attenuator in sour passion fruit / Adubação potássica como atenuante do estresse salino em maracujazeiro-azedo

Water scarcity associated with irregular rainfall in the semi-arid region of Northeastern Brazil stands out as a limiting factor for agricultural production. Thus, the use of waters with high concentration of salts is an alternative to expand irrigated agriculture in this region. In this context, this study evaluated the water status, intercellular electrolyte leakage, photosynthetic pigments, and gas exchange of 'BRS SC1' sour passion fruit as a function of irrigation with water of different levels of salinity and potassium doses. The experiment was carried out in pots adapted as drainage lysimeters under field conditions at the Experimental Farm of the Federal University of Campina Grande in São Domingos - PB, Brazil. The experimental design was randomized blocks, in a 5 × 4 factorial scheme, whose treatments were obtained by combining two factors: five levels of electrical conductivity of irrigation water - ECw (0.3, 1.1, 1.9, 2.7, and 3.5 dS m-1), associated with four potassium doses (60, 80, 100, and 120% of the recommendation), with three replicates. Water with electrical conductivity greater than 0.3 dS m-1 reduced the relative water content, chlorophyll a and chlorophyll b contents, CO2 assimilation rate, and instantaneous water use efficiency of 'BRS SC1' sour passion fruit plants. The estimated potassium dose of 85% of the recommendation (equivalent to 293 g per plant per year) mitigated the deleterious effects of salt stress on stomatal conductance, transpiration, internal CO2 concentration, and instantaneous carboxylation efficiency of passion fruit 'BRS SC1'.

Water scarcity associated with irregular rainfall in the semi-arid region of Northeastern Brazil stands out as a limiting factor for agricultural production. Thus, the use of waters with high concentration of salts is an alternative to expand irrigated agriculture in this region. In this context, this study evaluated the water status, intercellular electrolyte leakage, photosynthetic pigments, and gas exchange of 'BRS SC1' sour passion fruit as a function of irrigation with water of different levels of salinity and potassium doses. The experiment was carried out in pots adapted as drainage lysimeters under field conditions at the Experimental Farm of the Federal University of Campina Grande in São Domingos - PB, Brazil. The experimental design was randomized blocks, in a 5 × 4 factorial scheme, whose treatments were obtained by combining two factors: five levels of electrical conductivity of irrigation water - ECw (0.3, 1.1, 1.9, 2.7, and 3.5 dS m-1), associated with four potassium doses (60, 80, 100, and 120% of the recommendation), with three replicates. Water with electrical conductivity greater than 0.3 dS m-1 reduced the relative water content, chlorophyll a and chlorophyll b contents, CO2 assimilation rate, and instantaneous water use efficiency of 'BRS SC1' sour passion fruit plants. The estimated potassium dose of 85% of the recommendation (equivalent to 293 g per plant per year) mitigated the deleterious effects of salt stress on stomatal conductance, transpiration, internal CO2 concentration, and instantaneous carboxylation efficiency of passion fruit 'BRS SC1'.

Does silicon help to alleviate water deficit stress and in the recovery of Dipteryx alata seedlings? / O silício auxilia no alívio do estresse hídrico e na recuperação de mudas de Dipteryx alata?

Water deficit to causes serious problems in the growth and development of plants, impairing their metabolism. Thus, it is necessary to use agents that can mitigate plant damage. This study assesses the potential of silicon to mitigate water deficit stress in Dipteryx alata Vogel seedlings and to help in their recovery after the resumption of irrigation. The study analyzed four water regimes: (I) Continuous irrigation; (II) Water deficit without Si; (III) Water deficit + 0.75 mL Si; and (IV) Water deficit + 1.50 mL Si. Seedlings were evaluated in four periods: (1) (T0 ­ time zero) at the beginning of the experiment, before irrigation suspension; (2) (P0) when the photosynthetic rates (A) of seedlings under irrigation suspension reached values close to zero, period in which irrigation was resumed; (3) (REC) when A reached values ​​close to those of seedlings under continuous irrigation, characterizing the recovery period; and (4) (END) 45 days after REC, when seedlings were kept under continuous irrigation, similar to the control. Application of 0.75 mL Si alleviates damage to the photosynthetic apparatus of D. alata seedlings that remain longer under water deficit, and contributes to faster physiological recovery after the resumption of irrigation. D. alata seedlings have recovery potential after the stress period, regardless of Si application.

O déficit hídrico vem ocasionando sérios problemas no crescimento e desenvolvimento das plantas, fazendo com que ocorram danos no seu metabolismo. Assim, faz necessário o uso de agentes que possam mitigar os danos ocasionados nas plantas. Objetivamos neste estudo verificar o potencial do silício de mitigar os efeitos estressantes do déficit hídrico em mudas de Dipteryx alata Vogel, e auxiliar na recuperação após a retomada da irrigação. Foram estudados quatro regimes hídricos: (I) Irrigação contínua; (II): Déficit hídrico + Si 0; (III): Déficit hídrico + 0.75 mL de Si e (IV): Déficit hídrico + 1.50 mL de Si. As mudas foram avaliadas em quatro períodos: (1) (T0 ­ tempo zero) início do experimento, período antes de iniciar a suspensão da irrigação, (2) (F0) quando os valores da taxa fotossintética nas mudas sob suspensão da irrigação chegaram próximos à zero, quando ocorreu a retomada da irrigação; (3) (REC) quando os valores de A alcançaram valores próximos ao das mudas sob irrigação contínua, caracterizando o período de recuperação e (4) (FINAL) 45 dias após a REC quando as mudas foram mantidas sob irrigação contínua, semelhante ao controle. A aplicação de 0.75 mL de Si alivia os danos ao aparato fotossintético das mudas de D. alata por maior período sob déficit hídrico, e contribui na recuperação fisiológica de maneira mais rápida após a retomada da irrigação. As mudas de D. alata apresentam potencial de recuperação após o período de estresse, independente da aplicação de Si.

Attenuation of salt stress on the physiology and production of bell peppers by treatment with salicylic acid / Atenuação do estresse salino na fisiologia e produção de pimentões por tratamento com ácido salicílico

The use of saline water for irrigation in semi-arid regions has become a reality due to the water scarcity that occurs in most of the year. In this scenario, exogenous application of salicylic acid may be a strategy to mitigate the deleterious effects of salt stress on plants and ensure the production of socioeconomically important crops in the semiarid region of Northeast Brazil, such as bell pepper. Thus, this study examines the osmoprotective effect of salicylic acid on gas exchanges, chloroplast pigments and production components of 'All Big' bell pepper plants irrigated with water with different saline levels. The experiment was carried out in greenhouse conditions in Campina Grande - PB, Brazil. Treatments consisted of four levels of electrical conductivity on the irrigation water (0.8, 1.6, 2.4 and 3.2 dS m-¹) and four concentrations of salicylic acid (0, 1.2, 2.4 and 3.6 mM), which were distributed in a 4 × 4 factorial arrangement in a randomized block design with three replicates. Increases in irrigation water salinity from 0.8 dS m-¹ resulted in changes in gas exchange and total chlorophyll levels of 'All Big' bell pepper plants. The estimated salicylic acid concentration of 1.7 mM reduced the effects of salinity on stomatal conductance, transpiration, CO2 assimilation rate, instantaneous carboxylation efficiency, total chlorophyll and fruit diameters. Irrigation with water of 1.8, 0.8 and 1.6 dS m-¹ salinity associated with the estimated salicylic acid concentration of 1.6 mM increased the biosynthesis of chlorophylls a and b and the number of fruits, respectively, in bell pepper plants.

A utilização de água salina para irrigação, em regiões semiáridas, tem se tornado uma realidade em funçãoda escassez hídrica que ocorre na maior parte do ano. Assim, a aplicação exógena de ácido salicílico pode ser uma estratégia capaz de amenizar os efeitos deletérios do estresse salino sobre as plantas e garantira produção agrícola de culturas socioeconomicamente importantes no semiárido do Nordeste brasileiro, como o pimentão. Deste modo, objetivou-se com esta pesquisa avaliar a capacidade osmoprotetora do ácido salicílico sobre as trocas gasosas, pigmentos cloroplastídicos e componentes de produção de plantas de pimentão 'All Big' irrigadas com águas de diferentes níveis salinos. O experimento foi conduzido em condições de casa de vegetação, em Campina Grande-PB. Os tratamentos foram distribuídos no delineamento de blocos casualizados, em esquema fatorial 4 x 4, correspondendo a quatro níveis de condutividade elétrica da água de irrigação - CEa (0,8, 1,6; 2,4 e 3,2 dS m-¹) e quatro concentrações de ácido salicílico - AS (0; 1,2; 2,4 e 3,6 mM), com três repetições. O aumento da salinidade da água de irrigação a partir de 0,8 dS m-¹ resultou em alterações nas trocas gasosas e nos teores de clorofila total das plantas de pimentão 'All Big'. A concentração estimada em 1,7 mM de ácido salicílico amenizou os efeitos da salinidade sobre a condutância estomática, a transpiração, a taxa de assimilação de CO2, a eficiência instantânea da carboxilação, clorofila total e os diâmetros dos frutos. A irrigação com água de 1,8, 0,8 e 1,6dS m-¹, associada a concentração estimada de 1,6 mM de ácido salicílico incrementou respectivamente, a biossíntese de clorofila a e b, e o número de frutos em plantas de pimentão.

Cell damage, gas exchange, and growth of Annona squamosa L. under saline water irrigation and potassium fertilization / Dano celular, trocas gasosas e crescimento da Annona squamosa L. irrigada com águas salinas e fertilização potássica

The semi-arid region of Northeastern Brazil has water limitations in terms of both quantity and quality, with salt stress as a limiting factor for increasing yield in most crops. In this context, the present study aimed to evaluate cell damage, gas exchange, and growth of custard apple under salt stress and potassium fertilization. The research was carried out at the Experimental Farm of CCTA/UFCG, in São Domingos-PB, Brazil. A randomized block design was arranged in a 2 × 5 factorial scheme, with two levels of electrical conductivity of irrigation water (ECw; 1.3 and 4.0 dS m-1) and five potassium doses (10, 15, 20, 25, and 30 g of K2O per plant per year). Water salinity of 4.0 dS m-1 negatively affected the stem diameter and number of leaves in custard apple at 179 and 210 days after transplanting (DAT). The highest relative growth in stem diameter in the period of 179-245 DAT was obtained in plants irrigated with 4.0 dS m-1 water and fertilized with 20 g of K2O per plant. Potassium doses of up to 30 g of K2O resulted in a higher percentage of cell damage and relative water content in custard apple leaf tissue. Water saturation deficit decreased with the increase in K2O doses in plants irrigated with water of 1.3 dS m-1. Irrigation with 1.3 dS m-1 water and estimated K2O doses ranging from 16 to 22 g per plant resulted in an increase in stomatal conductance, transpiration, CO2 assimilation rate, and instantaneous carboxylation efficiency in custard apple plants at 210 DAT.(AU)

A região semiárida do Nordeste brasileiro apresenta limitações hídricas em termos quantitativos e qualitativos, tendo o estresse salino como um fator limitante para o aumento da produtividade na maioria das culturas. Neste contexto, o presente estudo teve como objetivo, avaliar o dano celular, as trocas gasosas e o crescimento da pinheira sob estresse salino e adubação potássica. A pesquisa foi realizada na Fazenda Experimental do CCTA/UFCG, em São Domingos-PB, Brasil. Foi utilizado o delineamento em blocos casualizados, arranjados em esquema fatorial 2 × 5 sendo dois níveis de condutividade elétrica da água de irrigação - CEa (1,3 e 4,0 dS m-1) e cinco doses de potássio (10, 15, 20, 25 e 30 g de K2O por planta por ano). A salinidade da água de 4,0 dS m-1 afetou de forma negativa o diâmetro de caule e o número de folhas da pinheira, aos 179 e 210 dias após o transplantio (DAT). O maior crescimento relativo em diâmetro de caule no período de 179-245 DAT foi obtido nas plantas irrigadas com água de 4,0 dS m-1 e adubação com 20 g de K2O por planta. Doses de potássio de até 30 g de K2O resultaram em maior percentual de dano celular e conteúdo relativo de água nos tecidos foliares da pinheira. O déficit de saturação hídrica diminuiu com o aumento nas doses de K2O nas plantas irrigadas com água de 1,3 dS m-1. Irrigação com água de 1,3 dS m-1 e doses estimadas de K2O variando de 16 a 22 g por planta resultaram em incremento na condutância estomática, transpiração, taxa de assimilação de CO2 e eficiência instantânea de carboxilação das plantas de pinheira, aos 210 DAT.(AU)

Attenuation of salt stress on the physiology and production of bell peppers by treatment with salicylic acid / Atenuação do estresse salino na fisiologia e produção de pimentões por tratamento com ácido salicílico

The use of saline water for irrigation in semi-arid regions has become a reality due to the water scarcity that occurs in most of the year. In this scenario, exogenous application of salicylic acid may be a strategy to mitigate the deleterious effects of salt stress on plants and ensure the production of socioeconomically important crops in the semiarid region of Northeast Brazil, such as bell pepper. Thus, this study examines the osmoprotective effect of salicylic acid on gas exchanges, chloroplast pigments and production components of 'All Big' bell pepper plants irrigated with water with different saline levels. The experiment was carried out in greenhouse conditions in Campina Grande - PB, Brazil. Treatments consisted of four levels of electrical conductivity on the irrigation water (0.8, 1.6, 2.4 and 3.2 dS m-¹) and four concentrations of salicylic acid (0, 1.2, 2.4 and 3.6 mM), which were distributed in a 4 × 4 factorial arrangement in a randomized block design with three replicates. Increases in irrigation water salinity from 0.8 dS m-¹ resulted in changes in gas exchange and total chlorophyll levels of 'All Big' bell pepper plants. The estimated salicylic acid concentration of 1.7 mM reduced the effects of salinity on stomatal conductance, transpiration, CO2 assimilation rate, instantaneous carboxylation efficiency, total chlorophyll and fruit diameters. Irrigation with water of 1.8, 0.8 and 1.6 dS m-¹ salinity associated with the estimated salicylic acid concentration of 1.6 mM increased the biosynthesis of chlorophylls a and b and the number of fruits, respectively, in bell pepper plants.(AU)

A utilização de água salina para irrigação, em regiões semiáridas, tem se tornado uma realidade em funçãoda escassez hídrica que ocorre na maior parte do ano. Assim, a aplicação exógena de ácido salicílico pode ser uma estratégia capaz de amenizar os efeitos deletérios do estresse salino sobre as plantas e garantira produção agrícola de culturas socioeconomicamente importantes no semiárido do Nordeste brasileiro, como o pimentão. Deste modo, objetivou-se com esta pesquisa avaliar a capacidade osmoprotetora do ácido salicílico sobre as trocas gasosas, pigmentos cloroplastídicos e componentes de produção de plantas de pimentão 'All Big' irrigadas com águas de diferentes níveis salinos. O experimento foi conduzido em condições de casa de vegetação, em Campina Grande-PB. Os tratamentos foram distribuídos no delineamento de blocos casualizados, em esquema fatorial 4 x 4, correspondendo a quatro níveis de condutividade elétrica da água de irrigação - CEa (0,8, 1,6; 2,4 e 3,2 dS m-¹) e quatro concentrações de ácido salicílico - AS (0; 1,2; 2,4 e 3,6 mM), com três repetições. O aumento da salinidade da água de irrigação a partir de 0,8 dS m-¹ resultou em alterações nas trocas gasosas e nos teores de clorofila total das plantas de pimentão 'All Big'. A concentração estimada em 1,7 mM de ácido salicílico amenizou os efeitos da salinidade sobre a condutância estomática, a transpiração, a taxa de assimilação de CO2, a eficiência instantânea da carboxilação, clorofila total e os diâmetros dos frutos. A irrigação com água de 1,8, 0,8 e 1,6dS m-¹, associada a concentração estimada de 1,6 mM de ácido salicílico incrementou respectivamente, a biossíntese de clorofila a e b, e o número de frutos em plantas de pimentão.(AU)

Experimental infection of commercial layers with wild or attenuated Salmonella Gallinarum mutant strains: anatomic pathology, total blood cell count and serum protein levels

The aim of the present study was to comparatively evaluate hemogram, blood serum components and anatomopathologic alterations in commercial layers experimentally challenged with an attenuated vaccine candidate strain (SG∆cobS∆cbiA) and other two pathogenic strains (SGDcobS and SGNalr) of Gallinarum (SG). In total, 280 commercial layers were randomly divided into 4 groups (G1, G2, G3 and G4). At five days of age, birds from groups G1 received approximately 107 colony forming units (CFU) of SGDcobS; meanwhile birds from group G2 and G3 received the same dose of SGNalr and SG∆cobS∆cbiA, respectively. Birds from G4 were not infected. At 24 hours before (DBI) and 24 hours after (1 DAI), and three (3 DAI), five (5 DAI), seven (7 DAI) ten (10 DAI), and fifteen (15 DAI) days after the infection, 10 birds of each group were humanely killed and blood samples collected to hematological and serum tests. Samples of liver, spleen, thymus, bursa of Fabricius, kidney and heart were also collected for the histological examination. Birds inoculated with SGDcobS and SGNalr showed similar alterations in hemogram, blood serum components and anatomopathologic exams. On the other hand, the exams of birds inoculated with SG∆cobS∆cbiA strain were similar to those of the uninfected birds. However, changes could be noticed in levels of uric acid and cholesterol during the course of the infection of birds from G3. Decrease in levels of light IgG 3 DAI was also observed in birds from this group. Pyknosis in kidney cells was a microscopic alteration found in birds from G3. Further studies must be done to verify if these alterations will not interfere in the performance of the vaccinate birds with SG∆cobS∆cbiA strain.(AU)

Experimental infection of commercial layers with wild or attenuated Salmonella Gallinarum mutant strains: anatomic pathology, total blood cell count and serum protein levels

The aim of the present study was to comparatively evaluate hemogram, blood serum components and anatomopathologic alterations in commercial layers experimentally challenged with an attenuated vaccine candidate strain (SGcobScbiA) and other two pathogenic strains (SGDcobS and SGNalr) of Gallinarum (SG). In total, 280 commercial layers were randomly divided into 4 groups (G1, G2, G3 and G4). At five days of age, birds from groups G1 received approximately 107 colony forming units (CFU) of SGDcobS; meanwhile birds from group G2 and G3 received the same dose of SGNalr and SGcobScbiA, respectively. Birds from G4 were not infected. At 24 hours before (DBI) and 24 hours after (1 DAI), and three (3 DAI), five (5 DAI), seven (7 DAI) ten (10 DAI), and fifteen (15 DAI) days after the infection, 10 birds of each group were humanely killed and blood samples collected to hematological and serum tests. Samples of liver, spleen, thymus, bursa of Fabricius, kidney and heart were also collected for the histological examination. Birds inoculated with SGDcobS and SGNalr showed similar alterations in hemogram, blood serum components and anatomopathologic exams. On the other hand, the exams of birds inoculated with SGcobScbiA strain were similar to those of the uninfected birds. However, changes could be noticed in levels of uric acid and cholesterol during the course of the infection of birds from G3. Decrease in levels of light IgG 3 DAI was also observed in birds from this group. Pyknosis in kidney cells was a microscopic alteration found in birds from G3. Further studies must be done to verify if these alterations will not interfere in the performance of the vaccinate birds with SGcobScbiA strain.

Safety and efficacy of a salmonella gallinarum ΔcobSΔcbiA strain with potential to prevent chicken infections by salmonella gallinarum and salmonella enteritidis

New vaccine design techniques have allowed the development of effective vaccine strains against Salmonella infections inwhich the risks of reversion to the wild type and virulence is null. The mutant strain Salmonella Gallinarum ΔcobSΔcbiA was previously shown to be avirulent in chickens. In this study, this strain was tested as a vaccine against Salmonella Gallinarum (SG) and S. Enteritidis (SE) infections, and its protection levels, safety and possible risks of reversion to virulence after vaccination of layers were evaluated. Birds were vaccinated at five days of age or at five and 25 days of age. At 45 days of age, brown and white layers were challenged with SG and SE wild strains, respectively. Two assays to test the possibility of reversion to virulence were performed. Five successive bacterial passages in brown layers were carried out in the first assay. In the second assay, brown layers received a ten-fold concentrated inoculum of the SGΔcobSΔcbiA strain and were evaluated for clinical signs and mortality. In both experiments, no birds that received the inoculation of the attenuated strain died. Additionally, the use of the mutant strain as a vaccine provided good protection levels against both challenge strains.(AU)

Safety and efficacy of a salmonella gallinarum cobScbiA strain with potential to prevent chicken infections by salmonella gallinarum and salmonella enteritidis

New vaccine design techniques have allowed the development of effective vaccine strains against Salmonella infections inwhich the risks of reversion to the wild type and virulence is null. The mutant strain Salmonella Gallinarum cobScbiA was previously shown to be avirulent in chickens. In this study, this strain was tested as a vaccine against Salmonella Gallinarum (SG) and S. Enteritidis (SE) infections, and its protection levels, safety and possible risks of reversion to virulence after vaccination of layers were evaluated. Birds were vaccinated at five days of age or at five and 25 days of age. At 45 days of age, brown and white layers were challenged with SG and SE wild strains, respectively. Two assays to test the possibility of reversion to virulence were performed. Five successive bacterial passages in brown layers were carried out in the first assay. In the second assay, brown layers received a ten-fold concentrated inoculum of the SGcobScbiA strain and were evaluated for clinical signs and mortality. In both experiments, no birds that received the inoculation of the attenuated strain died. Additionally, the use of the mutant strain as a vaccine provided good protection levels against both challenge strains.

A thymidine kinase-negative bovine herpesvirus 5 is highly attenuated for rabbits, but is neuroinvasive and establishes latent infection / Cepa do herpesvírus bovino tipo 5 defectiva na timidina quinase é altamente atenuada para coelhos mas é neuroinvasiva e estabelece infecção latente

Mutant viral strains deleted in non-essential genes represent useful tools to study the function of specific gene products in the biology of the virus. We herein describe an investigation on the phenotype of a bovine herpesvirus 5 (BoHV-5) recombinant deleted in the gene encoding the enzyme thymidine kinase (TK) in rabbits, with special emphasis to neuroinvasiveness and the ability to establish and reactivate latent infection. Rabbits inoculated with the parental virus (SV-507/99) (n=18) at a low titer (10(5.5)TCID50) shed virus in nasal secretions in titers up to 10(4.5)TCID50 for up to 12 days (average: 9.8 days [5-12]) and 5/ 16 developed neurological disease and were euthanized in extremis. Rabbits inoculated with the recombinant BoHV-5TKΔ at a high dose (10(7.1)TCID50) also shed virus in nasal secretions, yet to lower titers (maximum: 10(2.3)TCID50) and for a shorter period (average: 6.6 days [2-11]) and remained healthy. PCR examination of brain sections of inoculated rabbits at day 6 post-infection (pi) revealed a widespread distribution of the parental virus, whereas DNA of the recombinant BoHV-5TKΔ-was detected only in the trigeminal ganglia [TG] and olfactory bulbs [OB]. Nevertheless, during latent infection (52pi), DNA of the recombinant virus was detected in the TGs, OBs and also in other areas of the brain, demonstrating the ability of the virus to invade the brain. Dexamethasone (Dx) administration at day 65 pi was followed by virus reactivation and shedding by 5/8 rabbits inoculated with the parental strain (mean duration of 4.2 days [1 - 9]) and by none of seven rabbits inoculated with the recombinant virus. Again, PCR examination at day 30 post-Dx treatment revealed the presence of latent DNA in the TGs, OBs and in other areas of the brain of both groups. Taken together, these results confirm that the recombinant BoHV-5TKΔ is highly attenuated for rabbits. It shows a reduced ability to replicate in the nose but retains the ability [...](AU)

Cepas virais mutantes defectivas em genes não essenciais se constituem em ferramentas úteis para o estudo da função de proteínas virais na biologia dos vírus. Este estudo relata uma investigação do fenótipo, em coelhos, de uma cepa recombinante do herpesvírus bovino tipo 5 (BoHV-5) defectiva na enzima timidina quinase (TK), com ênfase para a neuroinvasividade e capacidade de estabelecer e reativar a infecção latente. Coelhos inoculados com o vírus parental (SV-507/99, n=18) em baixo título (10(5,5)TCID50) excretaram o vírus nas secreções nasais em títulos de até 10(4,5)TCID50/ mL por até 12 dias (média: 9,8 dias [5-12]) e 5/16 desenvolveram doença neurológica e morreram ou foram eutanasiados in extremis. Em contraste, coelhos inoculados com o recombinante BoHV-5TKΔ em alto título (10(7,1)TCID50) excretaram o vírus em títulos inferiores (máximo 10(2,3)TCID50/ mL), por um período menor (média: 6,6 days [2-11]) e permaneceram saudáveis. A realização de PCR em seções do encéfalo no dia 6 pós-infecção (pi) revelou uma ampla distribuição do DNA do vírus parental, enquanto o DNA do vírus recombinante foi detectado apenas nos gânglios trigêmeos [TGs] e nos bulbos olfatórios [OBs]. Não obstante, durante a infecção latente (52pi), o DNA do vírus recombinante foi detectado nos TGs, OBs e em outros locais do encéfalo, demonstrando que o vírus recombinante mantém a neuroinvasividade. Tratamento com dexamethasona (Dx) no dia 65 pi resultou em reativação e excreção viral por 5/8 dos coelhos inoculados com o vírus parental (duração média de 4,2 dias [1-9]) e por nenhum dos sete coelhos inoculados com o vírus recombinante. No entanto, PCR realizado no dia 30 pós-Dx revelou a presença de DNA latente do BoHV-5TKΔ nos TGs, OBs e em outras áreas do encéfalo. Esses resultados confirmam que o recombinante BoHV-5TKΔ é altamente atenuado para coelhos. A sua capacidade de replicação na mucosa nasal é reduzida, mas mantém a capacidade de invadir o encéfalo e estabelecer infecção [...](AU)

Experimental infection of calves with recombinants of bovine herpesvirus 5 defective in glycoprotein E (gE), thymidine kinase (TK) and both, gE/TK / Infecção experimental de bezerros com recombinantes do herpesvírus bovino tipo 5 defectivos na glicoproteína E (gE), timidina quinase (TK) e ambos, gE/TK

This article describes an investigation on the virulence/attenuation of bovine herpesvirus type 5 (BoHV-5) recombinants deleted in the genes encoding glycoprotein E (BoHV-5gEΔ), thymidine kinase (BoHV-5TKΔ), and both gE and TK (BoHV-5gEΔTKΔ). Seronegative calves (80 to 90 days-old) inoculated with the parental strain (SV-507/99, n=5) shed virus in nasal secretions for up to 15 days (average 10.8 days). Duration of virus shedding was 11 days for BoHV-5gΔ, 9.6 days for BoHV-5TKΔ and 6.2 days for BoHV-5gEΔTKΔ groups. The highest titers were observed between days 1 and 6 post-infection (pi) for SV-507/99 (10(6.8)TCID50/mL), 10(5.1)TCID50/mL (BoHV-5gEΔ), 10(5.9)TCID50/mL (BoHV-5TKΔ) and 10(4.7)TCID50/mL (BoHV-5gEΔTKΔ). Calves inoculated with the parental virus presented anorexia, profound apathy and loss of body condition. Two calves were euthanized in extremis on days 10 and 11 pi; infectious virus was recovered from several areas of the brain. In contrast, calves inoculated with the recombinants remained healthy and a few presented a mild and transient nasal secretion. Dexamethasone (Dx) administration at day 42 pi resulted in virus shedding by all controls calves (mean duration 3.7 days), by 2/5 of BoHV-5TKΔ calves (two days) and 2/5 of BoHV-5gEΔ (one day). No virus shedding was detected in BoHV-5gEΔTKΔ calves upon Dx treatment. PCR examination of brain sections of calves euthanized at day 30 post Dx treatment revealed the presence of latent viral DNA widely distributed in the brain of SV-507/99 calves. Latent viral DNA was detected in a few sections (3/30) of the brains of BoHV-5gEΔ calves and was not detected in the brains of calves inoculated with BoHV-5TKΔ and BoHV-5gEΔTKΔ. These results show that the single BoHV-5 mutants (gE and tk-deleted) are attenuated for calves and establish and/or reactivate latent infection inefficiently. The double mutant BoHV-5gEΔTKΔ is fully attenuated and appears not to establish or not reactivate efficiently from latent infection. Thus, these recombinants, especially the double mutant BoHV-5gEΔTKΔ, display an adequate phenotype for use in modified-live vaccine formulations.(AU)

Este artigo descreve uma investigação da virulência/atenuação de recombinantes do herpesvírus bovino tipo 5 (BoHV-5) com deleções nos genes da glicoproteína E (BoHV-5gEΔ), timidina quinase (BoHV-5TKΔ), e ambos gE e TK (BoHV-5gEΔTKΔ). Bezerros soronegativos (80-90 dias de idade) inoculados com o vírus parental SV-507/99 (n=5) excretaram o vírus em secreções nasais por até 15 dias (média de 10,8 dias). Nos animais inoculados com os recombinantes, a duração da excreção viral foi de 11 dias (BoHV-5gEΔ), 9,6 dias (BoHV-5TKΔ) e 6,2 dias (BoHV-5gEΔTKΔ). Os maiores títulos foram observados entre os dias 1 e 6 pós-inoculação (pi), sendo de 10(6,8)TCID50/mL para o SV-507/99, 10(5,1)TCID50/mL (BoHV-5gEΔ), 10(5,9)TCID50/mL (BoHV-5TKΔ) e 10(4,7)TCIΔ50/mL (BoHV-5gEΔTKΔ). Os bezerros inoculados com o vírus parental apresentaram anorexia e apatia; três deles mostraram apatia profunda e perda da condição corporal. Dois bezerros foram eutanasiados in extremis nos dias 10 e 11 pi, respectivamente e o vírus foi isolado de várias regiões do encéfalo. Já os bezerros inoculados com os recombinantes permaneceram saudáveis; alguns apresentaram uma secreção nasal serosa transitória. Administração de dexametasona (Dx) no dia 42 pi resultou em excreção viral por todos os bezerros inoculados com o vírus parental (duração média de 3,7 dias), por 2 de 5 bezerros dos grupos BoHV-5TKΔ (dois dias) e BoHV-5gEΔ (um dia). Os bezerros inoculados com o duplo mutante BoHV-5gEΔTKΔ não excretaram o vírus após o tratamento com Dx. Pesquisa de DNA viral por PCR no dia 30 pós-Dx revelou uma ampla distribuição do DNA do vírus parental no encéfalo; poucas seções (3/30) foram positivas no encéfalo dos animais do grupo BoHV-5gEΔ, e não detectou-se DNA latente no encéfalo dos animais dos grupos BoHV-5TKΔ e BoHV-5gEΔTKΔ. Esses resultados demonstram que os mutantes simples (gE and tk-deletados) são atenuados para bezerros e estabelecem e/ou reativam infecção latente ineficientemente. Já o duplo mutante BoHV-5gEΔTKΔ é atenuado e parece não estabelecer e/ou não reativar eficientemente a infecção latente. Portanto, os vírus recombinantes, e em especial o duplo mutante BoHV-5gEΔTKΔ apresentam um fenótipo compatível com a sua inclusão em vacinas vivas modificadas.(AU)

A recombinant bovine herpesvirus 5 defective in thymidine kinase and glycoprotein E is attenuated and immunogenic for calves / Cepa recombinante do herpesvírus bovino tipo 5 defectiva na timidina quinase e glicoproteína E é atenuada e imunogênica para bezerros

Bovine herpesvirus 5 (BoHV-5) is an important pathogen of cattle in South America and efforts have been made to produce safer and more effective vaccines. In addition to afford protection, herpesvirus vaccines should allow serological differentiation of vaccinated from naturally, latently infected animals. We previously reported the construction and characterization in vitro of a double mutant BoHV-5 (BoHV-5gE/TK􀀀Δ) lacking the genes encoding thymidine kinase (tk) for attenuation, and glycoprotein E (gE) as the antigenic marker, as a vaccine candidate strain (Brum et al. 2010a). The present article reports an investigation on the attenuation and immunogenicity of this recombinant in calves. In a first experiment, 80 to 90-day-old seronegative calves (n=6) inoculated intranasally with the recombinant (titer of 107.5TCID50) shed virus in low to moderate titers in nasal secretions for up to 6 days, yet did not develop any respiratory, systemic or neurological signs of infection. At day 30 post-infection (pi) all calves had BoHV-5 specific neutralizing (VN) antibodies in titers of 4 to 8 and were negative for anti-gE antibodies in a commercial ELISA test. Administration of dexamethasone (0.1mg/kg/day during 5 days) to four of these calves at day 42 pi did not result in virus shedding or increase in VN titers, indicating lack of viral reactivation. Secondly, a group of 8-month-old calves (n=9) vaccinated intramuscularly (IM) with the recombinant virus (107.5TCID50/animal) did not shed virus in nasal secretions, remained healthy and developed VN titers from 2 to 8 at day 42 post-vaccination (pv), remaining negative for gE antibodies. Lastly, 21 calves (around 10 months old) maintained under field conditions were vaccinated IM with the recombinant virus (titer of 107.3TCID50). All vaccinated animals developed VN titers from 2 to 16 at day 30 pv. A boost vaccination performed at day 240 pv resulted in a rapid and strong anamnestic antibody response, with VN titers reaching from 16 to 256 at day 14 post-booster. Again, serum samples remained negative for gE antibodies. Selected serum samples from vaccinated animals showed a broad VN activity against nine BoHV-5 and eight BoHV-1 field isolates. These results show that the recombinant virus is attenuated, immunogenic for calves and induces an antibody response differentiable from that induced by natural infection. Thus, the recombinant BoHV-5gE/TKΔ is an adequate candidate strain for a modified live vaccine.(AU)

Application of a bacterial extracellular polymeric substance in heavy metal adsorption in a co-contaminated aqueous system

The application of a bacterial extracellular polymeric substance (EPS) in the bioremediation of heavy metals (Cd, Zn and Cu) by a microbial consortium in a hydrocarbon co-contaminated aqueous system was studied. At the low concentrations used in this work (1.00 ppm of each metal), it was not observed an inhibitory effect on the cellular growing. In the other hand, the application of the EPS lead to a lower concentration of the free heavy metals in solution, once a great part of them is adsorbed in the polymeric matrix (87.12% of Cd; 19.82% of Zn; and 37.64% of Cu), when compared to what is adsorbed or internalized by biomass (5.35% of Cd; 47.35% of Zn; and 24.93% of Cu). It was noted an increase of 24% in the consumption of ethylbenzene, among the gasoline components that were quantified, in the small interval of time evaluated (30 hours). Our results suggest that, if the experiments were conducted in a larger interval of time, it would possibly be noted a higher effect in the degradation of gasoline compounds. Still, considering the low concentrations that were evaluated, it is possible that a real system could be bioremediated by natural attenuation process, demonstrated by the low effect of those levels of contaminants and co-contaminants over the naturally present microbial consortium.

A aplicação de uma substância polimérica extracelular (EPS) bacteriana na biorremediação de metais pesados (Cd, Zn e Cu) por um consórcio microbiano em um sistema aquoso co-contaminado com hidrocarbonetos foi estudada. Nas baixas concentrações usadas neste trabalho (1,00 ppm de cada metal), não foi observado um efeito inibitório no crescimento celular. Por outro lado, a aplicação da EPS bacteriana levou a uma menor concentração de metais livres em solução, uma vez que grande parte destes fica adsorvido na matriz polimérica (87,12% de Cd; 19,82% de Zn; e 37,64% de Cu) quando comparado ao que é adsorvido ou interiorizado pela biomassa (5,35% de Cd; 47,35% de Zn; e 24,93% de Cu). No pequeno intervalo de tempo avaliado (30 horas) e na baixa concentração de gasolina utilizada (0,1% (v/v)), foi percebido um aumento de 24% no consumo de etilbenzeno, entre os componentes da gasolina que foram quantificados. Nossos resultados sugerem que, se os experimentos fossem conduzidos em um intervalo de tempo maior, possivelmente poderia ter sido observado um maior efeito na degradação dos componentes da gasolina. Ainda, considerando as baixas concentrações avaliadas, é provável que um sistema real pudesse ser biorremediado pelo processo de atenuação natural, tendo em vista o baixo efeito desses níveis de contaminantes e co-contaminantes sobre o consórcio microbiano naturalmente presente.

Gamma-ray beam attenuation as an auxiliary technique for the evaluation of the soil water retention curve

The soil water retention curve is fundamental for the hydraulic characterization of a soil and has many applications in agricultural research as well as in practical agriculture. A new procedure for soil moisture and soil bulk density evaluation inside closed pressure chambers through gamma-ray beam attenuation is presented. The proposed procedure presents several advantages in relation to the traditional process: avoids the need of continuous sample manipulation; minimizes the problem of hysteresis; allows a more precise evaluation of soil moisture by taking into account changes of soil bulk density due to swelling or shrinking on addition or removal of water; allows frequent evaluation of soil moisture without the need of opening the pressure chamber; allows a more precise judgement of equilibrium; reduces drastically the time of the determination of the retention curve and allows easy automation of data acquisition by a computer.

A curva de retenção da água no solo é fundamental para a caracterização hídrica de um solo e tem muitas aplicações na pesquisa agrícola assim como na agricultura aplicada. Um novo procedimento para determinação da umidade e densidade do solo no interior de câmaras de pressão utilizadas para deter minação de curvas de retenção é apresentado. O procedimento proposto apresenta várias vantagens em relação ao processo tradicional, tais como: evita a necessidade de constante manipulação das amostras; minimiza o problema da histerese; permite uma determinação mais precisa da umidade por levar em conta possíveis alterações na densidade do solo devido à expansão e/ou contração durante a adição ou retirada de água; permite constante avaliação da umidade do solo sem a necessidade de abertura da câmara; permite um julgamento mais preciso do ponto de equilíbrio após cada período de extração; reduz drasticamente o tempo necessário para a determinação das curvas de retenção e permite fácil automação do sistema de aquisição de dados com ajuda de um computador.

Bioremediation of soil contaminated by diesel oil

Were evaluated natural attenuation, biostimulation and bioaugmentation on the degradation of total petroleum hydrocarbons (TPH) in soils contaminated with diesel oil. Bioaugmentation showed the greatest degradation in the light (C12 - C23) fractions (72.7%) and heavy (C23 - C40) fractions of TPH (75.2%) and natural attenuation was more effective than biostimulation. The greatest dehydrogenase activity was observed upon bioaugmentation of the Long Beach soil (3.3-fold) and the natural attenuation of the Hong Kong soil sample (4.0-fold). The number of diesel oil degrading microorganisms and heterotrophic population was not influenced by the bioremediation treatments. The best approach for bioremediation of soil contaminated with diesel oil is the inoculum of microorganisms pre-selected from their own environment.

Avaliou-se a degradação de hidrocarbonetos de petróleo (HP) em solos contaminados com óleo diesel através da atenuação natural, bioestimulação e bioaumentação. A bioaumentação apresentou a maior degradação da fração leve (72,6%) e da fração pesada (75,2%) de HP e a atenuação natural foi mais efetiva do que a bioestimulação. A maior atividade da dehidrogenase no solo Long Beach e Hong Kong foi observada nos tratamentos bioaumentação e atenuação natural, respectivamente. O número de microrganismos degradadores de diesel e a população de heterotróficos não foi influenciada pelas técnicas de biorremediação. A melhor performance para a biorremediação do solo contaminado com diesel foi obtida quando foram adicionados microrganismos pré-selecionados do ambiente contaminado.

Bioremediation of soil contaminated by diesel oil

Were evaluated natural attenuation, biostimulation and bioaugmentation on the degradation of total petroleum hydrocarbons (TPH) in soils contaminated with diesel oil. Bioaugmentation showed the greatest degradation in the light (C12 - C23) fractions (72.7%) and heavy (C23 - C40) fractions of TPH (75.2%) and natural attenuation was more effective than biostimulation. The greatest dehydrogenase activity was observed upon bioaugmentation of the Long Beach soil (3.3-fold) and the natural attenuation of the Hong Kong soil sample (4.0-fold). The number of diesel oil degrading microorganisms and heterotrophic population was not influenced by the bioremediation treatments. The best approach for bioremediation of soil contaminated with diesel oil is the inoculum of microorganisms pre-selected from their own environment.

Avaliou-se a degradação de hidrocarbonetos de petróleo (HP) em solos contaminados com óleo diesel através da atenuação natural, bioestimulação e bioaumentação. A bioaumentação apresentou a maior degradação da fração leve (72,6%) e da fração pesada (75,2%) de HP e a atenuação natural foi mais efetiva do que a bioestimulação. A maior atividade da dehidrogenase no solo Long Beach e Hong Kong foi observada nos tratamentos bioaumentação e atenuação natural, respectivamente. O número de microrganismos degradadores de diesel e a população de heterotróficos não foi influenciada pelas técnicas de biorremediação. A melhor performance para a biorremediação do solo contaminado com diesel foi obtida quando foram adicionados microrganismos pré-selecionados do ambiente contaminado.

Bioremediation of soil contaminated by diesel oil

Were evaluated natural attenuation, biostimulation and bioaugmentation on the degradation of total petroleum hydrocarbons (TPH) in soils contaminated with diesel oil. Bioaugmentation showed the greatest degradation in the light (C12 - C23) fractions (72.7%) and heavy (C23 - C40) fractions of TPH (75.2%) and natural attenuation was more effective than biostimulation. The greatest dehydrogenase activity was observed upon bioaugmentation of the Long Beach soil (3.3-fold) and the natural attenuation of the Hong Kong soil sample (4.0-fold). The number of diesel oil degrading microorganisms and heterotrophic population was not influenced by the bioremediation treatments. The best approach for bioremediation of soil contaminated with diesel oil is the inoculum of microorganisms pre-selected from their own environment.

Avaliou-se a degradação de hidrocarbonetos de petróleo (HP) em solos contaminados com óleo diesel através da atenuação natural, bioestimulação e bioaumentação. A bioaumentação apresentou a maior degradação da fração leve (72,6%) e da fração pesada (75,2%) de HP e a atenuação natural foi mais efetiva do que a bioestimulação. A maior atividade da dehidrogenase no solo Long Beach e Hong Kong foi observada nos tratamentos bioaumentação e atenuação natural, respectivamente. O número de microrganismos degradadores de diesel e a população de heterotróficos não foi influenciada pelas técnicas de biorremediação. A melhor performance para a biorremediação do solo contaminado com diesel foi obtida quando foram adicionados microrganismos pré-selecionados do ambiente contaminado.

Gamma-ray beam attenuation as an auxiliary technique for the evaluation of the soil water retention curve

The soil water retention curve is fundamental for the hydraulic characterization of a soil and has many applications in agricultural research as well as in practical agriculture. A new procedure for soil moisture and soil bulk density evaluation inside closed pressure chambers through gamma-ray beam attenuation is presented. The proposed procedure presents several advantages in relation to the traditional process: avoids the need of continuous sample manipulation; minimizes the problem of hysteresis; allows a more precise evaluation of soil moisture by taking into account changes of soil bulk density due to swelling or shrinking on addition or removal of water; allows frequent evaluation of soil moisture without the need of opening the pressure chamber; allows a more precise judgement of equilibrium; reduces drastically the time of the determination of the retention curve and allows easy automation of data acquisition by a computer.

A curva de retenção da água no solo é fundamental para a caracterização hídrica de um solo e tem muitas aplicações na pesquisa agrícola assim como na agricultura aplicada. Um novo procedimento para determinação da umidade e densidade do solo no interior de câmaras de pressão utilizadas para deter minação de curvas de retenção é apresentado. O procedimento proposto apresenta várias vantagens em relação ao processo tradicional, tais como: evita a necessidade de constante manipulação das amostras; minimiza o problema da histerese; permite uma determinação mais precisa da umidade por levar em conta possíveis alterações na densidade do solo devido à expansão e/ou contração durante a adição ou retirada de água; permite constante avaliação da umidade do solo sem a necessidade de abertura da câmara; permite um julgamento mais preciso do ponto de equilíbrio após cada período de extração; reduz drasticamente o tempo necessário para a determinação das curvas de retenção e permite fácil automação do sistema de aquisição de dados com ajuda de um computador.

Atenuação da radiação gama no estudo da distribuição de partículas em solo sob floresta e cana-de-açúcar

The use of a new soil particle size distribution analysis methodology is presented in connection with the study of particle size distribution in soils covered by natural forest and sugar cane crop cultivated during 16, 30 and 50 years, continuously. The technique is based on the attenuation of a gamma-ray beam by a conventional soil/water suspension under sedimentation. Results show a significant difference in clay content for the upper soil layer as a function of sugar cane cultivation.

É apresentado o uso de uma nova metodologia de análise da distribuição de tamanho de partículas de solo, em amostras de terra sob floresta nativa e submetido ao cultivo de cana-de-açúcar por 16, 30 e 50 anos consecutivos. A técnica para análise granulométrica do solo baseia-se na atenuação de um feixe de radiação gama, por uma suspensão convencional terra/água em sedimentação. Os resultados indicam uma diferença significativa dos teores de argila nas camadas superficiais do solo em função do cultivo com cana-de-açúcar.

Exact equations for soil particle-size analysis by gamma-ray attenuation

Soil particle-size analysis by gamma-ray attenuation was first suggested in 1992 and improved in 1997 by diminishing the measurement time and adapting it for automation. It is here demonstrated that when the mass attenuation coefficient FONT FACE="Symbol">m /font>w and the density Dw are replaced by FONT FACE="Symbol">m /font>s and Ds (where the subscripts w and s stand for water and solution) in the approximate equations used to estimate the concentration of suspended particles, they become exact. The demonstration is based by treating the dispersant and water solution as one single medium, instead of treating them as two media. In this way, six variables are reduced to only three. Physical considerations suggest that the precision in this analysis could be improved if other types of solutions and of photon energies would be used, so that the attenuation by the particles would differ more significantly from the attenuation by the solution.

A análise de tamanho de partículas de solo por atenuação da radiação gama foi primeiramente sugerida em 1992 e melhorada em 1997 diminuindo o tempo de medida e a adaptando para automação. É demonstrado aqui que quando se substitui o coeficiente de atenuação de massa FONT FACE="Symbol">m /font>w e a densidade Dw por FONT FACE="Symbol">m /font>s e Ds. (onde os índices w e s representam água e solução) nas equações aproximadas utilizadas para o cálculo da concentração dos sólidos suspensos, estas tornam-se exatas. A demonstração baseia-se em tratar a água e o dispersante como um único meio, ao invés de tratar a solução como dois meios, ou seja, água e dispersante. Assim, seis variáveis são reduzidas a somente três. Considerações físicas sugerem que a precisão nesta análise poderia aumentar se outros tipos de solução e energias de fótons fossem utilizadas, tais que a atenuação pelas partículas se diferenciasse tanto quanto possível da atenuação pela solução.