A terbenzimidazole that preferentially binds and conformationally alters structurally distinct DNA duplex domains: A potential mechanism for topoisomerase I poisoning

The terbenzimidazoles are a class of synthetic ligands that poison the human topoisomerase I (TOP1) enzyme and promote cancer cell death. It has been proposed that drugs of this class act as TOP1 poisons by binding to the minor groove of the DNA substrate of TOP1 and altering its structure in a manner that results in enzyme-mediated DNA cleavage. To test this hypothesis, we characterize and compare the binding properties of a 5-phenylterbenzimidazole derivative (5PTB) to the d(GA4T4C)2 and d(GT4A4C)2 duplexes. The d(GA4T4C)2 duplex contains an uninterrupted 8-bp A⋅T domain, which, on the basis of x-ray crystallographic data, should induce a highly hydrated “A-tract” conformation. This duplex also exhibits anomalously slow migration in a polyacrylamide gel, a feature characteristic of a noncanonical global conformational state frequently described as “bent.” By contrast, the d(GT4A4C)2 duplex contains two 4-bp A⋅T tracts separated by a TpA dinucleotide step, which should induce a less hydrated “B-like” conformation. This duplex also migrates normally in a polyacrylamide gel, a feature further characteristic of a global, canonical B-form duplex. Our data reveal that, at 20°C, 5PTB exhibits an ≈2.3 kcal/mol greater affinity for the d(GA4T4C)2 duplex than for the d(GT4A4C)2 duplex. Significantly, we find this sequence/conformational binding specificity of 5PTB to be entropic in origin, an observation consistent with a greater degree of drug binding-induced dehydration of the more solvated d(GA4T4C)2 duplex. By contrast with the differential duplex affinity exhibited by 5PTB, netropsin and 4′,6-diamidino-2-phenylindole (DAPI), two AT-specific minor groove binding ligands that are inactive as human TOP1 poisons, bind to both duplexes with similar affinities. The electrophoretic behaviors of the ligand-free and ligand-bound duplexes are consistent with 5PTB-induced bending and/or unwinding of both duplexes, which, for the d(GA4T4C)2 duplex, is synergistic with the endogenous sequence-directed electrophoretic properties of the ligand-free duplex state. By contrast, the binding to either duplex of netropsin or DAPI induces little or no change in the electrophoretic mobilities of the duplexes. Our results demonstrate that the TOP1 poison 5PTB binds differentially to and alters the structures of the two duplexes, in contrast to netropsin and DAPI, which bind with similar affinities to the two duplexes and do not significantly alter their structures. These results are consistent with a mechanism for TOP1 poisoning in which drugs such as 5PTB differentially target conformationally distinct DNA sites and induce structural changes that promote enzyme-mediated DNA cleavage.

Study of the 3-Hydroxy Eicosanoyl-Coenzyme A Dehydratase and (E)-2,3 Enoyl-Coenzyme A Reductase Involved in Acyl-Coenzyme A Elongation in Etiolated Leek Seedlings1

(R,S)-[1-14C]3-Hydroxy eicosanoyl-coenzyme A (CoA) has been chemically synthesized to study the 3-hydroxy acyl-CoA dehydratase involved in the acyl-CoA elongase of etiolated leek (Allium porrum L.) seedling microsomes. 3-Hydroxy eicosanoyl-CoA (3-OH C20:0-CoA) dehydration led to the formation of (E)-2,3 eicosanoyl-CoA, which has been characterized. Our kinetic studies have determined the optimal conditions of the dehydration and also resolved the stereospecificity requirement of the dehydratase for (R)-3-OH C20:0-CoA. Isotopic dilution experiments showed that 3-hydroxy acyl-CoA dehydratase had a marked preference for (R)-3-OH C20:0-CoA. Moreover, the very-long-chain synthesis using (R)-3-OH C20:0-CoA isomer and [2-14C]malonyl-CoA was higher than that using the (S) isomer, whatever the malonyl-CoA and the 3-OH C20:0-CoA concentrations. We have also used [1-14C]3-OH C20:0-CoA to investigate the reductant requirement of the enoyl-CoA reductase of the acyl-CoA elongase complex. In the presence of NADPH, [1-14C]3-OH C20:0-CoA conversion was stimulated. Aside from the product of dehydration, i.e. (E)-2,3 eicosanoyl-CoA, we detected eicosanoyl-CoA resulting from the reduction of (E)-2,3 eicosanoyl-CoA. When we replaced NADPH with NADH, the eicosanoyl-CoA was 8- to 10-fold less abundant. Finally, in the presence of malonyl-CoA and NADPH or NADH, [1-14C]3-OH C20:0-CoA led to the synthesis of very-long-chain fatty acids. This synthesis was measured using [1-14C]3-OH C20:0-CoA and malonyl-CoA or (E)-2,3 eicosanoyl-CoA and [2-14C]malonyl-CoA. In both conditions and in the presence of NADPH, the acyl-CoA elongation activity was about 60 nmol mg−1 h−1, which is the highest ever reported for a plant system.

The Arabidopsis CBF Gene Family Is Composed of Three Genes Encoding AP2 Domain-Containing Proteins Whose Expression Is Regulated by Low Temperature but Not by Abscisic Acid or Dehydration1

We have identified two genes from Arabidopsis that show high similarity with CBF1, a gene encoding an AP2 domain-containing transcriptional activator that binds to the low-temperature-responsive element CCGAC and induces the expression of some cold-regulated genes, increasing plant freezing tolerance. These two genes, which we have named CBF2 and CBF3, also encode proteins containing AP2 DNA-binding motifs. Furthermore, like CBF1, CBF2 and CBF3 proteins also include putative nuclear-localization signals and potential acidic activation domains. The CBF2 and CBF3 genes are linked to CBF1, constituting a cluster on the bottom arm of chromosome IV. The high level of similarity among the three CBF genes, their tandem organization, and the fact that they have the same transcriptional orientation all suggest a common origin. CBF1, CBF2, and CBF3 show identical expression patterns, being induced very rapidly by low-temperature treatment. However, in contrast to most of the cold-induced plant genes characterized, they are not responsive to abscisic acid or dehydration. Taken together, all of these data suggest that CBF2 and CBF3 may function as transcriptional activators, controlling the level of low-temperature gene expression and promoting freezing tolerance through an abscisic acid-independent pathway.

Abscisic Acid-Dependent and -Independent Expression of the Carrot Late-Embryogenesis-Abundant-Class Gene Dc3 in Transgenic Tobacco Seedlings1

We studied the expression of three promoter 5′ deletion constructs (−218, −599, and −1312) of the LEA (late embryogenesis abundant)-class gene Dc3 fused to β-glucuronidase (GUS), where each construct value refers to the number of base pairs upstream of the transcription start site at which the deletion occurred. The Dc3 gene is noted for its induction by abscisic acid (ABA), but its response to other plant hormones and various environmental stresses has not been reported previously for vegetative cells. Fourteen-day-old transgenic tobacco (Nicotiana tabacum L.) seedlings were exposed to dehydration, hypoxia, salinity, exogenous ethylene, or exogenous methyl jasmonate (MeJa). GUS activity was quantified fluorimetrically and expression was observed by histochemical staining of the seedlings. An increase in GUS activity was observed in plants with constructs −599 and −1312 in response to dehydration and salinity within 6 h of stress, and at 12 h in response to hypoxia. No increase in endogenous ABA was found in any of the three lines, even after 72 h of hypoxia. An ABA-independent increase in GUS activity was observed when endogenous ABA biosynthesis was blocked by fluridone and plants were exposed to 5 μL L−1 ethylene in air or 100 μm MeJa. Virtually no expression was observed in construct −218 in response to dehydration, salinity, or MeJa, but there was a moderate response to ethylene and hypoxia. This suggests that the region between −218 and −599 is necessary for ABA (dehydration and salinity)- and MeJa-dependent expression, whereas ethylene-mediated expression does not require this region of the promoter.

Overexpression of Nitrate Reductase in Tobacco Delays Drought-Induced Decreases in Nitrate Reductase Activity and mRNA1

Transformed (cauliflower mosaic virus 35S promoter [35S]) tobacco (Nicotiana plumbaginifolia L.) plants constitutively expressing nitrate reductase (NR) and untransformed controls were subjected to drought for 5 d. Drought-induced changes in biomass accumulation and photosynthesis were comparable in both lines of plants. After 4 d of water deprivation, a large increase in the ratio of shoot dry weight to fresh weight was observed, together with a decrease in the rate of photosynthetic CO2 assimilation. Foliar sucrose increased in both lines during water stress, but hexoses increased only in leaves from untransformed controls. Foliar NO3− decreased rapidly in both lines and was halved within 2 d of the onset of water deprivation. Total foliar amino acids decreased in leaves of both lines following water deprivation. After 4 d of water deprivation no NR activity could be detected in leaves of untransformed plants, whereas about 50% of the original activity remained in the leaves of the 35S-NR transformants. NR mRNA was much more stable than NR activity. NR mRNA abundance increased in the leaves of the 35S-NR plants and remained constant in controls for the first 3 d of drought. On the 4th d, however, NR mRNA suddenly decreased in both lines. Rehydration at d 3 caused rapid recovery (within 24 h) of 35S-NR transcripts, but no recovery was observed in the controls. The phosphorylation state of the protein was unchanged by long-term drought. There was a strong correlation between maximal extractable NR activity and ambient photosynthesis in both lines. We conclude that drought first causes increased NR protein turnover and then accelerates NR mRNA turnover. Constitutive NR expression temporarily delayed drought-induced losses in NR activity. 35S-NR expression may therefore allow more rapid recovery of N assimilation following short-term water deficit.

Characterization of a Gene for Spinach CAP160 and Expression of Two Spinach Cold-Acclimation Proteins in Tobacco1

The cDNA sequence for CAP160, an acidic protein previously linked with cold acclimation in spinach (Spinacia oleracea L.), was characterized and found to encode a novel acidic protein of 780 amino acids having very limited homology to a pair of Arabidopsis thaliana stress-regulated proteins, rd29A and rd29B. The lack of similarity in the structural organization of the spinach and Arabidopsis genes highlights the absence of a high degree of conservation of this cold-stress gene across taxonomic boundaries. The protein has several unique motifs that may relate to its function during cold stress. Expression of the CAP160 mRNA was increased by low-temperature exposure and water stress in a manner consistent with a probable function during stresses that involve dehydration. The coding sequences for CAP160 and CAP85, another spinach cold-stress protein, were introduced into tobacco (Nicotiana tabacum) under the control of the 35S promoter using Agrobacterium tumefaciens-based transformation. Tobacco plants expressing the proteins individually or coexpressing both proteins were evaluated for relative freezing-stress tolerance. The killing temperature for 50% of the cells of the transgenic plants was not different from that of the wild-type plants. As determined by a more sensitive time/temperature kinetic study, plants expressing the spinach proteins had slightly lower levels of electrolyte leakage than wild-type plants, indicative of a small reduction of freezing-stress injury. Clearly, the heterologous expression of two cold-stress proteins had no profound influence on stress tolerance, a result that is consistent with the quantitative nature of cold-stress-tolerance traits.

Products of Proline Catabolism Can Induce Osmotically Regulated Genes in Rice1

Many plants accumulate high levels of free proline (Pro) in response to osmotic stress. This imino acid is widely believed to function as a protector or stabilizer of enzymes or membrane structures that are sensitive to dehydration or ionically induced damage. The present study provides evidence that the synthesis of Pro may have an additional effect. We found that intermediates in Pro biosynthesis and catabolism such as glutamine and Δ1-pyrroline-5-carboxylic acid (P5C) can increase the expression of several osmotically regulated genes in rice (Oryza sativa L.), including salT and dhn4. One millimolar P5C or its analog, 3,4-dehydroproline, produced a greater effect on gene expression than 1 mm l-Pro or 75 mm NaCl. These chemicals did not induce hsp70, S-adenosylmethionine synthetase, or another osmotically induced gene, Em, to any significant extent. Unlike NaCl, gene induction by P5C did not depend on the normal levels of either de novo protein synthesis or respiration, and did not raise abscisic acid levels significantly. P5C- and 3,4-dehydroproline-treated plants consumed less O2, had reduced NADPH levels, had increased NADH levels, and accumulated many osmolytes associated with osmotically stressed rice. These experiments indicate that osmotically induced increases in the concentrations of one or more intermediates in Pro metabolism could be influencing some of the characteristic responses to osmotic stress.

Binding of the cellulose-binding domain of exoglucanase Cex from Cellulomonas fimi to insoluble microcrystalline cellulose is entropically driven.

Isothermal titration microcalorimetry is combined with solution-depletion isotherm data to analyze the thermodynamics of binding of the cellulose-binding domain (CBD) from the beta-1,4-(exo)glucanase Cex of Cellulomonas fimi to insoluble bacterial microcrystalline cellulose. Analysis of isothermal titration microcalorimetry data against two putative binding models indicates that the bacterial microcrystalline cellulose surface presents two independent classes of binding sites, with the predominant high-affinity site being characterized by a Langmuir-type Ka of 6.3 (+/-1.4) x 10(7) M-1 and the low-affinity site by a Ka of 1.1 (+/-0.6) x 10(6) M-1. CBDCex binding to either site is exothermic, but is mainly driven by a large positive change in entropy. This differs from protein binding to soluble carbohydrates, which is usually driven by a relatively large exothermic standard enthalpy change for binding. Differential heat capacity changes are large and negative, indicating that sorbent and protein dehydration effects make a dominant contribution to the driving force for binding.

Lyophilization-induced reversible changes in the secondary structure of proteins.

Changes in the secondary structure of some dozen different proteins upon lyophilization of their aqueous solutions have been investigated by means of Fourier-transform infrared spectroscopy in the amide III band region. Dehydration markedly (but reversibly) alters the secondary structure of all the proteins studied, as revealed by both the quantitative analysis of the second derivative spectra and the Gaussian curve fitting of the original infrared spectra. Lyophilization substantially increases the beta-sheet content and lowers the alpha-helix content of all proteins. In all but one case, proteins become more ordered upon lyophilization.

Sticky ions in biological systems.

Aqueous gel sieving chromatography on Sephadex G-10 of the Group IA cations (Li+, Na+, K+, Rb+, Cs+) plus NH4+ as the Cl- salts, in combination with previous results for the halide anions (F-, Cl-, Br-, I-) as the Na+ salts [Washabaugh, M.W. & Collins, K.D. (1986) J. Biol. Chem. 261, 12477-12485], leads to the following conclusions. (i) The small monovalent ions (Li+, Na+, F-) flow through the gel with water molecules attached, whereas the large monovalent ions (K+, Rb+, Cs+, Cl-, Br-, I-) adsorb to the nonpolar surface of the gel, a process requiring partial dehydration of the ion and implying that these ions bind the immediately adjacent water molecules weakly. (ii) The transition from strong to weak hydration occurs at a radius of about 1.78 A for the monovalent anions, compared with a radius of about 1.06 A for the monovalent cations (using ionic radii), indicating that the anions are more strongly hydrated than the cations for a given charge density. (iii) The anions show larger deviations from ideal behavior (an elution position corresponding to the anhydrous molecular weight) than do the cations and dominate the chromatographic behavior of the neutral salts. These results are interpreted to mean that weakly hydrated ions (chaotropes) are "pushed" onto weakly hydrated surfaces by strong water-water interactions and that the transition from strong ionic hydration to weak ionic hydration occurs where the strength of ion-water interactions approximately equals the strength of water-water interactions in bulk solution.

Immunolocalization of the mercurial-insensitive water channel and glycerol intrinsic protein in epithelial cell plasma membranes.

Two water channel homologs were cloned recently from rat kidney, mercurial-insensitive water channel (MIWC) and glycerol intrinsic protein (GLIP). Polyclonal antibodies were raised against synthetic C-terminal peptides and purified by affinity chromatography. MIWC and GLIP antibodies recognized proteins in rat kidney with an apparent molecular mass of 30 and 27 kDa, respectively, and did not cross-react. By immunofluorescence, MIWC and GLIP were expressed together on the basolateral plasma membrane of collecting duct principal cells in kidney. By immunohistochemistry, MIWC and GLIP were expressed on tracheal epithelial cells with greater expression of GLIP on the basal plasma membrane and MIWC on the lateral membrane; only MIWC was expressed in bronchial epithelia. In eye, GLIP was expressed in conjunctival epithelium, whereas MIWC was found in iris, ciliary body, and neural cell layers in retina. MIWC and GLIP colocalized on the basolateral membrane of villus epithelial cells in colon and brain ependymal cells. Expression of MIWC and GLIP was not detected in small intestine, liver, spleen, endothelia, and cells that express water channels CHIP28 or WCH-CD. These studies suggest water/solute transporting roles for MIWC and GLIP in the urinary concentrating mechanism, cerebrospinal fluid absorption, ocular fluid balance, fecal dehydration, and airway humidification. The unexpected membrane colocalization of MIWC and GLIP in several tissues suggests an interaction at the molecular and/or functional levels.

Efeito do estresse hídrico sobre a locomoção e morfologia de girinos de Leptodactylus fuscus e Physalaemus nattereri

Diversas espécies de anuros da família Leptodactylidae se reproduzem em corpos dágua sazonais, temporários e mantidos exclusivamente por chuvas. Em períodos de estiagem prolongada a poça pode secar completamente, ocasionando elevadas taxas de mortalidade de ovos e girinos dessas espécies, podendo exercer forte pressão seletiva na evolução de mecanismos de resistência e sobrevivência nas fases iniciais do desenvolvimento. Algumas espécies de girinos conseguem sobreviver cerca de cinco dias fora dágua o que pode proporcionar uma adaptação vantajosa, porque possibilita a sobrevivência dos girinos por um período que pode ser suficiente para a reincidência de novas chuvas e restabelecimento do corpo dágua. Apesar dessa capacidade de sobrevivência, pouco se sabe sobre as possíveis modificações que a desidratação pode causar na locomoção e na morfologia durante o desenvolvimento desses animais. O presente trabalho teve como objetivo avaliar o efeito do estresse hídrico: (1) no nível de sobrevivência e perda de massa corpórea; (2) no desempenho locomotor; (3) na morfologia externa (morfometria linear) e interna, analisando tanto o volume total quanto o volume visceral (estereologia); e (4) no tempo até metamorfose após o estresse. Utilizamos girinos de duas espécies de anuros, Leptodactylus fuscus (Leptodactylinae) e Physalaemus nattereri (Leiuperinae), ambas as espécies se reproduzem em corpos dágua temporários, em áreas com estação seca definida estando, portanto sujeitas as mesmas pressões seletivas. Além disso, as duas espécies apresentam modos reprodutivos diferentes, podendo apresentar diferentes graus de resistência ao estresse hídrico. Os girinos das duas espécies foram divididos em dois grupos, os que ficaram em água (grupo controle) e os que foram submetidos ao estresse hídrico (grupo tratamento), por três períodos de tempo (12, 24 e 72 horas). Houve diferenças significativas para valores de perda de massa entre os grupos controle e tratamento em ambas as espécies, sendo o grupo tratamento que mais perdeu massa corpórea em todos os períodos, além disso, quase metade dos girinos de P. nattereri morreram em 36 horas de estresse. Não houve diferenças significativas para os dados de desempenho locomotor e volume total entre os grupos testado para girinos de L. fuscus, mas houve diferenças morfometricas significantes nos componentes relacionados a cauda e no volume visceral, onde, o intestino do grupo tratamento foi menor que do controle. Já em P. nattereri, houve diferenças significativas entre os grupos testados para desempenho locomotor, volume total, morfometria da cauda e volume visceral, sendo o estomago e anexo do tratamento maior que do controle. Nossos resultados sugerem que a exposição ao estresse hídrico não afeta significativamente a morfologia e o desempenho locomotor dos girinos de L. fuscus. No entanto, girinos de P. nattereri apresentaram uma sensibilidade ao estresse hídrico prolongado, principalmente sobre o seu desempenho locomotor.

Avaliação do potencial antimicrobiano e antioxidante do extrato de jabuticaba (Myrciaria cauliflora) microencapsulado adicionado em linguiça frescal e mortadela

O objetivo do presente estudo foi utilizar o extrato de jabuticaba microencapsulado como corante e avaliar o seu potencial antimicrobiano e antioxidante em produtos cárneos embutidos do tipo linguiça frescal e mortadela, em substituição ao corante tradicionalmente utilizado carmim de cochonilha. Uma primeira etapa consistiu na avaliação in vitro da capacidade antioxidante e antimicrobiana do extrato de jabuticaba aquoso e microencapsulado. O extrato de jabuticaba foi obtido a partir do resíduo do despolpamento da fruta, com posterior desidratação (microencapsulação) por spray dryer, utilizando maltodextrina como agente carreador. A caracterização foi efetuada por determinação do teor de antocianinas e identificação destas por cromatografia líquida de alta eficiência (CLAE) e espectrometria de massas (MS), determinação da sua capacidade antioxidante pelos métodos Folin-Ciocalteu, capacidade redutora do ferro no plasma (FRAP) e capacidade antioxidante pelo radical DPPH. As características físicas avaliadas no extrato aquoso foram o valor de pH e o teor de sólidos solúveis. O potencial antimicrobiano foi determinado pelo método da concentração inibitória mínima (CIM) sobre as bactérias Staphylococcus aureus, Escherichia coli e Salmonella Enteritidis. O extrato de jabuticaba microencapsulado (EJM) foi utilizado para a elaboração de linguiça frescal e mortadela em duas diferentes concentrações: 2 e 4% de EJM para a linguiça frescal e 2% para mortadela. A linguiça frescal (à base de carne suína) e a mortadela (à base de carne bovina e carne mecanicamente separada de frango) foram avaliadas quanto à estabilidade durante armazenamento refrigerado a 1±1 e 4±1°C, por 15 e 56 dias, respectivamente. Os produtos foram caracterizados quanto à composição centesimal e foram realizadas análises físico-químicas, microbiológicas e sensoriais. Os resultados encontrados para a linguiça frescal confirmaram que o uso de 2% e 4% de EJM contribuíram para reduzir a oxidação lipídica durante os 15 dias de armazenamento e nas análises microbiológicas o EJM contribuiu para reduzir a contagem de microrganismos por quatro dias quando comparado com a linguiça controle (sem adição de EJM). A análise sensorial comprovou que 2% de EJM não comprometeu a maioria dos atributos sensoriais avaliados, com exceção da coloração mais escura. Recomenda-se, portanto, a utilização de 2% de EJM na produção de linguiça frescal. Nas mortadelas, os resultados não diferiram quando se comparou os produtos com 2% de EJM e sem adição do extrato (controle), porém, a utilização de 2% de EJM pode ser considerada uma alternativa interessante devido as demandas atuais por novas fontes de baixo custo e a utilização de pigmentos naturais que possam ser benéficos à saúde. Com estes resultados, pode-se dizer que o aproveitamento de cascas de jabuticaba oriundos do processamento da fruta, na forma de extrato microencapsulado, pode representar uma boa alternativa como corante natural, trazendo uma nova concepção da utilização de produtos mais saudáveis em linguiça frescal e mortadela.

Desempenho, mortalidade e escore fecal de bezerros aleitados com diferentes dietas líquidas e terapias de reidratação quando acometidos por diarreias

O objetivo deste trabalho foi avaliar o efeito da utilização de sucedâneo lácteo com alto conteúdo proteico de origem vegetal no desempenho e saúde de bezerros, e avaliar métodos de reidratação para o tratamento de diarreias. No primeiro estudo foram utilizados 33 bezerros da raça Holandês distribuídos nos tratamentos: 1) Alto volume e baixa proteína (AV/BP): 8 litros, 21,4% PB; 2) Alto volume e alta proteína (AV/AP): 8 litros, 23,7% PB e 3) Baixo volume e alta proteína (BV/AP): 6 litros, 23,7% PB. Os bezerros foram alojados em abrigos individuais, com livre acesso a água e concentrado. Não houve efeito dos tratamentos para o desempenho animal (P>0,05). Os tratamentos AV/BP e AV/AP resultaram em maior consumo de sucedâneo (P<0,05), mas não afetaram o consumo de concentrado nem o consumo total (P>0,05). O escore fecal foi maior (P>0,05) para animais nos tratamentos AV/AP e BV/AP. Os animais nos tratamentos AV/BP permaneceram maior número de dias em diarreia (P<0,05), em comparação aqueles aleitados com BV/AP, os quais tiveram menos dias com vida (P<0,05). A concentração de lactato foi maior (P<0,05) para animais nos tratamentos AV/BP e AV/AP enquanto a concentração de proteína total foi maior (P<0,05) nos tratamentos AV/BP e BV/AP. Sucedâneos com elevado conteúdo de proteína de origem vegetal afetam negativamente o desempenho de bezerros podendo levar o animal a morte. No segundo estudo foram comparados três soluções de hidratação oral quanto a sua eficiência em repor eletrólitos e água, além de manter o desempenho de bezerros. Foram utilizados 42 bezerros mestiços Holandês-Jersey, distribuídos nos tratamentos: 1) Soro comum, 2) Glutellac® e 3) Soro comum + Aminogut®. Os animais foram aleitados com 4 L/d de sucedâneo lácteo até a oitava semana de vida quando foram desaleitados de forma abrupta. As terapias de reidratação foram oferecidas quando os animais apresentavam escore fecal >= 3 na escala de 1 a 5. Não houve efeito das terapias de reidratação no desempenho nem em metabólitos sanguíneos (P>0,05). O consumo voluntário de água foi maior para os animais reidratados com Glutellac®, mas o consumo total maior para os animais reidratados com Soro comum. As concentrações de HCO3 e Na+ foram maiores para os animais no tratamento Glutellac® (P=0,088 e P=0,073 respectivamente), sendo a concentração de glicose também afetada pelo protocolo de hidratação (P<0,05). A concentração de HCO3 aumentou do primeiro para o segundo dia, a de K+ e glicose diminuíram do primeiro para o segundo dia, enquanto que o Beecf teve um comportamento variável segundo a terapia de reidratação utilizada. Houve efeito da interação tratamento x dia de avaliação apenas para a concentração de BUN (P<0,05). O pH, a concentração de Na+ e Beecf foram maiores em animais mais velhos, enquanto K+, hematócrito e hemoglobina, foram menores (P<0,05). O consumo voluntário de água foi maior em animais reidratados com Glutellac®, o que junto com a simplicidade de uso, representam as principais vantagens deste método de reidratação.

Avaliação da saúde e desempenho de bezerros leiteiros recebendo suplemento de colostro associado ao colostro materno

O objetivo deste estudo foi avaliar o efeito do suplemento de colostro (SC) associado ao colostro materno (CM) de alta ou média qualidade na saúde e desempenho de bezerros leiteiros. No primeiro experimento foram utilizados 44 machos da raça Holandesa, distribuídos nos tratamentos: 1) CM de alta qualidade fornecido em volume correspondente a 15% do peso ao nascer (PN); 2) CM de média qualidade (15%PN) e 3) CM de média qualidade (15% PN) + SC. O protocolo de colostragem afetou a concentração de proteína total nas primeiras 48 horas de vida e durante a fase de aleitamento (P<0,05). Em relação a saúde, o protocolo de colostragem não afetou o escore fecal, bem como o número de dias com diarreia, dias com febre e dias de hidratação (P>0,05); no entanto, os animais que receberam CM de alta qualidade foram medicados por um menor número de dias (P<0,05). O consumo e o desempenho não foram afetados pelos tratamentos (P>0,05), embora tenha ocorrido efeito de idade (P<0,0001). O fornecimento de suplemento de colostro associado ao CM de média qualidade não afetou a transferência de imunidade passiva, o desempenho ou o metabolismo dos animais durante o período de aleitamento. No segundo experimento foram utilizadas 80 fêmeas da raça Holandesa nos seguintes tratamentos: 1) CM de alta qualidade fornecido em volume correspondente a 15% PN; e 2) CM de alta qualidade (15% PN) + SC. Não houve diferença na qualidade do colostro ingerido pelos animais (P>0,05). Os parâmetros sanguíneos nas primeiras 24 horas não foram afetados pelos tratamentos (P>0,05). O consumo e o desempenho não foram afetados pela administração do suplemento de colostro (P>0,05), no entanto houve efeito de idade para ambos os parâmetros (P<0,0001). Em relação à saúde, o fornecimento adicional de Ig não afetou os dias com diarreia, dias com febre ou dias medicados (P>0,05). A taxa de mortalidade observada durante o experimento foi de 2,5%. O consumo adicional de Ig através do fornecimento de suplemento de colostro em associação ao colostro materno de alta qualidade não melhorou a imunidade, bem como não afetou o desempenho ou a saúde de bezerros leiteiros. No terceiro experimento, foram utilizadas 67 fêmeas da raça Holandesa ou mestiças Girolando nos seguintes tratamentos: 1) CM de alta qualidade fornecido em volume correspondente a 10% PN; e 2) CM de alta qualidade (10% PN) + SC. O consumo e o desempenho não foram afetados pelo SC (P>0,05). O escore fecal também não foi afetado (P>0,05), mas observou-se maior frequência de animais com escore de desidratação nas 2ª e 3ª semana de vida, quando os escores fecais foram mais altos para todos os tratamentos. Houve uma maior porcentagem de animais suplementados sendo medicados devido a diarreias na segunda semana de vida. Houve maior frequência dos animais do grupo não suplementado com escore 1 de descarga nasal nas semanas 3 a 7. O consumo adicional de imunoglobulinas através do fornecimento de suplemento de colostro em associação ao colostro materno de alta qualidade não afetou o desempenho ou a saúde dos animais.