Qualidade e quantidade do leite produzido por cabras da raça Saanen submetidas a estresse por três dias consecutivos

Estudou-se o efeito do estresse fisiológico, via administração pontual de ACTH, sobre os níveis de cortisol e a quantidade e qualidade do leite produzido. Para tanto, 12 cabras da raça Saanen foram alocadas em dois grupos experimentais: o grupo ACTH recebeu 0,6 UI de ACTH/kg PV, e o grupo placebo recebeu solução placebo. A produção de leite, os percentuais de proteína, gordura e lactose e a contagem de células somáticas (CCS) do leite foram mensurados antes, durante e após a administração de ACTH e do placebo. Simultaneamente à administração de ACTH e placebo por três dias consecutivos, foram realizadas colheitas de sangue para mensuração dos teores de cortisol. Nos tempos -30 e zero, ambos os grupos apresentaram concentrações basais de cortisol. O aumento nos níveis de cortisol foi significativo nos tempos 60min (grupo ACTH: 59,00±5,70 e grupo placebo: 5,23±1,37ng/mL) e 120min (grupo ACTH: 47,96±9,72 e grupo placebo: 4,38±1,14ng/mL), pois a concentração de cortisol foi maior no grupo ACTH. Os valores retornaram ao basal no tempo 300min. Não houve diferenças na produção leiteira entre os grupos ACTH e placebo, tampouco de proteína, gordura, lactose e CCS do leite dos grupos, portanto o estresse induzido por três dias consecutivos não trouxe prejuízos à produção.

Substituição do farelo de soja pela farinha de glúten de milho na alimentação de cabras leiteiras

Realizou-se um experimento com cabras Saanen, primíparas, pesando em média, 50 kg, alojadas em baias individuais de 3 m², objetivando avaliar a substituição da proteína do farelo de soja (FS) pela proteína da farinha de glúten de milho (FGM), na produção e composição do leite, consumo voluntário e níveis de uréia plasmática. O delineamento experimental foi o triplo quadrado latino 4x4, com quatro períodos de 21 dias, sendo 14 dias de adaptação à dieta e sete dias para colheita das amostras. As cabras foram alimentadas e ordenhadas pela manhã e tarde. Os níveis de substituição estudados foram 0, 10, 30 e 50% de FGM (base na proteína bruta). A substituição do farelo de soja pela farinha de glúten de milho não afetou o consumo (kg/dia e %PV) de matéria seca, proteína bruta e fibra em detergente ácido, mas houve efeito quadrático, para o consumo de fibra em detergente neutro (kg/dia e % PV). Houve efeito sobre os níveis de uréia plasmática (UP), que foram inferiores para os menores níveis de substituição, sendo os maiores valores de UP observados para o tratamento que possuía somente FS. A produção de leite decresceu linearmente com a inclusão da farinha de glúten de milho. Os níveis de substituição resultaram em decréscimos lineares na produção de gordura (kg/dia), no teor de gordura do leite (%) e no teor de sólidos totais. Houve efeito quadrático para produção de lactose (kg/dia), observando-se o menor valor estimado para o nível de substituição de 31,6%. Não foi observado efeito sobre a produção de proteína bruta no leite, cujo valor médio foi de 0,083 kg/dia. Os teores de proteina bruta, lactose e sólidos totais não sofreram efeito dos níveis de substituição, sendo os valores médios percentuais de 2,98; 4,35 e 11,51%, respectivamente.

Produção e composição do leite de vacas de quatro grupos genéticos submetidas a dois manejos alimentares no período de lactação

O objetivo deste trabalho foi avaliar a produção e composição do leite de vacas de corte das raças Charolês (C), Nelore (N), mestiças CN filhas de touros C e mestiças NC filhas de touros N, mantidas em pastagem nativa e submetidas a diferentes manejos no pré-desmame: suplementadas com farelo de arroz integral (0,7% do peso vivo) e que desmamaram aos 42 ou 63 dias pós-parto, ou não suplementadas e que desmamaram aos 63 dias. A idade das vacas variou de 3 a 12 anos, sendo agrupadas em quatro classes: primíparas, jovens, adultas e velhas. Os dados foram submetidos à análise de variância, cujo modelo estatístico incluiu os efeitos de manejo, grupo genético e idade da vaca, período e as interações entre esses fatores. Os teores de lactose e gordura não foram influenciados pelos fatores estudados. Houve interação significativa entre idade da vaca e período para a produção de leite e entre manejo e idade da vaca para o teor de proteína. Vacas suplementadas apresentaram maior produção de leite (3,85 contra 3,25 L/dia), teor de extrato seco total - EST (12,18 contra 11,83%) e teor de extrato seco desengordurado - ESD (8,75 contra 8,57%). A produção de leite decresceu com o avanço do período de lactação, sendo a queda mais acentuada nas vacas não suplementadas. Vacas CN produziram mais leite (4,17 L/dia) do que as puras, não diferindo das NC (3,76 L/dia). Vacas N apresentaram produção de leite similar (3,76 L/dia) às C (3,11 L/dia). No entanto, as primeiras apresentaram leite com melhor qualidade, incluindo proteína (3,16 contra 2,86%), EST (12,52 contra 11,46%) e ESD (8,87 contra 8,49%). Vacas adultas apresentaram maior teor de ESD, seguidas das vacas jovens, das primíparas e das velhas, com valores de 8,86; 8,62; 8,62; e 8,54%, respectivamente.

Consumo e digestibilidade total dos nutrientes e produção e composição do leite de vacas alimentadas com teores crescentes de proteína bruta na dieta contendo cana-de-açúcar e concentados

Objetivou-se avaliar o efeito de teores crescentes de PB na MS total da dieta (11,5; 13,0; 14,5 e 16,0%) sobre os consumos e a digestibilidade total dos nutrientes e a produção e composição do leite de vacas leiteiras no terço inicial da lactação. O experimento foi constituído de quatro períodos experimentais de 15 dias, sete para adaptação dos animais à dieta e oito para coleta de dados. Foram utilizadas oito vacas, manejadas em cochos individuais, distribuídas em dois quadrados latinos balanceados 4 × 4. Utilizou-se concentrado à base de farelo de algodão e fubá de milho (40%) e, como volumoso, cana-de-açúcar corrigida com 1% de uréia + sulfato de amônio (9:1), na proporção de 60% (% MS). Os consumos médios diários de nutrientes, exceto EE e CNF, diferiram à medida que os teores de PB da dieta aumentaram. Os teores de PB na dieta não afetaram os coeficientes de digestibilidade aparente total dos nutrientes. A produção de leite corrigida ou não para 3,5% de gordura e seus componentes (gordura, PB e lactose), expressa em g/dia, e o nitrogênio uréico no leite (NUL) tiveram efeito linear crescente, enquanto a eficiência de utilização de N sofreu efeito linear negativo dos teores de PB da dieta. A dieta com 16% PB resultou em valores numericamente maiores de consumo e produção do leite e de seus componentes se comparada às dietas com menores teores de PB.

Fertilidade do sêmen eqüino diluído, resfriado e transportado

Objetivou-se neste experimento estudar o efeito do transporte de sêmen equino, entre haras, sobre a fertilidade de éguas inseminadas. Setenta e seis fêmeas da raça Mangalarga Marchador foram distribuídas, ao acaso, com número desigual de repetições, para se estudarem os efeitos do sêmen in natura, transportado ou não, e do sêmen diluído, resfriado e transportado, sobre a fertilidade de éguas e diferentes características reprodutivas. O sêmen foi diluído no diluidor lactose-gema de ovo sem glicerina. As fêmeas foram inseminadas, em dias alternados, com sêmen de apenas um garanhão, com volumes/dose inseminante variáveis dependendo do número de fêmeas a serem inseminadas por dia com determinado ejaculado. Utilizou-se para o transporte do sêmen um contêiner especialmente projetado para este fim (MSP-1). As taxas de concepção ao primeiro ciclo e totais, a eficiência de prenhez, o número de ciclos/égua, ciclos/égua gestante, ciclo/prenhez e prenhez/ciclo, o número de IA/égua, o número de IA/égua gestante e o número de IA/égua vazia não foram influenciados pelos tratamentos. O diluidor de lactose gema, modificado pela retirada do glicerol de sua formulação, constitui uma opção para a diluição do sêmen a ser resfriado e transportado. O suceso do transporte do sêmen, diluído e resfriado, a pequenas distâncias, por período de tempo curto, depende mais de um bom diluídor que de um sofisticado contêiner de transporte.

Disturbances in beta-cell function in impaired fasting glycemia

In a cross-sectional study, we assessed beta-cell function and insulin sensitivity index (ISI) with hyperglycemic clamps (10 mmol/l) in 24 subjects with impaired fasting glycemia (IFG, fasting plasma glucose [FPG] between 6.1 and 7.0 mmol/l), 15 type 2 diabetic subjects (FPG >7.0 mmol/l), and 280 subjects with normal fasting glycemia (NFG, FPG <6.1 mmol/l). First-phase insulin release (0-10 min) was lower in IFG (geometric mean 541 pmol/l (.) 10 min; 95% confidence interval [CI] 416-702 pmol/l (.) 10 min) and in type 2 diabetes (geometric mean 376 pmol/l (.) 10 min; 95% CI 247-572 pmol/l (.) 10 min) than NFG (geometric mean 814 pmol/l (.) 10 min; 95% CI 759-873 pmol/l (.) 10 min) (P < 0.001). Second-phase insulin secretion (140-180 min) was also lower in IFG (geometric mean 251 pmol/l; 95% CI 198-318 pmol/l; P = 0.026) and type 2 diabetes (geometric mean 157 pmol/l; 95% CI 105-235 pmol/l; P < 0.001) than NFG (geometric mean 295 pmol/l; 95% CI 276-315 pmol/l): IFG and type 2 diabetic subjects had a lower ISI (0.15 +/- 0.02 and 0.16 +/- 0.02 mumol/kg fat-free mass [FFM]/min/ pmol/l, respectively) than NFG (0.24 +/- 0.01 mumol/kg FFM/min/pmol/l, P < 0.05). We found a stepwise decline in first-phase (and second-phase) secretion in NFG subjects with progressive decline in oral glucose tolerance (P < 0.05). IFG subjects with impaired glucose tolerance (IGT) had lower first-phase secretion than NFG subjects with IGT (P < 0.02), with comparable second-phase secretion and ISI. NFG and IFG subjects with a diabetic glucose tolerance (2-h glucose >11.1 mmol/l) had a lower ISI than their respective IGT counterparts (P < 0.05). We conclude that the early stages of glucose intolerance are associated with disturbances in beta-cell function, while insulin resistance is seen more markedly in later stages.

Production of transgalactosylated oligosaccharides (TOS) by galactosyltransferase activity from Penicillium simplicissimum

Ingestion of transgalactosylated oligosaccharides (TOS) and other non-digestible oligosaccharides (NDOs) induces a significant increase in Bifidobacterium, Lactobacillus and some desirable species of Streptococcus populations in the gut of human and other animals (prebiotic effect). This change in the intestinal flora is responsible for several beneficial physiological effects such as a decrease of putrefactive products in the feces, lower blood cholesterol content, higher Ca2+ absorption, a smaller loss of bone tissue in ovariotomized female rats and a lower incidence of colon cancer. beta-Galactosidase from Penicillium simplicissimum, a strain isolated from soil, showed high galactosyltransferase activity when incubated with a highly concentrated lactose solution. Optimum pH and temperature ranges for hydrolytic activity were 4.0-4.6 and 55-60 degrees C, respectively, for a lactose concentration of 5.0% (w/v). Maximal galactosyltransferase activity was obtained at pH 6.5 and 50 degrees C and TOS synthesis was positively associated with lactose concentration in the reaction medium. Thus, when 50 ml of a 60% (w/v) lactose solution was incubated with 26.6 U of beta-galactosidase under the best pH and temperature conditions for transferase activity, a final product with 30.5% TOS (183 mg ml(-1)), 27.5% residual lactose and 42.0% monosaccharides was obtained. (C) 1999 Elsevier B.V. Ltd. All rights reserved.

Avaliação de uma Técnica de Diluição e Transporte de Sêmen Eqüino, para as Condições Brasileiras

Ninety-seven Mangalarga Marchador mares were allocated in a randomized experimental design, with different number of replicates per treatments. The objective of this experiment was to study the effect of raw (T3) or diluted, cooled semen transported to the breeding farms 1 (T1) and 3 (T2), on mare fertility and different aspects of reproductive efficiency. The mares were inseminated, with semen of only one stallion (Farm 2), on alternate days using different inseminating volumes, according to number of inseminated mares per day using only one ejaculate. For transportation, semen diluted in a lactose-egg yolk modified extender and the MSP-2 container were used. There was no difference among treatments in relation to conception rates of mares inseminated with raw or diluted, cooled and transported semen. Regardless of the farm, the total fertility obtained with cooled semen was 80% (52/65). The following variables were not affected by the treatments: cycles/mare, cycles/pregnant mare, cycles/pregnancy, pregnancy/cycle, number of inseminations/mare, number of inseminations/pregnant mare, number of inseminations/open mare and efficiency of pregnancy. However, the breeding farm had an effect on water and semen temperature in the container, time between collection and insemination, according to time and distance covered. Based on the results, cooling and transporting equine semen, using an extender, lactose-egg yolk without glycerine and the MSP-2 container should be recommended for short distances. The use of raw semen immediately after collection is also recommended.

Haemagglutination induced by Bordetella pertussis filamentous haemagglutinin adhesin (FHA) is inhibited by antibodies produced against FHA(430-873) fragment expressed in Lactobacillus casei

Filamentous haemagglutinin adhesin (FHA) is an important virulence factor from Bordetella pertussis related to the adhesion and spread of the bacteria through the respiratory tract. Three distinct domains have been characterized in mature FHA, and among them, the FHA(442-863) fragment was suggested to be responsible for the heparin-binding activity. In this study, we cloned the gene encoding the HEP fragment (FHA(430-873)) in a Lactobacillus casei-inducible expression vector based on the lactose operon. The recombinant bacteria, transformed with the resulting construct (L. casei-HEP), were able to express the heterologous protein depending on the sugar added to the culture. Subcutaneous inoculation of L. casei-HEP in Balb/C mice, using the cholera toxin B subunit as adjuvant, induced systemic anti-HEP antibodies that were able to inhibit in vitro erythrocyte haemagglutination induced by FHA. This is the first example of a B. pertussis antigen produced in lactic acid bacteria and opens new perspectives for alternative vaccine strategies against whooping cough.

Peroxidase from peach fruit: Thermal stability

Peroxidase from peach fruit was purified 28.9-fold by DEAE-cellulose, Sephadex G-100 and hydroxylapatite chromatography. The purified enzyme showed only one peak of activity with an optimum pH of 5.0 and temperature of 40 degreesC. The calculated activation energy (Ea) for the reaction was 7.97 kcal/mol. The enzyme was heat-labile in the temperature range of 60 to 80 degreesC with a fast inactivation at 80 degreesC. PAGE of the inactivation course at 70 degreesC showed only one band of activity. Different sugars increased the heat stability of the activity in the following order: sucrose>lactose>glucose>fructose. Measurement of residual activity showed a stabilizing effect of sucrose at various temperature/sugar concentrations (10 to 40%, w/w) with the Ea for inactivation increasing with sucrose concentration from 0 to 20% (w/w). After inactivation at 70 degreesC and 75 degreesC the enzyme was able to be reactivated by up to 40% of the initial activity when stared at 30 degreesC.

Conversion of immunosuppressive monotherapy from cyclosporin A to tacrolimus reverses bone loss in rats

Tacrolimus is used for transplant patients with refractory graft rejection and those with intolerance to cyclosporin (CsA), without the disfiguring adverse effects frequently attributed to CsA therapy. Since we have shown that CsA-associated bone loss can also affect alveolar bone, the purpose of this study was to evaluate the effects of conversion of monotherapy from CsA to tacrolimus on alveolar bone loss in rats. Groups of rats were treated with either CsA (10 mg/kg/day, s.c.), tacrolimus (I mg/kg/day, s.c.), or drug vehicle for 60 and 120 days, and an additional group received CsA for 60 days followed by conversion to tacrolimus for a further 60-day period. Bone-specific alkaline phosphatase (BALP), tartrate-resistent acid phosphatase (TRAP-5b), calcium (Ca2+), interleukin (IL)-1 beta, IL-6, and tumor necrosis factor alpha (TNF-alpha) concentrations were evaluated in the serum. Analyses of bone volume, bone surface, number of osteblasts, and osteoclasts were performed. Treatment with CsA for either 60 or 120 days was associated with bone resorption, represented by lower bone volume and increased number of osteoclasts; serum BALP, TRAP-5b, IL-1 beta, IL-6, and TNF-alpha were also higher in these animals. After conversion from CsA to tacrolimus, all the altered serum markers returned to control values in addition to a significant increase of bone volume and a lower number of osteoclasts. This study shows that conversion from CsA to tacrolimus therapy leads to a reversal of the CsA-induced bone loss, which can probably be mediated by downregulation of IL-1 beta, IL-6, and TNF-alpha production.

Potentiometric determination of saccharin in commercial artificial sweeteners using a silver electrode

A simple, precise, rapid and low-cost potentiometric method for saccharin determination in commercial artificial sweeteners is proposed. Saccharin present in several samples of artificial sweeteners is potentiometrically titrated with silver nitrate solution using a silver wire as the indicator electrode, coupled to a titroprocessor. The best pH range was from 3.0 to 3.5 and the detection limit of sodium saccharin was 2.5 mg/ml. Substances normally found along with saccharin in several commercial artificial sweeteners such as maltodextrin, glucose, sucrose, fructose, aspartame, cyclamate, caffeine, sorbitol, lactose, nitrate, methyl- and n-propyl-p-hydroxybenzoate, benzoic, citric and ascorbic acids do not interfere even in significant amounts (e.g. 20 excess relative to saccharin). Chloride ion interferes when present in concentrations larger than 10 mg l(-1); this interference is eliminated with previous extraction of the sweetener from the aqueous medium with ethyl acetate. The results obtained by applying the proposed method compared very favorably with those given by the HPLC method recommended by the FDA. (C) 2003 Elsevier Ltd. All rights reserved.

Effect of maternal obesity on diabetes development in adult rat offspring

This study aimed to evaluate whether maternal obesity leads to the onset of diabetes in adult Wistar rats offspring. MSG solution neonatally administration induced obesity in rats (F(1)MSG group, n = 30); and saline solution was also administrated to control rats (F1CON group, n = 13). In 3rd month of age, both control and MS G groups were mated for offspring (generation FA named as F2CON, n = 28 and F(2)MSG groups, n = 15; and so both generations were studied until 7th month of life. Lee Index was measured for experimental obesity validation from 5th to 7th month. Glycemia was weekly determined during pregnancy and monthly from 3rd to 7th month. In the end of experimental period all rats were submitted to oral glucose tolerance test (OGTT), with estimation of total area under the curve (AUC); and insulin tolerance test (ITT). Rats were then anesthetized and killed. Data were statistically analyzed with significance level of p < 0.05. Lee Index has confirmed obesity in all MSG rats. Glycemic levels comparisons between generations showed significant maternal interference in control and MSG groups. OGTT analysis showed higher glycemia in obese rats (F(1)MSG) and their offspring (F(2)MSG) as compared to their respective controls; and MSG groups increased AUC from OGTT. As regards ITT, F(2)MSG showed higher glycemia at 30 and 120 min, suggesting a delay of insulin action decreasing. Although glucose intolerance and insulin resistance clinical conditions represent as a factors for type 2 Diabetes mellitus development, this experimental model proposal was not efficient to induce type 2 Diabetes mellitus, but for obesity developing, glucose intolerance and insulin resistance in successive generations of rats. (c) 2007 Elsevier B.V. All rights reserved.

Subclinical abnormalities of glucose metabolism in Brazilian women with a history of gestational diabetes mellitus

Background. To evaluate insulin release and insulin sensitivity in women with prior gestational diabetes mellitus (GDM) to gain a better understanding of type 2 diabetes pathogenesis.Methods. GDM women were individually matched for age, body mass index, and waist/hip ratio with those who were normal glucose tolerant in a previous pregnancy (NGT). All women presented with normal glucose tolerance. Twenty pairs were submitted to the oral glucose tolerance test (OGTT) with plasma glucose, insulin, and C-peptide determinations. of the 20 pairs, 18 participated in hyperglycemic (10.0 mmol/l) clamp experiments with frequent plasma glucose and insulin determinations, allowing us to calculate first- and second-phase insulin release and the insulin sensitivity index. GDM and NGT women were compared using Student's t-test, the Mann-Whitney U-test, Friedman's non-parametric test, and the two proportion test for independent groups.Results. GDM women showed higher glycosylated hemoglobin values; at OGTT, they showed late insulin peak with increased plasma insulin levels only during the second hour, and a similar plasma C-peptide response despite a higher plasma glucose curve; during hyperglycemic clamp procedures, they showed similar biphasic insulin release and insulin sensitivity index. Considering that a woman with previous GDM had a defect in insulin release and/or insulin sensitivity, if its magnitude was at least 25% lower than that of the matched NGT woman, 43.8% showed impairment of first-phase insulin release and 55.6% insulin resistance.Conclusions. GDM women showed some degree of glucose intolerance. It is therefore necessary to follow them for a longer time.

Potentiometric determination of alendronate in pharmaceutical formulations

A simple, precise, fast and low-cost potentiometric method for alendronate determination in pharmaceutical formulations has been proposed. Alendronate present in tablets at the known concentration was potentiometrically titrated in an aqueous solution using a combined glass electrode coupled with an autotitrator. Determination limit of alendronate was 20.0 mug mL(-1). In the presence of common excipients, such as lactose, microcrystalline cellulose, glucose, croscarmellose sodium, starch, talc, magnesium stearate and titanium dioxide no interferences were observed. The method proposed was validated by determining alendronate in various dosage forms. The average RSD was 0.75%.