Proteomic analysis of nitrate-dependent acetone degradation by Alicycliphilus denitrificans strain BC

Alicycliphilus denitrificans strain BC grows anaerobically on acetone with nitrate as electron acceptor. Comparative proteomics of cultures of A. denitrificans strain BC grown on either acetone or acetate with nitrate was performed to study the enzymes involved in the acetone degradation pathway. In the proposed acetone degradation pathway, an acetone carboxylase converts acetone to acetoacetate, an AMP-dependent synthetase/ligase converts acetoacetate to acetoacetyl-CoA, and an acetyl-CoA acetyltransferase cleaves acetoacetyl-CoA to two acetyl-CoA. We also found a putative aldehyde dehydrogenase associated with acetone degradation. This enzyme functioned as a -hydroxybutyrate dehydrogenase catalyzing the conversion of surplus acetoacetate to -hydroxybutyrate that may be converted to the energy and carbon storage compound, poly--hydroxybutyrate. Accordingly, we confirmed the formation of poly-?-hydroxybutyrate in acetone-grown cells of strain BC. Our findings provide insight in nitrate-dependent acetone degradation that is activated by carboxylation of acetone. This will aid studies of similar pathways found in other microorganisms degrading acetone with nitrate or sulfate as electron acceptor.

Electroquímica de sustancias de interés en los sistemas agroalimentario y de sanidad animal. Desarrollo de biosensores electroquímicos. Aplicaciones analíticas.Electrochemistry of substances of interest in agroalimentary and animal health systems. Development of electrochemical biosensors. Analytical applications.

Se estudiarán los mecanismos de reacción electroquímica de las micotoxinas (metabolitos tóxicos generados por hongos) citrinina (CIT), patulina (PAT) y moniliformina (MON), de los antioxidantes naturales alfa, beta, gama y delta tocoferoles, de los flavonoides fisetina (FIS), morina (MOR), luteolina (LUT), rutina (RUT), buteina (BUT), naringenina (NAR) y miricetina (MIR) y de las hormonas esteroides estradiol (EDIOL), estrona (EONA) y estriol (ETRIOL). Por otra parte, se implementarán técnicas electroanalíticas para la detección y cuantificación de estos sustratos en muestras de matrices naturales que los contengan. Se realizará el diseño y caracterización de biosensores enzimáticos a partir de peroxidasas y/o fosfatasa alcalina para la determinación de la micotoxina CIT y de los flavonoides y, por otro, de inmunosensores para las micotoxinas ocratoxina A (OTA) y PAT y hormonas. Para el anclaje de enzimas y/o anticuerpos, se estudiarán las propiedades de electrodos modificados por monocapas autoensambladas, nanotubos de carbono y partículas magnéticas. Se usarán las técnicas de voltamperometría cíclica, de onda cuadrada y de redisolución con acumulación adsortiva, espectroscopías de impedancia electroquímica, electrólisis a potencial controlado, uv-vis e IR, microbalanza de cristal de cuarzo y microscopías de alta resolución (SEM, TEM, AFM). La importancia de este proyecto apunta a la obtención de nuevos datos electroquímicos de los sustratos indicados y conocimientos relacionados con la aplicación de electrodos modificados en la preparación de biosensores y en el desarrollo de técnicas alternativas para la determinación de los analitos mencionados precedentemente. Electrochemical reaction mechanisms of mycotoxins (toxic metabolites generated by fungi) citrinin (CIT), Patulin (PAT) and moniliformin (MON), natural antioxidants alpha, beta, gamma and delta tocopherols, flavonoids fisetin (FIS), morin (MOR), luteolin (LUT), rutin (RUT), butein (BUT), naringenin (NAR), miricetin (MIR) and steroid hormones estradiol (EDIOL), estrone (EONA) and estriole (ETRIOL) will be explored. On the other hand, electroanalytical techniques for the detection and quantification of these substrates in samples of natural matrices will be implemented. The design and characterization of enzymatic biosensors from peroxidases and/or from alkaline phosphatase for the determination of CIT and flavonoids, and also of inmunosensors for ochratoxin A (OTA) and PAT and hormones will be performed. For the anchor of enzymes and/or antibody, properties of electrodes modified by self assembled monolayers, carbon nanotubes and magnetic particles will be explored. Cyclic, square wave and adsorptive stripping voltammetries, electrochemical impedance spectroscopy, controlled potential electrolysis, uv-vis and IR, quartz crystal microbalance and high-resolution microcopies (SEM, TEM, AFM) will be used. The importance of this project is aimed at obtaining new electrochemical data for the indicated substrates and knowledge on the application of modified electrodes in preparation of biosensors and in the development of alternative techniques for the determination of the above-mentioned analytes.

Untersuchung zur Nutzung der Vakuumtrocknungshärtung für die Herstellung und den Einsatz magnesiumsulfatgebundener Kerne für den Leichtmetallguss

Cylinder head casting; aluminum casting; inorganic bonded cores; vakuum; magnesium sulfate

Estudos sobre o «vermelhão» do algodoeiro (III)

In several cotton crops areas of the State of S. Paulo it was observed, during the years of 1948, 1949, and 1951, the appearance of a purple color of the leaves; the color appears in the opening of the bolls and was correlated with a decrease of production. The opinions concerning the cause of such abnormality were very different and sometimes contradictory; certain investigators attributed the disease to insect attack, others to bad climatic conditions whereas others to a potassium deficiency now called "fome de potássio" (potash hunger); our ideas on the subject is another one. We think that the disease is caused by lack of a suitable supply of magnesium. This opinion is largely based on the syntomatology found in the literature. To study the problem, several experiments were carried out, namely: 1. pot experiments using soil collected in areas where the disorder had appeared; 2. pot experiments controlling the water supply; 3. sand culture experiments omitting either potassium or magnesium; 4. leaf analysis of plant matrial collected troughout the Piracicaba County; 5. plot experiments with the varieties Texas, Express, and I.A. 817 Campinas. The first four experiments were discussed elsewhere. To study the point 5 an experiment was carried out, with the following treatments : 1 - NPKCaMg (no K added) - Mg supplied as MgSO4 (a soluble form); 2 -NPKCa (no Mg added); 3 -NPKCaMg (complete) - Mg supplied as MgSO4; 4 - NPKCaMg (complete) - Mg supplied as dolomitic limestone (a slightly soluble form) as a rate 2.5 higher than in the treatment 1 and 3. Organic matter as cottonseed meal was applied in the proportion of 500 kg per hectare. The experimental design was randomized blocks with 4 replications and the results can be summarized as follows: 1 the I.A 817 variety was the most strongly affected by the physiological disorder, with severe decrease in yield; 2. the disease occurred more frequently in the minus magnesium treatment; 3. dolomitic limestone is so effective as magnesium sulfate in the control of the disease as well in the raising of the yield; 4. in the minus K treatment it was observed a marked occurrence of the typical symptoms of potassium deficiency (cotton rust); 5. magnesium was actually, in the experimental conditions the responsible for the purple color (vermelhão) of the cotton leaves.

Contribuição ao estudo do «vermelhão» do algodoeiro (Gossipium herbaceum)

During the years 1948, 1949 and 1951 a disease occurred in the cotton crops of the state of S. Paulo Brazil (S. Am.), which caused a severe drop in yields. The abnormality was characterized by a typical reddish - purple color of the leaves, being by this reason, called "vermelhão", that is, reddening of the cotton plant. The disease was associated with a dry season. Among the several hypotheses raised to explain the causes of the disease were: insect attack, potassium deficiency - where from the name "potash hunger" was also given -, and magnesium deficiency: In order to study the problem the Department of Agricultural Chemistry of the College of Agriculture of the University of São Paulo, at Piracicaba, carried out a series of experiments as follows: 1. pot experiments in which soil of one of the affected regions was used ("terra roxa", a red-brownish soil derived from basalt); 2. pot-soil experiments varying the moisture supplied; 3. sand culture experiments omitting certain elements from the nutrient solutions; 4. field plot experiments, conducted on a sandy soil; three different varieties were employed: Texas, Express, and I.A. 817; magnesium was applied either as sulfate or dolomitic limestone. All the experiments were completed with suitable chemical analyses. The results can be summarized as follows: 1. in the first trial, the not properly manured pots (minus Mg), symptoms were registered which were similar to the symptoms observed in the field; it was possible to establish some differences among three different types of reddening: due to lack of K in the mixed fertilizers used, the characteristic cotton rust made its appearance, the red color in the leaves of the minus Mg plants was all alike that described in the current literature as a symptom of Mg-deficiency; in all the treatments ocurred a yellow-reddish color in the leaves associated with the latest stages of maturity; 2. in the second experiment it was verified that when the plants in the pots with soil were kept 75 per cent of the water holding capacity, no symptom of deficiency showed up; was true even for the plants not receiving neither K nor Mg; however, plants supplied with only 25 per cent of the water holding capacity showed, respectively, cotton rust in the minus K treatment and the red purplish color in the minus Mg series; 3. the sand culture experiment confirmed lack of Mg as the cause of "vermelhão", being potash deficiency the responsible for cotton rust; 4. in the field experiment, variety LA. 817 revealed to be the most sensitive to "vermelhão" when Mg was omitted from the fertilizers; symptoms of K deficiency appeared when no K was supplied; both magnesium sulfate and dolomitic limestone proved to be equally effective in the control of "vermelhão"; 5. the analyses of material collected both in the field as well in the pots revealed that leaf petiole in the most reliable part to indicate the K and Mg status of the plant; the variation in Mg content suffered by the plants showing different stages of "vermelhão was, quantitatively, at least as large as that in K content, however when one deals with K deficient plants, that is, plants showing the typical rust, no variation occurred in the Mg content, whereas K in the dry mater dropped from more than 1 per cent to less than half per cent. Then, the following general conclusions can be drawn: 1. Mg deficiency is the cause of "vermelhão" of cotton crops; 2. K deficiency also occurred, but in a lesser degree; 3. the climate conditions - especially the lack of rain influenced the soil dynamic of K, and especially Mg, bringing a severe reduction in their assimilability; 4. the "vermelhão" disease can be easily controlled upon additions either of magnesium sulfate or dolomitic limestone.

Estudos sôbre a alimentação mineral da mandioca (Manihot utilissima Pohl.)

1. The present work was carried out to study the effects of mineral nutrients in the yield as well as in the composition of cassava roots. The variety "Branca de Sta. Catarina" was grown by the sand culture method, the following treatments being used: N0 P0 K0, N0 P1 Kl, N1 P0 K1, N2 P1 K0, N2 P1 K1, N1 P2 K1, and N1 P1 K2, where the figures 0, 1, and 2 denote the relative proportion of a given element. The nutrients were given as follows: N = 35 grams of ammonium nitrate per pot loaded with 120 pounds of washed sand; P1 = 35 grams of monocalcium phosphate; Kl = 28 grams of sulfate of potash. Besides those fertilizers, each pot received 26 grams of magnesium sulfate and weekly doses of micronutrients as indicated by HOAGLAND and ARNON (1939). To apply the macronutrients the total doses were divided in three parts evenly distributed during the life cycle of cassava. 2. As far yield of roots and foliage are concerned, there are a few points to be considered: 2.1. the most striking effect on yield was verified when P was omitted from the fertilization; this treatment gave the poorest yields of the whole experiment; the need of that element for the phosphorylation of the starchy reserves explains such result; 2.2. phosphorus and nitrogen, under the experimental conditions, showed to be the most important nutrients for cassava; the effect of potassium in the weight of the roots produced was much less marked; it is noteworthy to mention, that in absence of potassium, the roots yield decreased whereas the foliage increased; as potassium is essential for the translocation of carbohydrates it is reasonable to admit that sugars produced in the leaves instead of going down and accumulate as starch in the roots were consumed in the production of more green matter. 3. Chemical analyses of roots revealed the following interesting points: 3.1. the lack of phosphorus brought about the most drastic reduction in the starch content of the roots; while the treatment N1 P1 K1 gave 32 per cent of starch, with NI PO Kl the amount found was 25 per cent; this result can be explained by the requirement of P for the enzymatic synthesis of starch; it has to be mentioned that the decrease in the starch content was associated with the remarkable drop in yield observed when P was omitted from the nutrient medium; 3.2. the double dosis of nitrogen in the treatment N2 P1 K1, gave the highest yields; however the increase in yield did not produce any industrial gain: whereas the treatment N1 P1 K1 gave 32 per cent of starch, by raising the N level to N2, the starch content fell to 24 per cent; now, considering the total amount of starch present in the roots, one can see, that the increase in roots yield did not compensate for the marked decrease in the starch content; that is, the amount of starch obtained with N1 P1 K1 does not differ statistically from the quantity obtained with N2 P1 K1; as far we know facts similar to this had been observed in sugar beets and sugar cane, as a result of the interaction between nitrogen and sugar produced; the biochemical aspect of the problem is very interesting: by raising the amount of assimilable nitrogen, instead of the carbohydrates polymerize to starch, they do combine to the amino groups to give proteinaceous materials; actually, it did happen that the protein content increased from 2.91 to 5.14 per cent.

Experiências de adubação mineral e orgânica com Capim Kikuyu (Pennisetum clandestinum Hochst.)

Kikuio grass (Pennisetum clandestinum Hochst) is beyond any doubt, a pasture very important for farm animals; since its chemical composition is very similar to that of alfalfa, the present field trial was carried out; a randomized block design with 8 treatments was selected as follows: 1 N - P - K - Ca - Mg (complete manuring) 2 N - P - K - Ca----- (without Mg) 3 N - P - K-------Mg (without Ca) 4 ----P - K - Ca - Mg (without N) 5 N------K - Ca Mg (without P) 6 N - P - Ca - Mg (without K) 7 organic matter (without mineral fertilizers) 8 control Nitrogen was applied as NaN03 (topdressed) and as ammonium sulfate; P2O5 was given as superphosphate associated to bonemeal; K2O was applied as muriate, CaO as "sambaquis" (oyster shells); MgO was given as MgSO4 (topdressed). The source of organic matter was farmyard manure. As far yields are concerned the following observations were made: 1. treatment n. 7 was superior to all others; 2. considering the mineral fertilizers, good responses were due to N and P2O5; 3. the control yield was exceedingly poor, being inferior to all the others treatments; The chemical analyses revealed that: 1. the protein content decreased accordingly to this order: 7, 6, 5 and 1; treatment 4 (without N) gave the lowest protein content; 2. treatment n. 4 produced the highest fat content; treatment no. 7 ranked second; no. 8 gave the lowest fat content; 3. crude fiber: highest - treatment 7; lowest - 8; 4. ashes: the ashes content was higher in treatment 5; proprobably because the most abundant element in the ashes is K, the ash content of treatment 6 (no K) was very low; 5. non nitrogenous substances (determined by difference) - high in treatment 8 and low in treatment 7; 6. mineral elements in the ashes - the element omitted from a given treatment was very low in the grasses therein obtained; this shows the relative poverty of the soil in that element. As general remark the Authors suggest the use of farmyard manure in the fertilization of Kikuio grass; farmyard manure could probably substitute wither green manure or compost.

Nitrificação e aproveitamento de alguns adubos nitrogenados

The rate of nitrification of several nitrogenous fertilizers (ammonium sulfate, nitre-chalk, ureia, and cottonseed meal) was studied in three soils, namely, "terra roxa legítima", a red soil derived from basalt, "terra roxa misturada", a soil also derived from basalt but with a higher proportion of sand, and "areito Corumbataí", a sandy soil. The effects of the following treatments on nitrification were considered: addition of limestone of micronutrients (Fe, Cu, Zn, Mn, and Mo), and inoculation with a suspension of spores of Aspergillus wentii, a heterotrophic nitrifier. The results showed that: in "terra roxa legítima" limestone had no influence on the nitrification rate, whereas the micronutrients estimulated the oxidation of nitre-chalk, cottonseed meal and urea; inoculation with A. wentii helped only the nitrification of ammonium sulfate and of the cottonseed meal; the latter, in all the treatments employed gave use to a smaller amount of nitrates; in "terra roxa misturada", all the fertilizers depending upon the treatments they were subjected to, presented maximum values for nitrification; limestone estimulated the oxidation of ammonium sulfate as well as the mineralization of the cottonseed meal; the addition of micronutrients helped the nitrification of all the fertilizers, except that of urea; inoculation showed a benefical influence on the nitrification of ammonium sulfate and cottonseed meal; in "arenito de Corumbatai", the amounts of nitrates produced was roughly the same for all the fertilizers investigated; limestone estimulated the nitrification of nitro-chalk, ammonium sulfate and cottonseed meal whilst the addition of micronutrients benefited only the latter two; the inoculation with A. wentii helped the oxidation of all the fertilizers. In order to study the availability of the various fertilizers above discussed, two plant growing experiments were carried cut, one in pots, using the three soil types and another one in the field, with "terra roxa misturada". In "arenito de Corumbatai" there was no significant difference in the yield both of straw and rice grains for none of the fertilizers: Chilean nitrate of soda was used as a control; ho marked agreement could be detected between the data concerning nitrification and the yield results. In "terra roxa legítima", ammonium sulfate won the competition and there was a good parallelism between nitrification and yield. In "terra roxa misturada", there was no statistical difference among the various fertilizers; the agreement between nitrification and yields was reasonable. In the field (corn), Chilean nitrate, ammonium sulfate and nitro-chalk were clearly beter than urea and cottonseed meal which did not differ from the minus nitrogen plots.

A absorção de nitrogênio, fósforo, potássio, cálcio, magnésio, enxôfre e silício pela cana de açúcar, Co 419, e o seu crescimento em função da idade

This paper describes the data obtained for the growth of sugar cane, Variety Co 419, and the amount and rate of absorption of nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, and silicon, according to the age of the plant, in the soil and climate conditions of the state of S. Paulo, Brazil. An experiment was installed in the Estação Experimental de Cana de Açúcar "Dr. José Vizioli", at Piracicaba, state of S. Paulo, Brazil, and the soil "tèrra-roxa misturada" presented the following composition: Sand (more than 0,2 mm)........................................................................ 8.40 % Fine sand (from 0,2 to less than 0,02 mm)................................................. 24.90 % Silt (from 0,02 to less than 0,002 mm)...................................................... 16.40 % Clay (form 0,002 mm and less)................................................................ 50.20 % pH 10 g of soil and 25 ml of distilled water)..................................................... 5.20 %C (g of carbon per 100 g of soil)................................................................. 1.00 %N (g of nitrogen per 100 g of soil)............................................................... 0.15 P0(4)-³ (me. per 100 g of soil, soluble in 0,05 normal H2SO4) ............................... 0.06 K+ (exchangeable, me. per 100 g of soil)....... 0.18 Ca+² (exchangeable, me. per 100 g of soil)...... 2.00 Mg+² (exchangeable, me. per 100 g of soil)...... 0.66 The monthly rainfall and mean temperature from January 1956 to August 1957 are presented in Table 1, in Portuguese. The experiment consisted of 3 replications of the treatments: without fertilizer and with fertilizer (40 Kg of N, from ammonium sulfate; 100 Kg of P(2)0(5) from superphosphate and 40 Kg K2 O, from potassium chloride). Four complete stools (stalks and leaves) were harvested from each treatment, and the plants separated in stalks and leaves, weighed, dried and analysed every month from 6 up to 15 months of age. The data obtained for fresh and dry matter production are presented in table 2, and in figure land 2, in Portuguese. The curves for fresh and dry matter production showed that fertilized and no fertilized sugar cane with 6 months of age presents only 5% of its total weight at 15 months of age. The most intense period of growth in this experiment is located, between 8 and 12 months of age, that is between December 1956 and April 1957. The dry matter production of sugar cane with 8 and 12 months of age was, respectively, 12,5% and 87,5% of the total weight at 15 months of age. The growth of sugar cane in relation to its age follows a sigmoid curve, according to the figures 1, 2 and 3. The increase of dry matter production promoted by using fertilizer was 62,5% when sugar cane was 15 months of age. The concentration of the elements (tables 4 and 5 in Portuguese) present a general trend of decreasing as the cane grows older. In the stalks this is true for all elements studied in this experiment. But in the leaves, somme elements, like sulfur and silicon, appears to increase with the increasing of age. Others, like calcium and magnesium do not show large variations, and finally a third group, formed by nitrogen, phosphorus and potassium seems to decrease at the beginning and later presents a light increasing. The concentration of the elements was higher in the leaves than in the stalks from 6 up to 15 months of age. There were some exceptions. Potassium, magnesium and sulfur were higher in the stalks than in the leaves from 6 up to 8 or 9 months of age. After 9 months, the leaves presented more potassium, magnesium and sulfur than the stalks. The percentage of nitrogen in the leaves was lower in the plants that received fertilizer than in the plants without fertilizer with 6, 7, 8, 10, 11 and 13 months of age. This can be explained by "dilution effect". The uptake of elements by 4 stools (stalks and leaves) of sugar cane according to the plant age is showed in table 6, in Portuguese. The absorption of all studied elements, nitrogen, phosphorus, potassium, calcium, magnesium, sulfur and silicon, was higher in plants that received fertilizer. The trend of uptake of nitrogen and potassium is similar to the trend of production of dry matter, that is, the maximum absorption of those two nutrients occurs between 9 and 13 months of age. Finaly, the maxima amounts of elements absorbed by 4 stools (stalks and leaves) of sugar cane plants that received fertilizer are condensed in the following table: Element Maximum absorption in grams Age of the plants in months Nitrogen (N) 81.0 14 Phosphorus (P) 6.8 15 Potassium (K) 81.5 15 Calcium (Ca) 19.2 15 Magnesium (Mg) 13.9 13 Sulfur (S) 9.3 15 Silicon (Si) 61.8 15 It is very interesting to note the low absorption of phosphorus even with 100 kg of P2O5 per hectare, aplied as superphosphate. The uptake of phosphorus was lower than calcium, magnesium and sulfur. Also, it is noteworthy the large amount of silicon absorbed by sugar cane.

Absorção de zinco pela cana de açúcar, Co 419, em função da idade

The status of zinc in sugar cane, variety Co 419, troughout its life cyle, was studid in samples cut monthly, from the 6th to 15th month, from an experiment carried on under the conditions of soil and climate prevailing in Piracicaba, State of São Paulo, Brazil. The experiment consisted of 6plots, 3 fertilized and 3 unfertilized. The fertilized ones received 40 kg of N (ammonium sulfate), 100 kg P2O5 (superphosphate) and 40 kg K2O (potassium cloride) per hectare, just before planting. The zinc content was determined by the Zincon method, after separation of zinc from other ions by means of the ion Exchange Resin III, Merck. The results obtained show that there was a tendency to decrease the zinc level in the stalks, whereas it kept more or less constant in the leaves; there was an exception in January, when the zinc level in the stalks had a sharp raise: 38-90-20 and 28-60-23 ppm for the fertilized an unfertilized treatments. There was a parallelism in the absorption of zinc by the plants from 4 hills of both treatments, through the whole - plantcycle but, the total amount taken up was higher with the fertilized plot due to its greater mass production.

A absorção de ferro pela cana de açúcar, Co 419, em função da idade

This paper describes the results obtained from the determination of iron in sugar cane according to the age of the plant, in the soil and climate conditions of the state of S. Paulo, Brazil. The iron was determined by 1-10- phenanthroline method, in samples cut monthly from 7th to 15th month from an experiment consisted de 3 plots fertilized with amonium sulfate, superphosphate and potassium cloride. The concentration of iron in the stalks and in the leaves varies according to the age of the plant. A ton of fresh stalks 15 months old contains 78,71 g of iron.

Absorção de molibdênio pela cana de açúcar variedade Co 419, em função da idade

In this paper the authors describe the results obtained from the determination of molybdenum in sugar cane plant, grown in soils and climate prevailing in Piracicaba, State of São Paulo, Brazil. The molybdenum was determined in samples cut monthly from the 8th to 14th month, from an experiment consisting of 6 plots, 3 fertilized and 3 unfertilized. The fertilized treatment received 40 kg N (ammonium sulfate) 100 kg P2O3, (superphosphate) and 40 kg K2O (potassium chloride) per hectare, just before planting. Molybdenum was determined by thiocyanate-stannous chloride method, using carbon tetrachloride-butyl alcohol misture, for extrating the colored complex. The results obtained show a parallelism in the absorption of molybdenum by the plants of both treatments. The concentration of molybdenum in the stalks have a tendency to decrease, where as it kept more or less constant in leaves, with a exception in the 14° month when it rised probable because of a migration of molybdenum of the stalks to the leaves. The total amount molybdenum taken up was higher with the fertilized plot due its greater mass prodution.

Transferência de imunidade passiva em equinos: comportamento imunológico do recém nascido

O objetivo deste trabalho foi observar o comportamento imunológico de potros recém-nascidos das raças Mangalarga e Anglo-Árabe no que se refere ao processo de aquisição de anticorpos maternos e sua correlação com os níveis de imunoglobulina do colostro. Foram utilizados 7 potros Anglo-Árabe e 6 potros Mangalarga para amostragem de sangue imediatamente após o nascimento (antes de qualquer ingestão de colostro), 24 e 48 horas, 5, 10, 15, 20, 25, 30, 40, 50, 60 e 70 dias após o nascimento, e suas respectivas mães cujo colostro foi amostrado imediatamente após a parição, antes da primeira mamada. A quantificação das imunoglobulinas séricas foi efetuada pelo método do ZST (Zinc Sulfate Turbidity) e para a análise dos resultados foram testados modelos matemáticos que estudam o processo em questão. Para a raça Mangalarga o modelo matemático foi: y = 23,9274 - 0,39766x + 6,4675 10-3 x², com r = 0,91 e P < 0,01 e para a raça Anglo Árabe foi y = 34,161 - 0,756062x + 0,015604x² - 1,013 10-3 x³, com r = 0,96 e P < 0,05. A concentração de IgG do colostro foi estimada com base na concentração de proteína total avaliada pelo método Micro-Kjeldhal. Este estudo permitiu concluir: (1) a presença de maior quantidade de IgG (imunoglobulina G) passiva no sangue dos potros, retardou o estabelecimento dos níveis normais de Ig (imunoglobulina G + M+A+E); (2) animais que adquiriram menor quantidade de IgG passiva apresentaram resposta mais intensa de produção endógena de Ig; (3) a concentração estimada de IgG no colostro apresentou correlação positiva com a concentração sérica dos potros no pico da absorção.

Lactacidemia: métodos de dosagem do ácido lático

A critical study of three methods for the determination of lactic acid (EDWARDS, MENDEL & GOLDSCHEIDER, MILLER & MUNTZ) is presented and some modifications are proposed. It was shown t hat more accurate results could be obtained with Edward's technic when an Iena glass filter is connected with the absorption tube. Before the dropping of the permanganate solution it is necessary to pass a current of air through the reaction flask to avoid the oxidation of the non-lactic acid substances which interfere with the reaction. The absorption tube must be maintained at 18°C during the destillation and the titration of the bisulphite binding aldehyde at 4°C. When the sample contains more than 5 mg it is useful to work with greater quantities of the bisulphite. More permanganate is consumed when the lactic acid concentration is higher. The sensivity of the method permits the titration of 0.04 mg to 5 mg of lactic acid in the sample. The calculated error of the method gave 0.018 % and the normal values for blood determined in 20 human cases averaged 10.30 mg per 100 ml (Table VI). MENDEL and GOLDSCHEIDER'S method was modified in the following details: Somogyis deproteinization was performed instead metaphosphoric acid as in the original method; to avoid the evaporation of the acetic aldehyde during the heating time with sulfuric acid a special glass stopped tube is proposed (Fig. 2). The reaction with sulfuric acid and veratrol is performed in an ice bath. Blood proteins precipitants were tried and Somogyi's lattest tecnic showed better results (Table V). Colorimetric readings were done in the PULFRICH photometer using filter S 53 and a 10 mm cup. The method is accurate within an error of 0.23 % and samples of 5 to 70 microg. could be easily determined. Normal values for human blood averaged 10.78 mg per 100 ml. More accurate results were obtained with the technic of MILLER & MUNTZ. Slight modifications were introduced: deproteinization with copper sulfate and sodium tungstate; satured p-hydroxydiphenyl solution according to KOENEMANN which is stable for 5 months when stored in the ice-box. Using the PULFRICH step-photometer the error is 0.17% with samples varying from 0.1 to 10 microg. of lactic acid. The filter employed was S 57 with the 5 mm cup. The method was adapted to 0.1 ml of blood. Normal values for human blood gave an average of 10.58 mg per 100 ml.

Sôbre u'a modificação do meio de Monteverde

It is well known that the culture media used in the presumptive diagnosis of suspiciuous colonies from plates inoculated with stools for isolation of enteric organisms do not always correctly indicate the major groups of enterobacteria. In an effort to obtain a medium affording more exact indications, several media (1-9) have been tested. Modifications of some of these media have also been tested with the result that a satisfactory modification of Monteverde's medium was finaly selected. This proved to be most satisfactory, affording, as a result of only one inoculation, a complete series of basic indications. The modification involves changes in the formula, in the method of preparation and in the manner of storage. The formulae are: A. Thymol blue indicator: NaOH 0.1/N .............. 34.4 ml; Thymol blue .............. 1.6 g; Water .................... 65.6 ml. B. Andrade's indicator. C. Urea and sugar solution: Urea ..................... 20 g; Lactose ................... 30 g; Sucrose ................... 30 g; Water .................... 100 ml. The mixture (C.) should be warmed slightly in order to dissolve the ingredients rapidly. Sterilise by filtration (Seitz). Keep stock in refrigeratior. The modification of Monteverde's medium is prepared in two parts. Semi-solid part - Peptone (Difco) 2.0 g; NaCl 0.5 g; Agar 0.5 g; Water 100.0 ml. Boil to dissolve the ingredients. Adjust pH with NaOH to 7.3-7.4. Boil again for precipitation. Filter through cotton. Ad indicators "A" 0.3 ml and "B" 1.0 ml. Sterilise in autoclave 115ºC, 15 minutes in amounts not higher than 200 ml. Just before using, add solution "C" asseptically in amounts of 10 ml to 200 ml of the melted semi-solid medium, maintained at 48-50ºC. Solid part - Peptone (Difco) 1.5 g; Trypticase (BBL) 0.5 g; Agar 2.0 g; Water 100,00 ml. Boil to dissolve the ingredients. Adjust pH with NaOH to 7.3-7.4. Boils again. Filter through cotton. Add indicators "A" 0.3 ml and "B" 1.0 ml; ferrous ammonium sulfate 0.02 g; sodiun thiosulfate 0.02 g. Sterilise in autoclave 115ºC, 15 minutes in amounts not higher than 200 ml. Just before using, add solution "C" asseptically in amounts of 10 ml to 200 ml of the melted solid medium, maintained at 48-50ºC. Final medium - The semi-solid part is dispensed first (tubes about 12 x 120 mm) in 2.5 ml amounts and left to harden at room temperature, in vertical position. The solid part is dispensed over the hardened semi-solid one in amounts from 2.0 ml to 2.5 ml and left to harden in slant position, affording a butt of 12 to 15 mm. The tubes of medium should be subjected to a sterility test in the incubator, overnight. Tubes showing spontaneous gas bubbles (air) should then be discarded. The medium should be stored in the incubator (37ºC), for not more than 2 to 4 days. Storage of the tubes in the ice-box produces the absorption of air which is released as bubbles when the tubes are incubated at 37ºC after inoculation. This fact confirmed the observation of ARCHAMBAULT & McCRADY (10) who worked with liquid media and the aplication of their observation was found to be essential to the proper working conditions of this double-layer medium. Inoculation - The inoculation is made by means of a long straight needle, as is usually done on the triple sugar, but the needel should penetrate only to about half of the height of the semi-solid column. Indol detection - After inoculation, a strip of sterelized filter papaer previously moistened with Ehrlich's reagent, is suspended above the surface of the medium, being held between the cotton plug and the tube. Indications given - In addition to providing a mass of organisms on the slant for serological invetigations, the medium gives the following indications: 1. Acid from lactose and/or sucrose (red, of yellowsh with strains which reduce the indicators). 2. Gas from lactose and/or sucrose (bubbles). 3. H[2]S production, observed on the solid part (black). 4. Motility observed on the semi-solid part (tubidity). 5. Urease production, observed on solid and semi-solid parts (blue). 6. Indol production, observed on the strip of filter paper (red or purplish). Indol production is not observed with indol positive strains which rapidly acidify the surface o the slant, and the use of oxalic acid has proved to give less sensitive reaction (11). Reading of results - In most cases overnight incubation is enough; sometimes the reactions appear within only a few hours of incubation, affording a definitive orientation of the diagnosis. With some cultures it is necessary to observe the medium during 48 hours of incubation. A description showing typical differential reaction follows: Salmonella: Color of the medium unchanged, with blackening of the solid part when H[2]S is positive. The slant tends to alkalinity (greenish of bluish). Gas always absent. Indol negative. Motility positive or negative. Shigella: Color of the medium unchanged at the beginning of incubation period, but acquiring a red color when the strain is late lactose/sucrose positive. Slant tending to alkalinity (greenish or purplish). Indol positive or negative. Motility, gas and H[2]S always negative. Proteus: Color of the medium generally changes entirely to blue or sometimes to green (urease positive delayed), with blackening of solid part when H[2]S is positive. Motility positive of negative. Indol positive. Gas positive or negative. The strains which attack rapidly sucrose may give a yellow-greenish color to the medium. Sometimes the intense blue color of the medium renders difficult the reading of the H[2]S production. Escherichiae and Klebsiellae: Color of the medium red or yellow (acid) with great and rapid production of gas. Motility positive or negative. Indol generally impossible to observe. Paracoli: Those lactose of sucrose positive give the same reaction as Esherichia. Those lactose or sucrose negatives give the same reactions as Salmonellae. Sometimes indol positive and H[2]S negative. Pseudomonas: Color of the medium unchanged. The slant tends to alkalinity. It is impossible to observe motility because there is no growth in the bottom. Alkaligenes: Color of the medium unchanged. The slant tends to alkalinity. The medium does not alter the antigenic properties of the strains and with the mass of organisms on the slant we can make the serologic diagnosis. It is admitted that this medium is somewhat more laborious to prepare than others used for similar purposes. Nevertheless it can give informations generally obtained by two or three other media. Its use represents much saving in time, labor and material, and we suggest it for routine laboratory work in which a quick presumptive preliminary grouping of enteric organisms is needed.