Para estudos da bioquímica da desnitrificação os desnitrificadores são cultivados em meio parcialmente sintético, com extrato de levedura ou de carne ou ainda peptona. Procurou-se então um meio de cultura completamente sintético no qual P. denitrificans pudesse desenvolver-se e fazer desnitrificação. Para tanto, células de um "strain" dessa bactéria foram crescidas durantes, 24 e 48 horas em tubos de ensaio contendo 10 ml de meio de cultura consistindo de succinato de sódio, nitrato de potássio, extrato de levedura o tampão de fosfato 1M, valôr pH 6,8. Dêsse meio (controle) 0,1 ml foi inoculado em 10 ml do meio de cultura em estudo, mantido então em condições parcialmente anaeróbicas. Após 24 horas uma alíquota foi retirada e suspensa em água destilada e a turbidês lida em espectrofotômetro Beckman, a 420 my. Os dados obtidos após longa série de ensaios permitiram concluir que a fim de se obter em 48 horas um crescimento da mesma ordem que o obtido com o meio controle, o meio de cultura sintético deve conter micronutrientes (Zn, Fe, Mn, Cu, Co, B e Mo, em EDTA), sulfato de sódio, sulfato de magnésio e ácido glutâmico, além de KNO3, succinato e tampão de fosfato.

The experiment described herein was performed in order to establish a completely synthetic medium for growing P. denitrificans cells. In order to obtain in 48 hr. a growth as good as that obtained with the control cultures (with yeast extract), the medium must be formed by micronutrients, the inorganic salts No2SO4, MgSO4 and (NH4)2SO4,and by an amino acid such as sodium glutamate. The KNO3 is for course needed in order to proporcionate the nitrate, and the medium must be buffered with 1M potassium phosphate buffer, pH 6.8. Succinate serves as a substrate. The role of glutamate is under investigation.

O método da diluição isotópica foi empregado com êxito para estudar a influência de doses crescentes de calcário na disponibilidade do fósforo do solo de cerrado. Os valores A foram determinados usando-se uma técnica simplificada; as amostras de solos de cerrado usadas no trabalho haviam sido submetidas prèviamente ao tratamento com diferentes dosagens de calcário em condições de campo. Tais amostras foram colhidas nas localidades de Orlandia, Pirassununga e Matão, todas elas no Estado de São Paulo. As seguintes conclusões puderam ser tiradas: (1) os três tipos de solos de cerrado apresentaram teores diferentes de fósforo assimilável; (2) nos casos das amostras de Pirassununga e Matão a calagem aumetou a disponibilidade do fósforo de modo linear; em Orlandia, entretanto, a dose mais alta de calcário diminuiu significativamente o teor de fósforo disponível.

The method of isotopic dilution was successfully used to study the influence of limestone in the availability of native phosphorus. A values were determined by a simplified technique using samples of cerrado soils previously submitted in the field to the treatment with different amounts of limestone. It has been found out: (1) the three types of cerrado soils had different amounts of available phosphorus; (2) liming increased linearly the A values in the case of two of the soils under study; in the third one, howerer, the response was linear also up to a point wherefrom a further increase in the aumount of limestone applied caused a significant drop in available phosphorus.

During the oxidation of the substrate, both Nitrosomonas and Nitrobacter have part of the energy made available as high energy phosphate, mamely ADP and ATP. This chemical energy is used to fix CO2. The nature of the reducing power is unknown at present. Active cells of Nitrobacter were shown to fix CO2 along the same pathway as found in higher plant photosynthesis. Sonic extracts of Nitrosomonas and Nitrobacter when incubated with NaH14CO3 and cofactors showed two ports of entry of CO2 into organic compounds one being, as expected, the carboxidismutase reaction. On protein basis an equivalent amount of CO2 was, however, incorporated via the oxaloacetic carboxylase reation. It is clear then that both micoorganisms possess typical autotrophic and heterotrophic mechanisms for the fixation of CO2 which is required for the primary synthesis of cell material.

Due to the great importance of coffee to the Brazilian economy, a good deal of the work carried out in the "Laboratório de Isótopos", E. E. A. "Luiz de Queiroz", Piracicaba, S. Paulo, Brazil, was dedicated to the study of some problems involving that plant. The first one was designed to verify a few aspects of the control of zinc deficiency which is common in many types of soils in Brazil. An experiment conducted in nutrient solution showed that the leaf absorption of the radiozinc was eight times as high as the root uptake; the lower surface of the leaves is particularly suited for this kind of absorption. Among the heavy metal micronutrients, only iron did not affect the absorption of the radiozinc; manganese, copper, and molybdenum brought about a decrease of fifty per cent in total uptake. In another pot experiment in which two soils typical of the coffee growing regions were used, namely, a sandy soil called "arenito de Bauru" and a heavy one, "terra roxa", only O.l and 0.2 per cent of the activity supplied to the roots was recovered", respectively. This indicates that under field conditions the farmer should not attempt to correct zinc deficiency by applying zinc salts to the soil: leaf sprays should be used wherever necessary. In order to find out the most suitable way to supply phosphatic fertilizers to the coffee plant, under normal farm conditions, an experiment with tagged superphosphate was carried out with the following methods of distribution of this material: (1) topdressed in a circular area around the trees; (2) placed in the bottom of a 15 cm deep furrow made around the plant; (3) placed in a semicircular furrow, as in the previous treatment; (4) sprayed directly to the leaves. It was verified that in the first case, circa 10 per cent of the phosphorus in the leaves came from the superphosphate; for the other treatments, the results ware, respectively: 2.4, 1.7, and 38.0 per cent. It is interesting to mention that the first and the last methods of distribution were those less used by the farmers; now they are being introduced in many coffee plantations. In a previous trial it was demonstrated that urea sprays were an adequate way to correct nitrogen deficiency under field conditions. An experiment was then set up in which urea-C14 was used to study the metabolism of this fertilizer in coffee leaves. In was verified that in a 9 hours period circa 95 per cent of the urea supplied to the leaves had been absorbed. The distribution of the nitrogen of the urea was followed by standard chemical procedures. On the other hand the fate of the carbonic moiety was studied with the aid of the radiochromatographic technique. Thus, the incorporation of C14 in aminoacids, sugars and organic acids was ascertained. Data obtained in this work gave a definite support to the idea that in coffee leaves, as in a few other higher plants, a mechanism similar to the urea cycle of animals does exist.