Inventário florístico de floresta natural na área da estrada Cuiabá-Porto Velho (BR - 364)

Foi feito um inventário de seis hectares de floresta de. terra firme na área de influência da Estrada Cuiabá-Porto Velho (BR-364). Destes seis hectares, dois foram feitos no Município de Jaru, vicinal 605 e quatro na área do Projeto Machadinho, vicinais MC-2, MA-9. A flonesta apresenta uma altura média de 15m e um total de 278 espécies diferentes, representadas por 2.235 indivíduos e 57 famílias nos seis hectares estudados. Os hectares I e II do Município de Jaru apresentaram, respectivamente, uma diversidade florística de 113 e 136 espécies de 10cm ou mais de diâmetro (DAP), enquanto que os hectares III, IV, V e VI da área do Projeto Machadinho apresentaram respectivamente, 103, 115, 122 e. 121 espécies. A espécie mais importante no hectare I é o breu manga (Tetragastris altíssima (Aubl.) Swartz) com o IVI (Índice de Valor de Importância) de 12,07, o que representa 4,0%do total; no hectare II, muiraquatiara (Astronium lecointelDucke) com o IVI 28,94, ou se,ja 9,6% do total; no hectare III, Macrolobiumsp., com o IVI de 28,94, representando 9,6% do total; no hectare IV, pau d'alho (Gallesia integrifolia(Sprenq.) Harms), com IVI de 39,41, representando 13,1% do total; no hectare V, tauari (Couratari macrospermaA.S. Smith), com IVI de 11,32, ou seja, 3,8% do total; no hectare VI, violeta Peltogyne catingae Ducke subsp. glabra(W. Rodr.) M.F. da Silva), com o IVI de 11,68, representando 3,9% do total. As seis famílias mais importantes em ordem de importância são: Leguminosae, Moraceae, Sapotaceae, Lecythidaceae, Bursenaceae e Pal mae. Os dados quantitativos da regeneração natural indicaram que as espécies que apre-sentaram maior número de indivíduos regenerando, foram: Duguetia flagellaresHuber e Maquirasp. no Município de Jaru e Coussareasp. e. Micrandrasp. na área do Projeto Macha dinho, em Ariquemes.

Notas sobre os gêneros Paloue Aublet e Paloveopsis Cowan (Leguminosae: Caesalpinioideae) no Brasil

RESUMOOs gêneros Paloue e Paloueopsis são representados na Amazônia brasileira respectivamente por Paloue brasiliensis Duche, Paloue guianensisAubl. Paloue induta Sandw. subsp. glabra W. Rodr. & Lima (nova subespécie), Paloue riparia Pulle e Paloueopsis emarginataCowan. Eles são aqui sinopticamente apresentados com uma chave para a identificação das respectivas espécies e complementados com comentários sobre o habitat e a distribuição geográfica de cada táxon.

Podridão-mole em plantas de cebolinha causada por Pectobacterium carotovorumsubsp. carotovorum em Roraima

A ocorrência de Pectobacterium carotovorum subsp. carotovorum (=Erwinia carotovora subsp. carotovora) em cebolinha (Allium fistulosum) é relatada pela primeira vez na região norte do Brasil. Até então sua ocorrência estava registrada apenas no Distrito Federal.

Efeito da exploração madeireira sobre o número de indivíduos férteis de três espécies arbóreas comerciais na Amazônia oriental

Estudos da ecologia reprodutiva de árvores são fundamentais para compreender os possíveis impactos da exploração madeireira e para subsidiar o aperfeiçoamento das práticas de manejo. Os objetivos desse trabalho foram: 1) estimar a proporção e o número de indivíduos reprodutivos por classe de diâmetro de Chrysophyllum lucentifolium subsp. pachycarpum, Lecythis lurida e Pseudopiptadenia psilostachya, três espécies madeireiras, em uma floresta em Paragominas (PA) e; 2) estimar o impacto da exploração de 90% dos indivíduos com DAP > 50 cm sobre o número de indivíduos reprodutivos das mesmas espécies no local. Durante uma estação reprodutiva de cada espécie, foram amostradas 80 árvores de L. lurida, 76 de P. psilostachya e 76 de C. lucentifolium. Foi estimado que 14,9% de todos os indivíduos férteis de C. lucentifolium, 35,9% de L. lurida e 72,4% de P. psilostachya tinham DAP > 50 cm no ano de amostragem. Assim, o corte de 90% dessas árvores causaria uma redução de 13,4%, 32,6 % e 65,2 % do número de indivíduos férteis de C. lucentifolium, L. lurida e P. psilostachya, respectivamente. Se as proporções de indivíduos férteis fossem constantes ao longo do tempo, para preservar metade dos indivíduos férteis de P. psilostachya seria necessário manter 30% e não 10% daqueles com DAP > 50 cm. Os resultados indicam que, adotando-se um único diâmetro mínimo de corte e retendo-se a mesma proporção de árvores acima desse diâmetro, o efeito em termos de porcentagem de indivíduos reprodutivos retirados da população pode variar entre espécies na ordem de aproximadamente cinco vezes.

Mimosoideae (Leguminosae) do litoral paraense

Este trabalho consiste no tratamento taxonômico de Mimosoideae nas restingas do litoral paraense, uma vez que estas são pouco conhecidas sob esse aspecto. Esse estudo abrangeu a análise do material proveniente de coletas e exsicatas dos herbários MG e IAN, além de literatura especializada. São apresentadas chave de identificação, descrições e ilustrações dos táxons, bem como dados adicionais sobre distribuição geográfica, comentários, período de floração e frutificação e hábitat das mesmas. O fruto foi o caráter de maior destaque na separação dos táxons. Os resultados evidenciaram a presença de sete espécies, três variedades e uma subespécie, distribuídas em seis gêneros. Inga thibaudiana subsp. thibaudiana e Mimosa candollei, são novos registros para o litoral do Pará. O gênero Inga Mill. foi o mais representativo com cinco espécies. Entada polystachya var. polystachya e principalmente Chloroleucon acacioides apresentaram distribuição mais ampla e a formação floresta de restinga foi o ecossistema que apresentou o maior número de espécie.

Chemical composition of the essential oils from two subspecies of Protium heptaphyllum

Qualitative and quantitative analyses of the volatile constituents from resin of Protium heptaphyllum (Aubl.) Marchand subsp. ulei (Swat) Daly (PHU), and Protium heptaphyllum (Aubl.) Marchand subsp. heptaphyllum (PHH), Burseraceae were performed using GC-MS and GC-FID. The resins were collected around the city of Cruzeiro do Sul, state of Acre, Brazil. Essential oils from the two subspecies were extracted by hydrodistillation with a yield of 8.6% (PHU) and 11.3% (PHH); the main components were terpinolene (42.31%) and p-cymene (39.93%) for subspecies ulei (PHU) and heptaphyllum (PHH), respectively.

Estudo taxonômico de Sapotaceae Juss. do litoral Paraense

Este trabalho consiste no tratamento taxonômico de Sapotaceae Juss. nas restingas do litoral paraense. Com auxilio de estereomicroscópio foram feitas as descrições morfológicas das partes vegetativas e reprodutivas. Utilizou-se literatura especializada, coleções identificadas por especialistas para confirmar as características diferenciais dos táxons e montada uma chave de identificação para as espécies. Foram descritas cinco espécies e quatro subespécies, distribuídas em quatro gêneros: Manilkara bidentata subsp. bidentata, M. bidentata subsp. surinamensis, M. triflora, M. paraensis; Micropholisvenulosa, M. gnaphaloclados; Pouteria ramiflora, P. reticulata subsp. reticulata and Pradosia schomburgkiana subsp. schomburgkiana. O gênero Manilkara Adans.foi o mais representativo com duas espécies e duas subespécies. A formação floresta de restinga foi o ecossistema que apresentou o maior número de táxons.

Bases para o estudo da genética de populações dos Hymenoptera em geral e dos Apinae sociais em particular

This paper deals with problems on population genetics in Hymenoptera and particularly in social Apidae. 1) The studies on populations of Hymenoptera were made according to the two basic types of reproduction: endogamy and panmixia. The populations of social Apinae have a mixed method of reproduction with higher percentage of panmixia and a lower of endogamy. This is shown by the following a) males can enter any hive in swarming time; b) males of Meliponini are expelled from hives which does not need them, and thus, are forced to look for some other place; c) Meliponini males were seen powdering themselves with pollen, thus becoming more acceptable in any other hive. The panmixia is not complete owing to the fact that the density of the breeding population as very low, even in the more frequent species as low as about 2 females and 160 males per reproductive area. We adopted as selection values (or survival indices) the expressions according to Brieger (1948,1950) which may be summarised as follows; a population: p2AA + ²pq Aa + q2aa became after selection: x p2AA + 2pq Aa + z q²aa. For alge-braics facilities Brieger divided the three selective values by y giving thus: x/y p2 AA + y/y 2 pq Aa + z/y q²aa. He called x/y of RA and z/y of Ra, that are survival or selective index, calculated in relation to the heterozygote. In our case all index were calculated in relation to the heterozygote, including the ones for haploid males; thus we have: RA surveval index of genotype AA Ra surveval index of genotype aa R'A surveval index of genotype A R'a surveval index of genotype a 1 surveval index of genotype Aa The index R'A ande R'a were equalized to RA and Ra, respectively, for facilities in the conclusions. 2) Panmitic populations of Hymenoptera, barring mutations, migrations and selection, should follow the Hardy-Weinberg law, thus all gens will be present in the population in the inicial frequency (see Graphifc 1). 3) Heterotic genes: If mutation for heterotic gene ( 1 > RA > Ra) occurs, an equilibrium will be reached in a population when: P = R A + Ra - 2R²a _____________ (9) 2(R A + Ra - R²A - R²a q = R A + Ra - 2R²A _____________ (10) 2(R A + Ra - R²A - R²a A heterotic gene in an hymenopteran population may be maintained without the aid of new mutation only if the survival index of the most viable mutant (RA) does not exced the limiting value given by the formula: R A = 1 + √1+Ra _________ 4 If RA has a value higher thah the one permitted by the formula, then only the more viable gene will remain present in the population (see Graphic 10). The only direct proof for heterotic genes in Hymenoptera was given by Mackensen and Roberts, who obtained offspring from Apis mellefera L. queens fertilized by their own sons. Such inbreeding resulted in a rapid loss of vigor the colony; inbred lines intercrossed gave a high hybrid vigor. Other fats correlated with the "heterosis" problem are; a) In a colony M. quadrifasciata Lep., which suffered severely from heat, the percentage of deths omong males was greater .than among females; b) Casteel and Phillips had shown that in their samples (Apis melifera L). the males had 7 times more abnormalities tian the workers (see Quadros IV to VIII); c) just after emerging the males have great variation, but the older ones show a variation equal to that of workers; d) The tongue lenght of males of Apis mellifera L., of Bombus rubicundus Smith (Quadro X), of Melipona marginata Lep. (Quadro XI), and of Melipona quadrifasciata Lep. Quadro IX, show greater variationthan that of workers of the respective species. If such variation were only caused by subviables genes a rapid increasse of homozigoty for the most viable alleles should be expected; then, these .wild populations, supposed to be in equilibrium, could .not show such variability among males. Thus we conclude that heterotic genes have a grat importance in these cases. 4) By means of mathematical models, we came to the conclusion tht isolating genes (Ra ^ Ra > 1), even in the case of mutations with more adaptability, have only the opor-tunity of survival when the population number is very low (thus the frequency of the gene in the breeding population will be large just after its appearence). A pair of such alleles can only remain present in a population when in border regions of two races or subspecies. For more details see Graphics 5 to 8. 5) Sex-limited genes affecting only females, are of great importance toHymenoptera, being subject to the same limits and formulas as diploid panmitic populations (see formulas 12 and 13). The following examples of these genes were given: a) caste-determining genes in the genus Melipona; b) genes permiting an easy response of females to differences in feeding in almost all social Hymenoptera; c) two genes, found in wild populations, one in Trigona (Plebéia) mosquito F. SMITH (quadro XII) and other in Melipona marginata marginata LEP. (Quadro XIII, colonies 76 and 56) showing sex-limited effects. Sex-limited genes affecting only males do not contribute to the plasticity or genie reserve in hymenopteran populations (see formula 14). 6) The factor time (life span) in Hymenoptera has a particular importance for heterotic genes. Supposing one year to be the time unit and a pair of heterotic genes with respective survival indice equal to RA = 0, 90 and Ra = 0,70 to be present; then if the life time of a population is either one or two years, only the more viable gene will remain present (see formula 11). If the species has a life time of three years, then both alleles will be maintained. Thus we conclude that in specis with long lif-time, the heterotic genes have more importance, and should be found more easily. 7) The colonies of social Hymenoptera behave as units in competition, thus in the studies of populations one must determine the survival index, of these units which may be subdivided in indice for egg-laying, for adaptive value of the queen, for working capacity of workers, etc. 8) A study of endogamic hymenopteran populations, reproduced by sister x brother mating (fig. 2), lead us to the following conclusions: a) without selection, a population, heterozygous for one pair of alleles, will consist after some generations (theoretically after an infinite number of generation) of females AA fecundated with males A and females aa fecundated with males a (see Quadro I). b) Even in endogamic population there is the theoretical possibility of the presence of heterotic genes, at equilibrium without the aid of new mutations (see Graphics 11 and 12), but the following! conditions must be satisfied: I - surveval index of both homozygotes (RA e Ra) should be below 0,75 (see Graphic 13); II - The most viable allele must riot exced the less viable one by more than is permited by the following formula (Pimentel Gomes 1950) (see Gra-fic 14) : 4 R5A + 8 Ra R4A - 4 Ra R³A (Ra - 1) R²A - - R²a (4 R²a + 4 Ra - 1) R A + 2 R³a < o Considering these two conditions, the existance of heterotic genes in endogamic populations of Hymenoptera \>ecames very improbable though not - impossible. 9) Genie mutation offects more hymenopteran than diploid populations. Thus we have for lethal genes in diploid populations: u = q2, and in Hymenoptera: u = s, being u the mutation ratio and s the frequency of the mutant in the male population. 10) Three factors, important to competition among species of Meliponini were analysed: flying capacity of workers, food gathering capacity of workers, egg-laying of the queen. In this connection we refer to the variability of the tongue lenght observed in colonies from several localites, to the method of transporting the pollen in the stomach, from some pots (Melliponi-ni storage alveolus) to others (e. g. in cases of pillage), and to the observation that the species with the most populous hives are almost always the most frequent ones also. 11) Several defensive ways used for Meliponini to avoid predation are cited, but special references are made upon the camouflage of both hive (fig. 5) and hive entrance (fig. 4) and on the mimetism (see list in page ). Also under the same heading we described the method of Lestrimelitta for pillage. 12) As mechanisms important for promoting genetic plasticity of hymenopteran species we cited: a) cytological variations and b) genie reserve. As to the former, duplications and numerical variations of chromosomes were studied. Diprion simile ATC was cited as example for polyploidy. Apis mellife-ra L. (n •= 16) also sugests polyploid origen since: a) The genus Melipona, which belongs to a" related tribe, presents in all species so far studied n = 9 chromosomes and b) there occurs formation of dyads in the firt spermatocyte division. It is su-gested that the origin of the sex-chromosome of Apis mellifera It. may be related to the possible origin of diplo-tetraploidy in this species. With regards to the genie reserve, several possible types of mutants were discussed. They were classified according to their survival indices; the heterotic and neutral mutants must be considered as more important for the genie reserve. 13) The mean radius from a mother to a daghter colony was estimated as 100 meters. Since the Meliponini hives swarm only once a year we may take 100 meters a year as the average dispersion of female Meliponini in ocordance to data obtained from Trigona (tetragonisca) jaty F. SMITH and Melipona marginata LEP., while other species may give different values. For males the flying distance was roughly estimated to be 10 times that for females. A review of the bibliography on Meliponini swarm was made (pg. 43 to 47) and new facts added. The population desity (breeding population) corresponds in may species of Meliponini to one male and one female per 10.000 square meters. Apparently the males are more frequent than the females, because there are sometimes many thousands, of males in a swarm; but for the genie frequency the individuals which have descendants are the ones computed. In the case of Apini and Meliponini, only one queen per hive and the males represented by. the spermatozoos in its spermateca are computed. In Meliponini only one male mate with the queen, while queens of Apis mellijera L. are fecundated by an average of about 1, 5 males. (Roberts, 1944). From the date cited, one clearly sees that, on the whole, populations of wild social bees (Meliponini) are so small that the Sewall Wright effect may become of great importance. In fact applying the Wright's formula: f = ( 1/aN♂ + 1/aN♀) (1 - 1/aN♂ + 1/aN♀) which measures the fixation and loss of genes per generation, we see that the fixation or loss of genes is of about 7% in the more frequent species, and rarer species about 11%. The variation in size, tergite color, background color, etc, of Melipona marginata Lep. is atributed to this genetic drift. A detail, important to the survival of Meliponini species, is the Constance of their breeding population. This Constance is due to the social organization, i. e., to the care given to the reproductive individuals (the queen with its sperm pack), to the way of swarming, to the food storage intended to control variations of feeding supply, etc. 14) Some species of the Meliponini are adapted to various ecological conditions and inhabit large geographical areas (e. g. T. (Tetragonisca jaty F. SMITH), and Trigona (Nanno-trigona testaceicornis LEP.) while others are limited to narrow regions with special ecological conditions (e. g. M. fuscata me-lanoventer SCHWARZ). Other species still, within the same geographical region, profit different ecological conditions, as do M. marginata LEP. and M. quadrifasciata LEP. The geographical distribution of Melipona quadrifasciata LEP. is different according to the subspecies: a) subsp anthidio-des LEP. (represented in Fig. 7 by black squares) inhabits a region fron the North of the S. Paulo State to Northeastern Brazil, ,b) subspecies quadrifasciata LEP., (marked in Fig. 7 with black triangles) accurs from the South of S. Paulo State to the middle of the State of Rio Grande do Sul (South Brazil). In the margined region between these two areas of distribution, hi-brid colonies were found (Fig. 7, white circles); they are shown with more details in fig. 8, while the zone of hybridization is roughly indicated in fig. 9 (gray zone). The subspecies quadrifasciata LEP., has 4 complete yellow bands on the abdominal tergites while anthidioides LEP. has interrupted ones. This character is determined by one or two genes and gives different adaptative properties to the subspecies. Figs. 10 shows certains meteorological isoclines which have aproximately the same configuration as the limits of the hybrid zone, suggesting different climatic adaptabilities for both genotypes. The exis-tance of a border zone between the areas of both subspecies, where were found a high frequency of hybrids, is explained as follows: being each subspecies adapted to a special climatic zone, we may suppose a poor adaptation of either one in the border region, which is also a region of intermediate climatic conditions. Thus, the hybrids, having a combination of the parent qualities, will be best adapted to the transition zone. Thus, the hybrids will become heterotic and an equilibrium will be reached with all genotypes present in the population in the border region.

Anatomia, dos nectános de algumas espécies da flora apícola

This paper deals with anatomical descriptions of some types of nectaries in 27 species of honey plants of Piracicaba, S. P. The material studied was divides in two groups: a) Extra-floral nectaries; b) Floral nectaries. Euphorbia pulcherrima, Willd; showed to belonging to the first group: its nectaries tissue consist of an epidermal layer of cell without stomata and with true gland, with subepidermal cells diferentiated by the thickness of the wall. Among the plants with floral nectaries, the following types has been listed, according the location of the nectary in the flower: 1 - with true glands: a) in sepals, Hibiscus rosa sinensis, L.; Dombeya Wallichii, Bth. e Hk; b) in the stamens tube, Antigonum leptopus, Hook e Arn.; 2 - on the receptacle with nectariferous tissue in the epidermal cell with: a) thickness wall with stomata, Prunus persical, L.; b) thin wall without stomata, Crotalaria paulinia, Shranck; Caesal-pinia sepiaria, Roxb; Aberia caffra; 3 - with a disc located in the receptacle with: epidermal: a) with stomata, Coffea arábica, L. var. semper florens; Citrus aurantifolia, Swing; Cinchona sp.; Pryrostegia ignea, Presl.; b) without stomata and with thin wall, Leojurus sibiricus, L.; Bactocydia unguis, Mart., Ipomoea purpurea, L.; Greviüea Thelemanniana, Hueg.; Dolichos lablab, L.; Vernonia polyanthes, Less., Montanoa bipinatifida, C. Koch., Eruca sativa, L. Brassica Juncea, Co; Eucalyptus tereticomis, Smith.; Eucalyptus rostrata, Schleche; Salvia splendens, Selow.; 4 - in the basal tissues of the ovary, Budleia brasiliensis, Jacq F.; Petrea subserrata, Cham.; 5 - in the base of stamens, Per sea americana, Mill. On the anatomical point of view, most of the types of nectary studied has external nectariferous tissues, located on the epidermal cells with thin periclinal wall and without stomata. The sub-epidermal layer were rich in sugar. Short correlation was found between the structure of the nectary and the amount of nectar secretion. So, in the nectary with true glands, in those with thin wall and without stomata on epidermal cells and in those with stomata, the secretion was higher than in the other types listed.

"Influencia da época de semeadura sôbre o rendimento do repôlho"

The author carried out a study on the behavior of cabbage for a period of two years (1952 and 1953). From the results obtained it was concluded that, cabbage, Brassica oleraceae, var. capitata, developed better during the cooler season of the year, that is, from April to June and, the yield decresed from the beginning to the end of the sowing time, that is, from April to Ocotber.

Técnica de réplica de carbono para o estudo da superfície de fôlhas de plantas ao microscópio eletrônico

O presente trabalho apresenta um método de réplica de superfície para o estudo da estrutura das fôlhas de vegetais ao microscípio eletrônico. O método permitiu a observação do mesmo tipo de estrutura observada por outros autores na superfície das fôlhas de couve (Brassica oleracea). O método permitiu observar que nas superfícies das fôlhas de café (Coffea arabica) a ultraestrutura característica e pràticamente desprovida do tipo de ornamentação encontrado nas fôlhas de couve.

Nutrição mineral de hortaliças: XII. deficiências de micronutrientes em couve-flor

Mudas de couve-flor (Brassica olerácea var. botrytes, cultivar Piracicaba precoce nº 1) foram transplantados aos 25 dias para vasos contendo silica. As mudas em número de quatro por tratamento foram submetidas as seguintes soluções: omissão de boro, cobre, ferro, manganês, molibdênio, zinco e solução completa. Todas as soluções sofreram uma purificação previa. Foram constatados e descritos os sintomas de carencia dos micronutrientes. Foi determinado a influência da omissão dêstes nutrientes sôbre o crescimento das plantas expresso em peso da materia seca. O quadro sintomatológico das deficiencias foi comprovado através de analise química das plantas.

Expression of mosquito active toxin genes by a Colombian native strain of the gram-negative bacterium Asticcacaulis excentricus

Mosquito control with biological insecticides, such as Bacillus sp. toxins, has been used widely in many countries. However, rapid sedimentation away from the mosquito larvae feeding zone causes a low residual effect. In order to overcome this problem, it has been proposed to clone the Bacillus toxin genes in aquatic bacteria which are able to live in the upper part of the water column. Two strains of Asticcacaulis excentricus were chosen to introduce the B. sphaericus binary toxin gene and B. thuringiensis subsp. medellin cry11Bb gene cloned in suitable vectors. In feeding experiments with these aquatic bacteria, it was shown that Culex quinquefasciatus, Aedes aegypti, and Anopheles albimanus larvae were able to survive on a diet based on this wild bacterium. A. excentricus recombinant strains were able to express both genes, but the recombinant strain expressing the B. sphaericus binary toxin was toxic to mosquito larvae. Crude protease A. excentricus extracts did not degrade the Cry11Bb toxin. The flotability studies indicated that the recombinant A. excentricus strains remained in the upper part of the water column longer than the wild type Bacillus strains.

Mycobacterium tuberculosis complex differentiation using gyrB-restriction fragment length polymorphism analysis

Mycobacterium tuberculosis complex (MTBC) members are causative agents of human and animal tuberculosis. Differentiation of MTBC members is required for appropriate treatment of individual patients and for epidemiological purposes. Strains from six MTBC species - M. tuberculosis, M. bovis subsp. bovis, M. bovis BCG, M. africanum, M. pinnipedii, and "M. canetti" - were studied using gyrB-restriction fragment length polymorphism (gyrB-RFLP) analysis. A table was elaborated, based on observed restriction patterns and published gyrB sequences. To evaluate applicability of gyrB-RFLP at Instituto Adolfo Lutz, São Paulo, Mycobacterial Reference Laboratory, 311 MTBC clinical isolates, previously identified using traditional methods as M. tuberculosis (306), M. bovis (3), and M. bovis BCG (2), were analyzed by gyrB-RFLP. All isolates were correctly identified by the molecular method, but no distinction between M. bovis and M. bovis BCG was obtained. Differentiation of M. tuberculosis and M. bovis is of utmost importance, because they require different treatment schedules. In conclusion, gyrB-RFLP is accurate and easy-to-perform, with potential to reduce time needed for conventional differentiation methods. However, application for epidemiological studies remains limited, because it cannot differentiate M. tuberculosis from M. africanum subtype II, and "M. canetti", M. africanum subtype I from M. pinnipedii, and. M. bovis from M. bovis BCG.

Application of PCR ribotyping and tDNA-PCR for Klebsiella pneumoniae identification

PCR analysis of 16S-23S internal transcribed spacer (PCR ribotyping) and tRNA intergenic spacer (tDNA-PCR) were evaluated for their effectiveness in identification of clinical strains of Klebsiella pneumoniae and differentiation with related species. For this purpose both methods were applied to forty-three clinical isolates biochemically identified as K. pneumoniae subsp. pneumoniae isolated from patients clinical specimens attended at five hospitals in three Brazilian cities. References strains of K. pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae, K. oxytoca, K. planticola and Enterobacter aerogenes were also analyzed. Both PCR methods showed specific patterns for each species. A conserved PCR ribotype pattern was observed for all clinical K. pneumoniae isolates, while differing from other related analyzed species. tDNA-PCR revealed five distinct patterns among the K. pneumoniae clinical isolates studied, demonstrating a predominant group with 90,6% of isolates presenting the same pattern of K. pneumoniae type strain. Both PCR-based methods were not able to differentiate K. pneumoniae subspecies. On the basis of the results obtained, both methods were efficient to differentiate the Klebsiella species analyzed, as well as E. aerogenes. Meanwhile tDNA-PCR revealed different tRNA arrangements in K. pneumoniae, suggesting intra-species heterogeneity of their genome organization, the polymorphism of the intergenic spacers between 16S and 23S rRNA genes appears to be highly conserved whithin K. pneumoniae clinical isolates, showing that PCR ribotyping can be an useful tool for identification of K. pneumoniae isolates.