Densidade e tamanho populacional de mamíferos cinegéticos em duas Unidades de Conservação do Estado do Rio de Janeiro, Brasil

A Mata Atlântica, apesar de ainda sofrer uma intensa devastação, abriga 261 espécies de mamíferos, sendo 73 endêmicos. Mamíferos de grande porte estão entre os mais vulneráveis à caça, perda de habitat e tráfico de animais. No Estado do Rio de Janeiro existem somente duas Reservas Biológicas de Mata Atlântica de baixada, a Reserva Biológica de Poço das Antas e a Reserva Biológica União. O presente estudo teve como objetivo avaliar a influência da prática da caça ilegal sobre a fauna de mamíferos nestas duas Unidades de Conservação. O levantamento populacional foi realizado utilizando o método de transecção linear e 375 quilômetros foram percorridos durante o período de dezembro de 2003 a janeiro de 2005. Os dados de estimativa de densidade populacional foram analisados no programa DISTANCE 5.0. Através de encontros visuais foram confirmadas 12 espécies durante o levantamento, sendo estas regularmente caçadas na região. As espécies que apresentaram maior densidade nas duas Unidades de Conservação foram o macaco-prego (Cebus nigritus Erxleben, 1777), o bugio (Alouatta guariba Lacépède, 1799), tatu-galinha (Dasypus novemcintus Linnaeus, 1758) e o tatu (Dasypus septemcinctus Linnaeus, 1758). As espécies mais raras ou ausentes foram a anta (Tapirus terrestris Brünnich, 1771), o veado (Mazama americana Rafinesque, 1817) e o queixada (Tayassu pecari Link, 1795). Evidências diretas e indiretas da ação da caça ilegal foram observadas nas duas áreas de estudo, indicando que a caça é uma prática comum nessas Reservas Biológicas. A sobrevivência a longo prazo dessas espécies é questionável, já que as populações remanescentes em fragmentos são pequenas e isoladas, o que as tornam muito susceptível à extinção mesmo sob uma baixa pressão de caça.

Didelphis albiventris como indutor de germinação de Rapanea ferruginea (Myrcinaceae) em área de Cerrado, Mato Grosso do Sul, Brasil

A endozoocoria é um dos processos mais importantes na dispersão de sementes em florestas tropicais, dependente em grande parte de aves e mamíferos, onde a passagem dos frutos pelo sistema digestório permite a escarificação química das sementes e sua germinação. Entre as diversas espécies consumidas por animais está Rapanea ferruginea Ruiz et Pav. (Myrsinaceae), uma árvore perenifólia, heliófila, higrófila e pioneira, cujos frutos são ingeridos pelo marsupial Didelphis albiventris Lund, um pequeno mamífero onívoro de hábito noturno. Levando-se em consideração a importância do conhecimento sobre os processos de mutualismo dispersivo, o objetivo neste experimento foi avaliar a germinação de sementes de R. ferruginea que passaram pelo sistema digestório de D. albiventris. As sementes foram submetidas a seis tratamentos: (1) Grupo-Controle - sementes sem tratamento; (2) Grupo Lixa - sementes escarificadas; (3) Grupo Sistema Digestório - sementes que passaram pelo sistema digestório dos animais; (4) Grupo pH 2; (5) Grupo pH 3; (6) Grupo pH4. Os resultados indicaram que o processo de escarificação foi necessário para a obtenção de maiores taxas e velocidades de germinação da espécie estudada. O tratamento com lixa alcançou resultados significativamente diferentes do Grupo Controle, porém inferiores aos demais tratamentos. O tratamento Grupo Sistema Digestório apresentou a maior taxa e velocidade de germinação. Estes resultados indicam que D. albiventris pode ser considerado um frugívoro indutor da taxa e velocidade de germinação de R. ferruginea.

Sobre duas espécies novas do gênero Culicoides latreille, 1809, do Brasil (Diptera, Ceratopogonidae)

São descritas as espécies Culicoides (Hoffmania) saintjusti sp. n. e Culicoides (Oecacta) macieli sp. n. (localidade tipo; Brasil, Rio de janeiro, Jacarepaguá, Pau da Fome) e discutidas sua posição taxionômica e suas relações com espécies afins.

Agar dilution method for susceptibility testing of Neisseria gonorrhoeae

The antibiotic susceptibilities of Neisseria gonorrhoeae isolates obtained from patients attending a clinic for sexually transmitted diseases in Tucumán, Argentina, were determined by the agar dilution method (MIC). 3.5% of the isolates produced ²-lactamase. A total of 96.5% of ²-lactamase negative isolates tested were susceptible to penicillin (MIC < 2 µgml-1); 14.03% of the tested isolates were resistant to tetracycline (MIC < 2 µgml-1), and 98% of the tested isolates were susceptible to spectinomycin (MIC < 64 µgml-1). The MICs for 95% of the isolates, tested for other drugs were: < 2 µgml-1 for cefoxitin, < 0.06 µgml-1 for cefotaxime, < 0.25 µgml-1 for norfloxacin, < 10 µgml-1 for cephaloridine, < 10 µgml-1 for cephalexin, and < 50 µgml-1 for kanamycin. Antibiotic resistance among N. gonorrhoeae isolates from Tucumán, Argentina, appeared to be primarily limited to penicillin and tetracycline, which has been a general use against gonorrhoeae in Tucumán since 1960. Periodic monitoring of the underlying susceptibility profiles of the N. gonorrhoeae strains prevalent in areas of frequent transmission may provide clues regarding treatment options and emerging of drug resistance.

Properties of ß-lactamase from Neisseria gonorrhoeae

ß-lactamase activity was studied in Neisseria gonorrhoeae strains. Optimum temperature was found to be 37°C. The enzyme was inactivated at temperatures higher than 60°C, but remained active during storage at low temperatures (4°C, -30°C and -70°C) for two months. Enzyme activity was observed within a pH range of 5.8-8.0, while the optimum pH was 7.0-7.2. Addition of Ni2+, Fe2+, Fe3+, Mn2+ and p-chloromercurybenzoate to the reaction buffer exerted a negative effect upon the activity, whereas Hg2+ and ethylene diamine tetra-acetic acid produced complete inhibition. These results would indicate the presence of -SH groups at the catalytic site of the enzyme.

In vitro Comparison of Disk Diffusion and Agar Dilution Antibiotic Susceptibility Test Methods for Neisseria gonorrhoeae

At present, most Neisseria gonorrhoeae testing is done with ß-lactamase and agar dilution tests with common therapeutic agents. Generally, in bacteriological diagnosis laboratories in Argentina, study of antibiotic susceptibility of N.gonorrhoeae is based on ß-lactamase determination and agar dilution method with common therapeutic agents. The National Committee for Clinical Laboratory Standards (NCCLS) has recently described a disk diffusion test that produces results comparable to the reference agar dilution method for antibiotic susceptibility of N.gonorrhoeae, using a dispersion diagram for analyzing the correlation between both techniques. We obtained 57 gonococcal isolates from patients attending a clinic for sexually transmitted diseases in Tucumán, Argentina. Antibiotic susceptibility tests using agar dilution and disk diffusion techniques were compared. The established NCCLS interpretive criteria for both susceptibility methods appeared to be applicable to domestic gonococcal strains. The correlation between the MIC's and the zones of inhibition was studied for penicillin, ampicillin, cefoxitin, spectinomycin, cefotaxime, cephaloridine, cephalexin, tetracycline, norfloxacin and kanamycin. Dispersion diagrams showed a high correlation between both methods.

Microbial flora variations in the respiratory tract of mice

A stable microbial system in the respiratory tract acts as an important defense mechanism against pathogenic microorganisms. Perturbations in this system may allow pathogens to establish. In an ecological environment such as the respiratory tract, there are many diverse factors that play a role in the establishment of the indigenous flora. In the present work we studied the normal microbial flora of different areas of the respiratory tract of mice and their evolution from the time the mice were born. Our interest was to know which were the dominant groups of microorganisms in each area, which were the first capable of colonizing and which dominated over time to be used as probiotic microorganisms. Our results show that Gram negative facultatively anaerobic bacilli and strict anaerobic microorganisms were the last ones to appear in the bronchia, while aerobic and Gram positive cocci were present in all the areas of the respiratory tract. The number of facultative aerobes and strict anaerobes were similar in the nasal passage, pharynx instilled and trachea, but lower in bronchia. The dominant species were Streptococcus viridans and Staphylococcus saprophyticcus, followed by S. epidermidis, Lactobacilli and S. cohnii I which were present on every studied days but at different proportions. This paper is the first part of a research topic investigating the protective effect of the indigenous flora against pathogens using the mice as an experimental model.

Population genetic analysis of Colombian Trypanosoma cruzi isolates revealed by enzyme electrophoretic profiles

Although Colombia presents an enormous biological diversity, few studies have been conducted on the population genetics of Trypanosoma cruzi. This study was carried out with 23 Colombian stocks of this protozoa analyzed for 13 isoenzymatic loci. The Hardy-Weinberg equilibrium, the genetic diversity and heterogeneity, the genetic relationships and the possible spatial structure of these 23 Colombian stocks of T. cruzi were estimated. The majority of results obtained are in agreement with a clonal population structure. Nevertheless, two aspects expected in a clonal structure were not discovered in the Colombian T. cruzi stocks. There was an absence of given zymodemes over-represented from a geographical point of view and the presumed temporal stabilizing selective phenomena was not observed either in the Colombian stocks sampled several times through the years of the study. Some hypotheses are discussed in order to explain the results found.

Biological characterization of Trypanosoma cruzi strains

Biological parameters of five Trypanosoma cruzi strains from different sources were determined in order to know the laboratory behaviour of natural populations. The parameters evaluated were growth kinetics of epimastigotes, differentiation into metacyclic forms, infectivity in mammalian cells grown in vitro and parasite susceptibility to nifurtimox, benznidazole and gentian violet. Differences in transformation to metacyclic, in the percentage of infected cells as well as in the number of amastigotes per cell were observed among the strains. Regarding to pharmacological assays, Y strain was the most sensitive to the three assayed compounds. These data demonstrate the heterogeneity of natural populations of T. cruzi, the only responsible of infection in humans.

Development of an enzyme-linked immunosorbent assay for detection of IgM antibodies to Babesia bigemina in cattle

A crude antigenic preparation of Babesia bigemina was used to develop an ELISA for the detection of IgM antibodies. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkerboard titrations. Negative sera from cattle imported from tick-free areas, serum samples collected from infected B. bigemina cattle were used to validate the test. The specificity was 94% and sensitivity of the Elisa 87.5%. Sera from 385 cattle deriving from areas free from tick-borne diseases, which were submitted to a preimmunization process, were screened by this technique. The Elisa detected seroconversion on the 14th day post-inoculation in animals either infested with Boophilus microplus ticks (infected with B. bigemina), or inoculated with B. bigemina infected blood. Antibody titers decreased after day 33; however, all animals remained positive until the end of the experiment (124 days). The ELISA described may prove to be an appropriate serological test for the detection of IgM antibodies against B. bigemina.

Relationship between biological behaviour and randomly amplified polymorphic DNA profiles of Trypanosoma cruzi strains

Once known some biological characteristics of six Trypanosoma cruzi strains, randomly amplified polymorphic DNA (RAPD) analysis was made. Cluster analysis by UPGMA (unweighted pair group method analysis) was then applied both to biological parameters and RAPD profiles. Inspection of the UPGMA phenograms indicates identical clusters, so supporting that usefulness of biological parameters to characterization of T. cruzi strains still remains.

A comparative study of preservation and storage of Haemophilus influenzae

The aim of this study was to compare the efficacy of conservation by freezing the strains of Haemophilus influenzae at -20ºC and -70ºC. Skim milk supplemented with glucose, yeast extract and glycerol allowed highest viability of H. influenzae both at -20ºC and -70ºC from the media analyzed. Trypticase soy broth and brain heart infusion broth supplemented with glycerol, allowed excellent recovery. Use of cotton swaps as supporting material, with or without addition of cryoprotective agents, did not modify H. influenzae viability after six months of storage. Concentration of the initial inoculum positively affected viability when stored at -20ºC. Initial concentration did not influence survival after storage at -70ºC. Thawing at room temperature should not exceed 3 h as to get highest survival percentage.