Epidemiological characterization of resistance and PCR typing of Shigella flexneri and Shigella sonnei strains isolated from bacillary dysentery cases in Southeast Brazil

Shigella spp are Gram-negative, anaerobic facultative, non-motile, and non-sporulated bacilli of the Enterobacteriaceae family responsible for "Shigellosis" or bacillary dysentery, an important cause of worldwide morbidity and mortality. However, despite this, there are very few epidemiological studies about this bacterium in Brazil. We studied the antibiotic resistance profiles and the clonal structure of 60 Shigella strains (30 S. flexneri and 30 S. sonnei) isolated from shigellosis cases in different cities within the metropolitan area of Campinas, State of São Paulo, Brazil. We used the following well-characterized molecular techniques: enterobacterial repetitive intergenic consensus, repetitive extragenic palindromic, and double-repetitive element-polymerase chain reaction to characterize the bacteria. Also, the antibiotic resistance of the strains was determined by the diffusion disk method. Many strains of S. flexneri and S. sonnei were found to be multi-resistant. S. flexneri strains were resistant to ampicillin in 83.3% of cases, chloramphenicol in 70.0%, streptomycin in 86.7%, sulfamethoxazole in 80.0%, and tetracycline in 80.0%, while a smaller number of strains were resistant to cephalothin (3.3%) and sulfazotrim (10.0%). S. sonnei strains were mainly resistant to sulfamethoxazole (100.0%) and tetracycline (96.7%) and, to a lesser extent, to ampicillin (6.7%) and streptomycin (26.7%). Polymerase chain reaction-based typing supported the existence of specific clones responsible for the shigellosis cases in the different cities and there was evidence of transmission between cities. This clonal structure would probably be the result of selection for virulence and resistance phenotypes. These data indicate that the human sanitary conditions of the cities investigated should be improved

Epidemiological Characterization Of Resistance And Pcr Typing Of Shigella Flexneri And Shigella Sonnei Strains Isolated From Bacillary Dysentery Cases In Southeast Brazil

Shigella spp are Gram-negative, anaerobic facultative, non-montile, and non-sporulated bacilli of the Enterobacteriaceae family responsible or "Shigellosis" or bacillary dysentery, an important cause of worldwide morbidity and mortality. However, despite this, there are very few epidemiological studies about this bacterium in Brazil. We studied the antibiotic resistance profiles and the clonal structure of 60 Shigella strains (30 S.flexneri and 30 S. sonnei) isolated from shigellosis cases in different cities within the metropolitan area of Campinas, State of São Paulo, Brazil. We used the following well-characterized molecular techniques: enterobacterial repetitive intergenic consensus, repetitive extragenic palindromic, and double-repetitive element-polymerase chain reaction to characterize the bacteria. Also, the antibiotic resistance of the strains was determined by the diffusion disk method. Many strains of S. flexneri and S. sonnei were found to be multi-resistant. S. flexneri strains were resistant to ampicillin in 83.3% of cases, chloramphenicol in 70.0%, streptomycin in 86.7%, sulfamethoxazole in 80.0%, and tetracycline in 80.0%, while a smaller number of strains were resistant to cephalothin (3.3%) and sulfazotrim (10.0%). S. sonnei strains were mainly resistant to sulfamethoxazole (100.0%) and tetracycline (96.7%) and, to a lesser extent, to ampicillin (6.7%) and streptomycin (26.7%). Polymerase chain reaction-based typing supported the existence of specific clones responsible for the shigellosis cases in the different cities and there was evidence of transmission between cities. This clonal structure would probably be the result of selection for virulence and resistance phenotypes. These data indicate that the human sanitary conditions of the cities investigated should be improved.402249258Murray, P.R., Rosenthal, K.S., Kobayashi, G.S., Pfaller, M.A., (1998) Medical Microbiology, , 3rd edn. St. Louis: Mosby-Year BookJawetz, E., Melnick, J.L., Adelber, E.A., (2000) Microbiologia Médica, , Rio de Janeiro: Guanabara KooganSansonetti, P.J., Rupture, invasion and inflammatory destruction of the intestinal barrier by Shigella, making sense of prokaryote-eukaryote cross-talks (2001) FEMS Microbiol Rev, 25, pp. 3-14Thong, K.L., Hoe, S.L., Puthucheary, S.D., Yasin, R.M., Detection of virulence genes in Malaysian Shigella species by multiplex PCR assay (2005) BMC Infect Dis, 5, p. 8Parsot, C., Shigella spp. and enteroinvasive Escherichia coli pathogenicity factors (2005) FEMS Microbiol Lett, 252, pp. 11-18Kopecko, D.J., Holcombe, J., Formal, S.B., Molecular characterization of plasmids from virulent and spontaneously occurring avirulent colonial variants of Shigella flexneri (1979) Infect Immun, 24, pp. 580-582Hale, T.L., Genetic basis of virulence in Shigella species (1991) Microbiol Rev, 55, pp. 206-224Rolland, K., Lambert-Zechovsky, N., Picard, B., Denamur, E., Shigella and enteroinvasive Escherichia coli strains are derived from distinct ancestral strains of E. coli (1998) Microbiology, 144 (PART 9), pp. 2667-2672Coimbra, R.S., Grimont, F., Grimont, P.A., Identification of Shigella serotypes by restriction of amplified O-antigen gene cluster (1999) Res Microbiol, 150, pp. 543-553Joklik, W.K., Willet, H.P., Amos, D.B., Wilfert, C.M., (1994) Microbiblogía, , Zinsser (Editor), Buenos Aires: Editora Médica PanamericanaShears, P., Shigelle infections (1996) Ann Trop Med Parasitol, 90, pp. 105-114Ochman, H., Whittam, T.S., Caugant, D.A., Selander, R.K., Enzyme polymorphism and genetic population structure in Escherichia Coli and Shigella (1983) J Gen Microbiol, 129, pp. 2715-2726Navia, M.M., Capitano, L., Ruiz, J., Vargas, M., Urassa, H., Schellemberg, D., Typing and characterization of mechanisms of resistance of Shigella spp. isolated from feces of children under 5 years of age from Ifakara, Tanzania (1999) J Clin Microbiol, 37, pp. 3113-3117Surdeanu, M., Ciudin, L., Pencu, E., Straut, M., Comparative study of three different DNA fingerprint techniques for molecular typing of Shigella flexneri strains isolated in Romania (2003) Eur J Epidemiol, 18, pp. 703-710Brenner, D.J., Fanning, G.R., Miklos, G.V., Steigerwalt, A.G., Polynucleotide sequence relatedness among Shigella species (1973) Int J Syst Bacteriol, 23, pp. 1-7Brenner, D.J., Fanning, G.R., Skerman, F.J., Falkow, S., Polynucleotide sequence divergence among strains of Escherichia coli and closely related organisms (1972) J Bacteriol, 109, pp. 953-965Brenner, D.J., Steigerwalt, A.G., Wathen, H.G., Gross, R.J., Rowe, B., Confirmation of aerogenic strains of Shigella boydii 13 and further study of Shigella serotypes by DNA relatedness (1982) J Clin Microbiol, 16, pp. 432-436Pupo, G.M., Karaolis, D.K., Lan, R., Reeves, P.R., Evolutionary relationships among pathogenic and nonpathogenic Escherichia coli strains inferred from multilocus enzyme electrophoresis and mdh sequence studies (1997) Infect Immun, 65, pp. 2685-2692Pupo, G.M., Lan, R., Reeves, P.R., Multiple independent origins of Shigella clones of Escherichia coli and convergent evolution of many of their characteristics (2000) Proc Natl Acad Sci U S A, 97, pp. 10567-10572Lee, T.M., Chang, C.Y., Chang, L.L., Chen, W.M., Wang, T.K., Chang, S.F., One predominant type of genetically closely related Shigelia sonnei prevalent in four sequential outbreaks in school children (2003) Diagn Microbiol Infect Dis, 45, pp. 173-181Liu, P.Y., Lau, Y.J., Hu, B.S., Shyr, J.M., Shi, Z.Y., Tsai, W.S., Analysis of clonal relationships among isolates of Shigella sonnei by different molecular typing methods (1995) J Clin Microbiol, 33, pp. 1779-1783Lima, A.A., Lima, N.L., Pinho, M.C., Barros Jr., E.A., Teixeira, M.J., Martins, M.C., High frequency of strains multiply resistant to ampicillin, trimethoprim-sulfamethoxazole, streptomycin, chloramphenicol, and tetracycline isolated from patients with shigellosis in northeastern Brazil during the period 1988 to 1993 (1995) Antimicrob Agents Chemother, 39, pp. 256-259Lima, A.A., Sidrim, J.J., Lima, N.L., Titlow, W., Evans, M.E., Greenberg, R.N., Molecular epidemiology of multiply antibiotic-resistant Shigella flexneri in Fortaleza, Brazil (1997) J Clin Microbiol, 35, pp. 1061-1065Lan, R., Alles, M.C., Donohoe, K., Martinez, M.B., Reeves, P.R., Molecular evolutionary relationships of enteroinvasive Escherichia coli and Shigella spp (2004) Infect Immun, 72, pp. 5080-5088Versalovic, J., Koeuth, T., Lupski, J.R., Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes (1991) Nucleic Acids Res, 19, pp. 6823-6831Friedman, C.R., Stoeckle, M.Y., Johnson Jr., W.D., Riley, L.W., Double-repetitive-element PCR method for subtyping Mycobacterium tuberculosis clinical isolates (1995) J Clin Microbiol, 33, pp. 1383-1384Ausubel, F.M., Brent, R., Kingston, R.E., Struhl, K., Moore, D.D., Smith, J.Á., (1988) Current Protocols in Molecular Biology, , Brooklyn: Greene Publishing AssociatesSambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular Cloning: A Laboratory Manual, , 2nd edn. New York: Gold Spring Harbor Laboratory PressYeh, F.C., Yang, R.C., Boyle, T., Popgene, Version 1.31. Microsoft Windows-Based Freeware for Population Genetic Analysis, , http://www.ualberta.ca/~fyeh/Almeida, M.T., Silva, R.M., Donaire, L.M., Moreira, L.E., Martinez, M.R., Enteropathogens associated with acute diarrheal disease in children (1998) J Pediatr, 74, pp. 291-298de Andrade, J.A., de Oliveira, J.O., Fagundes, N.U., Lethality in hospitalized infants with acute diarrhea: Risk factors associated with death (1999) Rev Assoc Med Bras, 45, pp. 121-127Fagundes-Neto, U., de Andrade, J.A., Acute diarrhea and malnutrition: Lethality risk in hospitalized infants (1999) J Am Coll Nutr, 18, pp. 303-308Fernandes, M.R., Trabulsi, L.R., Infectious resistance in pathogenic enteric organisms isolated in São Paulo, Brasil (Preliminary report) (1968) Rev Inst Med Trop São Paulo, 10, pp. 52-53Gomes, T.A., Rassi, V., MacDonald, K.L., Ramos, S.R., Trabulsi, L.R., Vieira, M.A., Enteropathogens associated with acute diarrheal disease in urban infants in São Paulo, Brazil (1991) J Infect Dis, 164, pp. 331-337Medeiros, L.C., Hillers, V.N., Kendall, P.A., Mason, A., Food safety education: What should we be teaching to consumers? (2001) J Nutr Educ, 33, pp. 108-113Piechaud, D., Szturm-Rubinsten, S., Pessoa, G., Diversity of resistance types among Shigelia in São Paulo (Brasil) (author's transl) (1974) Ann Microbiol, 125, pp. 581-584Zuliani, M.E., Trabulsi, L.R., In vitro sensitivity of 166 Shigella strains isolated in São Paulo, Brazil, to sulphadiazine and five antibiotics (1968) Rev Inst Med Trop São Paulo, 10, pp. 70-77Sears, C.L., Kaper, J.B., Enteric bacterial toxins: Mechanisms of action and linkage to intestinal secretion (1996) Microbiol Rev, 60, pp. 167-215Oplustil, C.P., Nunes, R., Mendes, C., Multicenter evaluation of resistance patterns of Klebsiella pneumoniae, Escherichia coli, Salmonenella spp and Shigella spp isolated from clinical specimens in Brazil: RESISTNET Surveillance Program (2001) Braz J Infect Dis, 5, pp. 8-12Casalino, M., Nicoletti, M., Salvia, A., Colonna, B., Pazzani, C., Calconi, A., Characterization of endemic Shigella flexneri strains in Somalia: Antimicrobial resistance, plasmid profiles, and serotype correlation (1994) J Clin Microbiol, 32, pp. 1179-118

In vitro evaluation of the cytotoxic and trypanocidal activities of Ampelozizyphus amazonicus (Rhamnaceae)

Ampelozizyphus amazonicus Ducke is a tree commonly found in the Amazon region and an extract of its stem bark is popularly used as an antimalarial and anti-inflammatory agent and as an antidote to snake venom. Ursolic acid; five lupane type triterpenes: betulin, betulinic acid, lupenone, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid, and 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid, and three phytosteroids: stigmasterol, sitosterol and campesterol, have been isolated from stem extracts of A. amazonicus Ducke. Their structures were characterized by spectral data including COSY and HMQC. In an in vitro biological screening of the isolated compounds, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid was cytotoxic against the SKBR-3 human adenocarcinoma cell line (1 to 10 mg/mL), while 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid exhibited cytotoxicity against both SKBR-3 human adenocarcinoma and C-8161 human melanoma tumor cell lines (>0.1 mg/mL). In the present study, different extracts and some fractions of this plant were also investigated for trypanocidal activity due to the presence of pentacyclic triterpenes. The triterpene classes are potent against Trypanosoma cruzi. The bioassays were carried out using blood collected from Swiss albino mice by cardiac puncture during the parasitemic peak (7th day) after infection with the Y strain of T. cruzi. The results obtained showed that A. amazonicus is a potential source of bioactive compounds since its extracts and fractions isolated from it exhibited in vitro parasite lysis against trypomastigote forms of T. cruzi at concentrations >100 µg/mL. Fractions containing mainly betulin, lupenone, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid, and 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid showed more activity than crude extracts

Brazilian Flora 2020: Leveraging the power of a collaborative scientific network

International audienceThe shortage of reliable primary taxonomic data limits the description of biological taxa and the understanding of biodiversity patterns and processes, complicating biogeographical, ecological, and evolutionary studies. This deficit creates a significant taxonomic impediment to biodiversity research and conservation planning. The taxonomic impediment and the biodiversity crisis are widely recognized, highlighting the urgent need for reliable taxonomic data. Over the past decade, numerous countries worldwide have devoted considerable effort to Target 1 of the Global Strategy for Plant Conservation (GSPC), which called for the preparation of a working list of all known plant species by 2010 and an online world Flora by 2020. Brazil is a megadiverse country, home to more of the world's known plant species than any other country. Despite that, Flora Brasiliensis, concluded in 1906, was the last comprehensive treatment of the Brazilian flora. The lack of accurate estimates of the number of species of algae, fungi, and plants occurring in Brazil contributes to the prevailing taxonomic impediment and delays progress towards the GSPC targets. Over the past 12 years, a legion of taxonomists motivated to meet Target 1 of the GSPC, worked together to gather and integrate knowledge on the algal, plant, and fungal diversity of Brazil. Overall, a team of about 980 taxonomists joined efforts in a highly collaborative project that used cybertaxonomy to prepare an updated Flora of Brazil, showing the power of scientific collaboration to reach ambitious goals. This paper presents an overview of the Brazilian Flora 2020 and provides taxonomic and spatial updates on the algae, fungi, and plants found in one of the world's most biodiverse countries. We further identify collection gaps and summarize future goals that extend beyond 2020. Our results show that Brazil is home to 46,975 native species of algae, fungi, and plants, of which 19,669 are endemic to the country. The data compiled to date suggests that the Atlantic Rainforest might be the most diverse Brazilian domain for all plant groups except gymnosperms, which are most diverse in the Amazon. However, scientific knowledge of Brazilian diversity is still unequally distributed, with the Atlantic Rainforest and the Cerrado being the most intensively sampled and studied biomes in the country. In times of “scientific reductionism”, with botanical and mycological sciences suffering pervasive depreciation in recent decades, the first online Flora of Brazil 2020 significantly enhanced the quality and quantity of taxonomic data available for algae, fungi, and plants from Brazil. This project also made all the information freely available online, providing a firm foundation for future research and for the management, conservation, and sustainable use of the Brazilian funga and flora