RESUMO
The microorganisms are the best source of extracellular enzymes since they allow an economical technology with low-resource consumption compared to animals and plants. The amylases are among the most important enzymes being the genus Bacillus one of the most investigated due to its ability to produce this enzyme. The objective of this study was to isolate and analyze the genetic diversity among bacteria of the genus Bacillus sp producer of amylase originated from the soil. To this end, soil samples were collected and submitted to the condition of extreme temperature. The serial dilution procedure followed by seeding on solid medium containing starch was used for isolation of strains that produce amylase. The microorganisms isolated were subjected to standard morphological methods for presumptive identification of the genus Bacillus. The PCR assay with the universal genetic marker 16S rDNA was used for confirmation of bacterial strain. All the 10 isolates presumptively identified as bacteria amplified a fragment of 370 bp corresponding to the 16S rDNA gene. The enzymatic activity was expressed as an enzymatic index (EI), after 24 h of incubation. All isolate producers of amylase exhibited EI ≥ 2.0. The determination of the genetic profile and the clonal relationship among the isolates were performed by the method of ERIC-PCR polymorphism. The isolates of Bacillus spp were divided into 2 groups (I and II). Through this method, the discriminatory capacity of this analysis of polymorphisms was verified in differing producer strains from those not producing amylase.
Assuntos
Amilases/metabolismo , Bacillus/enzimologia , Proteínas de Bactérias/metabolismo , Polimorfismo Genético , Microbiologia do Solo , Amilases/genética , Bacillus/genética , Bacillus/metabolismo , Proteínas de Bactérias/genética , Microbiologia Industrial/métodos , RNA Ribossômico 16S/genéticaRESUMO
The Staphylococcus aureus is the most common isolated microorganism in ruminant animal species diagnostic with clinical or subclinical mastitis. Dairy herds with these diseases can transfer S. aureus into the milk supply, which can lead to food poisoning in humans. The objective of this study was to evaluate the profile of antimicrobial susceptibility, the presence of femA gene, the genetic relationships among isolates of S. aureus obtained from milk originating from flocks diagnosed with subclinical mastitis in nine rural properties in the northern of Minas Gerais State. To this end, 498 samples of bovine milk tested positive for the California mastitis test (CMT) were subjected to morphological methods and biochemical patterns for microbiological presumptive identification of S. aureus. The PCR test with the genetic marker femA was used to confirm the species S. aureus. All the 26 isolates presumptively identified as S. aureus amplified a fragment of 132 bp corresponding to the femA gene. The profile of antimicrobial susceptibility was performed according to the disk-diffusion methodology and two isolates were susceptible to all the antibiotics tested. The drug multiresistence was found in 80.76% of the isolates. The determination of the genetic profile and the clonal relationship among the isolates was performed by the method of DNA RAPD-PCR polymorphism. The S. aureus isolates were divided into two groups with 26 distinct subgroups. The analysis of RAPD-PCR showed no genetic diversity among them, heterogeneous profile and absence of clonality.
Assuntos
Genótipo , Mastite Bovina/microbiologia , Leite/microbiologia , Fenótipo , Staphylococcus aureus/genética , Animais , Proteínas de Bactérias/genética , Bovinos , Resistência Microbiana a Medicamentos , Feminino , Mastite Bovina/diagnóstico , Polimorfismo Genético , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus/patogenicidadeRESUMO
Molecular studies of the evolutionary relationships among Leishmania species suggest the presence of high genetic variation within this genus, which has a direct effect on public health in many countries. The coexistence of species in a particular region can result in different leishmaniasis clinical forms and treatment responses. We aimed to standardize the kinetoplast DNA (kDNA) enterobacterial repetitive intergenic consensus (ERIC) sequence polymerase chain reaction (PCR) method for molecular epidemiological identification of Leishmania strains, and estimate existing inter-strain genomic differences and kDNA signatures using this technique. ERIC-PCR of genomic DNA revealed genetic polymorphisms between species, although some strains shared many DNA fragments. Leishmania guyanensis, L. amazonensis, and L. braziliensis clustered together in a dendrogram with similarities ranging from 42.0 to 61.0%, whereas L. chagasi grouped with these three species with a similarity of 28.0%. After amplification of kDNA, 780-bp bands were extracted from an agarose gel and purified for analysis of its genetic signature. kDNA ERIC-PCR electrophoretic patterns consisted of 100- to 600- bp fragments. Using these profiles, L. braziliensis and L. guyanensis grouped with a similarity of 26.0%, and L. amazonensis and L. chagasi clustered based on a similarity of 100%. The electrophoretic profiles and dendrograms showed that, for epidemiological identification by ERIC-PCR, genomic DNA had greater discriminatory power than kDNA did. More strains need to be analyzed to validate the kDNA ERIC-PCR method. The genomes of these strains should be sequenced for better epidemiological identification of Leishmania species.
Assuntos
Leishmania/classificação , Leishmania/genética , Leishmaniose/epidemiologia , Leishmaniose/parasitologia , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/normas , Sequência de Bases , DNA de Cinetoplasto/genética , Variação Genética , Humanos , Sequências Repetitivas DispersasRESUMO
Acinetobacter sp isolates deserve special attention once they have emerged globally in healthcare institutions because they display numerous intrinsic and acquired drug-resistance mechanisms. This study assessed the antibiotic susceptibility profile, the presence of the genetic marker blaOXA-23, and the clonal relationship among 34 nosocomial isolates of Acinetobacter spp obtained at a hospital in southeastern Brazil. Antibiotic sensitivity analysis was performed by the standard disc-diffusion method. All isolates were found to be extensively resistant to several drugs, but sensitive to polymyxin B. A polymerase chain reaction (PCR) assay was used to detect the blaOXA-23 gene, which is associated with carbapenem resistance. The genetic profile and the clonal relationship among isolates were analyzed via enterobacterial repetitive intergenic consensus (ERIC)-PCR. The Acinetobacter spp were divided into four groups with 22 distinct genetic subgroups. ERIC-PCR analysis revealed the genetic diversity among isolates, which, despite having a heterogeneous profile, displayed 100% clonality among 56% (19/34) of them.
Assuntos
Infecções por Acinetobacter/microbiologia , Acinetobacter/genética , Infecção Hospitalar/microbiologia , Acinetobacter/isolamento & purificação , Infecções por Acinetobacter/epidemiologia , Adolescente , Adulto , Idoso , Proteínas de Bactérias/genética , Criança , Pré-Escolar , Infecção Hospitalar/epidemiologia , Feminino , Genes Bacterianos , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Filogenia , Prevalência , Adulto Jovem , Resistência beta-Lactâmica , beta-Lactamases/genéticaRESUMO
Biotechnology industries that use recombinant DNA technology are potential sources for release of genetically modified organisms to the environment. Antibiotic-resistance marker genes are commonly used for recombinant bacteria selection. One example is the marker gene coding for ß-lactamase (bla) in plasmids found in Escherichia coli K-12. The aim of this study was to provide an approach to develop a molecular method for genetic marker detection in E. coli K-12 harboring bla genes from an industrial wastewater treatment effluent pond (IWTEP). For the detection of bla and Achromobacter lyticus protease I (api) genes in samples from IWTEP, we employed multiplex polymerase chain reaction (PCR) using E. coli K-12 genetic marker detection primers, previously described in the literature, and primers designed in our laboratory. The microbiological screening method resulted in 22 bacterial colony-forming units isolated from three different IWTEP harvesting points. The multiplex PCR amplicons showed that five isolates were positive for the bla gene marker and negative for the E. coli K-12 and api genes. The 16S rRNA regions of positive microorganisms carrying the bla gene were genotyped by the MicroSeq®500 system. The bacteria found were Escherichia spp (3/5), Chromobacterium spp (1/5), and Aeromonas spp (1/5). None of the 22 isolated microorganisms presented the molecular pattern of E. coli K-12 harboring the bla gene. The presence of microorganisms positive for the bla gene and negative for E. coli K-12 harboring bla genes at IWTEP suggests that the ampicillin resistance found in the isolated bacteria could be from microorganisms other than the E. coli K-12 strain harboring plasmid.
Assuntos
Resistência a Ampicilina/genética , Escherichia coli K12/genética , Marcadores Genéticos , Plasmídeos/genética , Águas Residuárias/microbiologia , Ampicilina/farmacologia , Antibacterianos/farmacologia , Brasil , Genes Bacterianos , Lagoas/microbiologia , RNA Ribossômico 16S , Serina Endopeptidases/genética , Instalações de Eliminação de Resíduos , Microbiologia da Água , Purificação da Água , beta-Lactamases/genéticaRESUMO
To investigate the effects of prolonged dietary sodium restriction on lipid metabolism, male rats weighing 35 to 40 g (just weaned) were fed either a low-salt (LSD) or a normal salt diet (NSD) and used in metabolic experiments after 1, 2, or 3 months of diet consumption. After 2 and 3 months on the diet, LSD rats showed increased amounts of lipid in carcass and retroperitoneal tissue. In both LSD and NSD, extending the feeding period from 2 to 3 months resulted in a marked reduction in the in vivo rates of adipose tissue fatty acid synthesis that was accompanied by increases in liver lipogenesis and in the activity of adipose tissue lipoprotein lipase (LPL). However, these increases were more marked in LSD rats. Thus, in vivo rates of liver fatty synthesis and LPL activity in LSD rats, which were already higher (by about 35% and 20%, respectively) than in controls after 2 months, attained levels 50% higher than those in NSD animals after another month on the diet. Brown adipose tissue (BAT) thermogenic capacity, estimated after 2 and 3 months by the tissue temperature response to norepinephrine (NE) injection and by guanosine diphosphate (GDP) binding to BAT mitochondria, did not change in controls, but was significantly reduced in LSD rats. This raises the possibility that a decrease in overall energy expenditure, together with an LPL-induced increased uptake of preformed fatty acids from the circulation, may account for the excessive lipid accumulation in LSD rats. Taken together, the data indicate that prolonged dietary sodium restriction exacerbates normal, age-related changes in white and BAT metabolism.
Assuntos
Tecido Adiposo/fisiologia , Envelhecimento/fisiologia , Dieta Hipossódica/efeitos adversos , Lipídeos/biossíntese , Fígado/metabolismo , Tecido Adiposo Marrom/enzimologia , Tecido Adiposo Marrom/metabolismo , Animais , Regulação da Temperatura Corporal/fisiologia , Peso Corporal/fisiologia , Ingestão de Alimentos/fisiologia , Ácidos Graxos/biossíntese , Glicerol/metabolismo , Guanosina Difosfato/metabolismo , Lipase Lipoproteica/biossíntese , Fígado/crescimento & desenvolvimento , Masculino , Mitocôndrias/metabolismo , Norepinefrina/farmacologia , Ratos , Ratos Wistar , Triglicerídeos/biossíntese , Vasoconstritores/farmacologiaRESUMO
Intravenous (IV) administration of angiotensin II (0.95 nmol/100 g body weight) produced a marked increase in plasma glucose of 20 h fasted rats. To investigate the possibility of a stimulation of gluconeogenesis, conscious unrestrained rats were continuously infused with [14C]bicarbonate, 60 microl/min (0.18 microCi/min), and label incorporation into circulating glucose was determined before and after angiotensin injection. The rate of 14C incorporation into blood glucose of fed rats increased significantly after angiotensin II administration, a 279% increase after 20 min (P < 0.01). In conclusion, the results of the present study show that the hyperglycemia induced by intravenous (IV) administration of angiotensin II is accompanied by an activation of gluconeogenesis, as evidenced by a rapid and marked increase in the rate of 14CO2 incorporation into circulating glucose.
Assuntos
Angiotensina II/administração & dosagem , Gluconeogênese/efeitos dos fármacos , Animais , Glicemia/metabolismo , Radioisótopos de Carbono , Hiperglicemia/sangue , Hiperglicemia/induzido quimicamente , Infusões Intravenosas , Cinética , Fígado/efeitos dos fármacos , Fígado/metabolismo , RatosRESUMO
Propoe-se um metodo para identificacao dos polos naturais de atracao medico-assistencial e suas respectivas areas de influencia, baseado no estudo da evasao e invasao de obitos de residentes ocorridos nas diversas localidades que compoem uma regiao. Tal metodo pressupoe que os falecimentos que se dao fora da sede domiciliar, salvo eventos acidentais, decorrem da busca de assistencia medica em localidades acessiveis e melhor aparelhadas.Quanto maior a evasao, menor o poder de fixacao da area evadida; quanto maior a invasao, maior o poder de atracao sobre a clientela. A forca de polarizacao calculada a partir desses dois atributos foi estudada nos 80 municipios da sexta Divisao Regional de Saude de Sao Paulo. O material foi constituido de 36.448 obitos de residentes certificados nos anos de 1972, 1974 e 1976, dos quais 3.930 aconteceram na regiao, porem fora do municipio de residencia. As maiores taxas de polarizacao foram observadas nos municipios mais providos de recursos. Os polos naturais identificados nesta pesquisa coincidiram, na sua maioria, com os polos definidos pela regionalizacao administrativa disposta por lei
Assuntos
Mortalidade , Assistência Médica , Programas Médicos Regionais , Regionalização da SaúdeAssuntos
Doenças Transmissíveis/epidemiologia , Doenças Parasitárias/epidemiologia , Infecções Bacterianas/epidemiologia , Infecções Bacterianas/mortalidade , Brasil , Doença de Chagas/epidemiologia , Doença de Chagas/mortalidade , Doenças Transmissíveis/mortalidade , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/mortalidade , Humanos , Doenças Parasitárias/mortalidadeRESUMO
An incidence rate of 1.66 was determined for Down's syndrome by a check of hospital birth records in the city of Ribeirão Preto for the year 1972, based on the premise that most births in the population residing in the urban area of Ribeirão Preto occur in hospitals. The prevalence rate was estimated as being 25.2. A search for affected individuals born before 1972 was carried out by physical examination of all persons receiving medical care or educational and social assistance from several city institutions. The prevalence value must be considered as an underestimate since the methods used allowed only for the detection of cases which were getting some kind of assistance.