ABSTRACT
Abstract The sugarcane in Brazil is passing through a management transition that is leading to the abolition of pre-harvest burning. Without burning, large amounts of sugarcane trash is generated, and there is a discussion regarding the utilization of this biomass in the industry versus keeping it in the field to improve soil quality. To study the effects of the trash removal on soil quality, we established an experimental sugarcane plantation with different levels of trash over the soil (0%, 50% and 100% of the original trash deposition) and analyzed the structure of the bacterial and fungal community as the bioindicators of impacts. The soil DNA was extracted, and the microbial community was screened by denaturing gradient gel electrophoresis in two different seasons. Our results suggest that there are no effects from the different levels of trash on the soil chemistry and soil bacterial community. However, the fungal community was significantly impacted, and after twelve months, the community presented different structures among the treatments.
Subject(s)
Soil Microbiology , Bacteria/isolation & purification , Saccharum/microbiology , Fungi/isolation & purification , Seasons , Soil/chemistry , Bacteria/classification , Bacteria/genetics , Brazil , Saccharum/growth & development , Biodiversity , Fungi/classification , Fungi/geneticsABSTRACT
Many studies have evaluated the effects of biochar application on soil structure and plant growth. However, there are very few studies describing the effect of biochar on native soil microbial communities. Microbial analysis of environmental samples requires accurate and reproducible methods for the extraction of DNA from samples. Because of the variety among microbial species and the strong adsorption of the phosphate backbone of the DNA molecule to biochar, extracting and purifying high quality microbial DNA from biochar-amended soil is not a trivial process and can be considerably more difficult than the extraction of DNA from other environmental samples. The aim of this study was to compare the relative efficacies of three commercial DNA extraction kits, the FastDNA® SPIN Kit for Soil (FD kit), the PowerSoil® DNA Isolation Kit (PS kit) and the ZR Soil Microbe DNA Kit MiniprepTM (ZR kit), for extracting microbial genomic DNA from sand treated with different types of biochar. The methods were evaluated by comparing the DNA yields and purity and by analysing the bacterial and fungal community profiles generated by PCR-DGGE. Our results showed that the PCR-DGGE profiles for bacterial and fungal communities were highly affected by the purity and yield of the different DNA extracts. Among the tested kits, the PS kit was the most efficient with respect to the amount and purity of recovered DNA and considering the complexity of the generated DGGE microbial fingerprint from the sand-biochar samples.
Subject(s)
Sequence Analysis, DNA/methods , Charcoal , Soil Microbiology , Polymerase Chain ReactionABSTRACT
The Atlantic Rainforest does not have a uniform physiognomy, its relief determines different environmental conditions that define the composition of its flora and fauna. Within this ecosystem, bromeliads that form tanks with their leaves hold water reservoirs throughout the year, maintaining complex food chains, based mainly on autotrophic and heterotrophic bacteria. Some works concluded that the water held by tank bromeliads concentrate the microbial diversity of their ecosystem. To investigate the bacterial diversity and the potential biotechnology of these ecosystems, tank bromeliads of the Neoregelia cruenta species from the Atlantic Rainforest in Brazil were used as models for this research. Bacteria isolated from these models were tested for production of bioactive compounds. DGGE of the water held by tank bromeliads was performed in different seasons, locations and sun exposure to verify whether these environmental factors affect bacterial communities. The DGGE bands profile showed no grouping of bacterial community by the environmental factors tested. Most of the isolates demonstrated promising activities in the tests performed. Collectively, these results suggest that tank bromeliads of the N. cruenta species provide important habitats for a diverse microbial community, suggesting that each tank forms a distinct micro-habitat. These tanks can be considered excellent sources for the search for new enzymes and/or new bioactive composites of microbial origin.
Subject(s)
Heterotrophic Bacteria , Bromeliaceae , Phytochemicals , Microbiota , Autotrophic ProcessesABSTRACT
Poribacterial clone libraries constructed for Aplysina fulva sponge specimens were analysed with respect to diversity and phylogeny. Results imply the coexistence of several, prevalently "intraspecific" poribacterial genotypes in a single sponge host, and suggest quantitative analysis as a desirable approach in studies of the diversity and distribution of poribacterial cohorts in marine sponges
Subject(s)
Environmental Microbiology , Genetic Variation , In Vitro Techniques , Phylogeny , Porifera , RNA, Bacterial/isolation & purification , Genotype , Methods , Evaluation Studies as TopicABSTRACT
This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.
ABSTRACT
This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.
Subject(s)
Biodiversity , Eukaryotic Cells/cytology , DNA, Bacterial , Environmental Microbiology , Elapidae/microbiology , In Vitro Techniques , Polymerase Chain Reaction/methods , Soil Microbiology , Methods , Guidelines as Topic , SoilABSTRACT
Arterial hypertension is one of the most prevalent health problems and one of the most important risk factors for cardiovascular and cerebrovascular disease. The purpose of this study was to identify if hypertensive patients are being treated in accordance to current recommendations (JNC-7) for this condition and to estimate the level of blood pressure control in the subjects at the moment of their participation in the study. A total of 138 patents with previous diagnosis of arterial hypertension were recruited from visitors to the Emergency Room at Dr. Pila Hospital in Ponce, Puerto Rico during the months of November 2003 through April 2004. Data was collected in the form of a questionnaire, administered by interview. The questionnaire included demographic data, blood pressure level, anthropometric variables, reported history of chronic diseases, use of antihypertensive agents and other variables. At the moment of their participation, 77 of the subjects had uncontrolled hypertension. Only 25 of subjects reported present use of diuretic agents, which are highly recommended as first line treatment for hypertension. The majority (54) of subjects were being treated with only one antihypertensive agent. The majority (58) of patients were not being treated in accordance with current recommendations (JNC-7) under the category of [quot ]compelling indications[quot ]. The results of this study could very well indicate tendencies and patterns of treatment in hypertensive patients of our community. This may warrant further research and inquiry for explanations
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Hypertension/drug therapy , Patient Compliance , Practice Guidelines as Topic , Emergency Service, Hospital , Hypertension/complications , Surveys and QuestionnairesABSTRACT
A 36 year old white female came to our service after having been evaluated on repetitive occasions in the past for a workup of gigantism and acromegalic features. Since childhood she had developed tall stature, frontal bossing, prominence of zygomatic bones, separated teeth, large hands and size 14 shoes. Human growth hormone and somatomedin serum levels had been normal on all occasions tested. Her past history was significant for primary amenorrhea and a 12 year history of hypertension. On physical examination BP was 140/100, height 6' 2[quot ], weight 2571 bs. Her phenotype was truly acromegalic. There was absence of axillary and pubic hair with no breast development. External genitalia was of female appearance. Laboratory evaluation showed increased FSH of 88 mlU/ml, increase LH of 65.6 mlU/ml and decreased E2 of 12.6 pg/ml. Other findings were low serum cortisol of 0.2 mg/dl, high ACTH of 344 pg/ml, low 17-Ketosteroids, high pregnenolone levels of 595 mg/dl, low 17-hydroxypregnenolone less than 10 ng/dl, very high aldosterone of 31 ng/dl and suppressed PRA of less than 0.1 ng/ml. A pelvic sonogram showed a right ovoid structure which could represent a gonad and failed to identify the uterus and left gonad. A bone densitometry showed a decrease bone mineral density compatible with osteoporosis. Chromosome study showed a karyotype of 46-XY. A diagnosis was made of congenital adrenal hyperplasia secondary to 17-alpha-hydroxylase deficiency in a genotypic male. Our patient was referred to the department of gynecology for surgical removal of the gonads. It is amazing how a patient with severe adrenal insufficiency can withstand 36 years of her life undiagnosed without going into an adrenal crisis. Her tall stature and acromegalic features were the striking signs confusing all physicians and delaying the correct diagnosis and appropriate treatment. There has been reported worldwide, nearly 120 cases with documented severe 17-alpha-hydroxylase deficiency. To our knowledge this is the first case identified in Puerto Rico of male pseudohermaphroditism secondary to 17-alpha-hydroxylase enzyme deficiency
Subject(s)
Humans , Male , Adult , Adrenal Hyperplasia, Congenital , Acromegaly/etiology , Disorders of Sex Development , Adrenal Hyperplasia, Congenital , Disorders of Sex Development , Steroids/blood , Hypertension/etiology , Gonadal Steroid Hormones/blood , Pituitary Hormones, Anterior/blood , Karyotyping , Osteoporosis/etiologyABSTRACT
Se estudiaron los perfiles cromatograficos de los aminoacidos libres asi como el contenido de proteinas (prot) y acido deoxirribonucleico (ADN) de la secrecion vaginal de mujeres normales y con cancer cervicouterino (Ca.Cu). La relacion ADN/prot fue mayor en las secreciones del grupo con Ca.Cu. (37 +/- 10) que en las del grupo normal (23 +/- 8), aunque la diferencia no fue significativa. La concentracion total de aminoacidos fue estadisticamente mayor (p < 0.001) en el grupo normal que en el grupo con Ca.Cu. (86.9 +/- 10.5 vs. 44.6 +/- 3.8) microgramo/mg prot.Los animoacidos que presentaron concentraciones (microgramo/mg ADN) significativamente diferentes en los grupos estudiados fueron: acido aspartico, acido glutamico, treonina, serina, metionina, isoleucina, tirosina, fenilalanina, lisina e histidina. Entre los aminoacidos que eluyen con el primer grupo de amortiguadores usados en la cromatografia, el acido aspartico, la asparagina y la glutamina desaparecieron practicamente en los casos con Ca.Cu., mientras que el acido glutamico experimento una notable disminucion. La concentracion de histidina y triptofano fue mas alta en las pacientes con Ca.Cu. que en las mujeres normales
Subject(s)
Humans , Female , Amino Acids , Uterine Cervical Neoplasms , Vaginal SmearsABSTRACT
En este trabajo se describe un metodo colorimetrico rapido y preciso para la determinacion del peso seco de muestras biologicas. El metodo consiste en hacer raccionar estas muestras con una solucion acida de dicromato durante 20 minutos a 92oC, y enseguida determinar la densidad optica a 630 nm del producto formado. Las ventajas de este procedimiento sobre el metodo estandar de pesada son: 1) Requiere un minimo de tiempo para la determinacion, 2) su mayor sensibilidad, exactitud y precision usando equipo estandar de laboratorio, 3) el metodo es insensible a compuestos inorganicos y, por lo tanto, puede ser aplicado a homogenizados de tejidos preparados en soluciones inorganicas. La unica desvantaja del metodo parece ser que el dicromato reacciona especificamente con grupos que contienen carbono y, por lo tanto, la participacion del nitrogeno u otros elementos en la composicion del material biologico daran proporcionalmente menor absorbancia
Subject(s)
Biological Products , ColorimetryABSTRACT
Se revisan las hipotesis biologicas mas importantes que tratan de explicar el mecanismo de carcinogenesis. La mayor parte postulan que la malignizacion es consecuencia de alteraciones en el material genetico celular. Por otro lado, se revisa la informacion obtenida a partir de la exposicion celular a cancerigenos quimicos, fisicos y virales. Los datos apoyan la idea de que la carcinogenesis se realiza mediante mutaciones somaticas. Se concluye que en el proceso de malignizacion intervienen factores externos e internos, y que la solucion al problema del cancer podria depender de los progresos futuros de la biologia y la genetica moleculares