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Echinodorus macrophyllus (Kunth.) Micheli (Alismataceae), known as chapéu-de-couro in Brazil, is popularly used to treat inflammatory diseases. We have previously demonstrated a significant reduction in the acute inflammation for the aqueous extract of E. macrophyllus (AEEm) and its ethanolic fraction (Fr20) and described that hydroxycinnamoyl derivatives present in SF1 (Fr20 subfraction) showed higher anti-inflammatory properties by mechanisms that include a reduction of TNF-α, IL-1β, CKCL1/KC, LTB4, and PGE2 levels in exudate. This work describes the acute toxicological effect of SF1 subfraction on SW mice treated orally for five days in the air pouch model by evaluating the hematological and biochemical determinations on the blood samples; the relative organ weight and its histopathological analysis; the liver genotoxicity assessment and the activity of liver enzymes from xenobiotic metabolism. Fr20 was earlier fractionated on the Sephadex LH-20 column, yielding mainly four subfractions, including SF1. The SF1 toxicity was evaluated in mice challenged with carrageenan on the air pouch inflammation model and orally treated for five days. The body weight was monitored daily, and the organs were weighed after the euthanasia. Hematological and biochemical determinations were carried out using specific commercial kits and following the protocols provided by the manufacturers. The organs were fixed, sectioned, processed for hematoxylin and eosin staining, and analyzed by light microscopy. Genotoxicity assessment was performed by the alkaline single-cell gel electrophoresis. Livers were processed for ethoxyresorufin-O-deethylase (EROD) and Glutathione S-transferase (GST) assays. SF1 exhibited low toxicity, as no significant discrepancy was observed in the relative weight of the body organs of mice. Moreover, the daily treatment with SF1 did not alter the number and percentage of red blood cells or hemoglobin concentration in the blood. The treatment with SF1 did not affect the creatinine concentration, but the 25 mg/kg dose reduced the plasma urea level and uric acid, suggesting its use in treating acute renal failure. The parameters analyzed did not present biochemical alterations indicative of liver disease. Regarding serum triglyceride and cholesterol levels, a significant decrease was detected in both parameters in mice treated with SF1. In addition, the histopathological analysis showed that inflammatory focus in the livers seemed more relevant in the control groups than in those treated. There were no significant changes in the renal or splenic tissues of animals treated with SF1. Treatment with SF1 also does not have a genotoxic effect on liver cells. Treatment with SF1 showed no toxicity in mice at doses equivalent to those recommended for humans, which provides evidence of the safety of the therapeutic use of this subfraction. [Display omitted] • Fr20 was fractionated, and the SF1 subfraction exhibited higher anti-inflammatory properties. • SF1 phytochemical analysis indicates the major presence of hydroxycinnamoyl derivatives. • Previously, SF1 showed reduced inflammatory response induced by carrageen on the air pouch model. • In this study, no significant toxicity was observed in mice after five days of SF1 treatment with all tested doses. • All results provide evidence of the safety of the therapeutic use of this subfraction. [ABSTRACT FROM AUTHOR]

Salmonella is one of the major causes of gastroenteritis worldwide in both humans and animals and one of the main agents involved in foodborne disease outbreaks. In this study, 70 raw kibbe samples from different commercial establishments were analyzed for Salmonella spp. The isolates were seroyped and tested for susceptibility to antimicrobial agents. Pulsed-field Gel Electrophoresis was carried out following the standard protocol of the PulseNet network. Fifteen (21.4%) samples were contaminated with Salmonella and S. Give was the prevalent serotype (46.7%). Similarity of 96.3% was observed among the S. Give isolates (n = 7), which indicates the possible spread of the same clone in the analyzed commercial establishments. S. Rissen and S. Typhimurium showed antimicrobial resistance. The detection of a significant percentage of contamination in raw kibbe and of the resistant strains indicates the risk that the consumption of this dish may represent. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate (1) the effect of a salivary substitute prepared using chamomile (Matricaria chamomilla L.) flower and flax (Linum usitatissimum L.) seed to relieve Primary burning mouth syndrome (BMS) symptoms, (2) their effect on the inhibition of matrix metallopeptidase 2 (MMP2) and MMP9 metalloproteinases, and (3) their potential cellular cytotoxic effect. Subjects: 40 women aging >40 years with diagnosis of primary BMS. Settings/Location: Center of Diagnosis of Diseases of the Mouth, Federal University of Pelotas, Brazil. Design: This was an open clinical trial where primary BMS patients used the homemade salivary. At the first appointment, after 30 and 60 days, the authors evaluated the pattern and intensity of BMS and xerostomia symptoms, and then determined and compared the unstimulated salivary flow rate (SFR), viscosity, and salivary pH. MMP2 and MMP9 activities in saliva and cytotoxicity were assessed using different concentrations of chamomile flower and flax seed separately. Interventions: Subjects used the homemade salivary substitute for 3 months and were instructed to rinse their mouth three to four times daily for 1 min. Outcome measures: A numeric rating scale to evaluate the intensity of burning sensation and xerostomia symptoms, salivary flow rate (SFR) to determine salivary volume, dynamic rheology technique for viscosity and a digital meter for salivary pH. MMP2 and MMP9 activities in saliva and cytotoxicity were assessed by zymography and cell viability assay respectively. Results: After treatment, severity of BMS symptoms decreased, the SFR increased, salivary viscosity decreased, and severity of xerostomia sensation (in patients who reported having this symptom) improved (p < 0.05). Chamomile flower and flax seed had no effect on inhibiting MMP2 and MMP9 activities, and neither showed cellular cytotoxic effects. Conclusion: This homemade salivary substitute is an economical, viable, easily manipulated, noncytotoxic, and a practical alternative to relieve BMS symptoms. [ABSTRACT FROM AUTHOR]

Background: Physiological values reflect the health condition and responses of individuals to handling in captivity. The aim of this study was to establish hematological and serum biochemistry parameters of clinically healthy animals of the Alouatta guariba clamitans subspecies. Methods: We collected blood samples from adult males and females kept at the Center for Biological Research of Indaial after chemical containment with 3.9 mg/kg of tiletamine hydrochloride and zolazepam. Results: Significant differences between males and females were found in the levels of erythrocytes, hemoglobin, hematocrit, lymphocytes, neutrophils, platelets, mean corpuscular hemoglobin concentration (MCHC), and gamma‐glutamyltranspeptidase (GGT). Conclusions: Our results suggest the existence of sexual dimorphism in some physiological parameters of A guariba clamitans. The parameters reported herein can be used as reference values for other populations kept under similar conditions. [ABSTRACT FROM AUTHOR]

Ethnopharmacological importance The effectiveness of aqueous extract of Bellucia dichotoma Cogn. (Melastomataceae) specimems collected in Santarém, PA, against some biological activities of Bothrops atrox venom (BaV) has been scientifically proven. Here, we analyzed the components and assessed the anti-snakebite potential of aqueous extracts of bark of B. dichotoma collected in Manaus, AM, (AEBd-MAO) and Santarém, PA, (AEBd-STM), both in Brazil. Materials and methods The phytochemical profiles of the aqueous extracts were identified using thin layer chromatography (TLC), and the concentrations of phenolics were determined by colorimetric assay. The inhibitory potential of the extracts was tested against the phospholipase A 2 , coagulant and gelatinolytic activities of BaV in vitro and its defibrinating and edema-inducing activities in vivo. Interaction between BaV and the extracts was investigated using SDS-Page electrophoresis and Western blotting. Extract cytotoxicity and antioxidant potential were assessed using the human fibroblast cell line MRC-5 and the DPPH assay in cell culture, respectively. Results While there was no difference between the phytochemical profiles of the extracts, AEBd-MAO had higher concentrations of total phenolics, total tannins and hydrolysable tannins. The extracts inhibited 100% of the phospholipase and coagulant activity of BaV when pre-incubated. Without pre-incubation, however, there was no reduction in phospholipase activity, although significant inhibition of coagulant activity was observed. In the doses used in folk medicine, without pre-incubation, both extracts inhibited 100% of the coagulant activity of BaV. In vivo, the extracts were unable to inhibit the defibrinating activity of the venom but were effective in inhibiting its edema-inducing activity. In the profiles of the extracts pre-incubated with BaV, not all the protein bands revealed by SDS-PAGE and Western blot were observed. Both extracts had a high antioxidant potential and neither had a cytotoxic effect. Conclusion Although the concentrations of phenolics in each extract were different, the anti-snakebite potential was similar for the concentrations of extract tested. Our findings are of importance for the quality control of this raw material, which, once tested in accordance with Brazilian National Health Surveillance Agency recommendations, may be suitable for use as a phytomedicine to complement treatment of the local effects induced by Bothrops venoms. [ABSTRACT FROM AUTHOR]

Aims: We aimed to investigate the impact of following a diet to induce weight loss (500 kcal deficit per day) over DNA damage and cardiometabolic risk factors in women with overweight/obesity diagnosed with polycystic ovary syndrome (PCOS). Methods: A study was conducted in Natal, RN, Brazil selecting overweight/obese (body mass index ≥ 25 and <39 kg/m²) women (18-35 years). The levels of DNA damage were assessed by a single cell gel electrophoresis. Repeated 24 h dietary recall questionnaires, anthropometry, biochemical profile and sex hormones were collected at baseline and after 12 weeks of intervention. Results: Women exhibiting a decrease in the markers of DNA damage: tail intensity (24.35 ± 5.86 -- pre diet vs. 17.15 ± 5.04 - post-diet; p < 0.001) and tail moment (20.47 ± 7.85 - pre diet vs. 14.13 ± 6.29 - post-diet; p < 0.002). Reduction of calorie intake, weight loss, decreased sexual hormone and cardiometabolic markers such as insulin, homeostasis model assessment of insulin resistance and low-density lipoprotein cholesterol were verified In the multivariate regression analysis, quantitative insulin sensitivity check index and progesterone were responsible for the variation markers in DNA damage before the diet, losing its influence upon diet. Conclusion: DNA damage and the impact of cardiometabolic risk factors decreased after the intervention in women with PCOS, indicating the relevance of a nutritional approach in this group of patients. [ABSTRACT FROM AUTHOR]

The prevalence of Salmonella spp. was investigated in 109 wild birds poached in the illegal wildlife trade in Rio de Janeiro; most of them are passerines from Thraupidae family and three from Psittacidae. One strain of Salmonella ser. Typhimurium and two strains of Salmonella ser. Panama were isolated from passerine species and all of them showed resistance to multiple antimicrobial drugs, like ampicillin, ceftriaxone, ceftiofur, tetracycline, gentamicin, nalidixic acid, ciprofloxacin, and enrofloxacin. PFGE showed 100% similarity among the Salmonella ser. Typhimurium strain isolated from a Temminck’s seedeater (Sporophila falcirostris) and the strains isolated from a human outbreak, in southern Brazil. The two Salmonella ser. Panama strains isolated from two chestnut-capped blackbirds (Chrysomus ruficapillus) present in the same catch showed the same clonal origin and have never been associated with epizooties and human outbreaks. Potential for dissemination of resistant Salmonella through situations offered by captive management and the isolation of the same strain from wild birds and human sources may become a problem for the conservation of natural populations and to public health. [ABSTRACT FROM AUTHOR]

In the New World, the leishmaniases are primarily transmitted to humans through the bites of Leishmania-infected Lutzomyia (Diptera: Psychodidae) phlebotomine sand flies. Any or both of two basic clinical forms of these diseases are endemic to several cities in Brazil—the American cutaneous leishmaniasis (ACL) and the American visceral leishmaniasis (AVL). The present study was conducted in the urban area of a small-sized Brazilian municipality (Jaboticatubas), in which three cases of AVL and nine of ACL have been reported in the last five years. Jaboticatubas is an important tourism hub, as it includes a major part of the Serra do Cipó National Park. Currently, no local data is available on the entomological fauna or circulating Leishmania. During the one-year period of this study, we captured 3,104 phlebotomine sand flies belonging to sixteen Lutzomyia species. In addition to identifying incriminated or suspected vectors of ACL with DNA of the etiological agent of AVL and vice versa, we also detected Leishmania DNA in unexpected Lutzomyia species. The expressive presence of vectors and natural Leishmania infection indicates favorable conditions for the spreading of leishmaniases in the vicinity of the Serra do Cipó National Park. [ABSTRACT FROM AUTHOR]

Hebanthe paniculata roots (formerly Pfaffia paniculata and popularly known as Brazilian ginseng) show antineoplastic, chemopreventive, and antiproliferative properties. Functional properties of these roots and their extracts are usually attributed to the pfaffosidic fraction, which is composed mainly by pfaffosides A-F. However, the therapeutic potential of this fraction in cancer cells is not yet entirely understood. This study aimed to analyze the antitumoral effects of the purified pfaffosidic fraction or saponinic fraction on the human hepatocellular carcinoma HepG2 cell line. Cellular viability, proliferation, and apoptosis were evaluated, respectively, by MTT assay, BrdU incorporation, activated caspase-3 immunocytochemistry, and DNA fragmentation assay. Cell cycle was analyzed by flow cytometry and the cell cycle-related proteins were analyzed by quantitative PCR and Western blot. The cells exposed to pfaffosidic fraction had reduced viability and cellular growth, induced G2/M at 48 h or S at 72 h arrest, and increased sub-G1 cell population via cyclin E downregulation, p27KIP1 overexpression, and caspase-3-induced apoptosis, without affecting the DNA integrity. Antitumoral effects of pfaffosidic fraction from H. paniculata in HepG2 cells originated by multimechanisms of action might be associated with cell cycle arrest in the S phase, by CDK2 and cyclin E downregulation and p27KIP1 overexpression, besides induction of apoptosis through caspase-3 activation. [ABSTRACT FROM AUTHOR]

Objective: to estimate survival, mortality and cause of death among users or not of hydroxyurea with sickle cell disease. Method: cohort study with retrospective data collection, from 1980 to 2010 of patients receiving inpatient treatment in two Brazilian public hospitals. The survival probability was determined using the Kaplan-Meier estimator, survival calculations (SPSS version 10.0), comparison between survival curves, using the log rank method. The level of significance was p=0.05. Results: of 63 patients, 87% had sickle cell anemia, with 39 using hydroxyurea, with a mean time of use of the drug of 20.0±10.0 years and a mean dose of 17.37±5.4 to 20.94±7.2 mg/kg/day, raising the fetal hemoglobin. In the comparison between those using hydroxyurea and those not, the survival curve was greater among the users (p=0.014). A total of 10 deaths occurred, with a mean age of 28.1 years old, and with Acute Respiratory Failure as the main cause. Conclusion: the survival curve is greater among the users of hydroxyurea. The results indicate the importance of the nurse incorporating therapeutic advances of hydroxyurea in her care actions. [ABSTRACT FROM AUTHOR]

Delftia acidovorans is an opportunistic agent in several types of infections, both in immunocompromised and immune-competent individuals; its resistance to aminoglycosides and polymyxin, choice drugs for empirical treatment of Gram-negative infections, is remarkable. We report the antimicrobial susceptibility and the genetic relatedness of 24 D. acidovorans strains recovered from tracheal aspirates of 21 adult inpatients hospitalized in an intensive care unit at a Brazilian hospital, from 2012 to 2013. All of the isolates were recovered as pure cultures and in counts above 1,000,000 CFU/mL. None of them were susceptible to polymyxin B, amikacin, gentamicin, or tobramycin; quinolones and trimethoprim-sulfamethoxazole presented varied activities against the isolates, while β-lactam resistance was not detected. Four clusters were verified in pulsed-field gel electrophoresis analysis, and a major pulsotype comprised 10 strains. A possible, but undetermined common source, can be responsible for this strain dissemination, underscoring the need of reinforcing the adherence to disinfection and infection control standard techniques. [ABSTRACT FROM AUTHOR]

Onychophorans (velvet worms) use an adhesive, protein-based slime secretion for prey capture and defence. The glue-like slime is ejected via a pair of modified limbs and the sticky threads entangle the victim. In this study, we analysed the protein composition of slime in twelve species of Onychophora from different parts of the world, including two species of Peripatidae from Costa Rica and Brazil and ten species of Peripatopsidae from Australia, using sodium dodecyl sulphate polyacrylamide gel electrophoresis. Our results revealed high intraspecific conservation in protein composition of slime in each species studied. In contrast, the protein profiles differ considerably in both number and position of bands between the species. We observed the highest number of differences (in 20 of 33 considered band positions) between a peripatid and a peripatopsid species, whereas the lowest number of differences (in four band positions) occurs between two closely related egg-laying species. The reconstructed maximum parsimony cladogram based on the electrophoretic characters largely reflects the phylogenetic relationships of the species studied, suggesting that the slime protein profiles contain useful phylogenetic information. Based on our findings, we suggest that the slime protein profiling is a valuable, non-invasive method for identifying the onychophoran species. Moreover, this method might help to discover potentially new species of Onychophora, given that the ~200 described species most likely underrepresent the actual diversity of the group. [ABSTRACT FROM AUTHOR]

Contents This study describes the reproductive parameters of Morada Nova rams, a breed of hair sheep from Brazil and with unique adaption to tropical environments. At 42 weeks of age, 15 rams were subjected to semen collection and, 1 week later, animals were slaughtered for collection of testes, epididymis and accessory sex glands. We conducted 2-D electrophoresis of seminal plasma proteins and major spots of stained gels were identified by LC- MS/ MS. Total RNA was isolated from testis, epididymis and vesicular glands and subjected to qPCR. At slaughter, scrotal circumference and testicular weight were 27.5 ± 0.5 cm and 109.5 ± 6.0 g, respectively. Seminiferous tubule (ST) diameter was 188.3 ± 4.0 μm and each testis contained 1.9 ± 0.1 Sertoli cells (×109). Each Sertoli cell supported 0.1 ± 0.01 A spermatogonia, 3.0 ± 0.2 pachytene spermatocytes and 7.7 ± 0.5 round spermatids/tubule cross section. Daily sperm production reached 5.6 × 106 cells/g of testis parenchyma. Testis size appeared as indicative of ST diameter and associated with epididymal measurements, as well as with the population of round spermatids and Sertoli cells/testis. Rams with heavier testes had greater daily sperm production and more Sertoli cells/testis. We detected 90.9 ± 9.6 spots per 2-D gel of seminal plasma. Major seminal proteins were identified as ram seminal vesicle proteins at 14 and 22 kDa, representing 16.2% and 12.8% of the total intensity of valid spots in the gels, respectively. Expression of both genes was greater in the vesicular glands as compared to testis and epididymis. Pixel intensity for those proteins in the 2-D gels was significantly correlated with seminal vesicle weight. This is the first description of the basic reproductive aspects of Morada Nova rams, including protein profiles of their seminal plasma. These findings will allow a better understanding of their reproductive physiology. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented regarding the molecular and clinical characterization of hemoglobin Maputo in Brazilian newborn screening.

Abstract: The natural Mg-vermiculite (VB) from Brazil and its Na-form (Na-VB) was studied after milling in ball, jet and vibrational mills. The morphology of vermiculite samples was studied using scanning electron microscopy (SEM) and the changes of vermiculite structures were monitored using X-ray diffraction (XRD) analysis. The particle size (PS) changes were characterized by the median particle size (d 50), volume-weighted mean diameter (d 43), mode diameter (d m) and span value. The surface changes of vermiculite samples were evaluated using ξ-potential measurements as a function of particle electrophoretic mobility (μ). The electrophoretic mobility (μ) of the VB and its milled samples was in the range of 1.16 to 1.96×10−4 m2/Vs and of the Na-VB and its milled samples were in range of 3.44 to 4.25×10−4 m2/Vs. The range of the difference in the ξ-potential of the VB was from −15.1 to −25.4mV and Na-VB was from −55mV to the −44.5mV. Samples containing small particles and homogeneous particle size distribution showed a low value of ξ-potential. [Copyright &y& Elsevier]

AbstractCynara scolymusL. (Asteraceae), popularly known as artichoke, has been widely used in herbal medicine for the treatment of hepatic diseases. The genotoxicity of C. scolymusL. leaf extract (LE) and the ability to modulate the genetic toxicity of the alkylating agent ethyl methanesulfonate (EMS) were assessed using the comet assay on Chinese hamster ovary cells. Genotoxicity was evaluated after 1- and 24-h treatments using four different LE concentrations: 0.62, 1.25, 2.5, and 5.0 mg/mL. Antigenotoxicity was assessed for pretreatment, simultaneous treatment, and post-treatment. All doses used led to a significant increase in the frequency of DNA damage, after exposure for 1 and 24 h. In the antigenotoxicity experiments, LE reduced the frequency of DNA damage induced by EMS in the simultaneous treatment only. However, the lowest dose was more protective than higher concentrations. Flavonoids and phenolic compounds are, probably, the C. scolymusconstituents responsible for its genotoxic and antigenotoxic effects. [ABSTRACT FROM AUTHOR]

Abstract: Objective: The aim of the present study was to investigate the protein profile of children with different levels of protein–energy undernutrition (PEU) through a proteomic approach of human whole saliva. Methods: Initially, saliva samples of children with mild, moderate, and severe PEU were collected and lyophilized. Saliva samples of healthy children were used as controls. Samples were analyzed for total protein using the Bradford method. Saliva samples were analyzed by two-dimensional electrophoresis according to their isoelectric point (pI) and their molecular weights (MWs). Results: Comparisons of protein bands among the healthy and mildly, moderately, and severely undernourished children showed significant differences in the MWs (P = 0.001) and pI values (P = 0.03). In total 159 spots were identified in the healthy children; 156, 168, and 221 spots were observed in mildly, moderately, and severely undernourished children, respectively. Mildly undernourished children presented with the spot with the highest MW of 293 kDa (pI = 7.77) and the lowest MW of 5 kDa (pI = 4.83). Moderately undernourished children were the only ones who did not present with a protein band with an MW of 30 kDa. The presence of a protein band with an MW of 123 kDa (pI = 516), possibly a cyclin-dependent protein kinase, was also observed only in this group. Conclusion: The protein profile in saliva varies according to the presence or absence of PEU, and these variations are specifically expressed in different grades of undernutrition. Thus, saliva may be an important diagnostic tool for the assessment of PEU. [Copyright &y& Elsevier]

The aim of this study was to investigate genetic polymorphism in Leishmania braziliensis population previously typed through isoenzyme electrophoresis, isolated from the same patient in two different moments: (A) before the beginning of treatment and (B) after treatment failure to meglumine antimoniate or reactivation after successful initial treatment. Fifteen pairs of isolates were assessed using the polymorphic molecular marker LSSP-PCR and following the phenetic analysis. The genetic profiles of the 30 samples were grouped in four clusters. Only two patients presented total identity in the A and B isolates. Most isolates presented similarity coefficients varying from 0.63 to 0.91. In this group of patients genetic polymorphisms could be observed indicating low similarity between the pairs of isolates. The results demonstrate the existence of genetic polymorphism between the samples isolated before treatment and after reactivation or treatment failure, suggesting a possible differentiation of the structure of the original parasite population which could be involved in the mechanisms of resistance to treatment or reactivation of lesions in the ATL. This phenomenon is important, although other factors also could be involved in this context and are discussed in this paper [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to investigate the naturally occurring bacteria from Culicidae larvae found in São Paulo state between 2006 and 2008 from the Tiete Ecological Park in the municipality of Caraguatatuba. Bacterial strains were obtained after surface sterilization of larvae followed by thermal treatment and incubation in bacteriological media under laboratory conditions. Identification was determined using cytomorphology, biochemical and physiological tests. Strains were characterized by qualitative evaluation of biological activity against Culicidae larvae, comparing protein profiles obtained by electrophoresis of crystal protoxins, electrophoresis of Lysinibacillus sphaericus isoenzymes, detection of the L2 fraction of Hemolytic BL enterotoxin and amplification of DNA using Multiplex-PCR to detect HBL, NHE, CytK, BceT and EntFM enterotoxins. The results obtained in this study demonstrated that the endosymbiotic bacterial diversity belonging to Bacillus and related genera associated with Culicidae larvae is limited to a number of species that does not vary throughout the seasons. [Copyright &y& Elsevier]

The Rodrigo de Freitas lagoon (RFL) is a tropical eutrophic coastal ecosystem located in the urban area of Rio de Janeiro, Brazil. This environment consists of freshwater but has communication with the ocean through a channel (Jardim de Alah's Channel). The aim of this study was to evaluate the influence of lagoon water on the nearby ocean using molecular and traditional microbiological methods. We hypothesised that due to the eutrophic low-salinity environment, the bacterioplankton community from the RFL would have a native 'brackish' composition influenced by both freshwater and marine phylotypes, and that bacterial phylotypes of this community would be detected in oceanic samples closer to the channel between the lagoon and the ocean. The cultivation and microscopy experiments clearly showed this influence. Bacterial cell counts revealed that the greater amounts of bacterial cells present in the lagoon increased the observed values seen at oceanic stations near the channel. The Denaturing gradient gel eletrophoresis community profiles also showed a clear influence of Rodrigo de Freitas lagoon waters on the adjacent beaches. The band patterns found for the stations near the channel showed that these communities were mixtures of the communities of the lagoon and sea, and as the distance from the channel increased, the samples became more similar to ocean bacterial communities. A 16S rRNA gene clone library was constructed using a sample acquired from the connection point between the lagoon and the ocean. Around 52% of the sequences in the library showed similarity to the genus Proteobacteria (1% Alpha, 21% Beta, 19% Gamma and 29% unclassified Proteobacteria), and the second most abundant genus was Bacteroidetes, with 15% of the total clones. The results showed that the structure of the bacterial community had both freshwater and marine characteristics. [ABSTRACT FROM AUTHOR]

Pseudorabies is a disease caused by Suid herpesvirus I (PrV) and is responsible for considerable economic losses in the swine industry. The PrV has only one serotype, but based on RFLP (restriction fragment length polymorphism) the virus was divided into four genotypes named I, II, III, IV. The classical methods for PrV genotyping usually require virus isolation. DNA purification enzyme restriction analysis and a long electrophoresis. The aim of this research was to describe a faster and more sensitive method to detect and genotype PrV based on nested-PCR and restriction enzyme analysis. Twenty PrV isolates from south and southeast regions of Brazil, and the standard strain Shope were grown in PK-15 cells and submitted to PCR for glycoprotein E gene amplification. Additionally were tested 75 clinical samples (swine brain), with 25 positives for virus isolation and seroneutralization, and 50 negatives from a flock free PR with negative results in seroneutralization test. There was 100% of agreement between results of nested-PCR and virus isolation and seroneutralization and all samples detected were classified as genotype II. The nested-PCR, combined with restriction enzyme analysis, was able to detect and genotype PrV in 1-2 days with a sensitivity of 10-1,3 TCID50 mL-1. It was faster than classical methods described in the literature that require at least 7 days to be completed. [ABSTRACT FROM AUTHOR]

Brazilian meat regulations allow the use of nonmeat extenders in certain meat products. Thus, the need for controlling the use of these ingredients with proper analytical methods is evident, in order to avoid illegal addition. In the present research, SDS-PAGE electrophoresis has been applied to pork enhanced with 1.5% of soy protein isolate and whey protein concentrate, through two different methods of protein extraction (urea 6M and Tris-HCl-SDS-mercaptoethanol), in the presence of positive and negative controls. Electrophoresis has revealed typical profiles, for whole-muscle and non-meat proteins, showing to be adequate to detect the addition of whey proteins in meat products. However, the soy proteins could not be detected, due to the overlapping of bands with meat proteins. [ABSTRACT FROM AUTHOR]

Abstract: Since the first report of the genus Leishmania, many species have been described. In Brazil, human leishmaniasis has been associated with eight Leishmania species, of which seven are responsible for cutaneous leishmaniasis (CL). In some endemic areas, CL is associated with only one species; however, in other areas, such as the Amazon, the etiology of CL can be assigned to many species. The multitude of highly similar Leishmania species in Brazil makes it difficult to develop an appropriate method of typing them. Most Leishmania species were first described based on epidemiological and biological characteristics, and these were later corroborated by Multilocus Enzyme Electrophoresis (MLEE), the gold standard technique for identifying Leishmania species. In an attempt to overcome the limitations of MLEE, many PCR-based methods have been developed and used for parasite identification. In the present study, we analyzed the sequence of the hsp70 gene in Leishmania species associated with human leishmaniasis in Brazil. This analysis led to the identification of restriction enzymes that could be used for PCR-RFLP-based identification. The results obtained were in complete agreement with those obtained by MLEE, suggesting that PCR-RFLP analysis of hsp70 could soon replace MLEE for routine Leishmania typing. [Copyright &y& Elsevier]

Aims: The genetic diversity of Beauveria bassiana was investigated by comparing isolates of this species to each other (49 from different geographical regions of Brazil and 4 from USA) and to other Beauveria spp. Methods and Results: The isolates were examined by multilocus enzyme electrophoresis (MLEE), amplified fragment length polymorphism (AFLP), and rDNA sequencing. MLEE and AFLP revealed considerable genetic variability among B. bassiana isolates. Several isolates from South and Southeast Brazil had high similarity coefficients, providing evidence of at least one population with clonal structure. There were clear genomic differences between most Brazilian and USA B. bassiana isolates. A Mantel test using data generated by AFLP provided evidence that greater geographical distances were associated with higher genetic distances. AFLP and rDNA sequencing demonstrated notable genotypic variation between B. bassiana and other Beauveria spp. Conclusion: Geographical distance between populations apparently is an important factor influencing genotypic variability among B. bassiana populations in Brazil. Significance and Impact of the Study: This study characterized many B. bassiana isolates. The results indicate that certain Brazilian isolates are considerably different from others and possibly should be regarded as separate species from B. bassiana sensu latu. The information on genetic variation among the Brazilian isolates, therefore, will be important to comprehending the population structure of B. bassiana in Brazil. [ABSTRACT FROM AUTHOR]

The aim of the present work is to identify the presence of Helicobacter pylori bacterium in samples of gastric mucosa fragments, obtained by gastric biopsy, from Brazilian patients with peptic ulcer and chronic gastritis and also to determine differences among the prevalent strains in these two diseases by urease C and urease B genes amplification utilizing nested polymerase chain reaction (PCR) and PCR. We encountered 17 genotyping patterns for urease C and 7 for urease B and, although no significant differences were found among the patterns encountered for both diseases, we found predominant groups for each disease. Typing methods of the products obtained by nested PCR and PCR show a functional scheme and are of great importance for epidemiologic studies and H. pylori strain characterization, in addition to allowing correlation among the several strains and their role in the diseases caused by this microorganism. [ABSTRACT FROM AUTHOR]

Aims: Sheep are important carriers of Shiga toxin-producing Escherichia coli (STEC) in several countries. However, there are a few reports about ovine STEC in American continent. Methods and Results: About 86 E. coli strains previously isolated from 172 healthy sheep from different farms were studied. PCR was used for detection of stx1, stx2, eae, ehxA and saa genes and for the identification of intimin subtypes. Restriction fragment length polymorphism (RFLP)–PCR was performed to investigate the variants of stx1 and stx2, and the flagellar antigen ( fliC) genes in nonmotile isolates. Five isolates were eae+ and stx−, and belonged to serotypes O128:H2/β-intimin (2), O145:H2/γ, O153:H7/β and O178:H7/ε. Eighty-one STEC isolates were recovered, and the stx genotypes identified were stx1c stx2d-O118 (46·9%), stx1c (27·2%), stx2d-O118 (23·4%), and stx1c stx2dOX3a (2·5%). Pulsed-field gel electrophoresis (PFGE) revealed 27 profiles among 53 STEC and atypical enteropathogenic Escherichia coli (EPEC) isolates. Conclusions: This study demonstrated that healthy sheep in São Paulo, Brazil, can be carriers of potential human pathogenic STEC and atypical EPEC. Significance and Impact of the Study: As some of the STEC serotypes presently found have been involved with haemolytic uraemic syndrome (HUS) in other countries, the important role of sheep as sources of STEC infection in our settings should not be disregarded. [ABSTRACT FROM AUTHOR]

Cold is one of the main problems for rice cultivation in Rio Grande do Sul because the vast majority of cultivars in use are of tropical origin. The occurrence of low temperatures combined with the susceptibility of the materials used can cause serious damage to the establishment of the crop reducing the plant stand. The objective of this study was to evaluate the physiological changes in different rice cultivars, when exposed to low temperature during germination. Seeds from four cultivars, BRS Agrisul, BRS Chuí, Lemont and Oro were used and evaluated regarding low temperature tolerance by the cold test. To evaluate the physiological changes, isoenzyme analyses of alcohol dehydrogenase and esterase were performed at temperatures of 25 °C (ideal for germination) and 13 °C (low temperature). The other characteristics evaluated were germination, germination speed, coleoptile length and dry weight at 25 °C and 13 °C. According to cold test results, seeds of the Lemont and Oro cultivars presented better tolerance to the low temperature. In the isoenzyme analysis, the cold caused an increase in the expression of the isoenzyme alcohol dehydrogenase and reduction in the esterase isoenzyme expression. For the other ratings, the cold caused damage to all the characteristics evaluated, especially in the BRS Agrisul and BRS Chuí cultivars. [ABSTRACT FROM AUTHOR]

Microorganisms can account for up to 60% of the fresh weight of marine sponges. Marine sponges have been hypothesized to serve as accumulation spots of particular microbial communities, but it is unknown to what extent these communities are directed by the organism or the site or occur randomly. To address this question, we assessed the composition of specific bacterial communities associated with Aplysina fulva, one of the prevalent sponge species inhabiting Brazilian waters. Specimens of A. fulva and surrounding seawater were collected in triplicate in shallow water at two sites, Caboclo Island and Tartaruga beach, Büzios, Brazil. Total community DNA was extracted from the samples using "direct" and "indirect" approaches. 16S rRNA-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analyses of the total bacterial community and of specific bacterial groups--Pseudomonas and Actinobacteria--revealed that the structure of these assemblages in A. fulva differed drastically from that observed in seawater. The DNA extraction methodology and sampling site were determinative for the composition of actinobacterial communities in A. fulva. However, no such effects could be gleaned from total bacterial and Pseudomonas PCR-DGGE profiles. Bacterial 16S rRNA gene clone libraries constructed from directly and indirectly extracted DNA did not differ significantly with respect to diversity and composition. Altogether, the libraries encompassed 15 bacterial phyla and the candidate division TM7. Clone sequences affiliated with the Cyanobacteria, Chloroflexi, Gamma- and Alphaproteobacteria, Actinobacteria, Bacteroidetes, and Acidobacteria were, in this order, most abundant. The bacterial communities associated with the A. fulva specimens were distinct and differed from those described in studies of sponge-associated microbiota performed with other sponge species. [ABSTRACT FROM AUTHOR]

Aims: Leptospirosis is a public health problem worldwide. Traditionally, microscopic agglutination test (MAT) and cross-agglutinin absorption test (CAAT) are used to identify leptospires. However, these techniques are laborious and time-consuming, requiring the maintenance of a collection of more than 200 reference strains and correspondent rabbit antisera. The purpose of this study was to evaluate the pulsed-field gel electrophoresis (PFGE) method for discrimination of Leptospira serovars. Methods and Results: Fourteen clinical isolates of Leptospira spp. were analysed by MAT before being characterized by PFGE. The isolates were compared with a library of 206 different reference Leptospira serovars. All the isolates gave clear profiles with high resolution. PFGE and MAT results were in agreement for all clinical isolates evaluated. Twelve isolates were classified as serovar Icterohaemorrhagiae/Copenhageni by PFGE. By MAT, these isolates were classified as serogroup Icterohaemorrhagiae with titres ranging from 3200 to 25 600. Two isolates were classified as serovar Canicola by PFGE, and as serogroup Canicola by MAT with titres higher than 3200. Conclusions: PFGE offers the advantages of simple, reliable and reproducible results. Significance and Impact of the Study: PFGE provides a convenient tool for the identification of clinical isolates. [ABSTRACT FROM AUTHOR]

Abstract: The theraphosid spider genus Vitalius contains several species found in southeastern Brazil. In this work, we used electrostimulation to obtain venom from Vitalius dubius and examined its general composition. Male spiders yielded significantly less (p < 0.05) venom (12.5 ± 0.7 mg of liquid/spider, n = 16; mean ± S.E.M.) than female spiders (25.5 ± 2.0 mg of liquid/spider, n = 11). However, when corrected for spider weight, males yielded slightly more venom (2.89 ± 0.16 mg/g vs. 2.45 ± 0.76 mg/g for males and females, respectively, p < 0.05). Venom yield correlated linearly with spider weight for spiders weighing up to ∼12–13 g, but decreased in very heavy females. There was a marked decrease in venom yield after the first milking. The protein concentration of pooled venom was 18.3 ± 2.4 mg/ml (n = 4) and accounted for 16.6 ± 4.7% of the dry venom weight. The venom contained high hyaluronidase activity (275 ± 24 TRU/mg of protein, n = 4), with a molecular mass of ∼45 kDa estimated by zymography. SDS-PAGE revealed a few proteins with molecular masses >14 kDa but showed two staining bands of peptides <14 kDa. The venom reacted in ELISA with affinity-purified IgG from commercial arachnidic antivenom. Immunoblotting with this IgG detected proteins of 30–140 kDa only. Fractionation of the venom by reverse-phase chromatography resulted in five major and eight minor peaks. [Copyright &y& Elsevier]

Fifty-one vancomycin-resistant enterococci samples isolated from different geographic regions in Brazil were studied. All the isolates harboured the vanA gene as demonstrated by PCR analysis, and in a majority of strains the gene was associated with a transferable plasmid of 70 kb. A single variant of the prototype Tn 1546 associated with common transferable vanA-containing plasmids has spread among the enterococcal strains circulating in Brazil. The VanA element integrity in these enterococci strains and the different pulsed-field gel electrophoresis patterns suggest horizontal transmission of the vancomycin resistance transposon in Brazilian strains. [ABSTRACT FROM AUTHOR]

Pelagic larvae are highly important for maintaining the gene flow among populations of sessile marine invertebrates. Colonial ascidians consist, exclusively, of brooding species, with lecithotrophic larvae that have a limited dispersal. As a result, there is a marked differentiation among populations. In this work, we used allozyme electrophoresis to access the genetic variation in four populations of Symplegma rubra, a colonial ascidian frequently found in the intertidal zone of Southeastern Brazilian coast. High variability was found at three of the four sites sampled, the exception being Praia Grande in the State of Rio de Janeiro. At this site, there was a great preponderance of clones, which possibly reflected the enclosed nature of the location and its low water circulation that reduce the dispersal capabilities of these animals. S. rubra did not conform to expectations for random mating (Hardy–Weinberg equilibrium): there was a deficit of heterozygotes that was more related to the small population size than to inbreeding processes, since F is analysis per locus revealed a deficiency of heterozygotes at only one locus— MDH*. The greatest variation in allele frequency was found for GPI-2*. Analyses of genetic variability revealed moderate differentiation among the populations ( F ST=0.051), which was unexpected for a species with a low dispersal capability. Rafting, a frequently underestimated means of dispersal, may be the main mode of gene flow in this species over large areas, since colonies of S. rubra are frequently seen growing on drift material and there is no evidence that the larva survive for a long time in the plankton. [ABSTRACT FROM AUTHOR]

Fifty fecal samples from calves with diarrhea, positive for group A rotavirus by enzyme immunoassay (EIA) and polyacrylamide gel electrophoresis (PAGE) were analyzed by a nested multiplex reverse transcription/polymerase chain reaction (Nested multiplex/RT-PCR) for identification of P and G genotypes. Samples were collected between 1996 and 1999 from eight dairy and/or beef cattle herds located in the Mato Grosso do Sul, Sa˜o Paulo, Goia´s and Parana´ States, Brazil. Complete genotyping was possible in 44 (88%) of the calf fecal samples. The VP7 gene could not be identified in six (12%) of the samples. Whilst the VP4 (P) gene was identified in 100% of samples. The genotypes of 35 (70%) samples corresponded to the NCDV-like (12%), UK-like (40%), B223-like (16%) and KN-4 (2%) reference rotavirus strains, which are commonly found in cattle. Mixed infections were detected in seven (14%) samples, and genotypes observed in two (4%) samples presented unusual combinations, carrying VP4 and VP7 gene of the bovine group A rotavirus (P[11],G8), in other one carrying a VP4 gene of bovine origin (P[1]) and a VP7 gene of swine origin (G5). [Copyright &y& Elsevier]

Severe epidemics of bacterial spot have been observed in central-west Brazil in fields of processing tomato. Several xanthomonads, Xanthomonas axonopodis pv. vesicatoria, X. vesicatoria, or X. gardneri, can cause the disease; therefore, attempts were made to identify the pathogen species present in this region. A total of 215 strains were obtained from 10 commercial areas in 1997, 1998, and 2000. The strains were characterized using pulsed-field gel electrophoresis (PFGE) and by their amylolytic and pectolytic activities. Representative strains from each PFGE haplotype then were tested for pathogenicity on tomato and pepper, carbon source utilization, and whole protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis, rRNA sequence comparisons also were performed. All strains recovered from six fields were classified as X. gardneri, whereas X. vesicatoria and X. axonopodis pv. vesicatoria also were detected in the remaining four fields. Strains of X. gardneri, which could be grouped into two PFGE haplotypes, were unable to hydrolyze starch and pectate and to utilize gentiobiose and maltose. They expressed the β protein of 27 kDa and were pathogenic on tomato but variable on pepper. This is the first report of outbreaks of bacterial spot on tomato caused by X. gardneri. [ABSTRACT FROM AUTHOR]

ABSTRACT Recent dolomitic sediment samples from Lagoa Vermelha, Brazil, were examined microscopically to study the process of bacterial fossilization in carbonate sediments. Bacteria-like bodies were intimately associated with carbonate mineral surfaces, and coatings on the former demonstrate the calcification of single bacterial cells. The bacterial fossilization process in Lagoa Vermelha sediments was simulated in the laboratory by cultivation of mixed and pure cultures of sulphate-reducing bacteria, which were isolated from the Lagoa Vermelha sediments. These cultures produced carbonate minerals that were studied to provide insight into the initiation of the fossilization process. In mixed culture experiments, bacterial colonies became calcified, whereas in pure culture experiments, single bacterial cells were associated with dolomite surfaces. Dolomite nucleated exclusively in bacterial colonies, intimately associated with extracellular organic matter and bacterial cells. Electrophoretic mobility measurements of the bacterial cells in electrolyte solutions demonstrated the specific adsorption of Ca2+ and Mg2+ onto the cell surfaces, indicating the role of the bacterial surface in carbonate nucleation and bacterial fossilization. The affinity of the cells for Mg2+ was related to the capability of the strains to mediate dolomite formation. Combined with sulphate uptake, which dissociates the [MgSO4 ]0 ion pair and increases the Mg2+ availability, the concentration of Mg2+ ions in the microenvironment around the cells, where the conditions are favourable for dolomite precipitation, may be the key to overcome the kinetic barrier to dolomite formation. These results demonstrate that bacterial fossilization is a consequence of the cell surface involvement in carbonate precipitation, implying that fossilized bacterial bodies can be used as a tool to recognize microbially mediated carbonates. [ABSTRACT FROM AUTHOR]

Thirty-one trypanosotomatid stocks were isolated from various sylvatic hosts and vectors in two different regions from the Parana´ State of Brazil. The stocks were analyzed by multilocus enzyme electrophoresis (MLEE) on cellulose acetate plates (22 genetic loci). All stocks were unambiguously attributed to Trypanosoma cruzi, and were found to be closely related to the formerly described zymodeme I (TC1 subgroup of T. cruzi). By comparison with other sylvatic cycles with similar sample sizes (Southern USA, Colombia, French Guiana), genetic variability among these stocks was very limited, with only two variable loci out of 22, and only three different multilocus genotypes. Population structure of T. cruzi in these cycles appears to correspond to a set of very closely related clonal genotypes. This very limited genetic variability could be due to a recent foundation of these populations (founder effect). The implications of an active Chagas sylvatic cycle in this area are discussed.Index Descriptors and Abbreviations: Trypanosoma cruzi; Chagas disease sylvatic cycle; Didelphis spp.; Panstrongylus megistus; Parana´ State, Brazil; MLEE (multilocus enzyme electrophoresis); Isoenzymes [Copyright &y& Elsevier]

Presents a study which examined the sympatric and allopatric populations of Crassostrea species occurring in the Brazilian coast through allozyme electrophoresis. Levels of genetic variation and population structure of C. brasiliana and C. rhizophorae; Methodology; Results and discussion.

Background: H19 is a strong candidate gene for influencing birth weight variation and is exclusively imprinted maternally. In an attempt to understand the relationship of this gene polymorphism with low birth weight children, we investigated association of H19/RsaI polymorphism with low birth weight and normal birth weight in children and their mothers. Objectives: The aim of our study was to establish the association between H19 gene polymorphism and LW in children born in Pernambuco, state of Brazil. Patients and Methods: It were selected 89 children, 40 low birth weight (LW) and 49 normal birth weight (NW) and 71 mothers (40 mothers of newborns NW and 31 mothers of newborns LW) attended at Dom Malan Hospital, Petrolina, Pernambuco - Brazil. Peripheral blood samples were collected from patients and genomic DNA was extracted and detected by electrophoresis agarose gel, stained by Blue Green Loading Dye. DNA PCR amplification was done using the primers H1 (sense) and H3 (antisense). PCR products were digested with RsaI and electrophoresed on agarose gel stained by ethidium bromide. Statistical analyses were performed using the program BioEstat version 5.0. Results: The RsaI polymorphism in the H19 gene showed that genotype frequencies did not differ statistically between low birth weight (AA = 12.5%, AB = 45%, BB = 42.5%) and control (AA = 8.6% AB = 36.73%, BB= 55.10% groups) and the allele frequencies were not significantly different (P = 0.2897). We also did not observe any association between maternal H19 allele polymorphism and low birth weight newborns (P =0.7799) or normal birth weight children (P = 0.8976). Conclusions: The small size of sample may be the explanation for these results; future studies with more patients are needed to confirm the effect of H19/RsaI polymorphism on birth weight of LW newborns. [ABSTRACT FROM AUTHOR]

Examines the genetic differentiation of the millipede Pycnotropis epiclysmus inhabiting in Amazonian forests in Manaus, Brazil. Analysis of the pPeP172 satellite DNA family identified in P. epicylsmus; Investigation of the enzyme variability of the populations; Use of cellulose acetate electrophoresis to examine the enzyme loci.

Electrophoretic surveys of 7794 individuals from different regions of Brazil and a study of subjects with hemolytic anemia have disclosed 9 putative G6PD variants in addition to the B, A. A-, and Mediterranean types. No variants were found among the 3739 Brazilian Indians tested. Four variants underwent DNA analysis. Three were identified with the Mediterranean, Seattle. and Anaheim types. but the fourth variant was previously undescribed and we propose to designate it G6PD São Borja337A. [ABSTRACT FROM AUTHOR]

Explores the allele distribution on the basis of eight short tandem repeats (STR) loci for direct descendants from Terena population in Brazil. Classifications of the Terenas; Extraction of DNA from buccal swab samples by organic extraction methods and ethanol precipitation; Performance of DNA quantification of each sample using the GeneQuant pro RNA/DNA/calculator; Details of the electrophoretical separation of STR-polymerase chain reaction products; Detection of the fluorescent signals using FMBIO II Fluorescent Scanner; Calculation of the relative allele frequencies and the observed heterozygosity rates for each locus.