Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers.

Paracoccidioidomycosis (PCM) is a life-threatening systemic fungal infection acquired after inhalation of Paracoccidioides propagules from the environment. The main agents include members of the P. brasiliensis complex (phylogenetically-defined species S1, PS2, PS3, and PS4) and P. lutzii. DNA-sequencing of protein-coding loci (e.g., GP43, ARF, and TUB1) is the reference method for recognizing Paracoccidioides species due to a lack of robust phenotypic markers. Thus, developing new molecular markers that are informative and cost-effective is key to providing quality information to explore genetic diversity within Paracoccidioides. We report using new amplified fragment length polymorphism (AFLP) markers and mating-type analysis for genotyping Paracoccidioides species. The bioinformatic analysis generated 144 in silico AFLP profiles, highlighting two discriminatory primer pairs combinations (#1 EcoRI-AC/MseI-CT and #2 EcoRI-AT/MseI-CT). The combinations #1 and #2 were used in vitro to genotype 165 Paracoccidioides isolates recovered from across a vast area of South America. Considering the overall scored AFLP markers in vitro (67-87 fragments), the values of polymorphism information content (PIC = 0.3345-0.3456), marker index (MI = 0.0018), effective multiplex ratio (E = 44.6788-60.3818), resolving power (Rp = 22.3152-34.3152), discriminating power (D = 0.5183-0.5553), expected heterozygosity (H = 0.4247-0.4443), and mean heterozygosity (H avp  = 0.00002-0.00004), demonstrated the utility of AFLP markers to speciate Paracoccidioides and to dissect both deep and fine-scale genetic structures. Analysis of molecular variance (AMOVA) revealed that the total genetic variance (65-66 %) was due to variability among P. brasiliensis complex and P. lutzii (PhiPT = 0.651-0.658, P < 0.0001), supporting a highly structured population. Heterothallism was the exclusive mating strategy, and the distributions of MAT1-1 or MAT1-2 idiomorphs were not significantly skewed (1:1 ratio) for P. brasiliensis s. str. (χ2 = 1.025; P = 0.3113), P. venezuelensis (χ2 = 0.692; P = 0.4054), and P. lutzii (χ2 = 0.027; P = 0.8694), supporting random mating within each species. In contrast, skewed distributions were found for P. americana (χ2 = 8.909; P = 0.0028) and P. restrepiensis (χ2 = 4.571; P = 0.0325) with a preponderance of MAT1-1. Geographical distributions confirmed that P. americana, P. restrepiensis, and P. lutzii are more widespread than previously thought. P. brasiliensis s. str. is by far the most widely occurring lineage in Latin America countries, occurring in all regions of Brazil. Our new DNA fingerprint assay proved to be rapid, reproducible, and highly discriminatory, to give insights into the taxonomy, ecology, and epidemiology of Paracoccidioides species, guiding disease-control strategies to mitigate PCM.

Trends in the molecular epidemiology and population genetics of emerging Sporothrix species.

Sporothrix (Ophiostomatales) comprises species that are pathogenic to humans and other mammals as well as environmental fungi. Developments in molecular phylogeny have changed our perceptions about the epidemiology, host-association, and virulence of Sporothrix. The classical agent of sporotrichosis, Sporothrix schenckii, now comprises several species nested in a clinical clade with S. brasiliensis, S. globosa, and S. luriei. To gain a more precise view of outbreaks dynamics, structure, and origin of genetic variation within and among populations of Sporothrix, we applied three sets of discriminatory AFLP markers (#3 EcoRI-GA/MseI-TT, #5 EcoRI-GA/MseI-AG, and #6 EcoRI-TA/MseI-AA) and mating-type analysis to a large collection of human, animal and environmental isolates spanning the major endemic areas. A total of 451 polymorphic loci were amplified in vitro from 188 samples, and revealed high polymorphism information content (PIC = 0.1765-0.2253), marker index (MI = 0.0001-0.0002), effective multiplex ratio (E = 15.1720-23.5591), resolving power (Rp = 26.1075-40.2795), discriminating power (D = 0.9766-0.9879), expected heterozygosity (H = 0.1957-0.2588), and mean heterozygosity (Havp  = 0.000007-0.000009), demonstrating the effectiveness of AFLP markers to speciate Sporothrix. Analysis using the program structure indicated three genetic clusters matching S. brasiliensis (population 1), S. schenckii (population 2), and S. globosa (population 3), with the presence of patterns of admixture amongst all populations. AMOVA revealed highly structured clusters (PhiPT = 0.458-0.484, P < 0.0001), with roughly equivalent genetic variability within (46-48 %) and between (52-54 %) populations. Heterothallism was the exclusive mating strategy, and the distributions of MAT1-1 or MAT1-2 idiomorphs were not significantly skewed (1:1 ratio) for S. schenckii (χ2 = 2.522; P = 0.1122), supporting random mating. In contrast, skewed distributions were found for S. globosa (χ2 = 9.529; P = 0.0020) with a predominance of MAT1-1 isolates, and regional differences were highlighted for S. brasiliensis with the overwhelming occurrence of MAT1-2 in Rio de Janeiro (χ2 = 14.222; P = 0.0002) and Pernambuco (χ2 = 7.364; P = 0.0067), in comparison to a higher prevalence of MAT1-1 in the Rio Grande do Sul (χ2 = 7.364; P = 0.0067). Epidemiological trends reveal the geographic expansion of cat-transmitted sporotrichosis due to S. brasiliensis via founder effect. These data support Rio de Janeiro as the centre of origin that has led to the spread of this disease to other regions in Brazil. Our ability to reconstruct the source, spread, and evolution of the ongoing outbreaks from molecular data provides high-quality information for decision-making aimed at mitigating the progression of the disease. Other uses include surveillance, rapid diagnosis, case connectivity, and guiding access to appropriate antifungal treatment.

The global epidemiology of emerging Histoplasma species in recent years.

Histoplasmosis is a serious infectious disease in humans caused by Histoplasma spp. (Onygenales), whose natural reservoirs are thought to be soil enriched with bird and bat guano. The true global burden of histoplasmosis is underestimated and frequently the pulmonary manifestations are misdiagnosed as tuberculosis. Molecular data on epidemiology of Histoplasma are still scarce, even though there is increasing recognition of histoplasmosis in recent years in areas distant from the traditional endemic regions in the Americas. We used multi-locus sequence data from protein coding loci (ADP-ribosylation factor, H antigen precursor, and delta-9 fatty acid desaturase), DNA barcoding (ITS1/2+5.8s), AFLP markers and mating type analysis to determine the genetic diversity, population structure and recognise the existence of different phylogenetic species among 436 isolates of Histoplasma obtained globally. Our study describes new phylogenetic species and the molecular characteristics of Histoplasma lineages causing outbreaks with a high number of severe outcomes in Northeast Brazil between 2011 and 2015. Genetic diversity levels provide evidence for recombination, common ancestry and clustering of Brazilian isolates at different geographic scales with the emergence of LAm C, a new genotype assigned to a separate population cluster in Northeast Brazil that exhibited low diversity indicative of isolation. The global survey revealed that the high genetic variability among Brazilian isolates along with the presence of divergent cryptic species and/or genotypes may support the hypothesis of Brazil being the center of dispersion of Histoplasma in South America, possibly with the contribution of migratory hosts such as birds and bats. Outside Brazil, the predominant species depends on the region. We confirm that histoplasmosis has significantly broadened its area of occurrence, an important feature of emerging pathogens. From a practical point of view, our data point to the emergence of histoplasmosis caused by a plethora of genotypes, and will enable epidemiological analysis focused on understanding the processes that lead to histoplasmosis. Further, the description of this diversity opens avenues for comparative genomic studies, which will allow progress toward a consensus taxonomy, improve understanding of the presence of hybrids in natural populations of medically relevant fungi, test reproductive barriers and to explore the significance of this variation.

Saccharomyces cerevisiae as a probiotic agent and a possible aflatoxin B1 adsorbent in simulated fish intestinal tract conditions / Saccharomyces cerevisiae como agente probiótico e possível adsorvente de aflatoxina B1 em condições simuladas do trato intestinal de peixes

The aim of this study was to evaluate in vitro the probiotic potential and absorption of Saccharomyces cerevisiae for the aflatoxin B1 in simulated fish intestinal tract conditions. Three yeast strains were used, two from brewery: S. cerevisiae RC1 and S. cerevisiae RC3 and one from a fish farming environment: S. cerevisiae A8L2. The selected yeasts were subjected to the following in vitro tests: homologous inhibition, self-aggregation, co-aggregation, antibacterial activity, gastrointestinal conditions tolerance and adsorption of AFB1. All S. cerevisiae strains showed good capability of self-aggregation and co-aggregation with pathogenic bacteria. All yeast strains were able to survive the gastrointestinal conditions. In acidic conditions, the factors (strain vs. time) had interaction (P=0.0317), resulting in significant variation among the strains tested in the time periods analyzed. It was observed that there was also interaction (P=0.0062) in intestinal conditions, with an increased number of cells in the 12-hour period for all strains tested. In the adsorption test, the A8L2 strain was statistically more effective (P<0.005) for both AFB1 concentrations evaluated in this study (10 and 25ng/mL). Thus, it was observed that the strains of S. cerevisiae have potential probiotic and adsorbent of AFB1.(AU)

Objetivou-se, com esta pesquisa, avaliar in vitro o potencial probiótico e adsorvente de Saccharomyces cerevisiae para aflatoxina B1 em condições simuladas do trato intestinal de peixes. Foram utilizadas três cepas de leveduras, sendo duas provenientes de cervejaria: S. cerevisiae RC1 e S. cerevisiae RC3, e uma de ambiente de piscicultura: S. cerevisiae A8L2. As leveduras selecionadas foram submetidas aos seguintes testes in vitro: inibição homóloga, autoagregação, coagregação, atividade antibacteriana, viabilidade às condições gastrointestinais e adsorção de AFB1. Todas as estirpes de S. cerevisiae mostraram boa capacidade de autoagregação e coagregação com bactérias patogênicas. Todas as estirpes de levedura foram capazes de sobreviver às condições gastrointestinais. Em condições ácidas, os fatores (cepa x tempo) tiveram interação (P=0,0317), resultando em variações significativas entre as cepas testadas nos períodos de tempo analisados. Observou-se que também houve interação (P=0,0062) em condições intestinais, havendo um aumento do número de células no período de 12h para todas as cepas avaliadas. No ensaio de adsorção, a estirpe A8L2 foi a mais eficaz estatisticamente (P<0,005), para as duas concentrações de AFB1 avaliadas neste estudo (10 e 25ng. mL-1). Dessa forma, conclui-se que as cepas de Saccharomyces cerevisiae possuem potencial probiótico e adsorvente de AFB1.(AU)

Saccharomyces cerevisiae as a probiotic agent and a possible aflatoxin B1 adsorbent in simulated fish intestinal tract conditions / Saccharomyces cerevisiae como agente probiótico e possível adsorvente de aflatoxina B1 em condições simuladas do trato intestinal de peixes

The aim of this study was to evaluate in vitro the probiotic potential and absorption of Saccharomyces cerevisiae for the aflatoxin B1 in simulated fish intestinal tract conditions. Three yeast strains were used, two from brewery: S. cerevisiae RC1 and S. cerevisiae RC3 and one from a fish farming environment: S. cerevisiae A8L2. The selected yeasts were subjected to the following in vitro tests: homologous inhibition, self-aggregation, co-aggregation, antibacterial activity, gastrointestinal conditions tolerance and adsorption of AFB1. All S. cerevisiae strains showed good capability of self-aggregation and co-aggregation with pathogenic bacteria. All yeast strains were able to survive the gastrointestinal conditions. In acidic conditions, the factors (strain vs. time) had interaction (P=0.0317), resulting in significant variation among the strains tested in the time periods analyzed. It was observed that there was also interaction (P=0.0062) in intestinal conditions, with an increased number of cells in the 12-hour period for all strains tested. In the adsorption test, the A8L2 strain was statistically more effective (P<0.005) for both AFB1 concentrations evaluated in this study (10 and 25ng/mL). Thus, it was observed that the strains of S. cerevisiae have potential probiotic and adsorbent of AFB1.(AU)

Objetivou-se, com esta pesquisa, avaliar in vitro o potencial probiótico e adsorvente de Saccharomyces cerevisiae para aflatoxina B1 em condições simuladas do trato intestinal de peixes. Foram utilizadas três cepas de leveduras, sendo duas provenientes de cervejaria: S. cerevisiae RC1 e S. cerevisiae RC3, e uma de ambiente de piscicultura: S. cerevisiae A8L2. As leveduras selecionadas foram submetidas aos seguintes testes in vitro: inibição homóloga, autoagregação, coagregação, atividade antibacteriana, viabilidade às condições gastrointestinais e adsorção de AFB1. Todas as estirpes de S. cerevisiae mostraram boa capacidade de autoagregação e coagregação com bactérias patogênicas. Todas as estirpes de levedura foram capazes de sobreviver às condições gastrointestinais. Em condições ácidas, os fatores (cepa x tempo) tiveram interação (P=0,0317), resultando em variações significativas entre as cepas testadas nos períodos de tempo analisados. Observou-se que também houve interação (P=0,0062) em condições intestinais, havendo um aumento do número de células no período de 12h para todas as cepas avaliadas. No ensaio de adsorção, a estirpe A8L2 foi a mais eficaz estatisticamente (P<0,005), para as duas concentrações de AFB1 avaliadas neste estudo (10 e 25ng. mL-1). Dessa forma, conclui-se que as cepas de Saccharomyces cerevisiae possuem potencial probiótico e adsorvente de AFB1.(AU)

P2X7 siRNA targeted to the kidneys increases klotho and delays the progression of experimental diabetic nephropathy.

Previous studies in our laboratory have suggested that P2X7 could contribute to the progression of diabetic nephropathy and modulated klotho expression. The aim of this study was to investigate if P2X7 receptor is related to the expression of klotho in the onset of diabetic nephropathy in rats. Seven-week-old male Wistar rats weighing 210 g were all uninephrectomized; two-third of the animals were induced to diabetes with 60 mg/kg streptozotocin i.v., and one-third received its vehicle (control rats). At 4th day of the fifth week of the protocol, half of the diabetic rats received a small interfering RNA targeting for P2X7 mRNA, and the other half received its vehicle. Euthanasia was made at the eighth week. Diabetic animals reproduced all classic symptoms of the disease; besides, they showed reduced renal function and low NO bioavailability; also, SOD1, SOD2, and catalase were increased, probably due to the oxidative stress which was elevated in this situation. Metabolic data of diabetic rats did not change by silencing P2X7 receptor. For the other hand, silencing P2X7 was able to contribute to balance oxidative and nitrosative profile, ultimately improving the renal function and increasing plasma and membrane forms of klotho. These findings suggest that the management of P2X7 receptor can benefit the kidneys with diabetic nephropathy. Further studies are needed to show the therapeutic potential of this receptor inhibition to provide a better quality of life for the diabetic patient.

Inhibition of the P2X7 receptor improves renal function via renin-angiotensin system and nitric oxide on diabetic nephropathy in rats.

AIM: To evaluate the effects of P2X7 receptor blockade on renin-angiotensin system (RAS) in rats with diabetic nephropathy (DN). MAIN METHODS: Wistar rats were unilaterally nephrectomized and received streptozotocin for diabetes mellitus (DM) induction; control animals (CTL) received the drug vehicle. The animals were submitted to P2X7 receptor silencing, forming the group (DM + siRNA). The animals were placed in metabolic cages for data collection and evaluation of renal function; at the end of the protocol, the kidney was removed for analysis of P2X7, renin, angiotensin-converting enzyme (ACE), ACE2, angiotensin, thiobarbituric acid reactive substance levels (TBARS), nitric oxide (NO) and qualitative histological. KEY FINDINGS: The metabolic profile was attenuated in DM + siRNA vs. DM and there was a significant improvement in creatinine, urea and proteinuria levels in the same group. Renin expression was significantly decreased in DM + siRNA vs. DM. ACE and ACE2 were significantly reduced in DM + siRNA vs. DM. TBARS levels were decreased and NO showed an increase in DM + siRNA vs. DM, both significant. All histological alterations were improved in DM + siRNA vs. DM. SIGNIFICANCE: Data have shown that although silencing of the P2X7 receptor did not decrease fasting glucose, it promoted an improvement in the metabolic profile and a significant recovery of renal function, revealing a protective action by the inhibition of this receptor. This effect must have occurred due to the inhibition of RAS and the increase of NO, suggesting that the use of P2X7 receptors inhibitors could be used as adjuvant therapy against DN progression.

CLA-supplemented diet accelerates experimental colorectal cancer by inducing TGF-ß-producing macrophages and T cells.

Conjugated linoleic acid (CLA) has been shown to activate the nuclear receptor PPAR-γ and modulate metabolic and immune functions. Despite the worldwide use of CLA dietary supplementation, strong scientific evidence for its proposed beneficial actions are missing. We found that CLA-supplemented diet reduced mucosal damage and inflammatory infiltrate in the dextran sodium sulfate (DSS)-induced colitis model. Conditional deletion of PPAR-γ in macrophages from mice supplemented with CLA diet resulted in loss of this protective effect of CLA, suggesting a PPAR-γ-dependent mechanism mediated by macrophages. However, CLA supplementation significantly worsened colorectal tumor formation induced by azoxymethane and DSS by inducing macrophage and T-cell-producing TGF-ß via PPAR-γ activation. Accordingly, either macrophage-specific deletion of PPAR-γ or in vivo neutralization of latency-associated peptide (LAP, a membrane-bound TGF-ß)-expressing cells abrogated the protumorigenic effect of CLA. Thus, the anti-inflammatory properties of CLA are associated with prevention of colitis but also with development of colorectal cancer.

P2X7 receptor and klotho expressions in diabetic nephropathy progression.

Diabetes mellitus is characterized by increased levels of reactive oxygen species (ROS), leading to high levels of adenosine triphosphate (ATP) and the activation of purinergic receptors (P2X7), which results in cell death. Klotho was recently described as a modulator of oxidative stress and as having anti-apoptotic properties, among others. However, the roles of P2X7 and klotho in the progression of diabetic nephropathy are still unclear. In this context, the aim of the present study was to characterize P2X7 and klotho in several stages of diabetes in rats. Diabetes was induced in Wistar rats by streptozotocin, while the control group rats received the drug vehicle. From the 1st to 8th weeks after the diabetes induction, the animals were placed in metabolic cages on the 1st day of each week for 24 h to analyze metabolic parameters and for the urine collection. Then, blood samples and the kidneys were collected for biochemical analysis, including Western blotting and qPCR for P2X7 and klotho. Diabetic rats presented a progressive loss of renal function, with reduced nitric oxide and increased lipid peroxidation. The P2X7 and klotho expressions were similar up to the 4th week; then, P2X7 expression increased in diabetes mellitus (DM), but klotho expression presented an opposite behavior, until the 8th week. Our data show an inverse correlation between P2X7 and klotho expressions through the development of DM, which suggests that the management of these molecules could be useful for controlling the progression of this disease and diabetic nephropathy.

Low validity of predictive equations for calculating resting energy expenditure in overweight and obese women with polycystic ovary syndrome.

BACKGROUND: Predictive equations are the main clinical tools for determining resting energy expenditure (REE). However, their adequate use in overweight and obese individuals is unclear. Thus, we investigated the best predictive equations for estimating REE in overweight and obese women with polycystic ovary syndrome (PCOS). METHODS: Eleven analyses were performed with prediction equations (pREE) based on anthropometric parameters in 30 overweight or obese women with PCOS without other chronic diseases. The measured REE (mREE) was calculated by indirect calorimetry. The validity of the equations was investigated by comparison, accuracy and agreement tests between pREE and mREE at both the individual and group level. RESULTS: Four analyses were similar to those of mREE, and smallest mean differences were observed for the World Health Organization/Food and Agriculture Organization of the United Nations/United Nations University (WHO/FAO/UNU) considering weight (W) [0.07 (1.13) MJ (16 [270] kcal)]. Individual accuracy was greater than 50% for Harris and Benedict, Müller and Lazzer equations. The percentage of REE underestimation ranged between 16.7% and 73.3%, whereas higher rates of overestimation were observed in the De Luis (66.7%) and Ireton-Jones (43.3%) equations. Mean bias at the group level was lowest in the WHO/FAO/UNU W and WHO/FAO/UNU considering weight and height (WH), Müller and Lazzer equations (-2.8 to 0.5). The WHO/FAO/UNU W and WHO/FAO/UNU WH formulas were optimal in individual agreement (33.3%). CONCLUSIONS: FAO/WHO/UNU W equations may estimate the REE in overweight and obese women with PCOS. However, the low individual accuracy and agreement in relation to mREE suggest caution regarding when to use the formula to perform an individual nutritional plan.

Moderate aerobic exercise on the recovery phase of gentamicin-induced acute kidney injury in rats.

INTRODUCTION: Acute kidney injury is a serious public health problem, especially in intensive care units, where patients may require dialysis support, resulting in 50% mortality. AIM: To evaluate the effects of moderate aerobic exercise on the recovery phase of acute kidney injury induced by gentamicin in rats. MAIN METHODS: Male adult Wistar rats were allocated into 4 groups: W10+R30, G10+R30, W10+EX30 and G10+EX30; W10 received water (gentamicin vehicle) and G10 received gentamicin for 10days; R30 remained resting and EX30 made exercise for 30days after gentamicin suspension. Training was performed on treadmill. Blood, 24h urine and kidneys were collected for renal function and oxidative stress, antioxidant, TGF-ß and histological analysis. KEY FINDINGS: Gentamicin treatment caused decreased renal function significant oxidative stress, reduced urinary nitric oxide and increased TGF-ß. G10+R30 presented partial recovery of metabolic data, renal function and lipoperoxidation levels, although they were still altered compared to W10+R30. Besides, we observed the presence of lymphomononuclear infiltrate in the kidneys of G10+R30. G10+EX30 vs G10+R30 showed additional improvement of all the mentioned parameters, showing at histology, regeneration of the tubule epithelium. SIGNIFICANCE: Our data suggest that moderate exercises could help in the recovery of metabolic parameters, renal function and structure on gentamicin-induced AKI, perhaps due to restoration of redox balance. This could protect the kidneys from further insults like challenges with nephrotoxic drugs or the aging per se.

Dacryocystitis due to Sporothrix brasiliensis: a case report of a successful clinical and serological outcome with low-dose potassium iodide treatment and oculoplastic surgery.

Sporothrix brasiliensis is the main species of the S. schenckii complex implicated in the zoonotic epidemics of sporotrichosis in Rio de Janeiro, Brazil. Epidemiological features have been already described, such as zoonotic transmission by cats and increased frequency of atypical clinical aspects. The involvement of the face by contact with cats is common in childhood; as a result, ophthalmic manifestations have increased. We report a case of acute dacryocystitis in a 9-year-old girl. A calmodulin-based molecular phylogeny was used to identify the agent as S. brasiliensis. This is a rare type of presentation, usually complicated with nasolacrimal duct occlusion. The patient was cured without sequelae after treatment with a low dose of saturated solution of potassium iodide and decompressive oculoplastic surgery. Therapeutic options and considerations of aetiological agents and serology are discussed.

Poor quality diet is associated with overweight status and obesity in patients with polycystic ovary syndrome.

BACKGROUND: The high rates of overweight status observed in women with polycystic ovary syndrome (PCOS) may reflect dietary intake, and so it is important to investigate diet quality and its relationship with the rates of overweight status and obesity among these patients. METHODS: A cross-sectional study was conducted in which 100 women with PCOS (Rotterdam criteria) were evaluated considering anthropometric and dietary data. The anthropometric evaluation included the measurement of weight, height, body mass index (BMI), waist circumference (WC), hip circumference and waist-hip ratio. Food intake data were collected from two 24-h dietary recalls to assess dietary patterns using the Brazilian Healthy Eating Index - Revised (BHEI-R). RESULTS: The anthropometric analysis indicated a high prevalence of overweight status, obesity and increased visceral fat (30.0%, 60.0% and 90.0%, respectively). The mean BHEI-R score was 56.1 ± 12.0 points (range 34.5-77.5 points). Diet quality was negatively correlated with obesity, which was evaluated by BMI (r = -0.248; P = 0.013) and WC (r = -0.278; P = 0.005). CONCLUSIONS: Dietary interventions focused on improvement of diet quality should be targeted to treat patients with PCOS because obesity in these women is associated with worsening endocrine, metabolic and reproductive functions.

Níveis de cloro da ração sobre parâmetros sanguíneos e ósseos de codornas italianas destinadas à produção de carne / Chlorine levels of diet on blood parameters and bone in meat production of Italian quail

Avaliaram-se diferentes níveis de cloro da ração sobre os parâmetros sanguíneos e o desenvolvimento ósseo de codornas destinadas à produção de carne. Foram utilizadas 384 codornas italianas de um a 49 dias de idade, distribuídas em delineamento inteiramente ao acaso, com seis tratamentos e oito repetições de oito aves. Os tratamentos consistiram de rações isonutrientes, exceto para o nível de cloro, que foram: 0,07; 0,12; 0,17; 0,22; 0,27 e 0,32%. Aos 49 dias, foram selecionadas duas aves por parcela para as coletas do sangue e das tíbias. Foram avaliados o pH e as concentrações de Na, Cl e K no sangue, bem como o comprimento, o diâmetro, o peso, o teor de matéria seca e cinzas, o índice de Seedor e a resistência à quebra e deformidade da tíbia. Os níveis de cloro da ração não influenciaram significativamente o pH, as concentrações de Na, Cl, K no sangue, assim como o comprimento, o diâmetro, o peso, o índice de Seedor, o teor de matéria seca e cinzas e a resistência à quebra e deformidade da tíbia. Conforme os resultados, o nível de cloro pode variar de 0,07 até 0,32%, sem alterar o pH e as concentrações de Na, Cl e K no sangue e sem prejuízos na qualidade óssea.

We evaluated different levels of chlorine in the feed on the blood parameters and bone development of quail for meat production. 384 Italian quails were used at 1-49 days of age, distributed in a completely randomized design with six treatments and eight replicates of eight birds. The treatments consisted of isonutrient diets, except for the level of chlorine, which were 0.07, 0.12, 0.17, 0.22, 0.27 and 0.32%. After 49 days, two birds were selected per plot for the collection of blood and tibia. The pH and the concentration of Na, K and Cl in the blood and the length, diameter, weight, dry matter and ash, Seedor index, breaking strength and deformity of the tibia were evaluated. The chlorine levels in the diet did not significantly influence the pH, the concentrations of Na, Cl, K blood, length, diameter, weight, Seedor index, dry matter and ash, crack resistance and deformity of the tibia. As the results, the level of chloride can range from 0.07 to 0.32%, without changing the pH and the concentrations of Na, K and Cl in the blood and without a loss in bone quality.

Níveis de cloro da ração sobre parâmetros sanguíneos e ósseos de codornas italianas destinadas à produção de carne / Chlorine levels of diet on blood parameters and bone in meat production of Italian quail

Avaliaram-se diferentes níveis de cloro da ração sobre os parâmetros sanguíneos e o desenvolvimento ósseo de codornas destinadas à produção de carne. Foram utilizadas 384 codornas italianas de um a 49 dias de idade, distribuídas em delineamento inteiramente ao acaso, com seis tratamentos e oito repetições de oito aves. Os tratamentos consistiram de rações isonutrientes, exceto para o nível de cloro, que foram: 0,07; 0,12; 0,17; 0,22; 0,27 e 0,32%. Aos 49 dias, foram selecionadas duas aves por parcela para as coletas do sangue e das tíbias. Foram avaliados o pH e as concentrações de Na, Cl e K no sangue, bem como o comprimento, o diâmetro, o peso, o teor de matéria seca e cinzas, o índice de Seedor e a resistência à quebra e deformidade da tíbia. Os níveis de cloro da ração não influenciaram significativamente o pH, as concentrações de Na, Cl, K no sangue, assim como o comprimento, o diâmetro, o peso, o índice de Seedor, o teor de matéria seca e cinzas e a resistência à quebra e deformidade da tíbia. Conforme os resultados, o nível de cloro pode variar de 0,07 até 0,32%, sem alterar o pH e as concentrações de Na, Cl e K no sangue e sem prejuízos na qualidade óssea.(AU)

We evaluated different levels of chlorine in the feed on the blood parameters and bone development of quail for meat production. 384 Italian quails were used at 1-49 days of age, distributed in a completely randomized design with six treatments and eight replicates of eight birds. The treatments consisted of isonutrient diets, except for the level of chlorine, which were 0.07, 0.12, 0.17, 0.22, 0.27 and 0.32%. After 49 days, two birds were selected per plot for the collection of blood and tibia. The pH and the concentration of Na, K and Cl in the blood and the length, diameter, weight, dry matter and ash, Seedor index, breaking strength and deformity of the tibia were evaluated. The chlorine levels in the diet did not significantly influence the pH, the concentrations of Na, Cl, K blood, length, diameter, weight, Seedor index, dry matter and ash, crack resistance and deformity of the tibia. As the results, the level of chloride can range from 0.07 to 0.32%, without changing the pH and the concentrations of Na, K and Cl in the blood and without a loss in bone quality.(AU)

Population structure of the bivalve Anomalocardia brasiliana, (Gmelin, 1791) in the semi-arid estuarine region of northeastern Brazil / Estrutura populacional do bivalve Anomalocardia brasiliana, (Gmelin, 1791) em região estuarina do semi-árido do nordeste brasileiro

For several decades, the bivalve mollusk Anomalocardia brasiliana has been extracted in estuarine areas in the semi-arid region of northeastern Brazil. However, information on both dynamics and abundance of their populations are missing. In this context, the present study, therefore, investigated several aspects of the population structure of A. brasiliana on beaches near the estuary of the Apodi River in Northeast Brazil. The aims were to determine the population density of A. brasiliana during different times of the year, to estimate population parameters, and to relate species density and distribution to the abiotic characteristics of the region. Sampling was performed from March/2007 and May/2008, we collected samples of sediments and mollusks along 180 m transects in the intertidal area on the beaches Barra and Pernambuquinho, Grossos - RN. At each sampling site, samples of sediments and mollusks were collected in the intertidal area along 180 m transects. The highest and lowest average densities on Pernambuquinho beach were 1148 (April/2007), and 100 individuals.m2 (May/ 2008), respectively. On Barra beach, the highest and lowest densities were 1813 (April/2007) and 951 individuals.m2, (November/2007), respectively. The densities of A. brasiliana on both beaches were significantly different only in the months of January, April and May/ 2008. Length of the individuals ranged from 1 to 28 mm, with most individuals measuring between 22 and 24 mm. Length growth rate parameters for A. brasiliana were L = 28.68 mm and k = 0.61 year1 on Barra beach, and L = 29.87 mm and k = 0.48 year1 on Pernambuquinho beach. The growth rate curves for A. brasiliana suggest the presence of three well defined cohorts, thus pointing to a continuous reproduction cycle with peak recruitment between October/2007 and March/2008. A. brasiliana density decreased predominantly during the heavy rains of 2008, probably due to an accumulation of sediments and decreasing salinity. This disturbance probably affected population of this species in the region.(AU)

O molusco bivalve Anomalocardia brasiliana há décadas vem sendo coletado em região estuarina do semi-árido do nordeste Brasileiro. No entanto, são escassas as informações sobre a dinâmica e abundância de suas populações na região. Neste contexto, o presente trabalho objetivou estudar aspectos da estrutura populacional de A. brasiliana em praias localizadas próximas à região de estuário do rio Apodi/Mossoró - RN no intuito de verificar a densidade populacional de A. brasiliana durante as diferentes épocas do ano; estimar os parâmetros populacionais e relacionar as características abióticas de região com a distribuição e densidade desta espécie. As coletas foram realizadas no período de março/2007 a maio/2008 nas praias de Barra e Pernambuquinho no município de Grossos - RN. Em cada ponto foram realizados transcectos de 180 metros na região de entremarés e coletadas amostras de sedimento e de moluscos. O maior valor médio de densidade na praia de Pernambuquinho foi de 1148 indivíduos/m2, no mês de abril/07 e o menor foi em maio/08 (100 indivíduos/m2). Na praia de Barra o maior valor foi de 1813 indivíduos/m2, no mês de abril/08 e o menor foi em novembro/07 com 951 indivíduos/m2. Os comprimentos variaram de 1 a 28 mm, sendo mais frequentes indivíduos com comprimentos entre 22 a 24 mm. Foram constatadas diferenças significativas entre as densidades de A. brasiliana em ambas as praias, apenas nos meses de janeiro, abril e maio de 2008. Os parâmetros que descrevem o crescimento em comprimento de A. brasiliana na praia de Barra foram: L = 28,68 mm e k = 0,61 ano1, enquanto que para a praia de Pernambuquinho foram: L = 29,87 mm e k = 0,48 ano1. As curvas de crescimento de A. brasiliana sugerem a presença de três coortes bem definidas, indicando uma reprodução contínua com picos de recrutamento ocorrerendo entre os meses de outubro/07 a março/08. Pode-se verificar uma diminuição da densidade de A. brasiliana, especialmente no período de chuvas intensas do ano de 2008, o que provavelmente esteve associado ao aporte de sedimentos e a diminuição da salinidade. Esta perturbação provavelmente afetou as populações desta espécie na região.(AU)

Population structure of the bivalve Anomalocardia brasiliana, (Gmelin, 1791) in the semi-arid estuarine region of northeastern Brazil / Estrutura populacional do bivalve Anomalocardia brasiliana, (Gmelin, 1791) em região estuarina do semi-árido do nordeste brasileiro

For several decades, the bivalve mollusk Anomalocardia brasiliana has been extracted in estuarine areas in the semi-arid region of northeastern Brazil. However, information on both dynamics and abundance of their populations are missing. In this context, the present study, therefore, investigated several aspects of the population structure of A. brasiliana on beaches near the estuary of the Apodi River in Northeast Brazil. The aims were to determine the population density of A. brasiliana during different times of the year, to estimate population parameters, and to relate species density and distribution to the abiotic characteristics of the region. Sampling was performed from March/2007 and May/2008, we collected samples of sediments and mollusks along 180 m transects in the intertidal area on the beaches Barra and Pernambuquinho, Grossos - RN. At each sampling site, samples of sediments and mollusks were collected in the intertidal area along 180 m transects. The highest and lowest average densities on Pernambuquinho beach were 1148 (April/2007), and 100 individuals.m2 (May/ 2008), respectively. On Barra beach, the highest and lowest densities were 1813 (April/2007) and 951 individuals.m2, (November/2007), respectively. The densities of A. brasiliana on both beaches were significantly different only in the months of January, April and May/ 2008. Length of the individuals ranged from 1 to 28 mm, with most individuals measuring between 22 and 24 mm. Length growth rate parameters for A. brasiliana were L = 28.68 mm and k = 0.61 year1 on Barra beach, and L = 29.87 mm and k = 0.48 year1 on Pernambuquinho beach. The growth rate curves for A. brasiliana suggest the presence of three well defined cohorts, thus pointing to a continuous reproduction cycle with peak recruitment between October/2007 and March/2008. A. brasiliana density decreased predominantly during the heavy rains of 2008, probably due to an accumulation of sediments and decreasing salinity. This disturbance probably affected population of this species in the region.

O molusco bivalve Anomalocardia brasiliana há décadas vem sendo coletado em região estuarina do semi-árido do nordeste Brasileiro. No entanto, são escassas as informações sobre a dinâmica e abundância de suas populações na região. Neste contexto, o presente trabalho objetivou estudar aspectos da estrutura populacional de A. brasiliana em praias localizadas próximas à região de estuário do rio Apodi/Mossoró - RN no intuito de verificar a densidade populacional de A. brasiliana durante as diferentes épocas do ano; estimar os parâmetros populacionais e relacionar as características abióticas de região com a distribuição e densidade desta espécie. As coletas foram realizadas no período de março/2007 a maio/2008 nas praias de Barra e Pernambuquinho no município de Grossos - RN. Em cada ponto foram realizados transcectos de 180 metros na região de entremarés e coletadas amostras de sedimento e de moluscos. O maior valor médio de densidade na praia de Pernambuquinho foi de 1148 indivíduos/m2, no mês de abril/07 e o menor foi em maio/08 (100 indivíduos/m2). Na praia de Barra o maior valor foi de 1813 indivíduos/m2, no mês de abril/08 e o menor foi em novembro/07 com 951 indivíduos/m2. Os comprimentos variaram de 1 a 28 mm, sendo mais frequentes indivíduos com comprimentos entre 22 a 24 mm. Foram constatadas diferenças significativas entre as densidades de A. brasiliana em ambas as praias, apenas nos meses de janeiro, abril e maio de 2008. Os parâmetros que descrevem o crescimento em comprimento de A. brasiliana na praia de Barra foram: L = 28,68 mm e k = 0,61 ano1, enquanto que para a praia de Pernambuquinho foram: L = 29,87 mm e k = 0,48 ano1. As curvas de crescimento de A. brasiliana sugerem a presença de três coortes bem definidas, indicando uma reprodução contínua com picos de recrutamento ocorrerendo entre os meses de outubro/07 a março/08. Pode-se verificar uma diminuição da densidade de A. brasiliana, especialmente no período de chuvas intensas do ano de 2008, o que provavelmente esteve associado ao aporte de sedimentos e a diminuição da salinidade. Esta perturbação provavelmente afetou as populações desta espécie na região.

Population structure of the bivalve Anomalocardia brasiliana, (Gmelin, 1791) in the semi-arid estuarine region of northeastern Brazil.

For several decades, the bivalve mollusk Anomalocardia brasiliana has been extracted in estuarine areas in the semi-arid region of northeastern Brazil. However, information on both dynamics and abundance of their populations are missing. In this context, the present study, therefore, investigated several aspects of the population structure of A. brasiliana on beaches near the estuary of the Apodi River in Northeast Brazil. The aims were to determine the population density of A. brasiliana during different times of the year, to estimate population parameters, and to relate species density and distribution to the abiotic characteristics of the region. Sampling was performed from March/2007 and May/2008, we collected samples of sediments and mollusks along 180 m transects in the intertidal area on the beaches Barra and Pernambuquinho, Grossos - RN. At each sampling site, samples of sediments and mollusks were collected in the intertidal area along 180 m transects. The highest and lowest average densities on Pernambuquinho beach were 1148 (April/2007), and 100 individuals.m-2 (May/ 2008), respectively. On Barra beach, the highest and lowest densities were 1813 (April/2007) and 951 individuals.m-2, (November/2007), respectively. The densities of A. brasiliana on both beaches were significantly different only in the months of January, April and May/ 2008. Length of the individuals ranged from 1 to 28 mm, with most individuals measuring between 22 and 24 mm. Length growth rate parameters for A. brasiliana were L∞ = 28.68 mm and k = 0.61 year-1 on Barra beach, and L∞ = 29.87 mm and k = 0.48 year-1 on Pernambuquinho beach. The growth rate curves for A. brasiliana suggest the presence of three well defined cohorts, thus pointing to a continuous reproduction cycle with peak recruitment between October/2007 and March/2008. A. brasiliana density decreased predominantly during the heavy rains of 2008, probably due to an accumulation of sediments and decreasing salinity. This disturbance probably affected population of this species in the region.

Effect of ischemic preconditioning on injuries caused by ischemia and reperfusion in rat intestine.

To study whether ischemic preconditioning (IPC) attenuated intestinal dysfunction caused by ischemia (I) and reperfusion (R), rats were underwent 60 minutes of I which was produced by occlusion of the superior mesenteric artery, and/or 120 minutes R. The IPC group had the I procedure previously stimulated for 5 minutes and the R for 10 minutes. IPC and sham groups were injected with saline solution (SS) via the femoral vein 5 minutes before the I and R, and for R. After I or I/R, 2-cm jejunal segments were mounted in an organ bath to study neurogenic contractions stimulated by electrical pulses or KCl using a digital recording system. Thin jejunal slices were stained with hematoxylin and eosin for optical microscopy. Compared with the sham group, jejunal contractions were similar in the IPC + I and the IPC + I/R groups, but reduced in the I + SS and the I/R + SS groups. The jejunal enteric nerves were damaged in the I + SS and the I/R + SS groups, but not in the IPC groups. These results suggested that ischemic preconditioning attenuated intestinal dysfunction caused by I and I/R.