Consumption of sugar-sweetened soft drinks and risk of metabolic syndrome and its components: results of the ELSA-Brasil study (2008-2010 and 2012-2014).

AIM: To estimate the association between consumption of sugar-sweetened soft drinks and unsweetened fruit juice with metabolic syndrome (MetS) and its components in participants of the Brazilian Longitudinal Adult Health Study (ELSA-Brasil) after 4 years of follow-up. METHODS: We used data from ELSA-Brasil cohort (N = 15,105). The sample consisted of 6,124 civil servants free of the MetS at baseline (35 to 74 years, both sexes). The consumption of sugar-sweetened soft drinks and unsweetened fruit juice was estimated by a food frequency questionnaire previously validated. The outcome was MetS and its components (Joint Interim Statement criteria). To test the association between beverage consumption at baseline (2008-2010) and MetS and its components at follow-up (2012-2014), we used Poisson regression models with robust variance adjusting for potential confounders. RESULTS: After 4-year follow-up, the higher consumption of sugar-sweetened soft drinks (≥ 1 serving/day = 250 mL/day) increased the relative risk of MetS (RR = 1.22; 95% CI 1.04-1.45), high fasting glucose (RR = 1.23; 95% CI 1.01-1.48), and high blood pressure (RR = 1.23; 95% CI 1.00-1.54). Moderate consumption of this beverage (0.4 to < 1 serving/day) increased the relative risk of high waist circumference (WC) (RR = 1.21; 95% CI 1.02-1.42). After adjustment for confounding variables, the consumption of unsweetened fruit juice was not associated with the MetS and its components. CONCLUSION: Higher sugar-sweetened soft drinks consumption was associated with a higher risk relative of MetS, high fasting glucose, and high blood pressure, while moderate consumption of this beverage increased the relative risk of high WC in Brazilian adults.

Cost-effectiveness analysis of grass pollen specific immunotherapy in children with allergic rhinitis compared to the standard of care symptomatic treatment in Portugal.

Summary: Background. Cost-effectiveness studies evaluating allergen immunotherapy (AIT) in children are scarce. We aim to compare the cost-effectiveness of subcutaneous (SCIT) and sublingual immunotherapy (SLIT) against standard-of-care (SOC) treatment in children with grass pollen allergic rhinitis. Methods. We created a Markov model to compare the three strategies over a 10-year horizon. SOC was the reference to calculate the incremental cost-effectiveness ratio (ICER). Deterministic and probabilistic sensitivity analysis were used to assess models' uncertainty. Results. We obtained an ICER of € 12,605 and € 6,318 for SLIT and SCIT, respectively. In sensitivity analysis, SCIT was more cost-effective than SLIT. Conclusions. AIT is cost-effective in children with grass pollen allergic rhinitis, especially for the subcutaneous route.

Diet quality, asthma and airway inflammation in school-aged children.

Summary: Background. Asthma is a major public health problem, with increasing prevalence in most countries, particularly among children. Poor dietary quality is also increasing in children, and evidence of the overall quality of children's food patterns effects on asthma is scarce. Methods. This cross-sectional analysis (660 children: 49.1% females, aged 7-12years) evaluated the association between diet quality and asthma (n = 56) and airway inflammation among school-aged children according to body mass index (BMI). Diet quality was assessed through the Healthy Eating Index (HEI)-2015, and categorized by tertiles. Higher scores represent a healthier diet. A questionnaire was used to enquire about self-reported medical diagnosis of asthma and asthma under medication. Lung function and airway reversibility were measured, and airway inflammation assessed measuring exhaled fractional nitric oxide (eNO). Two categories of BMI were considered: non-overweight/obese (p less than 85th), (n = 491), and overweight/obese (p ≥ than 85th), (n = 169). The associations between diet quality and asthma and airway inflammation were estimated using logistic regression models. Results. Non-overweight/obese children in 2nd tertile of HEI-2015 score had decreased odds of having eNO ≥ 35ppb (OR 0.43, 95%CI 0.19-0.98), medical diagnosis of asthma (OR 0.18; 95%CI 0.04-0.84), and asthma treatment (OR 0.12; 95%CI 0.01-0.95), compared to children in the 1st tertile. Conclusions. Our findings suggest that a higher diet quality associates with lower levels of airway inflammation and reduced prevalence of asthma among non-overweight/obese school-aged children.

Cloning and molecular properties of a novel luciferase from the Brazilian Bicellonycha lividipennis (Lampyridae: Photurinae) firefly: comparison with other firefly luciferases.

Several firefly luciferases eliciting light emission in the yellow-green range of the spectrum and with distinct kinetic properties have been already cloned, sequenced, and characterized. Some of them are currently being applied as analytical reagents and reporter genes for bioimaging and biosensors, and more recently as potential color tuning indicators of intracellular pH and toxic metals. They were cloned from the subfamilies Lampyrinae (Photinini: Photinus pyralis, Macrolampis sp2; Cratomorphini: Cratomorphus distinctus), Photurinae (Photuris pennsylvanica), Luciolinae (Luciola cruciata, L. lateralis, L. mingrelica, L. italica, Hotaria parvula), and Amydetinae (Amydetes vivianii) occurring in different parts of the world. The largest number has been cloned from fireflies occurring in Brazilian biomes. Taking advantage of the large biodiversity of fireflies occurring in the Brazilian Atlantic rainforest, here we report the cloning and characterization of a novel luciferase cDNA from the Photurinae subfamily, Bicellonycha lividipennis, which is a very common firefly in marshlands in Brazil. As expected, multialignements and phylogenetic analysis show that this luciferase clusters with Photuris pennsylvanica adult isozyme, and with other adult lantern firefly luciferases, in reasonable agreement with traditional phylogenetic analysis. The luciferase elicits light emission in the yellow-green region, has kinetics properties similar to other adult lantern firefly luciferases, including pH- and metal sensitivities, but displays a lower sensitivity to nickel, which is suggested to be caused by the natural substitution of H310Y.

Airborne Food Allergen and Aeroallergen Levels in Health Care Settings: An Unaccounted for but Potentially Relevant Source of Exposure?

BACKGROUND AND OBJECTIVE: Exposure to airborne allergens of biological origin is associated with the development and exacerbation of allergic asthma and rhinitis. Assessment of allergen exposure in health care facilities may improve monitoring of hygiene and surveillance of specific allergens that can cause symptoms in sensitized persons. Objective: To assess concentrations of airborne food and aeroallergens in various health care settings in Portugal. METHODS: Dust was vacuumed from primary health care centers in Lisbon and from the emergency department, day hospital, internal medicine ward, operating room, and outpatient clinic of a university central hospital in Porto. Samples were sieved, weighed, and extracted, and concentrations of Nbos d 5, Cor a 9, Gal d 2, Ara h 3, Ara h 6, Der p 1, Fel d 1, Can f 1, Bla g 2, Alt a 1, and Phl p 5 were determined using a multiplex array for allergens (MARIA). RESULTS: All airborne food and aeroallergens were found in at least 1 sampled area, except for Alt a 1. Levels of Der p 1 and Fel d 1 ranged from 13.0 µg/g to 971.0 µg/g and from 7.0 µg/g to 4618.8 µg/g, respectively. Higher levels of food allergens were found in the emergency department (Nbos d 5, 16 034.0 µg/g; and Cor a 9, 10 649.5 µg/g). CONCLUSIONS: Except for the operating room, exposure levels for dust mite and cat and dog dander in health care facilities were above the values associated with sensitization and allergic asthma or rhinitis symptoms in sensitized persons.

Mineral and natural films change the physical-chemical properties of grapes and modulate oviposition behaviour of Ceratitis capitata Wiedemann (Diptera: Tephritidae).

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann), is one of the main pests of fruit, worldwide, and the use of population suppression method with low environmental impact is an increasingly strong requirement of the consumer market. The aim of this study was to evaluate the effect of mineral and natural films on the physical-chemical properties of grapes (Vitis vinifera L.), cultivar Itália, and oviposition behaviour of C. capitata. Fruits were immersed in suspensions (100 and 200 g L-1) of mineral (kaolin Surround®WP, kaolin 607, kaolin 608, kaolin 611 and talc) and natural films (chitosan, cassava starch, potato starch and guar gum 5.0 g L-1) and distilled water (control). After drying, fruits were exposed to C. capitata pairs of males and females for 24 h in choice and non-choice tests; the number of punctures with and without eggs, eggs per fruit and behavioural response of fly to treated and untreated fruits were recorded. Results obtained in this study are promising, given the scientific evidence that films of mineral particles such as kaolin (Surround®, 607, 608 and 611) changed the firmness, luminosity, chroma and hue angle of grapes and reduced the oviposition of C. capitata. In addition, our results also showed that natural polymers do not deter C. capitata females, but rather seem to stimulate oviposition.

Systemic circulating inflammatory burden and periodontitis in adolescents.

OBJECTIVES: To analyze the association between systemic inflammatory burden of cardiovascular disease (CVD) risk and periodontitis in adolescents, including mediating pathways triggered by their common risk factors. MATERIALS AND METHODS: Using a population-based sample study (n = 405) of Brazilian adolescents (17-18 years old), direct and mediation pathways triggered by "Socioeconomic Status," "Adiposity," Smoking, and "Blood Pressure" were modelled for the association between the "Systemic Circulating Inflammatory Burden of CVD Risk" (IL-1ß, IL-6, IL-8, TNF-α) and the "Initial Periodontitis" (bleeding on probing (BoP), probing depth (PD) ≥ 4 mm, clinical attachment loss (CAL) ≥ 4 mm), both as continuous latent variables, using structural equation modeling. Sensitivity analysis was performed for the outcomes "Gingivitis" (visible plaque; BoP); "Moderate Periodontitis" (PD ≥ 5 mm and CAL ≥ 5 mm) and periodontitis (CDC-AAP case definition). RESULTS: Higher "Systemic Circulating Inflammatory Burden of CVD Risk" was directly associated with higher "Initial Periodontitis" (standardized coefficient [SC] = 0.178, P value < 0.001). Lower "Socioeconomic Status" (SC = - 0.022, P value = 0.015) and Smoking (SC = 0.030, P value = 0.021) triggered the "Initial Periodontitis", mediated by "Systemic Circulating Inflammatory Burden of CVD Risk". Sensitivity analysis showed a dose-response relationship between "Systemic Circulating Inflammatory Burden of CVD Risk" and "Moderate Periodontitis" (SC = 0.323, P value = 0.021). CONCLUSIONS: "Systemic Circulating Inflammatory Burden of CVD Risk" appeared as an underlying mechanism of early periodontal breakdown in adolescents, also triggered by social vulnerability and smoking. CLINICAL RELEVANCE: The association between periodontitis and CVD in adulthood seems to establish much earlier in life than had been previously studied, giving impetus to preventive approaches focused on their common risk factors.

Association between the density and type of trees around urban schools and exhaled nitric oxide levels in schoolchildren.

Summary: Greenspaces in school's neighbourhood represent an important environment to promote healthy development. The aim of this study was to assess the association between the density and type of trees around schools and exhaled Nitric Oxide (NO) levels in schoolchildren. Data on 845 children from 20 primary schools in Porto was analysed. Airway inflammation was assessed by measuring exhaled NO level. The density and type of trees were quantified within a 500 m buffer around schools. Associations were estimated using mixed-effect models. A significant association was observed between non-tree covered areas around schools and exhaled NO levels in schoolchildren (ß = - 1.42, 95% CI - 2.84, - 0.001). Our results suggested that the presence of trees in school neighbourhoods may play a role in the biological mechanisms underlying the complex links between environment and airway inflammation.

Frustrated Bearings.

In a bearing state, touching spheres (disks in two dimensions) roll on each other without slip. Here we frustrate a system of touching spheres by imposing two different bearing states on opposite sides and search for the configurations of lowest energy dissipation. If the dissipation between contacts of spheres is viscous (with random damping constants), the angular momentum continuously changes from one bearing state to the other. For Coulomb friction (with random friction coefficients) in two dimensions, a sharp line separates the two bearing states and we show that this line corresponds to the minimum cut. Astonishingly, however, in three dimensions intermediate bearing domains that are not synchronized with either side are energetically more favorable than the minimum-cut surface. Instead of a sharp cut, the steady state displays a fragmented structure. This novel type of state of minimum dissipation is characterized by a spanning network of slipless contacts that reaches every sphere. Such a situation becomes possible because in three dimensions bearing states have four degrees of freedom.

Atrial matrix remodeling in atrial fibrillation patients with aortic stenosis.

BACKGROUND: This study aimed to evaluate atrium extracellular matrix remodeling in atrial fibrillation (AF) patients with severe aortic stenosis, through histological fibrosis quantification and extracellular matrix gene expression analysis, as well as serum quantification of selected protein targets. METHODS: A posthoc analysis of a prospective study was performed in a cohort of aortic stenosis patients. Between 2014 and 2019, 56 patients with severe aortic stenosis submitted to aortic valve replacement surgery in a tertiary hospital were selected. RESULTS: Fibrosis was significantly increased in the AF group when compared to sinus rhythm (SR) patients (p = 0.024). Moreover, cardiomyocyte area was significantly higher in AF patients versus SR patients (p = 0.008). Conversely, collagen III gene expression was increased in AF patients (p = 0.038). TIMP1 was less expressed in the atria of AF patients. MMP16/TIMP4 ratio was significantly decreased in AF patients (p = 0.006). TIMP1 (p = 0.004) and TIMP2 (p = 0.012) were significantly increased in the serum of AF patients. Aortic valve maximum (p = 0.0159) and mean (p = 0.031) gradients demonstrated a negative association with serum TIMP1. CONCLUSIONS: Atrial fibrillation patients with severe aortic stenosis present increased atrial fibrosis and collagen type III synthesis, with extracellular matrix remodelling demonstrated by a decrease in the MMP16/TIMP4 ratio, along with an increased serum TIMP1 and TIMP2 proteins.

Dietary total antioxidant capacity is positively associated with muscular strength in cirrhotic outpatients: a cross-sectional study.

BACKGROUND: Cirrhosis is the end-stage of progressive fibrosis, in which oxidative stress and inflammation-related pathways can modulate the cellular and tissue events involved in the pathogenesis of liver fibrosis. Dietary intake of antioxidants has been suggested to protect against oxidative damage and related clinical complications. The present study aimed to investigate the potential association of the dietary total antioxidant capacity (dTAC) with anthropometric, functional and biochemical markers, as well as the severity of the disease, in cirrhotic outpatients. METHODS: Sixty-two outpatients (38 men and 24 women) with a mean (SD) age of 59.1 (9.9) years were evaluated. Dietary TAC was estimated from a food frequency questionnaire. Aetiology and severity of liver cirrhosis, lifestyle characteristics, occurrence of comorbidities and oedema, and anthropometric, functional and biochemical markers were all assessed. RESULTS: Cirrhotic outpatients with higher dTAC also had higher values of the hand-grip strength (P = 0.029) and arm muscle area (P = 0.027). After adjusting by sex, age, smoking and alcohol intake, the addition of 1 mmol day-1 of dTAC contributed to increase 0.552 kg f-1 in hand-grip strength (P < 0.05). The addition of one mmol day-1 of dTAC contributed to an arm muscle area increase 0.565 cm2 (P < 0.05) on average. CONCLUSIONS: The dTAC was positively associated with hand-grip strength and arm muscle area in cirrhotic outpatients. The implications of the present study are important in clinical practice because a diet rich in antioxidants may be an ally in the control of excessive reactive oxygen species production in cirrhotic outpatients with repercussion on muscle mass and strength.

CT-guided bone marrow aspirations and biopsies: retrospective study and comparison with blind procedures.

PURPOSE: To compare the pathology results of CT-guided and blind bone marrow aspirations and biopsies. METHODS: Ninety-eight consecutive CT-guided biopsies and 98 age- and gender-matched blind (non-CT-guided) posterior iliac crest bone marrow aspirations and biopsies performed in 2017 were reviewed for adequacy of core biopsies and aspirate smears. CT procedure images and CT abdomen/pelvis images were reviewed to evaluate anatomic features of the posterior ilium and soft tissues. Statistical analysis was performed using a T test, Fisher exact test, and Kruskal-Wallis test. RESULTS: There was no significant difference in the age and gender of the two groups (p > 0.05). However, the CT-guided group had a higher BMI (p = 0.0049) and posterior soft tissue thickness (p = 0.0016). More CT-guided biopsy samples (CT 93 (95%); blind 77 (79%); p = 0.0006) and aspirate smears (CT 90 (92%); blind 78 (80%); p = 0.042) were categorized as adequate. The CT-guided group had longer core lengths (CT 1.4 ± 0.6 (range 0.3-3.5) cm; blind 1.0 ± 0.60 (range 0-2.6) cm; p = 0.0001). Overall, 131/164 (80%) of the cases had at least one of the described features (slanted posterior ilium (angle > 30°), 30%; rounded posterior ilium, 20%; thick posterior ilium cortex, 13%; focal lesion in posterior ilium, 12%; prior procedure in posterior ilium, 5%; posterior soft tissue thickness > 3 cm, 40%). CONCLUSION: CT-guided bone marrow procedures were more likely to result in both adequate aspirate smears and biopsy samples and longer core lengths when compared with blind procedures.

Metabolic interactions in asthma.

Summary: Metabolomics can be used to explore altered metabolic pathways in asthma, giving insights into its pathophysiology. We aimed to review how metabolomics has been used to understand asthma by describing metabolic pathways under research and discussing clinical implications. The search was performed in PubMed, and studies published since 2000 using a metabolomics approach, were included. A total of 32 studies were analysed. Pathways related with cellular energy homeostasis, lipid metabolism and oxidative stress, immune and inflammatory processes and others were altered. Initial studies focused on biomarker discovery. But, metabolomics can be used to evaluate drug effects on specific pathways, to highlight pathways that can further develop in new targeted treatments and to identify differences according to asthma severity and phenotypes.

Long-term outcomes of transmesenteric portal vein recanalization for the treatment of chronic portal vein thrombosis after pediatric liver transplantation.

Portal vein thrombosis (PVT) may occur at any time following liver transplantation. We describe our experience with portal vein recanalization in cases of thrombosis after liver transplantation. Twenty-eight children (5%) out of 566 liver transplant recipients underwent portal vein recanalization using a transmesenteric approach. All children received left hepatic segments, developed PVT, and had symptoms or signs of portal hypertension. Portal vein recanalization was performed via the transmesenteric route in all cases. Twenty-two (78.6%) patients underwent successful recanalization and stent placement. They received oral anticoagulants after the procedure, and clinical symptoms subsided. Symptoms recurred due to portal vein restenosis/thrombosis in seven patients. On an intention-to-treat basis, the success rate of the proposed treatment was 60.7%. Only 17 out of 28 children with posttransplant chronic PVT retained stent patency (primary + assisted) at the end of the study period. In cases of portal vein obstruction, the transmesenteric approach via minilaparotomy is technically feasible with good clinical and hemodynamic results. It is an alternative procedure to reestablish the portal flow to the liver graft that can be performed in selected cases and a therapeutic addition to other treatment strategies currently used to treat chronic PVT.

The Warburg effect: persistence of stem-cell metabolism in cancers as a failure of differentiation.

Background: Two recent observations regarding the Warburg effect are that (i) the metabolism of stem cells is constitutive (aerobic) glycolysis while normal cellular differentiation involves a transition to oxidative phosphorylation and (ii) the degree of glucose uptake of a malignancy as imaged by 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) is associated with histologic measures of tumor differentiation. Combining these observations, we hypothesized that the high levels of glucose uptake observed in poorly differentiated cancers may reflect persistence of the glycolytic metabolism of stem cells in malignant cells that fail to fully differentiate. Patients and methods: Tumor glucose uptake was measured by FDG-PET in 552 patients with histologically diverse cancers. We used normal mixture modeling to explore FDG-PET standardized uptake value (SUV) distributions and tested for associations between glucose uptake and histological differentiation, risk of lymph node metastasis, and survival. Using RNA-seq data, we carried out pathway and transcription factor analyses to compare tumors with high and low levels of glucose uptake. Results: We found that well-differentiated tumors had low FDG uptake, while moderately and poorly differentiated tumors had higher uptake. The distribution of SUV for each histology was bimodal, with a low peak around SUV 2-5 and a high peak at SUV 8-14. The cancers in the two modes were clinically distinct in terms of the risk of nodal metastases and death. Carbohydrate metabolism and the pentose-related pathway were elevated in the poorly differentiated/high SUV clusters. Embryonic stem cell-related signatures were activated in poorly differentiated/high SUV clusters. Conclusions: Our findings support the hypothesis that the biological basis for the Warburg effect is a persistence of stem cell metabolism (i.e. aerobic glycolysis) in cancers as a failure to transition from glycolysis-utilizing undifferentiated cells to oxidative phosphorylation-utilizing differentiated cells. We found that cancers cluster along the differentiation pathway into two groups, utilizing either glycolysis or oxidative phosphorylation. Our results have implications for multiple areas of clinical oncology.

Mechanisms of exercise-induced bronchoconstriction in athletes: Current perspectives and future challenges.

The evidence of exercise-induced bronchoconstriction (EIB) without asthma (EIBwA ) occurring in athletes led to speculate about different endotypes inducing respiratory symptoms within athletes. Classical postulated mechanisms for bronchial obstruction in this population include the osmotic and the thermal hypotheses. More recently, the presence of epithelial injury and inflammation in the airways of athletes was demonstrated. In addition, neuronal activation has been suggested as a potential modulator of bronchoconstriction. Investigation of these emerging mechanisms is of major importance as EIB is a significant problem for both recreational and competitive athletes and is the most common chronic condition among Olympic athletes, with obvious implications for their competing performance, health and quality of life. Hereby, we summarize the latest achievements in this area and identify the current gaps of knowledge so that future research heads toward better defining the etiologic factors and mechanisms involved in development of EIB in elite athletes as well as essential aspects to ultimately propose preventive and therapeutic measures.

Effective anti-programmed death-1 therapy in a SUFU-mutated patient with Gorlin-Goltz syndrome.

We present the case of a 77-year-old male patient with more than 50 basal cell carcinomas on the head and upper trunk. The patient did not respond to several lines of treatment, including surgery, imiquimod, retinoids, itraconazole and therapy with the hedgehog inhibitor vismodegib. The patient responded well to off-label therapy with the anti-programmed death-1 antibody pembrolizumab after four infusions.

Low cytomegalovirus seroprevalence in early multiple sclerosis: a case for the 'hygiene hypothesis'?

BACKGROUND AND PURPOSE: Cytomegalovirus (CMV) infection has recently been associated with a lower multiple sclerosis (MS) susceptibility, although it remains controversial whether it has a protective role or is merely an epiphenomenon related to westernization and early-life viral infections. We aimed to evaluate whether CMV serostatus may differ in patients with early MS as compared with patients with non-early MS, analyzing the putative association of this virus with MS clinical course and humoral immune responses against other herpesviruses. METHODS: Multicentric analysis was undertaken of 310 patients with MS (early MS, disease duration ≤5 years, n = 127) and controls (n = 155), evaluating specific humoral responses to CMV, Epstein-Barr virus and human herpesvirus-6, as well as T-cell and natural killer (NK)-cell immunophenotypes. RESULTS: Cytomegalovirus seroprevalence in early MS was lower than in non-early MS or controls (P < 0.01), being independently associated with disease duration (odds ratio, 1.04; 95% confidence interval, 1.01-1.08, P < 0.05). CMV+ patients with MS displayed increased proportions of differentiated T-cells (CD27-CD28-, CD57+, LILRB1+) and NKG2C+ NK-cells, which were associated with a lower disability in early MS (P < 0.05). CMV+ patients with early MS had an age-related decline in serum anti-EBNA-1 antibodies (P < 0.01), but no CMV-related differences in anti-human herpesvirus-6 humoral responses. CONCLUSIONS: Low CMV seroprevalence was observed in patients with early MS. Modification of MS risk attributed to CMV might be related to the induction of differentiated T-cell and NK-cell subsets and/or modulation of Epstein-Barr virus-specific immune responses at early stages of the disease.

Short communication: Extended-spectrum AmpC-producing Escherichia coli from milk and feces in dairy farms in Brazil.

The AmpC enzyme is normally expressed constitutively in Escherichia coli, and its overproduction confers resistance to cefoxitin. A newly reported AmpC, the extended-spectrum AmpC (ESAC), is related to resistance to cefepime, a fourth-generation cephalosporin. This enzyme presents more flexibility in the active site due to insertions, replacements, and deletions on AA sequences. Many isolates producing ESAC were reported in human clinical isolates, but E. coli ESAC producers were reported in animals only in France. The animal E. coli strains can produce this enzyme and possibly disseminate it to human and production environments. In our study, 3 strains of E. coli from milk and feces bovine samples, collected in Rio de Janeiro, Brazil, were suspected to produce ESAC. After excluding other mechanisms of resistance, the gene was sequenced to verify ESAC characteristics. These strains presented replacement of AA in omega and R2 loops, suggesting ESAC production. This is the first report to study ESAC E. coli in dairy farms in Brazil.

Intrinsic reproductive isolating mechanisms in the maintenance of a hybrid zone between ecologically divergent subspecies.

Understanding factors involved in maintaining stable hybrid zones is important for predicting the ultimate fate of the interacting taxa, but the relative importance of mechanisms such as ecological selection and intrinsic reproductive isolation remains unclear. Most studies of reproductive isolation in hybrid zones have focused either on zones with strongly bimodal patterns in genotype or phenotype frequencies, with relatively strong isolation, or unimodal zones with relatively weak isolation, whereas less is known about more intermediate classes of hybrid zone. Here, we utilize a hybrid zone of this intermediate type occurring between northern and southern subspecies of Atlantic killifish, Fundulus heteroclitus, to identify isolating mechanisms playing a role in maintaining this type of zone. The two subspecies differ in environmental tolerance, and we found some evidence of microhabitat preference between subspecies within a small tidal creek at the centre of the hybrid zone. There was also an association between sex, mitochondrial genotype and habitat within this creek. Fertilization success did not differ between consubspecific and heterosubspecific crosses, but hatching success was significantly lower for crosses involving southern males and northern females, and crosses between southern females and northern males had altered developmental rates. Southern females and northern males showed patterns consistent with positive assortative mating. Together, these results indicate a role for a combination of factors including assortative mating and/or early hybrid inviability in the maintenance of this hybrid zone and suggest that hybrid zones with intermediate levels of reproductive isolation are likely to be maintained by multiple interacting isolating mechanisms.