Tropical grass digestibility assessed by the mobile bag and in vitro methods in horses.

This study aimed to compare the digestibility of tropical grasses by horses by the in vivo method using mobile nylon bags with the in vitro digestibility method using horse feces as a source of inoculum. Five horses were used in a 2 × 5 factorial design with randomized blocks featuring two methods (in vivo and in vitro) and five grasses: Tifton 85 hay (Cynodon spp.), sixweeks threeawn grass (Aristida adsencionis, Linn), Alexandergrass (Brachiaria plantaginea (Link) Hitchc.), capim-de-raiz (Chloris orthonoton, Doell), and Sabi grass (Urochloa mosambicensis). No difference (P>0.05) was found between the in vivo and in vitro methods regarding nutrient digestibility of Sabi grass and sixweeks threeawn. Tifton 85 was the only grass that showed differences (P<0.05) between the two methods concerning the apparent digestibility of all nutrients. Alexandergrass, Tifton 85, and capim-de-raiz exhibited the best digestibility of dry matter, neutral detergent fiber, acid detergent fiber, and organic matter by the mobile bag method compared to the in vitro method. Tifton 85 and capim-de-raiz had higher crude protein digestibility by the mobile bag method than by the in vitro method. The mean retention time of the mobile bags in the digestive tract of the horses was 43.69 h. The bags with samples of sixweeks threeawn and Sabi grass had shorter retention times than capim-de-raiz and Alexandergrass (P<0.0001). It is concluded that, for sixweeks threeawn and Sabi grass, digestibility in horses can be assessed using the in vitro method in place of the mobile nylon bag method.

Sleep deprivation induces late deleterious effects in a pharmacological model of Parkinsonism.

Parkinson's disease is a degenerative, chronic and progressive disease, characterized by motor dysfunctions. Patients also exhibit non-motor symptoms, such as affective and sleep disorders. Sleep disorders can potentiate clinical and neuropathological features and lead to worse prognosis. The goal of this study was to evaluate the effects of sleep deprivation (SD) in mice submitted to a progressive pharmacological model of Parkinsonism (chronic administration with a low dose of reserpine). Male Swiss mice received 20 injections of reserpine (0.1 mg/kg) or vehicle, on alternate days. SD was applied before or during reserpine treatment and was performed by gentle handling for 6 h per day for 10 consecutive days. Animals were submitted to motor and non-motor behavioral assessments and neurochemical evaluations. Locomotion was increased by SD and decreased by reserpine treatment. SD during treatment delayed the onset of catalepsy, but SD prior to treatment potentiated reserpine-induced catalepsy. Thus, although SD induced an apparent beneficial effect on motor parameters, a delayed deleterious effect on alterations induced by reserpine was found. In the object recognition test, both SD and reserpine treatment produced cognitive deficits. In addition, the association between SD and reserpine induced anhedonic-like behavior. Finally, an increase in oxidative stress was found in hippocampus of mice subjected to SD, and tyrosine hydroxylase immunoreactivity was reduced in substantia nigra of reserpine-treated animals. Results point to a possible late effect of SD, aggravating the deficits in mice submitted to the reserpine progressive model of PD.

Animal Approaches to Studying Risk Factors for Parkinson's Disease: A Narrative Review.

Despite recent efforts to search for biomarkers for the pre-symptomatic diagnosis of Parkinson's disease (PD), the presence of risk factors, prodromal signs, and family history still support the classification of individuals at risk for this disease. Human epidemiological studies are useful in this search but fail to provide causality. The study of well-known risk factors for PD in animal models can help elucidate mechanisms related to the disease's etiology and contribute to future prevention or treatment approaches. This narrative review aims to discuss animal studies that investigated four of the main risk factors and/or prodromal signs related to PD: advanced age, male sex, sleep alterations, and depression. Different databases were used to search the studies, which were included based on their relevance to the topic. Although still in a reduced number, such studies are of great relevance in the search for evidence that leads to a possible early diagnosis and improvements in methods of prevention and treatment.

Nitrogen demand and agro-environmental efficiency in Brazilian cereal production.

This paper aims to analyze the main factors that explain the demand for synthetic nitrogen fertilizers in Brazil, as well as the efficiency of their use. In addition, the research sought to relate the use of fertilizers with nitrous oxide (N2O) emissions. Demand was estimated using the two-stage least squares method (2SLS). Nitrogen use efficiency (NUE) was calculated using an agri-environmental index. The results indicated that demand of nitrogen fertilizers is positively affected by the price of cereal, cereal production and the number of fertilizers used in the past harvest. The calculated NUE presented an average value of 53% in the 1994-2018 period, indicating inefficient use of N. Emissions from nitrogen fertilization grew 59% for the same period. The increasing and / or inadequate rates of fertilizer use have resulted in agro-environmental inefficiency, that is, a decrease in NUE and an increase in N2O emissions. Public policies that guarantee more agricultural technical assistance and rational alternative forms of nitrogen use could contribute to optimizing the synthetic doses applied in production, minimizing adverse environmental effects without generating economic losses to farmers and Brazilian agricultural production.

Assessment of meteorological settings on air quality modeling system-a proposal for UN-SDG and regulatory studies in non-homogeneous regions in Brazil.

Air quality models are essential tools to meet the United Nations Sustainable Development Goals (UN-SDG) because they are effective in guiding public policies for the management of air pollutant emissions and their impacts on the environment and human health. Despite its importance, Brazil still lacks a guide for choosing and setting air quality models for regulatory purposes. Based on this, the current research aims to assess the combined WRF/CALMET/CALPUFF models for representing SO2 dispersion over non-homogeneous regions as a regulatory model for policies in Brazilian Metropolitan Regions to satisfy the UN-SDG. The combined system was applied to the Rio de Janeiro Metropolitan Area (RJMA), which is known for its physiographic complexity. In the first step, the WRF model was evaluated against surface-observed data. The local circulation was underestimated, while the prevailing observational winds were well represented. In the second step, it was verified that all CALMET three meteorological configurations performed better for the most frequent wind speed classes so that the largest SO2 concentrations errors occurred during light winds. Among the meteorological settings in WRF/CALMET/CALPUFF, the joined use of observed and modeled meteorological data yielded the best results for the dispersion of pollutants. This result emphasizes the relevance of meteorological data composition in complex regions with unsatisfactory monitoring given the inherent limitations of prognostic models and the excessive extrapolation of observed data that can generate distortions of reality. This research concludes with the proposal of the WRF/CALMET/CALPUFF air quality regulatory system as a supporting tool for policies in the Brazilian Metropolitan Regions in the framework of the UN-SDG, particularly in non-homogeneous regions where steady-state Gaussian models are not applicable.

Postoperative Sensitivity of Composite Replacement of Amalgam Restoration: A Randomized Clinical Trial.

PURPOSE: This randomized clinical trial aimed to evaluate the postoperative sensitivity of different resin composite/adhesive materials, placed either by an incremental or bulk-fill technique in posterior resin composite replacement of amalgam restorations. METHODS AND MATERIALS: A total of 47 patients with good overall health and at least four class I or class II amalgam restorations to be replaced participated in this study. The characteristics of 188 cavities were registered and randomly restored using incremental (Z350XT, 3M) or bulk-fill (Filtek Bulk Fill, 3M; Sonic Fill, Kerr; and Opus Bulk Fill, FGM) technique. The postoperative sensitivity was assessed using a Visual Analog Scale (0-100) after 24 hours, 7 days, and 30 days. Pain scores were temporally analyzed using Friedman test followed by Dunn post hoc test (α=0.05). The frequency of tests was calculated according to the frequency and percentage of the McNemar test. RESULTS: The restorative technique and the bulk-fill system used did not affect the postoperative sensitivity, except for Filtek Bulk Fill group, which presented less postoperative sensitivity after 24 hours (p=0.037). Regardless of the restorative material, the postoperative sensitivity decreased after 24 hours, and no differences were found after 7 and 30 days. CONCLUSIONS: After 1 week, the filling technique and the bulk-fill system have no influence on the postoperative sensitivity.

Rainfall and sea level drove the expansion of seasonally flooded habitats and associated bird populations across Amazonia.

Spatial arrangement of distinct Amazonian environments through time and its effect on specialized biota remain poorly known, fueling long-lasting debates about drivers of biotic diversification. We address the late Quaternary sediment deposition that assembled the world's largest seasonally flooded ecosystems. Genome sequencing was used to reconstruct the demographic history of bird species specialized in either early successional vegetation or mature floodplain forests. Sediment deposition that built seasonally flooded habitats accelerated throughout the Holocene (last 11,700 years) under sea level highstand and intensification of the South American Monsoon, at the same time as global increases in atmospheric methane concentration. Bird populations adapted to seasonally flooded habitats expanded due to enlargement of Amazonian river floodplains and archipelagos. Our findings suggest that the diversification of the biota specialized in seasonally flooded habitats is coupled to sedimentary budget changes of large rivers, which rely on combined effects of sea level and rainfall variations.

Morphological and cytochemical characterization of the peripheral blood cells of farmed streaked prochilod Prochilodus lineatus (Characiformes, Prochilodontidae) / [Caracterização morfológica e citoquímica das células do sangue periférico do curimatã Prochilodus lineatus (Characiformes, Prochilodontidae) em cultivo]

Morphological and cytochemical studies of peripheral blood cells of fish have improved the understanding of their functions and cell types. The present study performed the Morphological and cytochemical analysis of the peripheral blood of Prochilodus lineatus, Characiform native to South America, which has been gaining space in local aquaculture and as a species introduced in Asia. Our analysis provided information on the morphological and cytochemical characteristics of the leukocytes, for the formulation of hypothesis about their role in the immune system of the species. It was found that Prochilodus lineatus has morphological and cytochemical features in common with other fish species, mainly of the Characiformes order. However, we detected the presence of heterophils and PAS positive granulocytes simultaneously with neutrophils. We also found that heterophils and PAS positive granulocytes are very similar, both morphologically and cytochemically.(AU)

O estudo da morfologia e da citoquímica das células do sangue periférico dos peixes tem sido eficaz para o entendimento de suas funções e dos tipos celulares. Este estudo realizou a análise morfológica e citoquímica do sangue periférico de Prochilodus lineatus, caracídeo nativo da América do Sul que vem ganhando espaço na aquicultura local e como espécie introduzida na Ásia. Essa análise forneceu informações sobre a morfologia e as características citoquímicas dos leucócitos, visando a hipóteses sobre suas funções. Verificou-se que estas são semelhantes em vários aspectos a outras espécies, principalmente da ordem Characiformes. No entanto, neste estudo detectou-se a presença dos heterofilos e da célula granulocítica especial, simultaneamente à presença dos neutrófilos. Ainda, foi verificado que os heterofilos e a célula granulocítica especial são muito semelhantes morfológica e citoquimicamente.(AU)

Primary care-based interventions for treatment of obesity: a systematic review.

OBJECTIVE: The objective of this study was to synthesise evidence from primary care-based interventions for the treatment of obesity in adults and the elderly. STUDY DESIGN: Systematic review. METHODS: Eight electronic databases (MEDLINE, Lilacs, Embase, Psycinfo, Cochrane, WHOLIS, Open Gray and Scholar Google) were searched. There was no limitation on publication period; articles published in English, Portuguese or Spanish were included. The selection, data extraction and quality analyses were performed by three reviewers. RESULTS: A literature search retrieved 6464 publications, of which 5120 publications were excluded after reading the title/abstract and 293 after reading the full text. In total, 56 publications, representing 72 interventions were included. All studies were published between 2000 and 2020. Most studies were conducted in high-income countries. The mean duration of interventions was 11.5 months (SD: 7.5), ranging from 3 to 44 months. Most interventions were effective for body mass index reduction, weight loss and waist circumference change. CONCLUSION: Our study showed that most interventions were effective for outcomes analysed in adults and the elderly. We also found some literature gaps, such as the need to implement and evaluate obesity after intervention and the requirement to carry out more studies in low- and middle-income countries.

Parotid salivary mucocele in a companion rabbit (Oryctolagus cuniculus): a report on the clinical and laboratory characteristics and surgical treatment - case report / [Mucocele salivar parotídea em coelho doméstico (Oryctolagus cuniculus): relato de características clínico-laboratoriais e tratamento cirúrgico - relato de caso]

The present report aimed to describe the main characteristics of the parotid salivary mucocele in an adult male rabbit, which presented with a painless fluctuating mass with fluid content. Owing to the need for repeated fluid drainage, surgical excision was the chosen mode of treatment. Preoperative evaluation included radiography and blood analysis. The patient underwent surgical excision of the affected gland. Inhalation anesthesia was maintained by means of isoflurane and the cardio-respiratory parameters were monitored throughout the surgery. A bipolar electrocautery was used to assist in the incision and polyglactin 910 and nylon were the suture materials used in the wound closure technique. Post-operative medications included analgesics, anti-inflammatory agents and antibiotics. Histopathology of the excised glandular tissue revealed a mixed inflammatory infiltrate, fibroblasts, hemorrhage and hyperemia. The observations in this case suggest that surgical excision of the affected gland is the treatment of choice for the management of mucocele in companion rabbits, which has already been identified for other animals. Periodic acid-reactive Schiff staining confirmed the diagnosis of salivary mucocele. The complementary investigations helped to rule out the differential diagnosis and indicated a favorable prognosis, in view of the fact that the case was successfully resolved without complications or recurrence.(AU)

O presente relato teve como objetivo descrever as principais características da mucocele salivar parotídea em um coelho macho adulto, que apresentou uma massa flutuante indolor e com um conteúdo líquido em seu interior. Devido à necessidade de repetidas punções deste fluido, optou-se pela excisão cirúrgica como forma de tratamento. A avaliação pré-operatória incluiu radiografias e análises sanguíneas. O paciente foi submetido ao tratamento cirúrgico com excisão da glândula afetada. A anestesia inalatória foi mantida com isoflurano e os parâmetros cardio-respiratórios foram monitorados durante a cirurgia. Um eletrocautério bipolar foi utilizado para auxiliar na incisão e os fios poliglactina 910 e nylon foram utilizados na técnica de sutura. As medicações pós-operatórias incluíram agentes analgésicos, anti-inflamatórios e antibióticos. A histopatologia do tecido glandular removido revelou infiltrado inflamatório misto, fibroblastos, hemorragia e hiperemia. As observações neste caso sugerem que a remoção cirúrgica da glândula afetada seja o tratamento de escolha no manejo da mucocele em coelhos de companhia, o que já vem sendo identificado para outros animais. A coloração do ácido periódico reativo de Schiff confirmou o diagnóstico da mucocele salivar. Os exames complementares ajudaram a excluir as doenças que fazem parte do diagnóstico diferencial e indicaram um prognóstico favorável, tendo em vista o fato que este caso foi resolvido com sucesso, sem complicações ou recorrência.(AU)

Molecular evidence of bovine herpesvirus 1 and 5 in cattle with suspected rabies in Rio Grande do Sul state, Brazil.

Rabies and herpetic encephalitis are the main viral infections in bovines with neurological symptoms. Bovine rabies has a high prevalence in Central and South America, while bovine encephalitis associated with herpesvirus is especially important in South America. Viral isolation is the classical way to confirm herpesvirus infection, but molecular evidence of the presence of the virus in affected animals is gaining importance in the diagnosis of the disease in the laboratory. This study investigated the presence of herpesvirus type 1 and 5 (BoHV-1 and BoHV-5) in 182 encephalon of rabies-suspected cattle in Rio Grande do Sul state (RS), Brazil using multiplex real-time polymerase chain reaction (mRT-PCR). The rabies virus was investigated by direct fluorescent antibody assay and intracerebral suckling mouse inoculation. The genomes of BoHV-1 and BoHV-5 were detected in 17% of samples. BoHV-5 and BoHV-1 were detected in 100% and 19% of BoHV positive samples, respectively, indicating the circulation of the pathogens in cattle herds in RS. The high Ct values and the absence of isolation suggest viral latency. Coinfection of herpesvirus and the rabies virus was detected in 28% of samples, although no significant association between pathogens was observed. Rabies was detected in 57.7% of suspected samples, confirming the importance of the disease in the state. Concerning the method by which samples were conserved, no significant difference was observed between the number of positive results in frozen and refrigerated samples.

Ecosystem service valuation method through grey water footprint in partially-monitored subtropical watersheds.

The valuation of ecosystem services of pollution regulation in basins with partial monitoring does not have only one consolidated methodology which can be applied in all countries, biomes and across spatio-temporal scales. While different metrics can incorporate elements of uncertainty for decision makers, changes in land use, climate and sectoral demands in basins increase the need for the efficiency and complexity of valuation methods. Here, based on adapting a pre-existing method, we present a new ecosystem service valuation applied to river basins under different characteristics in the biomes of the Atlantic Forest and Brazilian savannah. Our assumptions of ecosystem service valuation concern an analogy based on willingness-to-pay for not marketable services, but adapted by data from the river basins' ecohydrological monitoring. First, the method depicts river ecosystem valuation with probabilistic criteria of both the water yield, as supply, and the grey Water Footprint (greyWF), as demand. Second, we introduced the comparison between water supply and demand carried out on the continuous flow regime and monitored loads in rivers with different land uses, sizes and biomes. Third, this new ecosystem service valuation method enabled us to quickly visualize the possible stages of sustainability concerning the Brazilian legal framework among different basins. The methodology was applied in 12 Brazilian river basins, with drainage areas between 17 and 26,500 km2, and changes in land use with variable percentages of urban (62-92%), forest (51-84%) and agriculture (51-89%) areas. The most polluted basins, with greyWF values far above those allowed, have the most significant, almost asymptotic valuation curves. Results range from a minimum reference value of 61 US$/ha/year for conservation, adapted from the Brazilian Water Producer, to US$ 330 for restoring high polluted basins. The results show the viability of this method and discuss further opportunities for water security, especially for climate change and non-stationary sectorial demands.

Structural differences of commercial and recombinant lipase B from Candida antarctica: An important implication on enzymes thermostability.

Lipase B from Candida antarctica (CalB) is the most widely used lipase, including in many industrial sectors, such as in biodiesel and pharmaceuticals production. CalB has been produced by heterologous expression using Pichia pastoris under PGK constitutive promoter (named LipB). Here, we have studied the structural features of commercial CalB and LipB enzymes using circular dichroism and fluorescence under different conditions. In the presence of denaturing agents CalB was more stable than LipB, in contrast, at increasing temperatures, LipB was more thermostable than CalB. Mass spectrometry data indicates that both enzymes have an insertion of amino acids related to α-factor yeast signal, however LipB enzyme showed the addition of nine residues at the N-terminal while CalB showed only four residues. Molecular modeling of LipB showed the formation of an amphipathic α-helix in N-terminal region that was not observed in CalB. This data suggests that this new α-helix possess could be involved in LipB thermostability. These results associated with new structural studies may provide information to the design of novel biocatalysts.

Incident hand OA is strongly associated with reduced peripheral blood leukocyte telomere length.

OBJECTIVE: To evaluate the relationship of telomere length to the prevalence and incidence of hand osteoarthritis in a longitudinal cohort. DESIGN: We conducted a cross-sectional and longitudinal analysis of data from a subset of participants in the Osteoarthritis Initiative (OAI) recruited between February 2004 and May 2006. 274 individuals were eligible for the study based on availability of both baseline and 48-month hand radiographs and peripheral blood leucocyte telomere length data. Mean telomere length of peripheral blood leukocytes (PBL)s from the DNA samples was determined using a validated quantitative polymerase chain reaction (PCR)-based assay, and hand radiographs were analyzed and graded using the Kellgren-Lawrence scale. RESULTS: In joint -level analyses, prevalent Interphalangeal Joint Osteoarthritis (IPJOA) was significantly associated with PBL telomere length in the baseline sample in unadjusted analyses (RR = 2.84; 95% CI:0.87-9.29) or in models adjusted for age, sex, and body mass index (aRR = 1.10; 95% CI: 0.96-1.27). The association in crude and adjusted analyses appeared slightly stronger with incident IPJOA, especially in the subset with normal hands at baseline (aRR = 1.62; 95% CI: 1.02-2.57). PBL telomere length was also associated with prevalent HOA at baseline (significant in unadjusted analysis: RR = 1.22; 95% CI 1.06-1.42), but not after adjusting for covariates: aRR = 1.12; 95% CI: 0.96-1.30). The magnitude of association was stronger for incident HOA, especially incident symptomatic HOA (aRR = 1.53; 95% CI: 1.09-2.15). CONCLUSIONS: In summary, the results of this exploratory analysis are confirmatory of previous work showing a cross-sectional relationship between telomere length and HOA and add to the field by demonstrating an even stronger association with incident IPJOA, both radiographic and symptomatic.

Risk factors and seroprevalence of hepatitis E evaluated in frozen-serum samples (2002-2003) of pregnant women compared with female blood donors in a Southern region of Brazil.

Hepatitis E has always been related to morbidity in pregnant women. Its epidemiology is not well understood in Brazil. Therefore, we tested sera from 209 pregnant women and 199 female blood donors, collected at a single center in Curitiba, Brazil. The Wantai assay was used for testing the anti-hepatitis E virus (anti-HEV), immunoglobulin G (IgG), and an in-house polymerase chain reaction process for testing HEV RNA. Anti-HEV was detected in 22.5% of the total group, 19% in the pregnant women group, and 26% in the blood donor group (P = 0.11), a much higher prevalence when compared with other studies in Brazil. Demographical analysis showed that 92.4% were born in the South Region of Brazil, 4.9% in the Southeast, and 2.7% were distributed over other regions of the country. With respect to their origin, 99% were from the South, 0.7% from the Southeast, and 0.2% from the Central-West regions. Income, education, race, number of pregnancies, and abortion did differ significantly when comparing both the groups (P < 0.001). Age >30 (P = 0.012) and the number (>3) of pregnancies (P = 0.008) were related to anti-HEV positivity. All anti-HEV IgG-positive females were HEV RNA negative. In conclusion, HEV positivity was found in one out of five young women, which showed an urgent need for further epidemiological studies in Brazil.

Using the method of triads in the validation of a food frequency questionnaire to assess the consumption of fatty acids in adults.

BACKGROUND: It is of great value to develop valid instruments to estimate food consumption; for this purpose, the triads method has been applied in validation studies of dietary intake to evaluate the correlation between three measurements [food frequency questionnaire (FFQ), reference method and biomarker]. The main aim of the present study was to validate a FFQ for Brazilian adults by means of the method of triads by estimating the ingestion of total fatty acids based on the level of saturation. METHODS: The present study enrolled 152 Brazilian adults of both sexes, who were residents in the city of Viçosa, Brazil. The ingestion of total saturated, monounsaturated, polyunsaturated, trans, linoleic and linolenic fatty acids was assessed by means of a FFQ, two food records, and biomarkers, which were detected by gas chromatography. The validation coefficients were calculated using the method of triads and concordance was determined by Kappa statistics. RESULTS: The FFQ was considered an adequate dietary method, because, based on the validation coefficients, it estimates the intake of total fat (0.84), as well as linolenic (0.90) and linoleic acids (0.31). A high concordance rate was confirmed for all nutrients assessed by the FFQ and food records. Regarding the biomarkers, linolenic acid and linoleic acid presented greater concordance. CONCLUSIONS: According to the validation coefficients, the FFQ precisely estimated total fat, linolenic acid and linoleic acid contents.

Efficacy of a mephentermine-based product as a vasopressor and a cardiac performance enhancer when given intramuscularly to cattle.

The goal of this study was to confirm the vasopressor and cardiac effects of POTENAY® INJETÁVEL (POT), a mephentermine-based product, given to cattle with induced vascular/cardiac depression. Ten healthy Holstein cattle (206 ± 13 kg) followed a randomized-complete-block design (RCBD) utilizing crossover study design. Each animal randomly received (1 ml/25 kg, IM) of either POT (n = 10) or volume-matched placebo control (0.9%NaCl, CP, n = 10). A subset of animals (n = 5) received POT first (day 0) while the remaining (n = 5) received CP; after a six-day washout period, cattle received the opposite compound. Animals were anesthetized and catheterized for systemic/left ventricular hemodynamic monitoring. Myocardial dysfunction/hypotension was induced by increasing the end-tidal isoflurane concentration until arterial blood pressure was 20% lower than at baseline and remained stable. Once the animal was determined to be hypotensive and hemodynamically stable, steady-state hypotensive baseline data (BL2) were acquired, and treatment with either POT or CP was given. Data were acquired post-treatment at every 15 min for 90 min. POT improved cardiac output (+68 L/min, ±14%, p < 0.05), MAP (+14 mmHg, ±4%, p < 0.05), HR (+22 bpm, ±8%, p < 0.05), and peak rates of ventricular pressure change during both systole (dP/dtmax : +37 mmHg/s ±13%, p < 0.05) and diastole (dP/dtmin : +31 mmHg/s, ±7%, p < 0.05). No improvements were noted following placebo-control administration. Results indicate that POT improves cardiac performance and systemic hemodynamics in cattle with induced cardiovascular depression when given as single intramuscular injection.

Phytosterol-enriched milk lowers LDL-cholesterol levels in Brazilian children and adolescents: Double-blind, cross-over trial.

BACKGROUND AND AIMS: Despite evidence of the lipid-lowering effect of plant sterols among adults with hypercholesterolemia, data regarding phytosterol use in children are limited. In this paper, we examined the effects of daily consumption of a phytosterol-enriched milk compound on the lipid profiles of Brazilian children and adolescents with dyslipidemia. METHODS AND RESULTS: This was a randomized, double blind, crossover clinical trial. Twenty eight dyslipidemics outpatients (aged 6-9 years) from an University Hospital were randomly allocated to control or intervention group. The intervention group received milk enriched with 1.2 g/day of plant sterol and the control group received the equivalent amount of skim milk during the period of 8 weeks. Changes from baseline in the mean lipid profile, including total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and triglyceride (TG) concentrations. Serum lipid profiles, glucose levels, dietary and anthropometric data were determined at weeks 0, 4, 8, 16, and 20. Details regarding the safety and tolerance of phytosterol were obtained, using an open-ended questionnaire. Intention-to-treat analysis were performed, using the proc mixed procedure in SAS. After 8 weeks, the mean concentrations of TC and LDL-C were significantly reduced in the intervention group as compared to the control group with reductions of 5.9% (p = 0.09) and 10.2% (p = 0.002), respectively. In addition, TG concentrations were reduced by 19.7% (p = 0.09). No serious side effects were reported during the study. CONCLUSION: Our results confirm that plant sterols are an effective and safe treatment of infantile dyslipidemia. TRIAL REGISTRATION: RBR-3h7f9k.

Characterization of microbiota in Arapaima gigas intestine and isolation of potential probiotic bacteria.

AIMS: The aim of this study was to determine the intestinal microbiota of pirarucu (Arapaima gigas) in different growth stages (adult and fingerlings) and to isolate and identify potential probiotic bacteria. METHODS AND RESULTS: High-throughput sequencing analysis of the intestinal contents revealed that the majority of sequences belonged to the Proteobacteria, Fusobacteria and Firmicutes phyla. At the genus level, the greatest number of sequences belonged to Bradyrhizobium in adult fish, while Cetobacterium was the most abundant in juvenile fish. Twenty-three lactic-acid bacteria (LABs) were isolated on MRS agar from healthy juvenile fish. The isolates were tested in vitro for probiotic properties. Two isolates (identified as strains of Lactococcus lactis subsp. lactis and Enterococcus faecium) displayed antagonism against all 10 pathogens tested, were nonhaemolytic and maintained good viability for at least 3 weeks when supplemented to fish diets. The presence of a number of antibiotic resistance genes (ARGs), conferring resistance to erythromycin, tetracycline and chloramphenicol, was investigated by PCR. CONCLUSIONS: The absence of ARGs investigated the potential to antagonize pathogens, and favourable growth and survival characteristics indicate that these autochthonous isolates have the potential to be considered probiotics, which will be studied in future in vivo experiments. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has demonstrated, for the first time, the normal microbiota in the A. gigas intestine during different life stages and the presence of LAB strains. It also demonstrated LAB antibiotic resistance and antagonistic behaviour against pathogens isolated from the same fish.