Estimation of genetic parameters for weight and length gains in tambaqui (Colossoma macropomum).

The goal of this study was to determine the genetic parameters for growth traits in tambaqui (Colossoma macropomum) fish of similar age and weight. The data set included monthly measurements of body weight and length from120 fish for a year (1,440 measurements). The study found that weight gain had a heritability estimate of 0.01, while length gain had a heritability estimate of 0.27. There was a high positive correlation (0.84) between weight and length gains. The genetic variance and heritability of body weight decreased with age. This study suggests that breeding programs should select for body weight and weight gain to promote genetic gain in tambaqui. Selecting younger animals can also help reduce costs while still achieving genetic gains.

Phylogenomic and expression analysis of Colossoma macropomum cyp19a1a and cyp19a1b and their non-classical role in tambaqui sex differentiation.

The genes coding for Cytochrome P450 aromatase (cyp19a1a and cyp19a1b) and estrogen (E2) receptors (esr1, esr2a and esr2b) play a conserved role in ovarian differentiation and development among teleosts. Classically, the "gonad form" of aromatase, coded by the cyp19a1a, is responsible for the ovarian differentiation in genetic females via ligation and activation of the Esr, which mediates the endocrine and exocrine signaling to allow or block the establishment of the feminine phenotype. However, in neotropical species, studies on the molecular and endocrine processes involved in gonad differentiation as well as on the effects of sex modulators are recent and scarce. In this study, we combined in silico analysis, real-time quantitative PCR (qPCR) assay and quantification of E2 plasma levels of differentiating tambaqui (Colossoma macropomum) to unveil the roles of the paralogs cypa19a1a and cyp19a1b during sex differentiation. Although the synteny of each gene is very conserved among characids, the genomic environment displays striking differences in comparison to model teleost species, with many rearrangements in cyp19a1a and cyp19a1b adjacencies and transposable element traces in both regulatory regions. The high dissimilarity (DI) of SF-1 binding motifs in cyp19a1a (DI = 10.06 to 14.90 %) and cyp19a1b (DI = 8.41 to 13.50 %) regulatory region, respectively, may reflect in an alternative pathway in tambaqui. Indeed, while low transcription of cyp19a1a was detected prior to sex differentiation, the expression of cyp19a1b and esr2a presented a large variation at this phase, which could be associated with sex-specific differential expression. Histological analysis revealed that anti-estradiol treatments did not affect gonadal sex ratios, although Fadrozole (50 mg kg-1 of food) reduced E2 plasma levels (p < 0,005) as well cyp19a1a transcription; and tamoxifen (200 mg kg-1 of food) down regulated both cyp19a1a and cyp19a1b but did not influence E2 levels. Altogether, our results bring into light new insights about the evolutionary fate of cyp19a1 paralogs in neotropical fish, which may have generated uncommon roles for the gonadal and brain forms of cyp19a1 genes and the unexpected lack of effect of endocrine disruptors on tambaqui sexual differentiation.

Infrared thermography as a technique for detecting subclinical bovine mastitis / Termografia infravermelha como técnica para detecção de mastite bovina subclinical

Infrared thermography is a non-invasive diagnostic tool for early detection of subclinical mastitis in pasture-raised dairy cows. The study was conducted on eight dairy farms, in Sena Madureira, Acre, in the Western Amazon, during the rainy season. One hundred and thirteen lactating crossbred cows were monitored, with measurements performed once day before milking, from 02:00 to 06:00 in the morning. The California Mastitis Test (CMT) was subsequently applied. Data processing was performed in the R programming language and evaluated by linear models. Our results show that cow udder surface temperature by infrared thermography showed significant differences (p<0.05) between the months of the year. Cow udder temperatures of thermographic images were significantly different (p<0.05) from CMT results for animals with subclinical mastitis. CMT results showed that 45 cows (39.8%) in January, 52 cows (46%) in February and 57 cows (50.4%) in March had subclinical mastitis. Infrared thermography is a technique that can be used for the early detection of subclinical mastitis in dairy herds in a grazing system, as it detected an increase in the udder surface temperature of the same cows that were positive for CMT.

Avaliou-se a termografia infravermelha como ferramenta diagnóstica, não invasiva, para detecção precoce da mastite subclínica em vacas leiteiras criadas a pasto. O estudo foi conduzido em oito propriedades leiteiras, em Sena Madureira, Acre, na Amazônia Ocidental, durante o inverno amazônico. Foram monitoradas 113 vacas mestiças em lactação, com mensurações realizadas uma vez por dia, sempre antes da realizadas das 2h às 6h da manhã. Em seguida, realizou-se exame de Califórnia Mastitis Test-CMT. O ordenha, processamento dos dados foi feito em linguagem de programação R, os quais foram avaliados por modelos lineares. Os resultados mostram que a temperatura da superfície do úbere das vacas avaliadas com a técnica da termografia infravermelha teve diferenças significativas (P<0,05) entre os meses do ano. Constatou-se alteração na na temperatura do úbere das vacas, captada por meio das imagens termográficas, e foi i significativo (P<0,05) o resultado do teste California Mastistis Test (CMT) para os animais que se encontravam com mastite subclínica. Com base nos resultados do CMT, foi possível constatar que 45 vacas (39,8%) em janeiro, 52 vacas (46%) em fevereiro e 57 vacas (50,4%) em março se encontravam com mastite subclínica. Conclui-se que a termografia infravermelha é uma técnica que pode ser utilizada para a detecção precoce de mastite subclínica em rebanho leiteiro em sistema de pastejo, pois foi capaz de detectar aumento na temperatura superficial do úbere das vacas, que resultaram positivas no teste de CMT.

Effects of thermal environment on dairy cattle under a grazing system in the Western Amazon, Brazil / Efeitos do ambiente térmico em bovinos leiteiros em sistema de pastejo na Amazônia Ocidental, Brasil

This study aimed to evaluate the thermal conditions to which animals are exposed and their effect on the comfort/discomfort of dairy herds in a grazing system in the municipality of Sena Madureira, Acre State, Brazil. Eight farms and a total of 113 lactating crossbred cows were evaluated. Dry-bulb temperature (DBT, °C), relative humidity (RH, %), black globe temperature (BGT, °C), and wind speed (v, km h¹) were measured in January, February, and March (rainy season) to calculate temperature-humidity index (THI), black globe-humidity index (GTWBI), and radiant heat load (RHL). Moreover, the animals were measured for the physiological variables: rectal temperature (RT), heart rate (HR), and respiratory rate (RR). Significant differences (P<0.05) were observed for THI, GTWBI, and RHL. THI reached a mean value of 80 on farm G in January. Farm F had a GTWBI value of 79 in February. RHL reached the highest value (510 W m²). RT and HR showed no differences among the analyzed months. Thus, the environmental conditions in which dairy cows are housed at milking time rarely promoted thermal discomfort, with farm G being the most affected by THI.

Objetivou-se avaliar as condições térmicas a que os animais estão submetidos, seu efeito no conforto/desconforto de rebanhos leiteiros em sistema de pastejo, no município de Sena Madureira-Acre. Foram avaliadas oito propriedades e o total de 113 vacas mestiças em lactação. As medições ocorreram nos meses: janeiro, fevereiro e março (inverno amazônico), mensurando-se: temperatura do bulbo seco (TBS, °C), umidade relativa do ar (UR, %) e temperatura de globo negro (TGN, °C), velocidade do vento (v, km/h) para cálculos dos índices da temperatura e umidade (ITU), índice da temperatura de globo e umidade (ITGU) e carga térmica radiante (CTR), além da temperatura retal (TR), frequência cardíaca (FC), frequência respiratória (FR). Entre as propriedades analisadas, houve diferenças significativas (P<0,05) para ITU, ITGU e CTR. Destaca-se que o ITU na propriedade G, em janeiro, atingiu o valor médio de 80. A propriedade F, em fevereiro, teve valor de 79 para o ITGU. A CTR apresentou carga elevada (510 W.m³). A TR e a FC não apresentaram diferença entre os meses analisados. Concluiu-se que o ambiente ao qual as vacas leiteiras estão alojadas na hora da ordenha encontraram-se poucas vezes em situação de desconforto térmico, sendo a propriedade, a mais afetada pelo ITU.

Evaluation of the biological properties of two experimental calcium silicate sealers: an in vivo study in rats.

AIM: To evaluate the biological properties of experimental sealers based on tricalcium silicate and dicalcium silicate, manipulated with polyethylene glycol (CE-1) and with the addition of calcium hypochlorite (CE-2) compared to AH Plus (AHP) and TotalFill BC Sealer (TBC). METHODOLOGY: The tissue reaction caused by the materials in the subcutaneous tissue of rats was evaluated after implantation of polyethylene tubes filled with the materials at 7, 15, 30 and 60 days. Sections were stained with haematoxylin and eosin (HE) for morphological analysis and to evaluated number of inflammatory cells/mm2 (ICs). Sections were used for immunohistochemical detection of interleukin-6 (IL-6) and osteocalcin (OC). The von Kossa method was used to identify calcium precipitation in the capsules. The data were submitted to anova and Tukey's tests, with 5% significance level. RESULTS: At 7 days, CE-1, CE-2 and AHP had higher numbers of ICs. AHP presented higher immunolabelling for IL-6. After 15 days, regarding IL-6, there was no difference between CE-2 and the control group. At 30 days, AHP exhibited the highest number of IC (P < 0.05) and CE-2 and the control group presented the lowest ICs and IL-6-positive cells. After 60 days, all materials exhibited decreases in ICs. CE-2, TBC and the control had the lowest values (P < 0.05). No significant difference was detected between CE-1 and TBC, and between CE-2 and control considering the immunoexpression of IL-6. In this period, AHP had the greatest number of IC and IL-6 (P < 0.05). In all periods, CE-1, CE-2 and TBC sealers had von Kossa-positive structures and OC-immunolabelled cells. CE-2 had higher number of OC-positive cells than the CE-1 and TBC sealers (P < 0.05), in all periods. OC immunolabelling was not observed in the capsules of AH Plus and the control groups. CONCLUSIONS: The experimental sealer and its association with calcium hypochlorite, in addition to TotalFill BC Sealer, were biocompatible and had bioactive potential.

Histomorphometric and immunohistochemical study shows that tricalcium silicate cement associated with zirconium oxide or niobium oxide is a promising material in the periodontal tissue repair of rat molars with perforated pulp chamber floors.

AIM: To evaluate the periodontium response to tricalcium silicate (TCS) with zirconium oxide (ZrO2 ) or niobium oxide (Nb2 O5 ) used in the sealing of perforated pulp chamber floors in rat maxillary molars. METHODOLOGY: In eighty rats, the perforations in right maxillary molars were filled with either TCS + ZrO2 , TCS + Nb2 O5 , White MTA (used as a gold standard material) or no repair material was placed (Sham Group, SG); the left molars of SG, were used as controls (CG). Sections of maxillary fragments following 7, 15, 30 and 60 days were used to evaluate the volume densities of inflammatory cells (VvIC) and fibroblasts (VvFb), width of the periodontal space, amount of collagen, number of osteoclasts and number of IL-6-immunostained cells. The data were subjected to two-way ANOVA followed by Tukey's test (P ≤ 0.05). RESULTS: At all periods, significant differences in VvIC were not detected among TCS + ZrO2, TCS + Nb2 O5 and MTA groups, which had values significantly lower (P < 0.05) than the SG. Significant differences in the number of IL-6-immunolabelled cells were not observed among TCS + ZrO2 , TCS + Nb2 O5 and MTA groups (P > 0.05) at 15, 30 and 60 days. At 7, 15 and 30 days, the number of osteoclast was significantly greater in TCS + ZrO2, TCS + Nb2 O5 and MTA (P < 0.05) than in the CG; no significant difference was detected after 60 days (P > 0.05). The width of the periodontal space and amount of collagen in TCS + ZrO2 and TCS + Nb2 O5 groups were similar to the CG at 30 and 60 days while SG specimens had a significant reduction (P < 0.05) in the amount of collagen and significant increase (P < 0.05) in the width of the periodontal space. CONCLUSIONS: TCS + ZrO2 and TCS + Nb2 O5 were associated with periodontium repair since these materials allowed the reestablishment of periodontal space width and collagen formation when used in the filling of uninfected perforations in the pulp chamber floor of maxillary rat molars. Furthermore, the significant reduction in the periodontal space of TCS + ZrO2 and TCS + Nb2 O5 specimens after 60 days confirmed that the experimental materials were associated with a more rapid recovery of the injured tissues than MTA.

Mast cells and immunoexpression of FGF-1 and Ki-67 in rat subcutaneous tissue following the implantation of Biodentine and MTA Angelus.

AIM: To compare the formation of fibrous capsules around Biodentine and MTA Angelus implants as well as the participation of fibroblast growth factor-1 (FGF-1) and mast cells in the tissue response to these endodontic materials. METHODOLOGY: Sixty polyethylene tubes filled with Biodentine or MTA, and empty tubes (control group) were implanted into the dorsal subcutaneous tissues of male rats. After 7, 15, 30 and 60 days, the specimens were embedded in paraffin and the number of fibroblasts and mast cells was quantified in the sections stained with Masson's trichrome or Alcian Blue, respectively. FGF-1 and Ki-67 were detected by immunohistochemistry, and the number of immunolabelled cells was computed. The collagen content was estimated in the picrosirius red-stained sections. The data were subjected to two-way ANOVA followed by Tukey's test (P ≤ 0.05). RESULTS: The capsules were associated with a significant increase (P < 0.0001) in the number of fibroblasts and mast cells, and in the collagen content over time. A significant decrease (P < 0.0001) in the immunoexpression of FGF-1 and Ki-67 was observed in all groups from the 7th-60th day. At 60 days, the number of fibroblasts (P = 0.0226) and the collagen content (P < 0.0001) were significantly greater in MTA than Biodentine specimens, while the greatest number of mast cells and FGF-1-immunolabelled cells was observed in Biodentine specimens (P < 0.0001). A significant difference in Ki-67 immunoexpression was not detected between specimens of Biodentine and MTA. CONCLUSIONS: The collagen-rich capsule formed slowly around Biodentine in comparison with MTA. FGF-1 and mast cells participated in capsule remodelling, stimulating fibroblast proliferation and subsequent collagen production, in response to subcutaneous implants.

Biocompatibility of mineral trioxide aggregate flow and biodentine.

AIM: To evaluate the influence of powder-to-gel ratio (0.19 g powder to 50 µL of gel, thick MTA Flow, and 0.06 g powder to 50 µL of gel, fluid MTA Flow) on biocompatibility of MTA Flow (Ultradent Products Inc., South Jordan, UT, USA, lot: 2015122901) and compare it with Biodentine (Septodont Inc., Saint-Maur-des-Fossés, France, lot: B18542A). METHODOLOGY: The materials were manipulated and inserted into polyethylene tubes for implantation in twenty rats. After 7, 15, 30 and 60 days, the specimens were removed and embedded in paraffin. Haematoxylin and eosin sections were used to count the number of inflammatory cells (IC) and fibroblasts mm-2 (Fb). In the Masson's trichrome-stained sections, the fibrous capsule thickness was measured; picrosirius red-stained sections were used for birefringent collagen quantification. The data were submitted to two-way ANOVA and Tukey test (P ≤ 0.05). RESULTS: A significantly lower number of IC and consequently higher number of Fb were observed in the capsules adjacent to thick MTA Flow at all periods, in comparison with other materials (P ≤ 0.05). At 60 days, the quantity of birefringent collagen was significantly greater in the tissue in contact with thick MTA Flow, when compared with fluid MTA Flow and Biodentine. CONCLUSIONS: Although thick MTA Flow induced a less intense inflammatory response, all evaluated materials are biocompatible because they allowed regression of this process after 60 days.

Anthropometric changes caused by hepatitis C treatment with interferon and ribavirin: which patients and which measures are affected?

BACKGROUND: Chronic hepatitis C causes chronic hepatic inflammation, which can lead to cirrhosis, terminal liver failure, and hepatocellular carcinoma. The treatment aims to achieve viral clearance, but the usage of pegylated interferon and ribavirin is linked to side effects such as severe weight loss, which can lead to complications and treatment discontinuation. The aim of this study was to investigate which anthropometric measures were more affected in patients with chronic hepatitis C during 24 weeks of treatment with pegy lated interferon and ribavirin. Then, the influence of age, sex, hepatic fibrosis stage, and ribavirin doses on each measure was also evaluated. METHODS: Seventy-six patients were included and their weight, triceps skin fold thickness, arm circumference, middle-arm muscle circumference, and corrected arm muscle area were measured before and after 24 weeks of treatment. Epidemiological data and liver biopsy findings were obtained from patients' records. The sample was divided into two groups: one with advanced hepatic fibrosis and another group with mild to moderate fibrosis. Comparisons into each group were made using Wilcoxon or paired tests. After that, a linear regression model was applied to estimate the anthropometric changes during the treatment according to age, sex, hepatic fibrosis stage, and ribavirin doses. RESULTS: The subjects suffered reductions of important anthropometric measures, mainly related to fat mass (p<0.001).Some decrease of fat-free mass was also observed in subjects with advanced fibrosis. The statistic model showed that age and sex were more associated with the anthropometric changes observed. CONCLUSIONS: In conclusion, the antiviral treatment caused loss of relevant anthropometric measures, and the model proposed was able to estimate some of them

Identification of Colletotrichum isolates from Capsicum chinense in Amazon.

Chili pepper (Capsicum chinense) is a great economic important culture on the State of Amazonas, and it represents, approximately, a production of 1.9 thousand tons per year. It is one of the hosts of Colletotrichum genus in the North region of Brazil. The aim of the study was to differentiate and to identify isolates of Colletotrichum collected from C. chinense in Amazon. Molecular characterization, using RFLP-PCR, ERIC-PCR and ISSR, was carried out initially for screening of morphologically similar isolates. Furthermore, phylogenetic analyses were performed using combined regions: Actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for the three isolates, INPA 2066, INPA 2286 and INPA 1858, plus superoxide dismutase (SOD2) for INPA 2066. We showed that the molecular markers were able to distinguish the isolates of Colletotrichum studied and these results were confirmed with the phylogenetic analyses, three different occurrences of Colletotrichum species (C. siamense, C. scovillei and C. brevisporum) causing anthracnose in C. chinense in the State of Amazonas. This study represents the first report of the species C. siamense and C. scovillei in this host in Brazil.

Zirconium oxide and niobium oxide used as radiopacifiers in a calcium silicate-based material stimulate fibroblast proliferation and collagen formation.

AIM: To evaluate the influence of the addition of microparticulate (micro) and nanoparticulate (nano) zirconium oxide (ZrO2 ) and niobium pentoxide (Nb2 O5 ) to a calcium silicate-based cement (CS) on the subcutaneous healing process in rats compared with MTA Angelus™. METHODOLOGY: In each rat, two polyethylene tubes filled with the following materials: (i) MTA; (ii) CS + ZrO2 micro; (iii) CS + ZrO2 nano; (iv) CS + Nb2 O5 micro or (v) CS + Nb2 O5 nano were implanted subcutaneously; empty polyethylene tubes were used in the Control group. After 7, 15, 30 and 60 days, the specimens (n = 5 per group in each period) were fixed and embedded in paraffin. Masson's trichrome sections were used to obtain the volume density of the inflammatory cells (VvIC) and fibroblasts (VvFb). The sections were also stained with Picrosirius-red to calculate the birefringent collagen content. Fibroblast growth factor-1 (FGF-1) was detected by immunohistochemistry, and the number of immunolabelled cells was obtained. The data were subjected to two-way anova followed by Tukey's test (P ≤ 0.05). RESULTS: At all periods, the VvIC was significantly lower (P < 0.001) in all the CS and Control groups than in the MTA group. At all periods, the VvFb was reduced significantly (P = 0.023) in the MTA group in comparison with the other groups. In addition, the number of immunolabelled cells in the capsules of the CS groups was significantly higher (P < 0.001) than in the MTA group at all time-points. CONCLUSIONS: The experimental materials (CS + ZrO2 and CS + Nb2 O5 ) induced fibroblast proliferation and accelerated the regression of the inflammatory reaction. However, the addition of nanoparticulate radiopacifiers did not improve the biological properties of a calcium silicate-based cement when compared to microparticulate agents.

Influence of a commercial hatchery thermal environmental on the heat loss of fertile broiler eggs

The objectives of this study were to monitor the thermal environment of different hatchery locations during the transfer of fertile eggs from the setter and to the hatcher, to measure egg heat loss, and to determine its effects on hatchery results. In total, 1,728 fertile eggs of Cobb broiler breeders were divided into two treatments. In treatment 1 (T1), after 19 days of incubation, eggs were removed from the incubator and transferred to the hatcher in aninsulated box, and in treatment 0 (T0), eggs were transferred with no thermal insulation (T0). The duration of egg transfer was 10 minutes. Eggs were photographed using a thermographic camera at the exit of the setter, arrival at and exit from the candling room, and arrival at the hatcher. Based on the thermographic images, egg heat loss between these locations was calculated. At hatch, total hatchability, hatchability of fertile eggs, and hatchling weight were recorded and compared between T0 and T1. The temperature and relative humidity of the corridor between the setter and the candling room, of the candling room, of the corridor between candling roomand the hatcher were monitored using data loggers. The results indicated that T1 eggs lost 0.15 kJ less heat than T0 eggs during transfer. However, hatchability and hatchling weight were not affected by transfer treatment during the studies period.

Influence of a commercial hatchery thermal environmental on the heat loss of fertile broiler eggs

The objectives of this study were to monitor the thermal environment of different hatchery locations during the transfer of fertile eggs from the setter and to the hatcher, to measure egg heat loss, and to determine its effects on hatchery results. In total, 1,728 fertile eggs of Cobb broiler breeders were divided into two treatments. In treatment 1 (T1), after 19 days of incubation, eggs were removed from the incubator and transferred to the hatcher in aninsulated box, and in treatment 0 (T0), eggs were transferred with no thermal insulation (T0). The duration of egg transfer was 10 minutes. Eggs were photographed using a thermographic camera at the exit of the setter, arrival at and exit from the candling room, and arrival at the hatcher. Based on the thermographic images, egg heat loss between these locations was calculated. At hatch, total hatchability, hatchability of fertile eggs, and hatchling weight were recorded and compared between T0 and T1. The temperature and relative humidity of the corridor between the setter and the candling room, of the candling room, of the corridor between candling roomand the hatcher were monitored using data loggers. The results indicated that T1 eggs lost 0.15 kJ less heat than T0 eggs during transfer. However, hatchability and hatchling weight were not affected by transfer treatment during the studies period.(AU)

Post-transcriptional silencing of the SGE1 gene induced by a dsRNA hairpin in Fusarium oxysporum f. sp cubense, the causal agent of Panama disease.

Fusarium oxysporum f. sp cubense (Foc), the causal agent of Panama disease, is responsible for economic losses in banana crops worldwide. The identification of genes that effectively act on pathogenicity and/or virulence may contribute to the development of different strategies for disease control and the production of resistant plants. The objective of the current study was to analyze the importance of SGE1 gene expression in Foc virulence through post-transcriptional silencing using a double-stranded RNA hairpin. Thirteen transformants were selected based on different morphological characteristics, and sporulation in these transformants was significantly reduced by approximately 95% (P < 0.05) compared to that of the wild-type strain. The relative SGE1 expression levels in the transformant strains were reduced by 27 to 47% compared to those in the wild-type strain. A pathogenicity analysis revealed that the transformants were able to reach the rhizomes and pseudostems of the inoculated banana plants. However, the transformants induced initial disease symptoms in the banana plants approximately 10 days later than that by the wild-type Foc, and initial disease symptoms persisted even at 45 days after inoculation. These results indicate that the SGE1 gene is directly involved in the virulence of Foc. Therefore, SGE1 may be a potential candidate for host-induced gene silencing in banana plants.

Impacts of stocking on the genetic diversity of Colossoma macropomum in central Amazon, Brazil.

Tambaqui (Colossoma macropomum) is the main fish species farmed on a commercial scale in northern Brazil. In view of the current scenario of Brazilian aquaculture, studies on the genetic improvement and reproductive management of captive tambaqui are crucial in identifying the genetic variability of broodstocks and devising management practices. Genetic diversity of three tambaqui broodstocks in western Amazon was evaluated using molecular markers. Fin samples were collected from 89 fish; 38 from Balbina, 30 from a hatchery in Rio Preto da Eva, and 21 from the experimental farm of the Federal University of Amazonas (UFAM). Ten primers were used for the analysis of diversity and genetic structure. Of the 152 bands produced, 146 were polymorphic. The proportion of polymorphic loci showed little variation among the three stocks. The lowest and highest rates were found in the Rio Preto da Eva (80.92%) and Balbina (85.53%) stocks, respectively. Heterozygosity (H) and Shannon (I) indices were similar among the stocks; the lowest values were found in Balbina (H = 0.279 and I = 0.419), and the highest in UFAM (H = 0.294 and I = 0.439). Following analysis of the genetic structure and relationship, the sample was divided into two groups, with the Balbina stock clearly deviating from the others. The results suggest that, to increase genetic variability, molecular information may be used instead of replacement of wild breeders. The groups characterized here can be used in genetic improvement programs with other tambaqui broodstocks from different areas of South America.

Virulence and genetic diversity among isolates of Mycosphaerella fijiensis in two regions of Brazil.

Black sigatoka, caused by the fungus Mycosphaerella fijiensis (anamorphic stage: Paracercospora fijiensis), was first detected in Brazil in early 1998 in the Benjamin Constant and Tabatinga municipalities in the State of Amazonas, near to where the borders of Brazil, Colombia, and Peru converge. Understanding how cultivars react to the pathogen, and characterizing the genetic variability of isolates from two distant and distinct banana-producing regions, are important for determining the virulence of M. fijiensis. In the present study, the genetic diversity of 22 M. fijiensis isolates was assessed using simple sequence repeats (SSR) markers, and their virulence was determined following inoculation on three different banana tree cultivars. All 22 isolates caused symptoms of the disease in the Maçã and Prata Comum cultivars 45 days after inoculation, and at least two virulence groups were identified for the Maçã and Prata Comum cultivars. For the D'Angola cultivars, two virulence groups were observed only after 60 days post-inoculation, and three of the isolates were not virulent. Using SSR markers, the isolates from two different regions of Brazil were placed into two genetic groups, both genetically distant from the Mf 138 isolate collected in Leticia, Colombia. There was no evidence of correlation between the virulence groups and the genetic diversity groups. These results demonstrate variability in virulence between isolates as measured by the severity of black sigatoka in the analyzed cultivars.

Lower values of handgrip strength and adductor pollicis muscle thickness are associated with hepatic encephalopathy manifestations in cirrhotic patients.

Hepatic encephalopathy (HE) is a late complication of liver cirrhosis and is clearly associated with poor outcomes. Chronic liver insufficiency leads to progressive muscle wasting, impairing ammonia metabolism and thus increasing the risk for HE. Given the association between lean mass and adductor pollicis muscle thickness (APMT), it has been used to predict outcome and complications in many conditions, but not yet in cirrhotic patients. Therefore, this article aimed to study the association between HE manifestations and measures related to muscle mass and strength. This cross-sectional study included 54 cirrhotic outpatients with HE varying from subclinical to grade II according to the West-Haven criteria, who were submitted to neuropsychometric tests, electroencephalogram, brain Single Photon Emission Computed Tomography (SPECT), anthropometric measurements, handgrip strength (HGS) and dual energy X-ray absorptiometry exam (DXA). Multiple logistic regression analysis was performed to investigate the association between body composition measures and HE grade. Analysis of the area under the receiver operator characteristic (AUROC) curve revealed the values related to neurological manifestations (HE grades I and II). Reductions in APMT and HGS were associated with higher HE grades, suggesting a big impact caused by the loss of muscle mass and function on HE severity. The link between HE manifestations and anthropometric measures, namely APMT and HGS, point to a significant relation concerning skeletal muscles and the neurological impairment in this population.

SHORT-COMMUNICATION Validation of reference genes for real-time quantitative PCR in tambaqui (Colossoma macropomum).

Tambaqui, Colossoma macropomum, is the main native freshwater fish in Brazilian aquaculture. Therefore, intensive research pressure has been applied to the species to support new technologies for tambaqui farming. Molecular biology represents a tool that can be used to investigate every field of applied biology, from fish physiology to the effects of climate change. Based on the importance of reference genes for the relative or absolute quantification of gene transcripts, we cloned and sequenced three candidate reference genes in tambaqui (18S ribossomal RNA - 18s, glyceraldehyde-3-phosphate dehydrogenase - gapdh, and actin beta - ß-actin), and validated a set of primers for each gene for use in real-time quantitative PCR. The results were evaluated by RefFinder, which indicated that ß-actin is the most suitable reference gene for tambaqui among those studied, followed by 18s.

In vivo evaluation of the inflammatory response and IL-6 immunoexpression promoted by Biodentine and MTA Angelus.

AIM: To evaluate the inflammatory process induced by Biodentine and mineral trioxide aggregate (MTA) in rat subcutaneous tissues. METHODOLOGY: A polyethylene tube filled with Biodentine (n = 20) or MTA (n = 20) was placed into the dorsal subcutaneous of forty male rats; in the control group (CG; n = 20), empty tubes were implanted. After 7, 15, 30 and 60 days, the polyethylene tubes surrounded by connective tissue were fixed and embedded in paraffin. The number of inflammatory cells was estimated in HE-stained sections; numerical density of interleukin-6 (IL-6)-immunolabelled cells was also performed. The differences amongst the groups were analysed statistically by Tukey's test (P ≤ 0.05). RESULTS: A high number of inflammatory cells and IL-6-positive cells were observed at 7 days, in all groups; however, in the Biodentine group, the number of inflammatory cells and IL-6-immunolabelled cells was significantly higher (P ≤ 0.05) in comparison with the other groups at 7 and 15 days. In the capsules of animals from all groups, a gradual and significant reduction (P ≤ 0.05) of these parameters was seen over time. At 60 days, the capsules exhibited numerous fibroblasts and bundles of collagen fibres; in addition, the number of IL-6-positive cells was not significantly different amongst Biodentine, MTA and control groups. CONCLUSIONS: There was a significant regression in the inflammatory reaction in the capsules indicating, therefore, that Biodentine is a biocompatible material.