Optimization of management procedures for sea urchin (paracentrotus lividus) artificial reproduction

Aquaculture industry has a huge contribution for the world food production, and is extremely necessary to develop and optimize the production techniques, especially not to depend totally on wild stocks. In the case of the sea urchin Paracentrotus lividus, the demand increase of their roe is provoking a drastically decrease of their wild stocks, particularity in the recent years. This demand for sea urchin roe is arousing interest for the industry for a new profitable product and for the recovering and preserve wild populations. In this sense, new research needs to guarantee optimization of the techniques used to reduce the effort and the production cost. The selection of breeders is a very important aspect for sea urchin industry, since the success of the reproduction and quality of their product are dependent of the breeders. In addition to the high cost of acquiring wild specimens, it is not viable in the long term. Therefore, is important to study the possibility of use breeders from captive generations. The optimization of in vitro reproduction techniques is crucial for an efficiency production. The time that gametes need to be in contact for the best fertilization rate and the ratio of spermatozoa per oocyte are two of the principal aspects to be considered in an optimization of an in vitro reproduction protocol. Through preservation and cryopreservation technology is possible to promote alternatives techniques to contribute for the resource management efficiency of P. lividus. The objectives of this work are to study the hypothesis of using male wild sea urchins from different locations and captive generations specimens as breeders, optimize in vitro protocols using different contact times and different spermatozoa to oocyte ratio, and compare cryopreservation and preservation techniques for the storage of the sperm. To study the possibility of use captive generations as breeders were evaluated the sperm quality of each case, for this evaluation were analyzed the viability and motility, being used two approaches for the last one (mean values and subpopulation). The optimization of fertilization outcomes was evaluated counting the number of fertilized eggs using a counting chamber (Sedgwick) in optical microscope. Moreover, the comparation of the preservation and cryopreservation techniques was analyzed the motility by mean values, viability and DNA fragmentation (comet assay). The motility analysis were assessed by CASA system, viability through staining techniques and counting at a fluorescent microscope, DNA fragmentation was quantified the DNA tail. Significant differences were observed (p<0.05) in breeders selection, revealing the domestication seems to have a positive effect in P. lividus sperm quality. The in vitro spermatozoa to oocyte ration trial reveled significant differences between all ratios, where the 20 000 spermatozoa to oocyte ratio had the higher value. Contact times chosen differed negatively from the control. The preservation technique had higher viability in the first 6 days comparing to cryopreservation, although had no damage in DNA were detected in both. The present study revealed that domestication seems to be a good step to optimize the breeders sperm quality. With the higher spermatozoa to oocyte ratio used is possible to reach almost the total egg fertilization percentage. The preservation at 4ºC is a good technique to preserve the sperm quality until 6 days, cryopreservation seems to be better from there if the protocol is optimized.A indústria de aquacultura tem uma enorme contribuição para a produção de alimentos a nível mundial, e é extremamente necessário para desenvolver e otimizar as técnicas de produção, especialmente para não depender totalmente nos stocks selvagens. No caso do ouriço do mar Paracentrotus lividus, o aumento da procura das suas ovas tem provocado uma descida drástica dos stocks selvagens, particularmente nos últimos anos. Esta procura pelas ovas de ouriço do mar tem despertado o interesse da indústria para a produção de um novo produto lucrativo e para a recuperação e preservação das populações selvagens. Neste sentido, os futuros estudos devem garantir a otimização das técnicas usadas para reduzir o esforço e o custo da produção. A seleção dos progenitores é um aspeto muito importante para a indústria do ouriço do mar, visto que o sucesso da reprodução e a qualidade do produto final é dependente dos mesmos. Além do elevado custo da aquisição de espécimes selvagens, não é viável a longo termo a sua captura no meio natural. Portanto, o estudo da possibilidade de utilizar progenitores descendentes dos reproduzidos em cativeiro é bastante importante. A otimização das técnicas de reprodução in vitro é crucial para uma reprodução eficiente. O tempo que os gametas necessitam de estar em contacto para a melhor taxa de fertilização e o rácio de espermatozoides por oócito são dois dos principais aspetos a considerar na otimização de um protocolo de reprodução in vitro. Através de tecnologias de criopreservação e preservação é possível utilizar técnicas alternativas que podem contribuir para um maneio de recursos mais eficiente no caso do P. lividus. Os objetivos deste trabalho englobam o estudo da hipótese do uso de espécimes de ouriços do mar macho capturados em diferentes localizações geográficas e gerações de cativeiro como progenitores, otimização de protocolos in vitro usando tempos de contato e rácios espermatozoide para oócito diferentes, e comparar técnicas de criopreservação e preservação para o armazenamento do sémen. Para estudar a possibilidade do uso de espécimes selvagens e de diferentes gerações de cativeiro como progenitores foi avaliada a qualidade do sémen de cada caso, nesta avaliação foi analisada a viabilidade e a motilidade dos espermatozoides, tendo sido usadas duas abordagens diferentes para a última (valores médios e subpopulações). A otimização das técnicas de fertilização foi avaliada através da contagem do número de ovos fertilizados usando uma camara de contagem (Sedwick) num microscópio ótico. Por fim, na comparação das técnicas de criopreservação e preservação foi analisada a motilidade através de valores médios, a viabilidade e a fragmentação do ADN (teste de cometas). A análise da motilidade foi efetuada usando o sistema CASA (Computer-assisted sperm analysis), viabilidade através de técnicas de coloração fluorescente e contagens num microscópio de fluorescência, a fragmentação do ADN avaliada através da quantificação cauda do ADN. Foram observadas diferenças significativas (p<0.05) na seleção dos progenitores, revelando que a domesticação parece ter um efeito positivo na qualidade do sémen de P. lividus. Os ensaios in vitro com diferentes rácios de espermatozoide por oócito mostraram diferenças significativas em todos os rácios, sendo que o rácio de 20 000 espermatozoides por oócito teve a maior percentagem de fertilização. Os tempos de contacto escolhidos diferenciaram negativamente do controlo. Comparando com a técnica de criopreservação, a técnica de preservação teve maior viabilidade nos primeiros 6 dias, no entanto não foi detetado nenhum dano no ADN em ambas as técnicas. Este trabalho revelou que a domesticação parece ser um bom passo para a otimização da qualidade do sémen dos progenitores de P. lividus. Com o maior rácio de espermatozoides por oócito foi possível atingir quase a percentagem total de ovos fertilizados. A preservação a 4ºC é uma boa técnica para preservar a qualidade do sémen até aos 6 dias, depois deste tempo, a criopreservação parece ser melhor, contundo o protocolo necessita de ser otimizado.Queria expressar o meu agradecimento a alguns projetos que foram fundamentais para a realização deste trabalho. Dois projetos da EPPO (Estação Piloto de Piscicultura de Olhão /IPMA - Instituto Português do Mar e da Atmosfera), OURIÇAQUA (FA_05_2017_009) e DIVERSIAQUA II (MAR2020-P02M01-0656P), e ao projeto do Centro de Ciências do Mar (CCMAR), Universidade do Algarve denominado por EBB European Blue Biobank (EBB-EAPA_501/2016 (Interreg Atlantic Area)

Atypical presentation of colon cancer: perforation with hepatic fistulization

One of the known and rare complications of the colon cancer is perforation, with fistulas or abscesses formation. In this paper it is reported the case of a 61 year old man who presented himself with pain and a palpable mass in the right upper abdominal quadrant in association with fever and weight loss. The radiological investigation showed findings compatible with perforated colon cancer and hepatic fistulization. An emergent surgery was performed and confirmed the radiologic suspicion

Morgagni Hernia Associated with Hiatus Hernia, a Rare Case

The simultaneous occurrence of two separate non-traumatic diaphragmatic hernias is extremely rare. We report a case of an old man with two diaphragmatic hernias (Morgagni and Hiatal hernias) and we also review the clinical and imagiologic features (Radiographic and Computed Tomography) of Morgagni and hiatal herniation

Contents of ethyl carbamate in sugar cane and manioc spirits: part II

Herein, we report the concentration of ethyl carbamate (EC) and copper in 380 samples of sugar-cane spirit and 45 samples of manioc spirit as determined by GC-MS and FAAS respectively. The cyanide content determined spectrophotometrically is reported for the manioc spirit. Sugar cane spirit produced by alembic distillation (70,0 µg L-1) shown a lower content of EC than samples produced by column distillation (270 µg L-1). No simple correlation between the content of EC and copper for sugar cane spirit as well among the concentration of EC, copper, and cyanide for manioc spirit could be observed.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)CNPqFAPES

Comparação entre a água de coco em pó (ACP®) e o Tris como diluidores na criopreservação do sêmen de cães

The present study compared powder coconut water (ACP®) and Tris extenders on canine semen cryopreservation by classic evaluation and thermoresistance test. Five stud dogs were submitted to two semen collections by digital manipulation. Sperm fractions were analyzed regarding to its color, volume, sperm concentration, motility, vigor and morphology. Semen samples were divided into two aliquots: the first one was extended in Tris and second one in ACP®. Both extenders contained 20% egg yolk. Samples were cooled to 5ºC, glycerol added (6%), packaged in 0.25mL straws, frozen in nitrogen vapors and finally stored in liquid nitrogen. One week later, thaw was performed at 38ºC per 1min in water bath and new evaluations of sperm motility, vigor and morphology were conducted. Samples were kept at 38ºC in water bath and evaluated at 15 and 30min after thaw. No significant difference was observed between extenders throughout cryopreservation or thawing procedures, as well as during thermoresistance test, concerning to characteristics evaluated. Tris and ACP® were efficient in conserving 50% mobile spermatozoa and 70% morphologically normal sperm after thaw. Thus, ACP® can be used as an alternative extender for canine semen cryopreservation.O presente trabalho comparou a água de coco em pó (ACP®) com o diluidor Tris na criopreservação do sêmen canino através da avaliação clássica e do teste de termorresistência. Cinco reprodutores caninos foram submetidos a duas coletas de sêmen através de manipulação digital. As frações espermáticas foram avaliadas quanto à sua coloração, volume, concentração, motilidade, vigor e morfologia espermática. As amostras de sêmen foram divididas em duas alíquotas: a primeira foi diluída em Tris e a segunda em ACP®. Ambos os diluidores continham 20% de gema de ovo. As amostras foram refrigeradas, adicionadas de glicerol (6%), envasadas em palhetas de 0,25mL, criopreservadas em vapores de nitrogênio, e finalmente armazenadas em nitrogênio líquido. Após uma semana, foi realizada a descongelação a 38ºC por 1min em banho-maria, sendo realizadas novas avaliações de motilidade, vigor e morfologia espermática. As amostras foram mantidas a 38ºC no banho-maria e avaliadas aos 15 e 30 min após descongelação. Não foram observadas diferenças significativas entre os dois diluidores no decorrer dos procedimentos de criopreservação e descongelação, bem como durante o teste de termorresistência, em relação às características avaliadas. O Tris e o ACP® foram eficientes em conservar 50% de espermatozóides móveis e 70% de espermatozóides morfologicamente normais após a descongelação. Assim, o ACP® pode ser utilizado como diluidor alternativo para a criopreservação do sêmen canino

Multivariate analyses of UV-Vis absorption spectral data from cachaca wood extracts: a model to classify aged Brazilian cachacas according to the wood species used

Multivariate analyses of UV-Vis spectral data from cachaca wood extracts provide a simple and robust model to classify aged Brazilian cachacas according to the wood species used in the maturation barrels. The model is based on inspection of 93 extracts of oak and different Brazilian wood species by a non-aged cachaca used as an extraction solvent. Application of PCA (Principal Components Analysis) and HCA (Hierarchical Cluster Analysis) leads to identification of 6 clusters of cachaca wood extracts (amburana, amendoim, balsamo, castanheira, jatoba, and oak). LDA (Linear Discriminant Analysis) affords classification of 10 different wood species used in the cachaca extracts (amburana, amendoim, balsamo, cabreuva-parda, canela-sassafras, castanheira, jatoba, jequitiba-rosa, louro-canela, and oak) with an accuracy ranging from 80% (amendoim and castanheira) to 100% (balsamo and jequitiba-rosa). The methodology provides a low-cost alternative to methods based on liquid chromatography and mass spectrometry to classify cachacas aged in barrels that are composed of different wood species.FAPESPCAPESCNP

A bibliometric analysis of research productivity on parasitic infections in children during the 15-year period / Uma análise bibliométrica da produtividade da investigação sobre infecções parasitárias em crianças durante o período de 15 anos

Parasitic infections in children (PIC) represent an important public health issue regarding that children are more vulnerable to these infections and its consequences. Many countries have neglected PIC in their health agendas and budgets, which has hindered scientific production about it. In this sense, bibliometric analyzes have been useful to identify the panorama of scientific production for many diseases and thus assist in decision-making. Thus, this review aims to analyze the brazilian and worldwide scientific production of PIC between 2006 and 2020. The terms “Parasitic diseases” and “Children” were used for search in Scopus database. The variables were organized and simple linear regression was applied using Graphpad Prism 7. Values of p&lt;0.05 were considered statistically significant. was USA (18.5%), UK (8.9%) and Brazil (7.8%) were the countries that performed more scientific research on IPC. Among Brazil´s federal unities, São Paulo (34.3%), Rio de Janeiro (32%) and Minas Gerais (17.3%) published more scientific papers on PIC. The Brazilian research were mostly performed by public institutions, especially FIOCRUZ, USP and UFMG which published 19%, 16% and 10.6% of total Brazilian scientific production about PIC, respectively. There was a significant increase in scientific production about PIC over the 15 years analyzed, especially about schistosomiasis, giardiasis and trichuriasis. In addition, scientific journals from developed countries such as London School of Hygiene &amp; Tropical Medicine and Plos Neglected Tropical Diseases were the main publishers of papers about PIC. Scientific production on PIC increased between 2006 and 2020 with participation of developed countries (USA and UK) and developing countries (Brazil). This increase in scientific production seems to be driven by the increase in studies on giardiasis, trichuriasis and especially schistosomiasis

Transcranial Direct Current Stimulation on Parkinson’s Disease: Systematic Review and Meta-Analysis

Background: Clinical impact of transcranial direct current stimulation (tDCS) alone for Parkinson’s disease (PD) is still a challenge. Thus, there is a need to synthesize available results, analyze methodologically and statistically, and provide evidence to guide tDCS in PD. Objective: Investigate isolated tDCS effect in different brain areas and number of stimulated targets on PD motor symptoms. Methods: A systematic review was carried out up to February 2021, in databases: Cochrane Library, EMBASE, PubMed/MEDLINE, Scopus, and Web of science. Full text articles evaluating effect of active tDCS (anodic or cathodic) vs. sham or control on motor symptoms of PD were included. Results: Ten studies (n=236) were included inmeta-analysis and 25 studies (n=405) in qualitative synthesis. The most frequently stimulated targets were dorsolateral prefrontal cortex and primary motor cortex. No significant effect was found among single targets on motor outcomes: Unified Parkinson’s Disease Rating Scale (UPDRS) III – motor aspects (MD = −0.98%, 95% CI = −10.03 to 8.07, p = 0.83, I² = 0%), UPDRS IV – dyskinesias (MD = −0.89%, CI 95%= −3.82 to 2.03, p = 0.55, I² = 0%) and motor fluctuations (MD = −0.67%, CI 95% = −2.45 to 1.11, p = 0.46, I² = 0%), timed up and go – gait (MD = 0.14%, CI 95% = −0.72 to 0.99, p = 0.75, I² = 0%), Berg Balance Scale – balance (MD = 0.73%, CI 95% = −1.01 to 2.47, p = 0.41, I² = 0%). There was no significant effect of single vs. multiple targets in: UPDRS III – motor aspects (MD = 2.05%, CI 95% = −1.96 to 6.06, p = 0.32, I² = 0%) and gait (SMD = −0.05%, 95% CI = −0.28 to 0.17, p = 0.64, I² = 0%). Simple univariate meta-regression analysis between treatment dosage and effect size revealed that number of sessions (estimate = −1.7, SE = 1.51, z-score = −1.18, p = 0.2, IC = −4.75 to 1.17) and cumulative time (estimate = −0.07, SE = 0.07, z-score = −0.99, p = 0.31, IC = −0.21 to 0.07) had no significant association. Conclusion: There was no significant tDCS alone short-term effect on motor function, balance, gait, dyskinesias or motor fluctuations in Parkinson’s disease, regardless of brain area or targets stimulated

Comparative untargeted metabolome analysis of ruminal fuid and feces of Nelore steers (Bos indicus).

We conducted a study to identify the fecal metabolite profle and its proximity to the ruminal metabolism of Nelore steers based on an untargeted metabolomic approach. Twenty-six Nelore were feedlot with same diet during 105 d. Feces and rumen fuid were collected before and at slaughter, respectively. The metabolomics analysis indicated 49 common polar metabolites in the rumen and feces. Acetate, propionate, and butyrate were the most abundant polar metabolites in both bio-samples. The rumen presented signifcantly higher concentrations of the polar compounds when compared to feces (P< 0.05); even though, fecal metabolites presented an accentuated representability of the ruminal fuid metabolites. All fatty acids present in the ruminal fuid were also observed in the feces, except for C20:2n6 and C20:4n6. The identifed metabolites ofer information on the main metabolic pathways (higher impact factor and P< 0.05), as synthesis and degradation of ketone bodies; the alanine, aspartate and glutamate metabolisms, the glycine, serine; and threonine metabolism and the pyruvate metabolism. The fndings reported herein on the close relationship between the ruminal fuid and feces metabolic profles may ofer new metabolic information, in addition to facilitating the sampling for metabolism investigation in animal production and health routines