Effect of functional diets on intestinal microbiota and resistance to Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease (AHPND) of Pacific white shrimp (Penaeus vannamei).

AIMS: The present study evaluated the effect of four functional diets and a reference diet on the survival and intestinal bacterial community of shrimp Penaeus vannamei infected with acute hepatopancreatic necrosis disease (AHPND). METHODS AND RESULTS: After 42 days of feeding trail, shrimp were inoculated with a Vibrio parahaemolyticus (CIB-0018-3) carrying the plasmid encoding for the PirAB toxins responsible for AHPND. After 120 h postinfection (hpi), shrimp fed with a diet containing 2% of a mix with Curcuma longa and Lepidium meyenii (TuMa) and a diet containing 0.2% of vitamin C (VitC) showed a significantly higher survival (85%) compared to the remaining treatments (50%-55%) (p < 0.05). Infected shrimp fed with TuMa diet, showed a significant reduction of Vibrionales, and VitC diet promoted an increase of Alteromonadales. CONCLUSIONS: Our findings suggest that the TuMa diet conferred protection against AHPND and could be attributed to a combined effect of antibacterial properties against Vibrionales, and promoting a desirable bacterial community in the shrimp intestine, while the VitC diet protection could be attributed to their antioxidant capacity and in a lower proportion to a bacterial modulation in shrimp gut. SIGNIFICANCE AND IMPACT OF THE STUDY: Acute hepatopancreatic necrosis disease is a devastating disease that significantly affects aquaculture production of shrimps. Therefore, the use of functional diets that promote resistance to AHPND represents a valuable tool to reduce the mortality of farmed shrimp.

Adaptive mitochondrial response of the whiteleg shrimp Litopenaeus vannamei to environmental challenges and pathogens.

In most eukaryotic organisms, mitochondrial uncoupling mechanisms control ATP synthesis and reactive oxygen species production. One such mechanism is the permeability transition of the mitochondrial inner membrane. In mammals, ischemia-reperfusion events or viral diseases may induce ionic disturbances, such as calcium overload; this cation enters the mitochondria, thereby triggering the permeability transition. This phenomenon increases inner membrane permeability, affects transmembrane potential, promotes mitochondrial swelling, and induces apoptosis. Previous studies have found that the mitochondria of some crustaceans do not exhibit a calcium-regulated permeability transition. However, in the whiteleg shrimp Litopenaeus vannamei, contradictory evidence has prevented this phenomenon from being confirmed or rejected. Both the ability of L. vannamei mitochondria to take up large quantities of calcium through a putative mitochondrial calcium uniporter with conserved characteristics and permeability transition were investigated in this study by determining mitochondrial responses to cations overload. By measuring mitochondrial swelling and transmembrane potential, we investigated whether shrimp exposure to hypoxia-reoxygenation events or viral diseases may induce mitochondrial permeability transition. The results of this study demonstrate that shrimp mitochondria take up large quantities of calcium through a canonical mitochondrial calcium uniporter. Neither calcium nor other ions were observed to promote permeability transition. This phenomenon does not depend on the life cycle stage of shrimp, and it is not induced during hypoxia/reoxygenation events or in the presence of viral diseases. The absence of the permeability transition phenomenon and its adaptive meaning are discussed as a loss with biological advantages, possibly enabling organisms to survive under harsh environmental conditions.

Contribuições da prática de pilates na aptidão física e na força de preensão manual de idosos / Elderly physical fitness and manual grip strength improvement as a result of pilates

O objetivo deste estudo foi verificar a influência do Pilates de solo na aptidão física e na força de preensão manual de idosos. Participaram do estudo 11 idosos, nove mulheres e dois homens, com média de idade de 68,73 anos (DP= 6,06). Estes realizaram 34 sessões de exercícios do Pilates de Solo, em 17 semanas, duas vezes por semana, com duração de 60 minutos cada sessão. No início da intervenção foi aplicada a ficha diagnóstica em forma de entrevista individual. Antes e após a intervenção foram aplicados os seguintes instrumentos: medidas antropométricas, preensão manual e a bateria de testes físicos para idosos (Senior Fitness Test ­ SFT). Quanto às aptidões físicas verificou-se diferença significativa após o programa de Pilates de solo na força dos membros superiores e inferiores, flexibilidade de membros inferiores, agilidade/equilíbrio dinâmico e resistência aeróbia. Não houve diferença na flexibilidade de membros superiores e na força de preensão manual. Concluiu-se neste estudo que o Método Pilates de solo influenciou na melhora da força, flexibilidade de membros inferiores, agilidade/equilíbrio dinâmico e resistência aeróbia, demonstrando ser um método que proporciona benefícios na aptidão física dos idosos.

The purpose of this study was to verify the influence of Mat Pilates in physical fitness and manual grip strength of the elderly. Eleven elderly individuals - 9 women and two men - with mean age of 68.73 years (SD = 6.06) participated in the study. They were submitted to 34 sessions of Mat Pilates exercises twice a week for 17 weeks, with 60-minute sessions. At the beginning of the intervention, a diagnostic form was applied in the form of an individual interview. Before and after the intervention, the following instruments were applied: anthropometric measures, manual gripping and a set of physical tests for the elderly (Senior Fitness Test - SFT). Regarding physical fitness, there was a significant difference after the Pilates program on the upper and lower limb strength, lower limb flexibility, dynamic agility/balance, and aerobic resistance. No differences were found in upper limb flexibility and manual grip strength. It can be concluded that the Mat Pilates Method influenced the improvement of the strength, flexibility of lower limbs, dynamic agility/balance and aerobic resistance, proving to be a method that provides benefits in the physical fitness of the elderly.

The white spot syndrome virus hijacks the expression of the Penaeus vannamei Toll signaling pathway to evade host immunity and facilitate its replication.

The white spot syndrome virus (WSSV), the most lethal pathogen of shrimp, is a dsDNA virus with approximately a 300,000 base pairs and contains approximately 180-500 predicted open reading frames (ORFs), of which only 6% show homology to any known protein from other viruses or organisms. Although most of its ORFs encode enzymes for nucleotide metabolism, DNA replication, and protein modification, the WSSV uses some of its encoded proteins successfully to take control of the metabolism of the host and avoid immune responses. The contribution of the shrimp innate immune response to prevent viral invasions is recognized but yet not fully understood. Thus, the role of several components of Toll pathway of the shrimp Penaeus vannamei against WSSV has been previously described, and the consequential effects occurring through the cascade remain unknown. In the current study the effects of WSSV over various components of the shrimp Toll pathway were studied. The gene expression of Spätzle, Toll, Tube, Cactus and Dorsal was altered after 6-12 h post inoculation. The expression of LvToll3, LvCactus, LvDorsal, decreased ~4.4-, ~3.7- and ~7.3-fold at 48, 24 and 48 hpi, respectively. Furthermore, a remarkable reduction (~18-fold) in the expression of the gene encoding LvCactus in WSSV infected specimens was observed at 6 hpi. This may be a sophisticated strategy exploited by WSSV to evade the Toll-mediated immune action, and to promote its replication, thereby contributing to viral fitness.

Fibrobroncoscopía en pacientes onco-hematológicoscon infiltrados pulmonares, estudio transversal / Fibrobronchoscopy in onco-hematological patients with pulmonary infiltrates, cross-sectional study

Introducción: Las enfermedades infecciosas y masas pulmonares son una causa importante de mortalidad en los pacientes onco-hematológicos. El objetivo del presente estudio fue determinar la sensibilidad y especificidad de la fibrobroncoscopía (FBC) en el diagnóstico temprano de pacientes hemato-oncológicos. Métodos: El presente estudio descriptivo transversal se realizó en pacientes onco-hematológicos con hallazgos anormales en imágenes de tórax que fueron sometidos a FBC en el Instituto Oncológico de Solca-Guayaquil entre el periodo enero 2014 -diciembre 2016. Resultados: Ingresaron al estudio 98 pacientes: 54 mujeres (55.1%), de 61 ±18 años, 39 (39.8 %) con tumores sólidos, 76 (77.6 %) con síntomas respiratorios, 88 (89.8 %) con alta sospecha de lesión maligna. La sensibilidad del diagnóstico con líquido bronquial fue del 62.5 %; con cepillado, 88 % y del esputo post FBC fue de 14.3 %. La especificidad del diagnóstico con líquido bronquial fue del 89.7 %; con cepillado, 73.9 % y del esputo post FBC fue de 93.3 %. Conclusiones: En el presente estudio existe mayor sensibilidad con el cepillado bronquial comparativamente al líquido bronquial y el esputo post FBC para el diagnóstico de lesiones malignas

Introduction:Infectious diseases and lung masses are an important cause of mortality in onco-hematological patients. The aim of this study was to determine the sensitivity and specificity of Fiber Optic Bronchoscopy (FOB) in the early diagnosis of haemato-oncological patients. Methods:The present descriptive cross-sectional study was performed in onco-hematological patients with abnormal findings in chest images that were submitted to BCF in the Oncology Institute of Solca-Guayaquil between the period January 2014 to December 2016. Results: 98 patients, 54 women (55.1 %), 61 ± 18 years old, 39 (39.8 %) with solid tumors, 76 (77.6 %) with respiratory symptoms, 88 (89.8%) with high suspicion of malignant injury, entered the study. The sensitivity of the diagnosis with bronchial fluid was 62.5 %, with brushing 88 % and sputum post FOB was 14.3 %. The specificity of the diagnosis with Bronchial fluid was 89.7 %, with Brush 73.9 % and sputum post FOB was 93.3 %. Conclusions: In the present study there is greater sensitivity with Bronchial Brush comparatively to bronchial fluid and sputum post FOB.

Selection and validation of candidate reference genes for quantitative real-time PCR studies in the shrimp Penaeus vannamei under viral infection.

The decapod Penstyldensovirus 1 (PstDV-1) represents one of the most serious threats for penaeid shrimp farming. Studies aimed at defining relevant molecular effects of this virus over its host are imperative in the attempt to increase our understanding of its pathogenesis. Unfortunately, few studies have focused on the definition of the expression profile of reference genes in shrimp challenged with a pathogen. As a result, there are no studies on the selection of reference genes for the normalization of target gene expression changes yielding reliable data of the effects following PstDV-1 infection in shrimp. Therefore, the aim of the present study was to evaluate and validate the appropriateness of four candidate reference genes (ef1-α, gapdh, rpl8 and ß-tubulin) for their use as reference genes to normalize qPCR data in gene expression studies of PstDV-1-shrimp interactions. By analyzing the expression profile of those genes, gapdh was validated as a suitable reference gene to normalize expression data gathered from a PstDV1-challenge, while ef1-α, ß-tubulin, and rpl8 were identified as unstably expressed during the infectious process. The suitability of gapdh as a common reference gene in studies of host gene response to viral infections is underlined.

High Occurrence of the Decapod Penstyldensovirus (PstDV1) Detected in Postlarvae of Penaeus vannamei Produced in Commercial Hatcheries of Mexico.

The decapod penstyldensovirus (PstDV1) is a widely spread shrimp pathogen that causes high mortalities in the shrimp Penaeus stylirostris, while in P. vannamei, it has been associated with induction of the runt deformity syndrome. Using shrimp post-larvae (PL, stages PL13-PL21) collected from 16 commercial hatcheries from Mexico, and a sensitive PCR protocol for its detection, a survey of the PstDV1 prevalence in larvae was undertaken. A high overall prevalence of PstDV1 (49.5 %) in shrimp PL from the studied hatcheries was found. This study reveals that PstDV1 occurs persistently in PL populations, which may have significant implications for its dispersal.

An improved validated SYBR green-based real-time quantitative PCR assay for the detection of the Penaeus stylirostris densovirus in penaeid shrimp.

The Penaeus stylirostris densovirus (PstDV) (also known as infectious hypodermal and hematopoietic necrosis virus, IHHNV), one of the major shrimp pathogens, has a worldwide distribution in farmed and wild shrimp populations. Outbreaks of IHHNV have been associated with substantial economic losses which are accompanied by a negative social impact. Current diagnostic PCR tests may result in false-positive results as several parts of PstDV genome may be endogenized in the nuclear genome of the shrimp P. stylirostris. A one-step qPCR SYBR-Green based quantitative real-time polymerase chain reaction (qPCR) assay to detect different isolates of the IHHNV in shrimp samples was developed. The detection limit of the assay was 81 viral copies of targeted DNA per reaction. The specificity of the assay was evaluated by melting curve analysis, which showed that the IHHNV product generated a single melt peak at 81.4±0.044°C. The assay was more sensitive than conventional PCR. The standardized PCR was shown to be highly sensible, specific, robust, and reproducible, which makes it an economical and powerful tool for both diagnostic applications and general research of IHHNV.

Prevalence of the infectious hypodermal and hematopoietic necrosis virus in shrimp (Penaeus vannamei) broodstock in northwestern Mexico.

The Penaeus stylirostris densovirus (PstDNV or IHHNV) is the smallest of the known shrimp viruses. It causes severe mortalities in juveniles and sub-adults of the blue shrimp Penaeus stylirostris, while specimens of the white shrimp Penaeus vannamei infected by this virus exhibit reduced growth rates and negative effects on the feed-conversion rate (FCR). To date, no descriptive epidemiological surveys on the prevalence of this virus in shrimp broodstock have been performed. In this study, the prevalence of IHHNV in broodstock of the white shrimp P. vannamei from hatcheries on the northwest of Mexico region was estimated. Prevalence vary across different regions from high (63%) to low (6%) in shrimp broodstock. Several factors, as transport of pathogens by human activities, or the absence or implementation of ineffective biosecurity measures, may explain the observed differences. To the best of our knowledge, the present study is the first to examine the prevalence of IHHNV on broodstock.

The endemic copepod Calanus pacificus californicus as a potential vector of white spot syndrome virus.

The susceptibility of the endemic copepod Calanus pacificus californicus to white spot syndrome virus (WSSV) was established by the temporal analysis of WSSV VP28 transcripts by quantitative real-time PCR (qRT-PCR). The copepods were collected from a shrimp pond located in Bahia de Kino Sonora, Mexico, and challenged per os with WSSV by a virus-phytoplankton adhesion route. Samples were collected at 0, 24, 48 and 84 h postinoculation (hpi). The VP28 transcripts were not detected at early stages (0 and 24 hpi); however, some transcript accumulation was observed at 48 hpi and gradually increased until 84 hpi. Thus, these results clearly show that the copepod C. pacificus californicus is susceptible to WSSV infection and that it may be a potential vector for the dispersal of WSSV. However, further studies are still needed to correlate the epidemiological outbreaks of WSSV with the presence of copepods in shrimp ponds.

Experimental evidence of metabolic disturbance in the white shrimp Penaeus vannamei induced by the Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV).

The Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) is a single-stranded DNA virus that infects several penaeid shrimp species, provoking economic losses in farmed shrimp populations estimated at several million of dollars. Furthermore, IHHNV has historically been considered an important threat for wild shrimp populations, but its real measurable impact remains unknown. Currently no treatments are available against IHHNV, and research to develop potential antiviral strategies depends on a detailed understanding of the viral life cycle. However, the exact pathophysiological events underlying the development of metabolic changes in IHHNV-infected shrimp are still unknown. Thus, the biochemical changes caused by the IHHNV infection in plasma and hepatopancreas of the economically important shrimp species Penaeus vannamei were evaluated. Glucose, lactate, total protein, glycogen, triacylglycerides, cholesterol, and total lipids were measured in healthy and IHHNV-infected shrimp. Significant changes were observed in energy substrates (glucose, lactate, triacylglycerides and cholesterol), in plasma and hepatopancreas. These changes may indicate a temporal sequestration of the host-cell metabolic pathways by the virus to maximize its replication and propagation.

Experimental infection and detection of necrotizing hepatopancreatitis bacterium in the American lobster Homarus americanus.

Necrotizing hepatopancreatitis bacterium (NHPB) is an obligated intracellular bacteria causing severe hepatopancreatic damages and mass mortalities in penaeid shrimp. The worldwide distribution of penaeid shrimp as alien species threatens the life cycle of other crustacean species. The aim of the experiment was to evaluate the possibility of experimentally infecting the American lobster (Homarus americanus) with NHPB extracted from shrimp hepatopancreas. Homogenates from infected shrimp were fed by force to lobsters. Other group of lobsters was fed with homogenates of NHPB-free hepatopancreas. After the 15th day from initial inoculation, the presence of NHPB was detected by polymerase chain reaction in feces and hepatopancreas from lobsters inoculated with infected homogenates. Necrotized spots were observed in the surface of lobster hepatopancreas. In contrast, lobsters fed on NHPB-free homogenates resulted negative for NHPB. Evidence suggests the plasticity of NHPB which can infect crustacean from different species and inhabiting diverse latitudes. Considering the results, the American lobster could be a good candidate to maintain available NHPB in vivo.

[Burnout syndrome in medical staff affiliated to a tertiary care hospital]. / Síndrome de agotamiento profesional en personal médico de un hospital de tercer nivel.

INTRODUCTION: the burnout syndrome (BS) is common among physicians, affecting their behavior, personality and professional goals; thus deteriorating the quality of their performance. OBJECTIVE: to determine the BS prevalence among physicians affiliated to a tertiary care hospital and its associated risk factors. MATERIAL AND METHODS: The study was conducted through a cross-sectional design. The sampling was nonprobabilistic. Participating physicians were interviewed using the Maslach Burnout Inventory and the Beck's depression inventory. The analysis included ascertainment of the BS prevalence and use of chi(2) and logistic regression analysis. The local Internal Review Board and the Ethics Committee approved the project and all participants gave their informed consent. RESULTS: 166 physicians participated; the average age was 42 years, most were men. The response rate was 87.4 %. The prevalence of depression with more than 10 points in the Beck inventory was 18.6 % (95 % CI: 12-24). Depression in the last two years was 38.6%; the prevalence of BS was 26%; 15. 2 % scored above 27 in emotional exhaustion and 19.3 % scored more than 10 points in depersonalization. The risk of being depressed due to BS was five times higher (OR 5.163; 95 % CI 2.130-12.5). Working in the night shift, living without a permanent couple and being depressed were the main risk factors for BS. CONCLUSIONS: BS is common among physicians and is associated with organizational and personal variables.

Tentativa de suicídio e adesão ao tratamento: um estudo descritivo em hospital geral / Attempted suicide and treatment compliance: a descriptive study in the general hospital

Dados sócio-demográficos e clínicos foram coletados de forma padronizada em uma amostra consecutiva de 156 casos de tentativa de suicídio (TS) atendidos no pronto-socorro de um hospital geral universitário. Um programa de atendimento para esses pacientes incluiu a constituição de uma equipe multidisciplinar e estratégias para o tratamento ambulatorial. A adesão ao tratamento foi verificada após três meses da TS. Foram atendidos 110 (70,5 por cento) mulheres e 46 (29,5 por cento) homens, apresentando idade média de 27 anos e com 2/3 dos casos abaixo de 30 anos. Geralmente a TS foi ato impulsivo, com baixa intencionalidade suicida. Excesso de medicamentos e ingestão de venenos foram utilizados em 60 por cento e 20 por cento dos casos, respectivamente. Desentendimento com pessoa próxima ocorrido no último mês foi relatado por 70 por cento dos pacientes, funcionando como desencadeante da TS. Alguns pacientes já haviam passado por tratamento (35 por cento) e internação (15 por cento) psiquiátricos. O índice médio de comparecimento à primeira consulta ambulatorial foi de 55 por cento, sendo maior quando havia facilidade de transporte (65-78 por cento) e história de tratamento (68 por cento) e internação (79 por cento) psiquiátricos. Após três meses da TS, o abandono do tratamento chegou a 59 por cento. Estratégias para se aumentar a adesão através do correio não foram eficazes. Ao final desse período, os casos que permaneciam em tratamento em nosso serviço eram em sua maioria de transtornos psiquiátricos estabelecidos, semelhantes aos de um ambulatório psiquiátrico geral

Tentativa de suicidio e adesao ao tratamento: um estudo descritivo em hospital geral

Dados socio-demograficos e clinicos foram coletados de forma padronizada em uma amostra consecutiva de 156 casos de tentativa de suicidio (TS) atendidos no pronto-socorro de um hospital geral universitario. Um programa de atendimento para esses pacientes inclui a constituicao de uma equipe multidisciplinar e estrategias para o tratamento ambulatorial. A adesao ao tratamento foi verificada apos tres meses da TS. Foram atendidos 110(70,5 por cento) mulheres e 46(29,5 por cento) homens, apresentando idade media de 27 anos e com 2/3 dos casos abaixo de 30 anos. Geralmente a TS foi ato impulsivo, com baixa intencionalidade suicida. Excesso de medicamentos e ingestao de venenos foram utilizados em 60 por cento e 20 por cento dos casos, respectivamente. Desentendimento com pessoa proxima ocorrido no ultimo mes foi relatado por 70 por cento dos pacientes, funcionando como desencadeante da TS. Alguns pacientes ja haviam passado por tratamento (35 por cento) e internacao (15 por cento) psiquiatricos. O indice medio de comparecimento a primeira consulta ambulatorial foi de 55 por cento, sendo maior quando havia facilidade de transporte (65-78 por cento) e historia de tratamento (68 por cento) e de internacao (79 por cento) psiquiatricos. Apos tres meses da TS, o abandono do tratamento chegou a 59 por cento. Estrategias para se aumentar a adesao atraves do correio nao foram eficazes. Ao final desse periodo, os casos que permaneciam em tratamento em nosso servico eram em sua maioria de transtornos psiquiatricos estabelecidos, semelhantes aos de um ambulatorio psiquiatrico geral.