Práctica deportiva e Inteligencia Emocional en jóvenes. Revisión sistemática / Prática desportiva e Inteligência Emocional em Jovens. Revisão sistemática / Sports practice and Emotional Intelligence in young people. Systematic review

La práctica deportiva y la Inteligencia Emocional en jóvenes se están convirtiendo en constructos muy estudiados en los últimos años. La evidencia disponible revela que, tanto la práctica deportiva como la Inteligencia Emocional, producen beneficios físicos y psicológicos. No obstante, los estudios que analizan la relación entre Inteligencia Emocional y práctica deportiva aún siguen siendo escasos. El objetivo de este estudio se centró en verificar si existe algún tipo de relación entre ambas variables mediante la revisión del estado de la cuestión en este ámbito, permitiendo de esta manera abrir nuevas líneas de investigación sobre esta temática. Se seleccionaron tres bases de datos (WOS, Scopus y PubMed) para la extracción de documentos científicos, analizando los documentos obtenidos con la metodología PRISMA y un análisis de la calidad de la muestra final obtenida. Finalmente fueron revisados 38 artículos. Además, se llevó a cabo una evaluación del contenido de los artículos, realizando un análisis descriptivo de la información mediante el uso de tablas de contingencia y sus índices estadísticos. Los resultados, en concordancia con la literatura examinada, reflejaron la existencia de una asociación positiva entre la Inteligencia Emocional y la práctica deportiva. (AU)

The practice of sports and emotional intelligence in young people have become very much studied constructs in recent years. The aim of this study focused on verifying whether there is any kind of relationship between the two variables by reviewing the state of the art in this field, thus allowing new lines of research to be opened up on this subject. Three databases (WOS, Scopus, and PubMed) were selected for the extraction of scientific documents, analyzing the documents obtained with the PRISMA methodology and an analysis of the quality of the final sample obtained, finally 38 articles were selected. An analysis of the content of the articles was carried out and a descriptive analysis of the information was performed using comparative tables and their statistical indices. The results showed the existence of a positive association between emotional intelligence and sports practice. (AU)

A prática do desporto e a inteligência emocional nos jovens tornaram-se trmas muito estudadas nos últimos anos. O objectivo deste estudo centrou-se em verificar se existe algum tipo de relação entre as duas variáveis através da revisão do estado da arte neste campo, permitindo assim a abertura de novas linhas de investigação sobre este assunto. Foram seleccionadas três bases de dados (WOS, Scopus e PubMed) para a extracção de documentos científicos, analisando os documentos obtidos com a metodologia PRISMA e uma análise da qualidade da amostra final obtida, onde foram incluidos 38 artigos. Foi realizada uma análise do conteúdo dos artigos e uma análise descritiva da informação, utilizando tabelas de comparação e os seus índices estatísticos. Os resultados mostraram a existência de uma associação positiva entre a inteligência emocional e a prática desportiva. (AU)

Characterization of lesions of nutritional congenital goitre in cattle.

There are few studies that classify and characterize the morphometric and immunohistochemical features of goitre in bovine thyroid glands (TGs). We investigated 39 bovine TGs (fetuses [9], stillbirths [18], neonates [12]) born to dams with low T4 hormone levels and no iodine supplementation and 10 (fetuses [3], stillbirths [3], neonates [4]) born to dams with normal T4 levels and supplemented with iodine. Body weight (BW), TG weight (TGW), TGW:BW ratio and histological lesions were determined. The TGs were classified histopathologically as normal gland (G0), mild goitre (G1), moderate goitre (G2) or severe goitre (G3). Various morphological and morphometric parameters were calculated from microscopic images using image analysis software. Immunohistochemistry was performed to detect proliferating cell nuclear antigen (PCNA). There were significant differences in the TGW:BW ratio among groups (P <0.05): 0.3 ± 0.1 in G0, 0.5 ± 0.3 in G1, 0.8 ± 0.3 in G2 and 1.3 ± 0.7 in G3. In G0, large homogeneous follicles with eosinophilic colloid were seen. In the groups with lesions (G1, G2 and G3), heterogeneity in follicle shape and size, height and area of thyroid follicular cells, height of thyroid follicular epithelium and PCNA immunolabelling were directly related to histopathological grade, with significant differences among groups (P <0.001), gradually increasing from G1 to G3 compared with G0. The TGW:BW ratio and histological grade were positively correlated (P = 0.008).

Detection of a novel microsporidium with intranuclear localization in farmed Penaeus vannamei from Latin America.

Microsporidia are emerging intracellular parasites of most known animal phyla in all ecological niches. In shrimp aquaculture, the microsporidium Enterocytozoon hepatopenaei (EHP) is a major cause of concern inflicting tremendous losses to shrimp producers in southeast Asia. During a histopathological examination of Penaeus vannamei samples originating in a country from Latin America presenting slow growth, we observed abnormal nuclei in the epithelial cells of the hepatopancreas. A PCR screening of the samples using DNA isolated from paraffin embedded tissues for the SSU rRNA gene of EHP provided a 149 bp amplicon. In situ hybridization using the SSU rRNA gene probe provided a positive signal in the nuclei instead of the cytoplasm. Sequence analysis of the SSU rRNA gene product revealed a 91.3 %, 89.2 % and 85.4 % sequence identity to Enterocytozoon bieneusi, E. hepatopenaei and Enterospora canceri respectively. Furthermore, phylogenetic analysis revealed the newly discovered microsporidium clustered with E. bieneusi. Considering the intranuclear location of the novel microsporidium and the differences in the sequence of the SSU rRNA, we tentatively consider this parasite a new member of the genus Enterospora sp. The pathogenicity and distribution of the shrimp Enterospora sp. are currently unknown. Our future efforts are focused on the characterization and development of diagnostic tools for this parasite to understand if it acts as an emergent pathogen that might require surveillance to prevent its spread.

An efficient strategy for evaluating new non-invasive screening tests for colorectal cancer: the guiding principles.

OBJECTIVE: New screening tests for colorectal cancer (CRC) are rapidly emerging. Conducting trials with mortality reduction as the end point supporting their adoption is challenging. We re-examined the principles underlying evaluation of new non-invasive tests in view of technological developments and identification of new biomarkers. DESIGN: A formal consensus approach involving a multidisciplinary expert panel revised eight previously established principles. RESULTS: Twelve newly stated principles emerged. Effectiveness of a new test can be evaluated by comparison with a proven comparator non-invasive test. The faecal immunochemical test is now considered the appropriate comparator, while colonoscopy remains the diagnostic standard. For a new test to be able to meet differing screening goals and regulatory requirements, flexibility to adjust its positivity threshold is desirable. A rigorous and efficient four-phased approach is proposed, commencing with small studies assessing the test's ability to discriminate between CRC and non-cancer states (phase I), followed by prospective estimation of accuracy across the continuum of neoplastic lesions in neoplasia-enriched populations (phase II). If these show promise, a provisional test positivity threshold is set before evaluation in typical screening populations. Phase III prospective studies determine single round intention-to-screen programme outcomes and confirm the test positivity threshold. Phase IV studies involve evaluation over repeated screening rounds with monitoring for missed lesions. Phases III and IV findings will provide the real-world data required to model test impact on CRC mortality and incidence. CONCLUSION: New non-invasive tests can be efficiently evaluated by a rigorous phased comparative approach, generating data from unbiased populations that inform predictions of their health impact.

Outlier Vehicle Trajectory Detection Using Deep Autoencoders in Santiago, Chile.

In the last decade, a large amount of data from vehicle location sensors has been generated due to the massification of GPS systems to track them. This is because these sensors usually include multiple variables such as position, speed, angular position of the vehicle, etc., and, furthermore, they are also usually recorded in very short time intervals. On the other hand, routes are often generated so that they do not correspond to reality, due to artifacts such as buildings, bridges, or sensor failures and where, due to the large amount of data, visual analysis of human expert is unable to detect genuinely anomalous routes. The presence of such abnormalities can lead to faulty sensors being detected which may allow sensor replacement to reliably track the vehicle. However, given the reliability of the available sensors, there are very few examples of such anomalies, which can make it difficult to apply supervised learning techniques. In this work we propose the use of unsupervised deep neural network models based on stacked autoencoders to detect anomalous routes in vehicles within Santiago de Chile. The results show that the proposed model is capable of effectively detecting anomalous paths in real data considering validation given by an expert user, reaching a performance of 82.1% on average. As future work, we propose to incorporate the use of Long Short-Term Memory (LSTM) and attention-based networks in order to improve the detection of anomalous trajectories.

A laboratory challenge model for evaluating enyterocytozoon hepatopenaei susceptibility in selected lines of pacific whiteleg shrimp Penaeus vannamei.

Here we report for the first time a laboratory challenge model for Enterocytozoon hepatopenaei (EHP) to determine the difference of two Specific Pathogen Free (SPF) lines of Penaeus vannamei shrimp. These lines were experimentally challenged using EHP-infected fecal strings as inoculum. Real-time PCR and histopathology assays were performed to confirm EHP infection and evaluate differences in EHP susceptibility in the two genetic lines screened. Although the histopathology of the hepatopancreas tissue showed EHP lesions in both challenged groups, the histological lesions were more pronounced in one of the SPF lines. Quantitative PCR results revealed that animals displaying less hepatocellular damage have lower EHP load compared to animals displaying more pronounced pathological changes. There was no significant difference in final survival at 36 days post-infection in these lines with survival ranging between 80 and 100%. The data showed that mortality as an endpoint metric is not a suitable parameter to determine genetic susceptibility to EHP. Instead, histopathological changes in hepatopancreas, EHP load of the same tissue, and growth retardation would be better metrics to screen EHP susceptibility in P. vannamei. The results show the feasibility of screening genetic lines of P. vannamei for EHP resistance/tolerance using fecal string as an inoculum and, assessing histopathological changes, EHP load, and weight as indicators of resistance.

Effect of flunixin meglumine on pain-related behaviours following cautery disbudding in dairy goat kids.

The aim of this research was to evaluate the physiological and behavioural parameters in Saanen goat kids treated with flunixin meglumine compared with no analgesic treatment after cautery disbudding using a device fueled by propane gas. At 7 days of age, 30 goat kids were randomly allocated to three groups: Sham (Sh) control, (i.e., simulating disbudding); Disbudding (Di), using thermal cauterization; Disbudding + Flunixin (DiFl), thermal cauterization + flunixin meglumine (2.2 mg/kg body weight, 15 min before treatment). Each kid was video-recorded for 30 min before and after treatment to evaluate the frequency of head shaking, head scratching, body shaking, grooming, head rubbing, jumping, running, and bleating. Blood samples were taken 30 min after disbudding to evaluate serum cortisol concentrations and white blood cell count. Heart rate (HR) and respiratory rate (RR) were measured using a stethoscope. Liveweight of goat kids was recorded at birth, -24 h and 7 days after treatment, and at 25 days of age. The mean frequency of head shaking and head scratching was higher (p = 0.0001) after disbudding in Di (37.5 ± 47.8 and 33.32 ± 31.2) group compared to Sh (3.42 ± 3.58 and 2.2 ± 1.8) and DiFl, (4.02 ± 2.76 and 4.42 ± 3.72) groups. The frequency of jumping was higher (p = 0.022) for Di (3.74 ± 2.24) than that of DiFl (0.39 ± 0.92). The remaining behaviours did not show differences (p > 0.05). HR and RR were higher (HR: p < 0.0048; RR: p < 0.035) in group Di (HR: 156 ± 13.6; RR: 66 ± 14.8) than in Sh (HR: 138 ± 8.48; RR: 55.6 ± 5.4) and in DiFl (HR: 136 ± 6.38; RR: 52.8 ± 4.13). No differences were detected between live weight, serum cortisol and white blood cell count data (p > 0.05). Results show that flunixin meglumine was effective at reducing pain-related behaviours when given at the time of disbudding.

Implementation of a virtual multicenter gastrointestinal tumor board to reduce cancer disparities in Argentina.

Given the increasing complexity of cancer care, multidisciplinary tumor boards have become essential in daily clinical oncology practice. The Project Extension for Community Healthcare Outcomes (ECHO) initiative developed an innovative telementoring model using a "hub and spoke" design consisting of a team of experts (hub) that offers a full service to multiple participants (the spokes) during regularly scheduled sessions discussing patients' clinical cases. The Alexander Fleming Cancer Institute in Buenos Aires was the first hub in Latin America to implement Project ECHO for gastrointestinal tumors. In our 3-year experience, 80 patients from 37 centers were evaluated within Project ECHO and a range of three to five cases were discussed in each meeting. From our perspective, the impact of this novel approach was a remarkable strategy to reduce care disparities by equalizing access to high-quality medical knowledge in a multidisciplinary environment for medical discussions. Additionally, it was shown to have a cost-effective impact directly on the patients and the local health system, since relevant costs were saved after unnecessary treatments, studies and travel expenses were avoided.

Current status of infection with infectious hypodermal and hematopoietic necrosis virus (IHHNV) in the Peruvian and Ecuadorian shrimp industry.

Infection with infectious hypodermal and hematopoietic necrosis virus (IHHNV) is a crustacean disease that caused large-scale mortality in Penaeus stylirostris, deformity and growth retardation in Penaeus vannamei and Penaeus monodon. We surveyed the presence of IHHNV in three major shrimp-producing regions in Ecuador, namely Guayas, El Oro, and Esmeralda. The data show that IHHNV is endemic (3.3-100% prevalence) to shrimp farms in these regions. The whole genome sequences of representative circulating IHHNV genotypes in Ecuador and Peru showed that these genotypes formed a separate cluster within the Type II genotypes and were divergent from other geographical isolates of IHHNV originating in Asia, Africa, Australia, and Brazil. In experimental bioassays using specific pathogen-free (SPF) P. vannamei, P. monodon, and P. stylirostris and representative IHHNV isolates from Ecuador and Peru, the virus did not cause any mortality or induce clinical signs in any of the three penaeid species. Although IHHNV-specific Cowdry type A inclusion bodies were histologically detected in experimentally challenged P. vannamei and P. monodon and confirmed by in situ hybridization, no such inclusions were observed in P. stylirostris. Moreover, P. vannamei had the highest viral load, followed by P. monodon and P. stylirostris. Based on IHHNV surveillance data, we conclude that the currently farmed P. vannamei lines in Ecuador are tolerant to circulating IHHNV genotypes. The genome sequence and experimental bioassay data showed that, although the currently circulating genotypes are infectious, they do not induce clinical lesions in the three commercially important penaeid species. These findings suggest a potentially evolving virus-host relationship where circulating genotypes of IHHNV co-exist in equilibrium with P. vannamei raised in Peru and Ecuador.

Does Roux-en-Y Gastric Bypass Really Cure Gastroesophageal Reflux Disease? Analysis of Objective Data.

Background: Laparoscopic Roux-en-Y gastric bypass (LRYGB) is considered the treatment of choice for obesity with gastroesophageal reflux disease (GERD). There are few reports showing objective data based on esophageal function tests (EFTs). The aim of our study was to evaluate the influence of LRYGB on GERD. Methods: Candidates for laparoscopic sleeve gastrectomy (LSG) underwent preoperative esophageal manometry (EM) and 24-hour pH monitoring. Based on the negative influence of LSG on GERD, patients with abnormal pH were offered LRYGB. Those patients repeated EFTs, esophagogastroduodenoscopy, and symptom questionnaire 1 year after surgery. Results: Two hundred fifty LSG candidates underwent preoperative EFTs; 38% were redirected to LRYGB due to abnormal pH and 13 (18%) completed EFTs postoperatively. In ten women, age: 40 ± 7 years, body mass index: 41 ± 1 kg/m2. EM: lower esophageal sphincter (LES) length increased from 2.6 to 2.9 cm (P = not statistically significant [NS]), and LES pressure decreased from 15 to 14.2 mmHg (P = NS). Preoperatively, LES was normotensive in 12 (92%) patients and postoperatively in 11 (85%) (P = NS). DeMeester score decreased from 35.7 to 11 (P < .001). Postoperatively, 9 (69%) patients resolved their GERD, 3 (23%) improved, and 1 (8%) remained the same (P < .001). Symptoms decreased significantly after surgery. Two patients (15%) had Grade A esophagitis. One of them was able to resolve it, while the other 1 remained the same. Conclusions: Our preliminary data showed that after LRYGB, LES pressure remained the same and DeMeester score decreased, while 69% of patients resolved their GERD. Therefore, LRYGB seems to be an excellent option for obesity and GERD.

Experimental reproduction of White Feces Syndrome in whiteleg shrimp, Penaeus vannamei.

White Feces Syndrome (WFS) is an emergent disease of penaeid shrimp (Penaeus monodon and P. vannamei) that is identified by the presence of floating white fecal strings on pond water in grow-out ponds. Although the clinical manifestations of WFS are well defined, the underling etiology remains obscure. WFS has been associated with several enteric pathogens, including Enterocytozoon hepatopenaei (EHP). The association is based on studies that found areas where WFS has been reported, the prevalence and severity of EHP infection are high. In this study, we describe an experimental reproduction of WFS in P. vannamei pre-infected with EHP and challenged with a unique isolate of Vibrio parahaemolyticus isolated from the gastrointestinal tract of a shrimp displaying WFS. Upon laboratory challenge, shrimp displaying white fecal strings and white discoloration of the gastrointestinal tract were analyzed by histopathology, in-situ hybridization and quantitative PCR. Histological analysis confirmed the lesions of EHP and septic hepatopancreatic necrosis in the hepatopancreas of shrimp exposed to both pathogens. Quantitative PCR showed shrimp infected with both EHP and V. parahaemolyticus had a significantly higher load of EHP compared to shrimp infected with EHP alone. This is the first demonstration of experimental reproduction of WFS under laboratory conditions when animals are infected with EHP and V. parahaemolyticus concurrently. The data revealed a synergistic relation between EHP and V. parahaemolyticus isolate that led to the manifestation of WFS. We propose the gross signs of WFS can be used as an indicator of the presence of EHP infection in association with a particular strain of an enteric Vibrio spp. in countries where EHP is endemic.

Differentially Expressed Genes in Hepatopancreas of Acute Hepatopancreatic Necrosis Disease Tolerant and Susceptible Shrimp (Penaeus vannamei).

Acute hepatopancreatic necrosis disease (AHPND) is a lethal disease in marine shrimp that has caused large-scale mortalities in shrimp aquaculture in Asia and the Americas. The etiologic agent is a pathogenic Vibrio sp. carrying binary toxin genes, pirA and pirB in plasmid DNA. Developing AHPND tolerant shrimp lines is one of the prophylactic approaches to combat this disease. A selected genetic line of Penaeus vannamei was found to be tolerant to AHPND during screening for disease resistance. The mRNA expression of twelve immune and metabolic genes known to be involved in bacterial pathogenesis were measured by quantitative RT-PCR in two populations of shrimp, namely P1 that showed susceptibility to AHPND, and P2 that showed tolerance to AHPND. Among these genes, the mRNA expression of chymotrypsin A (ChyA) and serine protease (SP), genes that are involved in metabolism, and crustin-P (CRSTP) and prophenol oxidase activation system 2 (PPAE2), genes involved in bacterial pathogenesis in shrimp, showed differential expression between the two populations. The differential expression of these genes shed light on the mechanism of tolerance against AHPND and these genes can potentially serve as candidate markers for tolerance/susceptibility to AHPND in P. vannamei. This is the first report of a comparison of the mRNA expression profiles of AHPND tolerant and susceptible lines of P. vannamei.

Revisión sistemática sobre los efectos de la actividad física durante el embarazo / Systematic review on the effects of physical activity during pregnancy

Existen dudas sobre los efectos de la actividad física durante el periodo de gestación. El objetivo del presente estudio esrealizar una revisión sistemática para conocer la efectividad de la actividad física durante el embarazo, así como para deter-minar cuáles son las más beneficiosas. Para ello, se han empleado diversas bases de datos, como son Scopus, Scielo, Elsevier,ScienceDirect, ResearchGate, Springer Link, World Wide Science, MedlinePlus, Dialnet y Google Scholar, utilizando las palabrasclaves “actividad física” y “embarazo”. Los documentos seleccionados han sido 79, entre los cuales destacan artículos y tesisen español e inglés publicadas entre 2004 y 2020. Para llevar a cabo el análisis descriptivo, se han extraído ocho variables:objetivos de la investigación, edad media de las embarazadas, instrumentos de recogida de datos, planificación de las ac-tividades físicas realizadas, periodo gestacional en el que comienzan las actividades físicas, parto más frecuente producido,intensidad de las actividades y resultados del estudio. Tras ello, se han combinado entre sí, con la finalidad de obtener unosresultados. Estos indican que los estudios tienen como principal objetivo evaluar el efecto de la actividad física sobre la saludde la madre en mujeres mayores de 30 años y que el periodo en el que más se pone en práctica es del primer y segundotrimestre en adelante. Se puede concluir que la actividad física tiene efectos muy positivos en las gestantes, no alterando lasalud de la madre y del bebé. Del mismo modo, las más beneficiosas son la realización de actividades físicas programadascon una intensidad moderada.(AU)

There are many queries regarding about the effects of physical activity during the gestation period. The aim of this studyis to conduct a systematic review in order to find out the effectiveness of physical activity during pregnancy, as well as todetermine which are the most beneficial. For this purpose, several databases have been used, such as Scopus, Scielo, Elsevier,ScienceDirect, ResearchGate, Springer Link, World Wide Science, MedlinePlus, Dialnet and Google Scholar using the keywords“physical activity” and “pregnancy”. The documents selected were 79, including articles and theses in Spanish and English pu-blished between 2004 and 2020. To carry out the descriptive analysis, eight variables have been extracted: research objectives,mean age of pregnant women, data collection tools, planning of physical activities performed, gestational period in whichphysical activities begin, the most frequent mode of delivery, intensity of activities and results of the study. After that, theyhave combined with each other, in order to obtain outcomes. The outcomes indicate that the main objective of the studiesis to evaluate the effect of physical activity on the health of the mother over thirty years of age and that the period in whichit is most commonly put into practice is from the first and second trimesters onwards. In conclusion, physical activity hasvery positive effects on pregnant women, as it does not alter the health of either the mother or the child. Likewise, the mostbeneficial are activities programs with moderate intensity.(AU)

Photosensitization Induced by Heterophyllaea pustulata in Goats: Sequential Development of Skin Lesions.

Five adult Saanen goats received a single oral dose of Heterophyllaea pustulata containing 42.25 µg/kg rubiadin (anthraquinone) and 3 adult goats were untreated controls. All goats were exposed to sunlight and sequential ear skin biopsies were collected before treatment and at 32 hours, 3 days, 8 days, and 15 days after treatment. Changes at 32 hours after dosing included epidermal spongiosis, single cell death and acantholysis, an increased BAX/BCL-2 protein ratio, and dermal edema. Lesions at day 3 included epidermal and adnexal necrosis, crust formation, and acanthosis. Acanthosis, hyperkeratosis, and dermal fibrosis and neovascularization were present at day 15. The pro-apoptotic (BAX)/anti-apoptotic (BCL-2) protein ratio increased at 32 hours, whereas epidermal and dermal PCNA immunolabeling increased between days 8 and 15 after treatment. The cutaneous lesions were consistent with sunlight-induced damage, and the occurrence in treated but not control goats indicates photosensitization.

Co-Training for Visual Object Recognition Based on Self-Supervised Models Using a Cross-Entropy Regularization.

Automatic recognition of visual objects using a deep learning approach has been successfully applied to multiple areas. However, deep learning techniques require a large amount of labeled data, which is usually expensive to obtain. An alternative is to use semi-supervised models, such as co-training, where multiple complementary views are combined using a small amount of labeled data. A simple way to associate views to visual objects is through the application of a degree of rotation or a type of filter. In this work, we propose a co-training model for visual object recognition using deep neural networks by adding layers of self-supervised neural networks as intermediate inputs to the views, where the views are diversified through the cross-entropy regularization of their outputs. Since the model merges the concepts of co-training and self-supervised learning by considering the differentiation of outputs, we called it Differential Self-Supervised Co-Training (DSSCo-Training). This paper presents some experiments using the DSSCo-Training model to well-known image datasets such as MNIST, CIFAR-100, and SVHN. The results indicate that the proposed model is competitive with the state-of-art models and shows an average relative improvement of 5% in accuracy for several datasets, despite its greater simplicity with respect to more recent approaches.

Development of a Recombinase Polymerase Amplification (RPA) assay for acute hepatopancreatic necrosis disease (AHPND) detection in Pacific white shrimp (Penaeus vannamei).

Acute hepatopancreatic necrosis disease (AHPND) is currently the most important bacterial disease of shrimp that has caused enormous losses to the shrimp industry worldwide. The causative agent of AHPND are Vibrio spp. Carrying plasmids containing the pirA and pirB genes which encode binary toxins, PirAB. Currently, AHPND is mostly diagnosed by PCR-based platforms which require the use of sophisticated laboratory instrumentation and are not suitable for a point-of-care diagnostics. Therefore, the availability of an alternative method based on isothermal amplification would be suitable for AHPND detection outside a laboratory setting and extremely useful at a pond side location. Isothermal amplification is based on the nucleic acid amplification at a single temperature and does not require the use of a thermal cycler. In this study, we developed an isothermal Recombinase Polymerase Amplification (RPA) assay for AHPND detection targeting both pirA and pirB genes, simultaneously and evaluated the specificity and sensitivity of the assay. The assay could detect AHPND without any cross-reaction with other microbial pathogens and Specific Pathogen Free (SPF) shrimp. The limit of detection of the assay was 5 copies of pirAB genes. To evaluate the reliability of the assay in detecting AHPND, DNA from Penaeus vannamei shrimp displaying acute and chronic infection were analyzed by the RPA assay and the results were compared with SYBR Green real-time PCR assay. While there was a 100% conformity between the two assay while detecting acute phase infection, RPA appeared to be more sensitive in detecting chronic phase infection. The data suggest that RPA assay described here would be a reliable method in detecting AHPND outside a standard laboratory setting.

A Mutant of Vibrio parahaemolyticus pirABVP (+) That Carries Binary Toxin Genes but Does Not Cause Acute Hepatopancreatic Necrosis Disease.

Vibrio parahaemolyticus carrying binary toxin genes, pirAB, is one of the etiological agents causing acute hepatopancreatic necrosis disease (AHPND) in shrimp. This disease has emerged recently as a major threat to shrimp aquaculture worldwide. During a routine PCR screening of AHPND-causing V. parahaemolyticus strains, an isolate tested PCR positive for pirB (R13) and another isolate tested positive for both the pirA and pirB (R14) genes. To evaluate the pathogenicity of these isolates, specific pathogen-free (SPF) Penaeus vannamei were experimentally challenged. For both R13 and R14 isolates, the final survival rate was 100% at termination of the challenge, whereas the final survival with the AHPND-causing V. parahaemolyticus was 0%. The nucleotide sequence of the plasmid DNA carrying the binary toxin genes revealed that R13 contains a deletion of the entire pirA gene whereas R14 contains the entire coding regions of both pirA and pirB genes. However, R14 possesses an insertion upstream of the pirA gene. In R14, mRNA for both pirA and pirB genes could be detected but no cognate proteins. This shows that the genome of AHPND-causing V. parahaemolyticus is highly plastic and, therefore, detection of the pirA and pirB genes alone by DNA-PCR is insufficient as a diagnostic test for AHPND.

Evidences supporting Enterocytozoon hepatopenaei association with white feces syndrome in farmed Penaeus vannamei in Venezuela and Indonesia.

White feces syndrome (WFS) is an emerging and poorly described disease characterized by the presence of floating white fecal strings in shrimp (Penaeus monodon and P. vannamei) grow-out ponds. WFS has been associated with several pathogens, including Enterocytozoon hepatopenaei. This association is based on the fact that in areas where E. hepatopenaei has been reported, there was also a high WFS prevalence. E. hepatopenaei is an emerging pathogen that has affected cultured shrimp in Indonesia, Vietnam, China, Thailand, and India. In 2016, we reported the presence of E. hepatopenaei in farmed P. vannamei in Venezuela. In this study, we describe the first case of WFS in Venezuela associated with E. hepatopenaei. The white fecal strings and shrimp displaying white feces along the gastrointestinal tract observed in this study were similar to the gross signs found in WFS-impacted P. vannamei in SE Asian countries. Furthermore, we describe a strong association between WFS and E. hepatopenaei in the samples obtained from Venezuela and Indonesia. Quantification of E. hepatopenaei in WFS-affected ponds, ponds with a history of WFS, and ponds with no WFS showed that E. hepatopenaei loads were significantly higher in WFS-affected ponds. Furthermore, these findings constitute the first report of WFS being associated with E. hepatopenaei in farmed shrimp in Latin America. Additionally, we propose that the gross signs of WFS such as floating whitish fecal strings can be used as an indicator of the presence of E. hepatopenaei in countries where E. hepatopenaei is endemic.

Genome reconstruction of white spot syndrome virus (WSSV) from archival Davidson's-fixed paraffin embedded shrimp (Penaeus vannamei) tissue.

Formalin-fixed paraffin-embedded (FFPE) tissues are a priceless resource for diagnostic laboratories worldwide. However, DNA extracted from these tissues is often not optimal for most downstream molecular analysis due to fragmentation and chemical modification. In this study, the complete genome of white spot syndrome virus (WSSV) was reconstructed from ~ 2-year-old archived Davidson's-fixed paraffin-embedded (DFPE) shrimp tissue using Next Generation Sequencing (NGS). A histological analysis was performed on archived DFPE shrimp tissue and a sample showing a high level of WSSV infection was selected for molecular analysis. The viral infection was further confirmed by molecular methods. DNA isolated from DFPE and fresh frozen (FF) tissues were sequenced by NGS. The complete genome reconstruction of WSSV (~ 305 kbp) was achieved from both DFPE and FF tissue. Single nucleotide polymorphisms, insertion and deletions were compared between the genomes. Thirty-eight mutations were identified in the WSSV genomes from the DFPE and FF that differed from the reference genome. This is the first study that has successfully sequenced the complete genome of a virus of over 300 kbp from archival DFPE tissue. These findings demonstrate that DFPE shrimp tissue represents an invaluable resource for prospective and retrospective studies, evolutionary studies and opens avenues for pathogen discovery.

Acute hepatopancreatic necrosis disease (VPAHPND), a chronic disease in shrimp (Penaeus vannamei) population raised in latin America.

In Latin American shrimp farming, acute hepatopancreatic necrosis disease (AHPND) does not cause the acute mortalities observed in SE Asia. Herein we report for the first time a new phase of infection of AHPND, a chronic phase based on two experimental AHPND-challenge trials using shrimp lines from Latin America. Three shrimp lines of Penaeus vannamei were challenged with a highly pathogenic strain of Vibrio parahaemolyticus causing AHPND (VPAHPND). PCR and histopathology assays were used for confirmation of AHPND in the trials. The first study was to compare survival between the lines. A follow-up trial was conducted to document hepatopancreas heterotrophic bacterial count and to measure the expression of VPAHPND binary toxin genes (pirAB genes) at 24 h.p.i. One of the Latin American shrimp lines, APE1, had significantly higher survival than recorded for the other two lines (APE2 & APE3) and the specific-pathogen-free positive control line. Histopathology showed typical AHPND acute and terminal phase lesions in VPAHPND challenged groups, although destructive cellular changes were more pronounced in the SPF line. Histopathology of animals surviving AHPND revealed a unique chronic phase of infection that resembles septic hepatopancreatic necrosis (SHPN), recognized as diagnostic of digestive tract vibriosis. Data to support our finding, including a quantitative RT-PCR assay, confirmed the expression of pirAB genes and the differential hepatopancreas heterotrophic plate count (HPC) among the different lines challenged. The results explain in part why the shrimp industry in some Latin American countries continues to grow despite the presence of AHPND. In addition, the biology and pathology of AHPND resistant/tolerant shrimp appear to be quite unique in this Latin American shrimp population.