Experimental Infection and the Effects of Temperature on the Pathogenicity of the Infectious Spleen and Kidney Necrosis Virus in Juvenile Nile Tilapia (Oreochromis niloticus).

The infectious spleen and kidney necrosis virus (ISKNV) is one of the most important emerging viral pathogens for Nile tilapia (Oreochromis niloticus) farming. While prevalent worldwide, it has recently been detected in Brazil. However, despite the importance of the virus and the affected fish species, there are no scientific data on the effects of water temperature on disease pathogenesis in Nile tilapia. In the present study, we conducted two trials using juvenile Nile tilapia over a 15-day period. In trial 1, an experimental infection model was developed based on the intraperitoneal inoculation of active viral homogenates (4.3 × 104 virus fish-1), while control fish were similarly inoculated with inactivated viral homogenates. In trial 2, the fish were maintained at different water temperatures (26, 28, 30, 32, and 34 °C) and then infected with ISKNV. For virus detection, kidney and spleen samples were collected and analyzed by qPCR. Our results show that the disease was successfully reproduced in experimental conditions with active homogenates, with the first signs of the disease appearing on the third day after infection. In addition, a significant reduction in mortality was observed in the groups maintained at higher temperatures (>30 °C). This suggests that a treatment of the disease with non-lethal hyperthermia can be used to control the symptoms and mortality of ISKNV-infected Nile tilapia juveniles.

From Platelet-Rich Plasma to Mesenchymal Stem Cells: Cartilage, Tendon, and Bone Regeneration With Orthobiologics.

The use of orthobiologics such as platelet-rich plasma, bone marrow aspirate, and stem cells has been proposed as a biologic augmentation for treatment of various conditions of cartilage, tendon, and bone. Although the published evidence is not conclusive, the safety of these treatments and benefits in improving the biologic condition of treated tissues have been confirmed. Osteoarthritis, rotator cuff injuries, and osteonecrosis of the femoral head are three common musculoskeletal conditions associated with the use of orthobiologics in patients with cartilage, tendon, and bone injuries. When reviewing the use of platelet-rich plasma, bone marrow aspirate, and mesenchymal stem cells in patients with these conditions, there is evidence of high safety and positive, but variable, efficacy. Recent studies have shown promising results and have paved the way for research being conducted at many specialized centers around the world.

Susceptibility Profile and Epidemiological Cut-Off Values Are Influenced by Serotype in Fish Pathogenic Streptococcus agalactiae.

Streptococcus agalactiae is a major health concern in tilapia farming worldwide. In contrast to the availability of susceptibility profile results, interpretative criteria for disk diffusion assays and the influence of serotypes on resistance profiles are not available. To address this, sixty isolates (thirty of each serotype, Ib and III) were evaluated using the disk diffusion assay against six antibiotics, and the epidemiological cut-off value (ECV) was calculated. All the isolates were classified as non-wild type (NWT) for sulfamethoxazole (SUT) and norfloxacin (NOR). The inhibition zones for oxytetracycline (OXY) and doxycycline (DOX) were largely distinct; all serotype Ib and III isolates were classified as wild-type (WT) and NWT, respectively. The results for serotype III of fish group B Streptococcus (GBS) were comparable to the NWT tetracycline profile of human GBS available in EUCAST, suggesting the presence of resistance mechanisms in these fish isolates. The calculation of the cut-off wild type (COWT) values for OXY and DOX was appropriate for both serotypes. Differences between the distribution of florfenicol (FLO) and amoxicillin (AMO) were found, and we attribute this to the faster growth rate of serotype III, which promotes smaller inhibition zones. Therefore, using separate COWT for each serotype is necessary. In conclusion, the serotype of fish GBS affects its susceptibility profile, and it is recommended to use serotype-specific COWT values as interpretative criteria for disk diffusion assays against FLO and AMO.

Sodium and lithium exert differential effects on the central carbon metabolism of Debaryomyces hansenii through the glyoxylate shunt regulation.

Debaryomyces hansenii is a halotolerant/halophilic yeast usually found in salty environments. The yeast accumulated sodium at high concentrations, which improved growth in salty media. In contrast, lithium was toxic even at low concentrations and its presence prevented cell proliferation. To analyse the responses to both cations, metabolite levels, enzymatic activities and gene expression were determined, showing that NaCl and LiCl trigger different cellular responses. At high concentrations of NaCl (0.5 or 1.5 M) cells accumulated higher amounts of the intermediate metabolites glyoxylate and malate and, at the same time, the levels of intracellular oxoglutarate decreased. Additionally, 0.5 M NaCl increased the activity of the enzymes isocitrate lyase and malate synthase involved in the synthesis of glyoxylate and malate respectively and decreased the activity of isocitrate dehydrogenase. Moreover, transcription of the genes coding for isocitrate lyase and malate synthase was activated by NaCl. Also, cells accumulated phosphate upon NaCl exposure. None of these effects was provoked when LiCl (0.1 or 0.3 M) was used instead of NaCl. Lithium induced accumulation of higher amounts of oxoglutarate and decreased the concentrations of glyoxylate and malate to non-detectable levels. Cells incubated with lithium also showed higher activity of the isocitrate dehydrogenase and neither increased isocitrate lyase and malate synthase activities nor the transcription of the corresponding genes. In summary, we show that sodium, but not lithium, up regulates the shunt of the glyoxylic acid in D. hansenii and we propose that this is an important metabolic adaptation to thrive in salty environments.

Therapeutic efficacy of enrofloxacin in treatment of Francisella orientalis infections in juvenile Nile tilapia (Oreochromis niloticus L.).

Outbreaks of infections by Francisella orientalis represent one of the main obstacles to Nile tilapia (Oreochromis niloticus L.) farming. It is responsible for acute mortality in fingerlings and juveniles. The main control measure available is oral antibiotic therapy. This study compared the therapeutic efficacy of the antibiotics enrofloxacin and oxytetracycline, the most commonly used antimicrobial, against francisellosis in juvenile Nile tilapia (O. niloticus). Fish were challenged with a virulent isolate of F. orientalis and treated with medicated feed containing one of two doses of oxytetracycline (100 or 300 mg/kg of live weight (LW)) or 10 mg/kg of LW of enrofloxacin. The positive and negative control groups received feed without antibiotics; the negative control group was unchallenged. The results showed that enrofloxacin at a dose of 10 mg/kg of LW is effective against francisellosis in juvenile Nile tilapia (O. niloticus). Treatment with oxytetracycline did not eliminate the pathogen from the infected host, and the surviving fish became carriers. Enrofloxacin was able to cure the fish of infection with F. orientalis. This study suggests that enrofloxacin is a better option for treating francisellosis in Nile tilapia (O. niloticus L.). It controls mortality and avoids the carrier state in the fish, thus reducing the possibility of recurrence in the affected batches.

How we approach conservative treatment of retinoblastoma in South America in the era of local ocular treatments: A consensus of the Grupo America Latina de Oncologia Pediatrica (GALOP).

Local therapies are increasingly used for ocular preservation in retinoblastoma. In middle-income countries, these techniques pose specific challenges mostly related to more advanced disease at diagnosis. The Grupo de America Latina de Oncología Pediátrica (GALOP) developed a consensus document for the management of conservative therapy for retinoblastoma. Intra-arterial chemotherapy (OAC) is the preferred therapy, except for those with less advanced disease or age younger than 6 months. OAC allowed for a reduction in the use of external beam radiotherapy in our setting. Intravitreal chemotherapy is the preferred treatment for vitreous seeding. Enucleation is the treatment of choice for eyes with advanced disease.

First report of Crassiphiala sp. (Trematoda: Diplostomidae) as an etiological agent of black spot disease in commercial ornamental fish from Brazil.

Ornamental fish are becoming increasingly popular, but the lack of knowledge regarding their various diseases is a major challenge. Skin diseases commonly found in freshwater fish include black spot disease (BSD), which is characterized by melanin deposits around the metacercariae of some trematode species. Since BSD remains poorly understood, this study describes an outbreak of BSD in Etroplus maculatus raised in outdoor ponds at a Brazilian fish farm. Metacercariae samples were collected, examined, and subjected to molecular phylogenetic analysis. The parasites were conspecific to an unnamed species, Crassiphiala lineage 5, recently found in Brazilian birds (Megaceryle torquata). Sequences obtained for longifurcate cercariae of the planorbid snail Biomphalaria straminea from the same region were identical to our metacercariae of Crassiphiala sp. These results suggest that Biompahalaria snails are likely an intermediate host of this parasite on farms where E. maculatus was found to be infected. We provide the first molecular evidence that Crassiphiala are the causative agents of BSD in fish from Brazil. Combatting snails and preventing access of fish-eating birds to outdoor ponds are strategies to control this disease in ornamental fish farms.

The benefits of contributing to the citizen science platform iNaturalist as an identifier.

As the number of observations submitted to the citizen science platform iNaturalist continues to grow, it is increasingly important that these observations can be identified to the finest taxonomic level, maximizing their value for biodiversity research. Here, we explore the benefits of acting as an identifier on iNaturalist.

Development of an inactivated whole cell vaccine through immersion for immunoprophylaxis of Francisella orientalis infections in Nile tilapia (Oreochromis niloticus L.) fingerlings and juveniles.

Francisella orientalis infections, known as francisellosis, are one of the most important diseases affecting the production of Nile tilapia, causing high mortality rates in the most susceptible fish stages: fingerlings and juveniles. Antibiotic therapy is the method of choice for treating the disease, as there are no commercially available vaccines. In this study, we developed an inactivated whole-cell vaccine using an isolate of F. orientalis in combination with the aqueous adjuvant Montanide IMS 1312 VG, which was administered to Nile tilapia through immersion. Two immunization trials (1 and 2) were conducted with fish at the fingerling and juvenile stages. For each trial, five different experimental groups were established: a complete vaccine (bacterin in combination with aqueous adjuvant), bacterin, aqueous adjuvant, and positive and negative controls. Thirty days after vaccination, an experimental challenge was performed through intraperitoneal injection of the same F. orientalis isolate. As a result, the vaccinated fingerlings were the only group in which mortality and progression of clinical signs of francisellosis were statistically significantly reduced, although relative percentage of survival (RPS) was low at 50%. In the juvenile group, RPS was higher at 63%, but not statistically significant. Nevertheless, an RPS of only 50% is acceptable for using vaccines in the field. The bacterin and adjuvant treatments alone were not effective, showing an RPS of 37% and 0%, respectively. Post-vaccination mortality was observed in the group exposed only to the adjuvant, which may indicate excessive immune stimulation at this stage. Interestingly, the immune response elicited by the vaccine was unable to eliminate the pathogen from the host; therefore, the surviving animals became carriers. Although the immune response elicited by the vaccine was unable to eliminate the pathogen from the host, this vaccine formulation could be a viable alternative for use in the field and serve as another means of controlling the mortality caused by the pathogen. Our study provides the first report of vaccination, using immersion, against francisellosis at the most susceptible stages of farmed Nile tilapia. Future studies should address the efficiency of immersion vaccines under field conditions.

Diversity and potential genetic relationships amongst Brazilian Brachyspira hyodysenteriae isolates from cases of swine dysentery.

The aim of this study was to evaluate genetic diversity, distribution, evolution and population structure of Brazilian Brachyspira hyodysenteriae strains isolated from swine. Multilocus Sequence Typing (MLST) analysis using seven housekeeping genes was applied to 46 isolates obtained from outbreaks of swine dysentery that occurred between 2011 and 2015 in the states of Minas Gerais, São Paulo, Mato Grosso, Rio Grande do Sul and Santa Catarina. Historical isolates from Rio Grande do Sul obtained in 1998 were also included in the study. An independent international profile of the global sequences deposited in the B. hyodysenteriae database was used for comparisons with the Brazilian strains. All isolates from 2011 to 2015 were classified into nine sequence type (STs) and divided into four clonal complexes. These findings indicated genetic relationships among the B. hyodysenteriae from different Brazilian states, among historical strains isolated in 1998 and from recent outbreaks, and relatedness with global isolates. Seven STs were unique and, to date, only reported in Brazil.

ACUTE NECROTIZING AND EOSINOPHILIC MYOCARDITIS IN A CHIMPANZEE (PAN TROGLODYTES).

Cardiac disease is of importance in captive chimpanzee (Pan troglodytes) health. Here we report an eosinophilic and necrotizing myocarditis in a 17-y-old chimpanzee with no previous history of cardiac disease that progressed to death within 48 h. Toxic and infectious causes were ruled out. The chimpanzee had eosinophilia at different occasions in previous years. The animal had a severe, diffuse, and acute monophasic necrotizing myocarditis, with a moderate lymphoplasmacytic infiltrate that was rich in eosinophils. Ante- and postmortem investigations are compatible with an unusual eosinophilic myocarditis with clinical evolution and morphology comparable with human eosinophilic myocarditis secondary to hypereosinophilic syndrome.

Inovação tecnológica no Brasil: desafios e insumos para políticas públicas / Innovación tecnológica en Brasil: desafíos e insumos para las políticas públicas / Technological innovation in Brazil: challenges and inputs for public policies

Resumo Este artigo oferece uma breve reflexão sobre a natureza do investimento em pesquisa e desenvolvimento (P&D) no Brasil. Seu objetivo é proporcionar alguns insumos para avançar no debate sobre esse tema na sociedade brasileira. Desde 1999, o Brasil tem aumentado de maneira consistente o seu investimento em P&D, considerado um dos insumos para inovação e produtividade. Porém, tal esforço tem gerado resultados limitados. Esses resultados limitados não parecem refletir mera insuficiência de investimentos em inovação no Brasil, mas a maneira e a eficácia de sua implementação.

Resumen Este artículo ofrece una breve reflexión sobre la naturaleza de la inversión en investigación y desarrollo (I&D) en Brasil. Su objetivo es brindar algunos insumos para avanzar en el debate sobre este tema en la sociedad brasileña. Desde 1999, Brasil ha aumentado constantemente su inversión en I&D, considerada uno de los insumos para la innovación y la productividad. Sin embargo, tal esfuerzo ha dado resultados limitados. Dichos resultados no parecen reflejar la mera insuficiencia de las inversiones en innovación en Brasil, sino la forma y efectividad de su implementación.

Abstract: This article offers a brief reflection on the nature of research and development (R&D) investment in Brazil, contributing to advancing the debate on this topic. R&D is one of the bases of innovation and productivity, and since 1999 Brazil has consistently increased investment in this area. However, this effort has yielded limited results, suggesting that supplying sufficient resources must be accompanied by effective implementation strategies.

Total mercury in commercial fishes and estimation of Brazilian dietary exposure to methylmercury.

BACKGROUND: Mercury, in particular its most toxic form methylmercury, poses a risk to public health. Dietary methylmercury exposure is mainly by fish, and it can vary with fish contamination and by dietary habits of the population. This study aimed to quantify total mercury levels in different fish from Brazil and to estimate Brazilian exposure to methylmercury by fish consumption. METHODS: Total mercury occurrence was investigated in 18 different fish species by atomic absorption spectrometry with thermal decomposition and gold amalgamation. Dietary exposure to methylmercury was estimated by a deterministic method for different groups considering consumption by sex, different Brazilian geographical regions and habitat (rural or urban). RESULTS: Carnivorous fish showed higher levels of mercury (0.01 to 0.93 mg/kg) compared to non-strictly carnivorous fish (<0.01 to 0.30 mg/kg). Farmed fishes showed significantly lower levels compared to wild fish. Mean Brazilian fish consumption achieves FAO/WHO health recommendation of about two portions of fish per week. However, there is a large difference between fish consumption at urban and rural homes and among Brazilian geographic regions. These differences in consumption impacted estimated methylmercury intake that was higher in the Northern (1.85 µg/kg bw week) and in the Northeastern (0.72 µg/kg bw week) regions and also by rural population (1.08 µg/kg bw week). These values were compared with the toxicological reference dose for neurotoxicity of 1.6 µg/kg bw week. CONCLUSION: Even though total levels of mercury in fish were lower than Brazilian and international legislations, in the Northern Brazilian region methylmercury intake overpassed the toxicological reference dose for neurotoxicity and in rural areas it achieved 68% of this reference dose.

Da Bíblia de Gutenberg à COVID-19 / De la Biblia de Gutenberg a la COVID-19 / From the Gutenberg Bible to COVID-19

Resumo O mundo após a COVID-19 será diferente do mundo de antes, não apenas do ponto de vista dos desafios econômicos, mas também na perspectiva da geopolítica. A globalização tenderá a ser revertida, com claras implicações para a posição dos vários países. O fenômeno da expansão do uso generalizado da tecnologia da informação marcará a dinâmica de vários países, inclusive do Brasil. A academia precisa refletir sobre estratégias de longo prazo para criar empregos e sustentar a empregabilidade enquanto a estrutura da economia se modifica.

Resumem El mundo después de la COVID-19 será diferente del anterior, no solo desde el punto de vista de los desafíos económicos, sino también desde la perspectiva de la geopolítica. La globalización tenderá a revertirse, con claras implicaciones para la posición de los distintos países. El fenómeno de expansión del uso generalizado de la tecnología de la información caracterizará la dinámica de varios países, inclusive de Brasil. La academia necesita reflexionar sobre estrategias a largo plazo para crear empleos y mantener la empleabilidad mientras la estructura de la economía se modifica.

Abstract The world after the COVID-19 pandemic will change, from the perspective of both geopolitical and economic challenges. There will be a tendency for globalization to reverse, affecting the international position of several countries. The phenomenon of expanding the heavy use of information technology will mark the dynamics of many countries, including Brazil. Therefore, academia has to reflect on long-term strategies to create and maintain jobs while the economic structure changes.

Giant aneurysm of the right coronary artery, report of a case and review of the literature.

Coronary artery aneurysms are described as a localized dilatation that exceeds the normal diameter by 1.5 times. This is a rare condition; its incidence varies from 0.3% up to 5.3% of all coronary angiographies. Those aneurysms that exceed 4 times the diameter of a normal artery are considered giant aneurysms, which are even more uncommon, presenting between 0.02% and 0.2% of all cases. There is controversy regarding its pathophysiology, however, up to 50% of the cases are related to atherosclerosis. They are diagnosed more frequently between the sixth and seventh decade of life. The main clinical manifestations are related to ischemic heart disease. Regarding their treatment, there is no general consensus towards its management in adult patients. The options are medical, surgical or percutaneous treatment. We report the presence of a giant aneurysm of the right coronary artery and giant ectasia of the left coronary system with active thrombosis in a man with history of an abdominal aortic aneurysm, with endovascular treatment and a non-ST segment elevation myocardial infarction with no reperfusion strategy, who required a coronary computed tomography, identifying the anatomical characteristics of this disease.

Los aneurismas de las arterias coronarias se definen como una dilatación localizada que excede el diámetro normal en 1.5 veces. Esta es una condición poco frecuente, su incidencia varía del 0.3 hasta el 5.3% de las angiografías coronarias. Los aneurismas que exceden cuatro veces el diámetro del vaso normal se consideran gigantes. Estos son aún más raros y se presentan en el 0.02 a 0.2% de todos los casos. Existe controversia en cuanto a su fisiopatología, sin embargo, hasta el 50% de los casos se relacionan con la aterosclerosis. Se diagnostican más frecuentemente entre la sexta y séptima décadas de vida. Las principales manifestaciones clínicas están relacionadas con la cardiopatía isquémica. Respecto a su tratamiento, no existe un consenso del manejo en los pacientes adultos, las opciones son: médico, quirúrgico o intervencionismo. Reportamos la presencia de un aneurisma gigante de la coronaria derecha y ectasia gigante del sistema izquierdo con trombosis activa en un hombre con antecedentes de un aneurisma en la aorta abdominal, tratado por vía endovascular, e infarto agudo al miocardio sin elevación del segmento ST no reperfundido. Requirió de estudio de angiotomografía coronaria, el cual permitió la identificación de las características anatómicas de esta enfermedad.

Aneurisma gigante de la arteria coronaria derecha, reporte de un caso y revisión de la literatura / Giant aneurysm of the right coronary artery, report of a case and review of the literature

Resumen Los aneurismas de las arterias coronarias se definen como una dilatación localizada que excede el diámetro normal en 1.5 veces. Esta es una condición poco frecuente, su incidencia varía del 0.3 hasta el 5.3% de las angiografías coronarias. Los aneurismas que exceden cuatro veces el diámetro del vaso normal se consideran gigantes. Estos son aún más raros y se presentan en el 0.02 a 0.2% de todos los casos. Existe controversia en cuanto a su fisiopatología, sin embargo, hasta el 50% de los casos se relacionan con la aterosclerosis. Se diagnostican más frecuentemente entre la sexta y séptima décadas de vida. Las principales manifestaciones clínicas están relacionadas con la cardiopatía isquémica. Respecto a su tratamiento, no existe un consenso del manejo en los pacientes adultos, las opciones son: médico, quirúrgico o intervencionismo. Reportamos la presencia de un aneurisma gigante de la coronaria derecha y ectasia gigante del sistema izquierdo con trombosis activa en un hombre con antecedentes de un aneurisma en la aorta abdominal, tratado por vía endovascular, e infarto agudo al miocardio sin elevación del segmento ST no reperfundido. Requirió de estudio de angiotomografía coronaria, el cual permitió la identificación de las características anatómicas de esta enfermedad.

Abstract Coronary artery aneurysms are described as a localized dilatation that exceeds the normal diameter by 1.5 times. This is a rare condition; its incidence varies from 0.3% up to 5.3% of all coronary angiographies. Those aneurysms that exceed 4 times the diameter of a normal artery are considered giant aneurysms, which are even more uncommon, presenting between 0.02% and 0.2% of all cases. There is controversy regarding its pathophysiology, however, up to 50% of the cases are related to atherosclerosis. They are diagnosed more frequently between the sixth and seventh decade of life. The main clinical manifestations are related to ischemic heart disease. Regarding their treatment, there is no general consensus towards its management in adult patients. The options are medical, surgical or percutaneous treatment. We report the presence of a giant aneurysm of the right coronary artery and giant ectasia of the left coronary system with active thrombosis in a man with history of an abdominal aortic aneurysm, with endovascular treatment and a non-ST segment elevation myocardial infarction with no reperfusion strategy, who required a coronary computed tomography, identifying the anatomical characteristics of this disease.

Identification and Characterization of Escherichia coli, Salmonella Spp., Clostridium perfringens, and C. difficile Isolates from Reptiles in Brazil.

Considering the increasing popularity of reptiles as pets and their possible role as reservoirs of pathogenic microorganisms, the aim of this study was to isolate Escherichia coli, Salmonella spp., Clostridium perfringens, and C. difficile strains from reptiles in Brazil and to characterize the isolated strains. The characterization was based on phylogenetic typing of E. coli, identification of virulence genes of E. coli, C. perfringens, and C. difficile, serotyping of Salmonella spp., ribotyping and MLST of C. difficile and antimicrobial susceptibility test of pathogenic strains. Cloacal swabs were collected from 76 reptiles, of which 15 were lizards, 16 chelonians, and 45 snakes, either living in captivity, in the wild, or as companion animals. E. coli was isolated from 52 (68.4%) reptiles, of which 46 (88.4%) were characterized as phylogroup B1. The virulence factor CNF1 of E. coli was found in seven (9.2%) sampled animals, whereas the gene of EAST1 was found in isolates from two (2.6%) reptiles. Three isolates positive for CNF1 were resistant to cephalothin, one of which was also resistant to ciprofloxacin, trimethoprim/sulfamethoxazole, and chloramphenicol, being then classified as multidrug resistant strain (MDR). Salmonella enterica was identified in 26 (34.2%) reptiles, of which 13 belonged to the subspecies enterica. Serotypes such as S. Mbandaka, S. Panama, S. Infantis, S. Heidelberg, and S. Anatum were identified. One isolate of S. enterica subsp. houtenae was resistant to cephalothin and ciprofloxacin. C. perfringens type A was isolated from six (7.8%) animals. C. difficile was isolated from three (3.9%) reptiles. Two of these isolates were toxigenic and classified into ribotypes/MLST 081/ST9 and 106/ST42, which have been previously reported to infect humans. In conclusion, reptiles in Brazil can harbor toxigenic C. difficile and potentially pathogenic E. coli and Salmonella enterica subsp. enterica, thus representing a risk to human and animal health.