In vitro Trypanocidal Activity, Genomic Analysis of Isolates, and in vivo Transcription of Type VI Secretion System of Serratia marcescens Belonging to the Microbiota of Rhodnius prolixus Digestive Tract

Serratia marcescens is a bacterium with the ability to colonize several niches, including some eukaryotic hosts. S. marcescens have been recently found in the gut of hematophagous insects that act as parasite vectors, such as Anopheles, Rhodnius, and Triatoma. While some S. marcescens strains have been reported as symbiotic or pathogenic to other insects, the role of S. marcescens populations from the gut microbiota of Rhodnius prolixus, a vector of Chagas’ disease, remains unknown. Bacterial colonies from R. prolixus gut were isolated on BHI agar. After BOX-PCR fingerprinting, the genomic sequences of two isolates RPA1 and RPH1 were compared to others S. marcescens from the NCBI database in other to estimate their evolutionary divergence. The in vitro trypanolytic activity of these two bacterial isolates against Trypanosoma cruzi (DM28c clone and Y strain) was assessed by microscopy. In addition, the gene expression of type VI secretion system (T6SS) was detected in vivo by RT-PCR. Comparative genomics of RPA1 and RPH1 revealed, besides plasmid presence and genomic islands, genes related to motility, attachment, and quorum sensing in both genomes while genes for urea hydrolysis and type II secretion system (T2SS) were found only in the RPA1 genome. The in vitro trypanolytic activity of both S. marcescens strains was stronger in their stationary phases of growth than in their exponential ones, with 65–70 and 85–90% of epimastigotes (Dm28c clone and Y strain, respectively) being lysed after incubation with RPA1 or RPH1 in stationary phase. Although T6SS transcripts were detected in guts up to 40 days after feeding (DAF), R. prolixus morbidity or mortality did not appear to be affected. In this report, we made available two trypanolytic S. marcescens strains from R. prolixus gut to the scientific community together with their genomic sequences. Here, we describe their genomic features with the purpose of bringing new insights into the S. marcescens adaptations for colonization of the specific niche of triatomine guts. This study provides the basis for a better understanding of the role of S. marcescens in the microbiota of R. prolixus gut as a potential antagonist of T. cruzi in this complex system

Measurement of the cosmic ray spectrum above 4×10184{\times}10^{18} eV using inclined events detected with the Pierre Auger Observatory

A measurement of the cosmic-ray spectrum for energies exceeding $4{\times}10^{18}$ eV is presented, which is based on the analysis of showers with zenith angles greater than $60^{\circ}$ detected with the Pierre Auger Observatory between 1 January 2004 and 31 December 2013. The measured spectrum confirms a flux suppression at the highest energies. Above $5.3{\times}10^{18}$ eV, the "ankle", the flux can be described by a power law $E^{-\gamma}$ with index $\gamma=2.70 \pm 0.02 \,\text{(stat)} \pm 0.1\,\text{(sys)}$ followed by a smooth suppression region. For the energy ($E_\text{s}$) at which the spectral flux has fallen to one-half of its extrapolated value in the absence of suppression, we find $E_\text{s}=(5.12\pm0.25\,\text{(stat)}^{+1.0}_{-1.2}\,\text{(sys)}){\times}10^{19}$ eV.Comment: Replaced with published version. Added journal reference and DO

In vitro Trypanocidal Activity, Genomic Analysis of Isolates, and in vivo Transcription of Type VI Secretion System of Serratia marcescens Belonging to the Microbiota of Rhodnius prolixus Digestive Tract

Submitted by Sandra Infurna ([email protected]) on 2019-04-29T17:31:35Z No. of bitstreams: 1 FabioFaria_Mota_etal_IOC_2019.pdf: 4237942 bytes, checksum: d580927bcebfc83c0dcafba666d7274b (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2019-04-29T17:41:25Z (GMT) No. of bitstreams: 1 FabioFaria_Mota_etal_IOC_2019.pdf: 4237942 bytes, checksum: d580927bcebfc83c0dcafba666d7274b (MD5)Made available in DSpace on 2019-04-29T17:41:25Z (GMT). No. of bitstreams: 1 FabioFaria_Mota_etal_IOC_2019.pdf: 4237942 bytes, checksum: d580927bcebfc83c0dcafba666d7274b (MD5) Previous issue date: 2019Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Computacional e Sistemas. Rio de Janeiro, RJ. Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ. Brasil.Universidade Federal Fluminense. Instituto Biomédico. Departamento de Microbiologia e Parasitologia. Laboratório de Enteropatógenos, Microbiologia Veterinária e de Alimentos. Niterói, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Instituto Nacional de Ciência e Tecnologia em Inovação em Doenças Negligenciadas. Laboratório de Modelagem de Sistemas Biológicos. Rio de Janeiro, RJ. Brasil.Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ. Brasil.Serratia marcescens is a bacterium with the ability to colonize several niches, including some eukaryotic hosts. S. marcescens have been recently found in the gut of hematophagous insects that act as parasite vectors, such as Anopheles, Rhodnius, and Triatoma. While some S. marcescens strains have been reported as symbiotic or pathogenic to other insects, the role of S. marcescens populations from the gut microbiota of Rhodnius prolixus, a vector of Chagas' disease, remains unknown. Bacterial colonies from R. prolixus gut were isolated on BHI agar. After BOX-PCR fingerprinting, the genomic sequences of two isolates RPA1 and RPH1 were compared to others S. marcescens from the NCBI database in other to estimate their evolutionary divergence. The in vitro trypanolytic activity of these two bacterial isolates against Trypanosoma cruzi (DM28c clone and Y strain) was assessed by microscopy. In addition, the gene expression of type VI secretion system (T6SS) was detected in vivo by RT-PCR. Comparative genomics of RPA1 and RPH1 revealed, besides plasmid presence and genomic islands, genes related to motility, attachment, and quorum sensing in both genomes while genes for urea hydrolysis and type II secretion system (T2SS) were found only in the RPA1 genome. The in vitro trypanolytic activity of both S. marcescens strains was stronger in their stationary phases of growth than in their exponential ones, with 65-70 and 85-90% of epimastigotes (Dm28c clone and Y strain, respectively) being lysed after incubation with RPA1 or RPH1 in stationary phase. Although T6SS transcripts were detected in guts up to 40 days after feeding (DAF), R. prolixus morbidity or mortality did not appear to be affected. In this report, we made available two trypanolytic S. marcescens strains from R. prolixus gut to the scientific community together with their genomic sequences. Here, we describe their genomic features with the purpose of bringing new insights into the S. marcescens adaptations for colonization of the specific niche of triatomine guts. This study provides the basis for a better understanding of the role of S. marcescens in the microbiota of R. prolixus gut as a potential antagonist of T. cruzi in this complex system

CORTICOSTEROIDS USE IN NEUROLOGY: HOW DOGS ARE REFERRAL TO A NEUROLOGY SERVICE?

Corticosteroids are drugs widely used in veterinary neurology due to their recognized analgesic and anti-inflammatory effects. However, their use in high doses and long terms may result in undesirable effects and disorders in other systems. This study describes the prevalence of dogs with neurological diseases referred to a veterinary neurology reference service with previous prescription of corticosteroids. For the retrospective study 284 medical records of dogs were evaluated in the period between August 2017 and April 2019. Of these, 194 (68%) patients already had medical care by another veterinary, and, 100 (52%) had previous prescription for the corticosteroid use. After the diagnosis, the dose was suspended in 28% of the cases, adjusted in 41% and maintained in 31%. Dogs with immunosuppressive doses prescription were the most affected, and the medication was adjusted or suspended in 71% and 21% of the cases, respectively. In conclusion, the therapy for neurological diseases is still strongly associated with the indiscriminate use of corticosteroids, without a dose correlation with the desired effect

Miastenia gravis adquirida em cão – relato de caso

Doenças neuromusculares são incomuns na rotina clínica de cães e gatos e o seu diagnóstico é um desafio para o Médico Veterinário. Desta forma, este trabalho relata o diagnóstico clínico e terapêutico em uma cadela sem raça definida de quatro anos com histórico de claudicação e paresia intermitente, inicialmente em membros pélvicos evoluindo para membros torácicos, apresentando sinais clínicos de ataxia e tetraparesia sobretudo após exercício. Em virtude do histórico e sinais clínicos apresentados suspeitou-se de miastenia gravis adquirida, optando-se pelo diagnóstico terapêutico, por ser mais simples, rápido e acessível, utilizando o anticolinesterásico neostigmina. A paciente apresentou melhora significativa no quadro clínico após administração do referido anticolinesterásico por via intramuscular, retornando à atividade deambulatória normal após administração do fármaco por via intravenosa, sendo possível assim o diagnóstico da doença. Foi instituído tratamento utilizando o brometo de piridostigmina (2,5mg/Kg, duas vezes ao dia) associado ao corticosteroide em dose imunossupressora (0,5mg/Kg, duas vezes ao dia), ambos por via oral, por um período de seis meses, observando-se remissão completa dos sinais clínicos sem recidivas até o presente momento

Käytännön kosteikkosuunnittelu

Maatalouden vesiensuojelua edistetään monin tavoin. Ravinteita ja eroosioainesta sisältäviä valumavesiä pyritään puhdistamaan erilaisissa kosteikoissa. Tämä opas on kirjoitettu avuksi pienimuotoisten kosteikkojen perustamiseen. Oppaassa esitetään käytännönläheisesti kosteikon toteuttamisen eri vaiheet paikan valinnasta suunnitteluun ja rakentamiseen. Vuonna 2010 julkaistun painoksen tiedot on saatettu ajantasalle. Julkaisu on toteutettu osana Tehoa maatalouden vesiensuojeluun (TEHO) -hanketta ja päivitetty TEHO Plus -hankkeen toimesta. Oppaan toivotaan lisäävän kiinnostusta kosteikkojen suunnitteluun ja edelleen niiden rakentamiseen