Assignment of gene function has been a crucial, laborious, and time-consuming step in genomics. D... more Assignment of gene function has been a crucial, laborious, and time-consuming step in genomics. Due to a variety of sequencing platforms that generates increasing amounts of data, manual annotation is no longer feasible. Thus, the need for an integrated, automated pipeline allowing the use of experimental data towards validation of in silico prediction of gene function is of utmost relevance. Here, we present a computational workflow named AnnotaPipeline that integrates distinct software and data types on a proteogenomic approach to annotate and validate predicted features in genomic sequences. Based on FASTA (i) nucleotide or (ii) protein sequences or (iii) structural annotation files (GFF3), users can input FASTQ RNA-seq data, MS/MS data from mzXML or similar formats, as the pipeline uses both transcriptomic and proteomic information to corroborate annotations and validate gene prediction, providing transcription and expression evidence for functional annotation. Reannotation of t...
COVID-19 has assumed significant and lasting proportions worldwide. Following initial cases in th... more COVID-19 has assumed significant and lasting proportions worldwide. Following initial cases in the Western mesoregion, the State of Santa Catarina (SC), southern Brazil, was heavily affected as a whole by the pandemic in early 2021. This study aimed to evaluate the dynamics of the SARS-CoV-2 virus spreading patterns in the SC state through March 2020 to April 2021 using genomic surveillance. During this period, 23 distinct variants, including two VOCs (Beta and Gamma) were identified, among which, the Gamma and related lineages were predominant in the second pandemic wave within SC. However, a regionalization of P.1-like-II in the Western region was observed, concomitant to the increase in cases, mortality, and case fatality rate (CFR) index. This is the first evidence of the regionalization of the SARS-CoV-2 transmission in the and highlight the importance of tracking variants, dispersion and their impact of SARS-CoV-2 on the public health system in Brazilian states.
Trypanosoma rangeli e um parasito nao patogenico que infecta mamiferos nas Americas, incluindo hu... more Trypanosoma rangeli e um parasito nao patogenico que infecta mamiferos nas Americas, incluindo humanos. Em razao da sua semelhanca morfologica e antigenica com Trypanosoma cruzi , agente etiologico da Doenca de Chagas, a coexistencia desses parasitas em humanos pode levar a erros no diagnostico da doenca. Em infeccoes causadas por T. cruzi , o isotipo IgG se apresenta em elevados titulos, porem, a variacao desse isotipo em T. rangeli e desconhecido . Avaliou-se, neste trabalho, a influencia dos anticorpos da classe IgG na sororeatividade cruzada desses parasitos em camundongos durante a infeccao experimental. Parasitos foram cultivados in vitro ( T. rangeli Choachi e T. cruzi Y) utilizando meio LIT e as formas tripomastigotas foram obtidas em cultura com meio DMEM e culturas envelhecidas, respectivamente. Aproximadamente 10 3 formas tripomastigotas foram inoculadas por via intraperitoneal e os antissoros obtidos por meio de coletas durante 64 dias. Mediante o immunoblotting foi obse...
<p>The blue lines represent regions of homology between the contigs. Annotated genes and ot... more <p>The blue lines represent regions of homology between the contigs. Annotated genes and other sequence characteristics are indicated by colored boxes. Arrows indicate sense transcription. <b>A</b>. Comparison between Scaffold Tr 61 (4,000–53,457 nt) and TcChr27-P (794,000–850,241 nt). <b>B</b>. Comparison between Scaffold Tr 115 (136,482–164,482 nt) and TcChr33-S (975,000–1,041,172 nt). Contig ends were oriented in the 5′ to 3′ direction according to the TriTrypDB assemblies of <i>T. cruzi</i> scaffolds. The accession numbers of the annotated sequences in the <i>T. cruzi</i> scaffolds (TriTrypDB) are displayed below the sequences.</p
Flebotomineos sao dipteros com habitos hematofagos, vetores de agentes etiologicos de leishmanios... more Flebotomineos sao dipteros com habitos hematofagos, vetores de agentes etiologicos de leishmanioses. O estudo objetivou identificar as especies de flebotomineos em municipios do oeste de Santa Catarina e verificar a possivel infeccao destas por Leishmania infantum . As coletas foram realizadas em sete pontos em: Joacaba (1-27° 9'43.77"S; 51°31'36.54"O, 2-27° 9'31.59"S; 51°30'45.35"O, 3-27°10'6.41"S; 51°30'14.04"O), Herval d’Oeste (4-27°10'14.21"S; 51°29'23.21"O), Lacerdopolis (5-27°16'24.23"S; 51°33'35.92"O), Herval Velho (6-27°18'14.31"S; 51°28'21.48"O) e Ouro (7-27°21'9.20"S; 51°41'24.11"O), com auxilio de armadilhas luminosas do tipo Falcao, entre 09/2015 e 03/2016. Os flebotomineos coletados foram preparados e identificados. As femeas, exceto Brumptomyia , tiveram o abdomen e a cabeca separados para a identificacao e o restante do corpo preparado para rea...
Clostridioides difficile BI/NAP1/ribotype 027 is an epidemic hypervirulent strain found worldwide... more Clostridioides difficile BI/NAP1/ribotype 027 is an epidemic hypervirulent strain found worldwide, including in Latin America. We examined the genomes and exoproteomes of two multilocus sequence type (MLST) clade 2 C. difficile strains considered hypervirulent: ICC-45 (ribotype SLO231/UK[CE]821), isolated in Brazil, and NAP1/027/ST01 (LIBA5756), isolated during a 2010 outbreak in Costa Rica. C. difficile isolates were cultured and extracellular proteins were analyzed using high-performance liquid chromatography-tandem mass spectrometry. Genomic analysis revealed that these isolates shared most of the gene composition. Only 83 and 290 NAP1/027 genes were considered singletons in ICC-45 and NAP1/027, respectively. Exoproteome analysis revealed 197 proteins, of which 192 were similar in both strains. Only five proteins were exclusive to the ICC-45 strain. These proteins were involved with catalytic and binding functions and indirectly interacted with proteins related to pathogenicity. ...
O Trypanosoma rangeli e um parasita hemoflagelado, muito similar geneticamente e antigenicamente ... more O Trypanosoma rangeli e um parasita hemoflagelado, muito similar geneticamente e antigenicamente com o Trypanosoma cruzi, agente etiologico da Doenca de Chagas. Porem, se tem conhecimento de varios genes exclusivos de cada especie, e no T. rangeli, alguns destes genes codificam proteinas hipoteticas, que ainda nao foram caracterizadas. Neste contexto, o objetivo deste estudo foi expressar uma proteina hipotetica de superficie do T. rangeli, para posterior avaliacao desta em diagnosticos sorologicos especificos. Primeiramente foi realizada busca no genoma para se obter uma proteina hipotetica especifica do T. rangeli, sendo selecionada a proteina hipotetica KC544871. Com posse da sequencia de nucleotideos, um par de iniciadores especificos para amplificar a sequencia que codifica a proteina de interesse a ser estudada foi construido. A amplificacao deste gene foi realizada atraves de PCR, gerando o produto esperado de 795pb. Apos foi realizado o processo de clonagem em vetor pGEM T-e...
We aimed to find out the sand fly species in five municipalities along the Peixe River in the wes... more We aimed to find out the sand fly species in five municipalities along the Peixe River in the west of Santa Catarina state, Brazil, and verify the presence of females naturally infected with Leishmania ( Leishmania ) infantum Nicolle . Phlebotomine were collected using a Falcao light trap and identified through morphology. A total of 149 specimens of 10 phlebotomine species were identified, four of which were already reported with natural Leishmania spp. infection. However, using amplification of L . ( L .) infantum kDNA through multiplex polymerase chain reaction (PCR) assay, no natural infection was observed in these specimens. The region is non-endemic for leishmaniasis and the low amount of collected specimens may have contributed to the negative results. However, the simultaneous presence of phlebotomine species reported in the literature as naturally infected with Leishmania spp. and a recent report of dogs naturally infected with L . ( L .) infantum demonstrate the need to ...
Diversas areas naturais sao rapidamente transformadas, provocando um desequilibrio ambiental, lev... more Diversas areas naturais sao rapidamente transformadas, provocando um desequilibrio ambiental, levando muitos animais silvestres a viverem e se adaptarem as condicoes urbanas. Com abundância de alimentos, ausencia de predadores e presenca de abrigo, individuos de Hydrochoerus hydrochaeris popularmente conhecidos como capivara, estao cada vez mais presentes nestes ambientes. Sendo assim, adquirem parasitas mais facilmente, podendo vir a transmitir patogenias ao homem, se tornando um problema a saude publica. O objetivo desse estudo e avaliar a presenca de parasitas em fezes de H. hydrochaeris ocorrentes no municipio de Joacaba nas margens do Rio do Peixe – SC. Os pontos de coletas foram divididos em: (P1) e (P4) sendo areas residenciais, (P2) area hospitalar e (P3) e (P5) areas comerciais. No periodo de janeiro a maio foram coletadas duas amostras de cada ponto, totalizando 50, nas quais 47 (94%) apresentaram resultados positivos para um ou mais parasitas. As analises das amostras for...
O Trypanosoma cruzi, agente causador da doenca de Chagas, possui em comum com o Trypanosoma range... more O Trypanosoma cruzi, agente causador da doenca de Chagas, possui em comum com o Trypanosoma rangeli, causador da rangeliose, antigenos que comprometem o diagnostico sorologico diferencial da doenca de Chagas, podendo haver resultados falsos positivos nos exames sorologicos para esta doenca. Deste modo, o presente estudo avaliou um peptideo TKPPPKEGEEYG da proteina GP63 de T. rangeli, como possivel marcador diferencial para o diagnostico sorologico desta doenca. Antissoros policlonais anti-Tc e anti-Tr foram obtidos atraves da infeccao experimental de camundongos Swiss e antissoros anti-peptideos pela imunizacao de camundongos Swiss com 50 μg de cada peptideo. Os ensaios de ELISA foram realizados em placas sensibilizadas com 10 μg/poco do peptideo. Ja os ensaios de imunoblotting foram realizados com 25μg de proteinas soluveis de formas epimastigotas destes parasitos. Os antissoros foram avaliados nas diluicoes de 1:50 a 1:400. Ensaios de ELISA utilizando o peptideo como antigeno resu...
Canine visceral leishmaniasis (CVL) caused by Leishmania (Leishmania) infantum is transmitted by ... more Canine visceral leishmaniasis (CVL) caused by Leishmania (Leishmania) infantum is transmitted by phlebotomine sandflies and a major zoonotic disease in Brazil. Due to the southward expansion of the disease within the country and the central role of dogs as urban reservoirs of the parasite, we have investigated the occurrence of CVL in two municipalities Erval Velho and Herval d’Oeste in the Midwest region of Santa Catarina state. Peripheral blood samples from 126 dogs were collected in both cities and tested for anti-L. infantum antibodies by indirect enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence reaction (IIF) and for the presence of parasite DNA by polymerase chain reaction (PCR) in peripheral blood. From examined dogs, 35.71% (45/126) were positive for at least one of the three tests and two (1.6%) were positive in all performed tests. Twelve dogs (9.5%) were positive for both ELISA and IIF, while 21 dogs were exclusively positive for ELISA (16.7%), an...
Trypanosoma rangeli is a non-virulent hemoflagellate parasite infecting humans, wild and domestic... more Trypanosoma rangeli is a non-virulent hemoflagellate parasite infecting humans, wild and domestic mammals in Central and Latin America. The share of genotypic, phenotypic, and biological similarities with the virulent, human-infective T. cruzi and T. brucei, allows comparative studies on mechanisms of pathogenesis. In this study, investigation of the T. rangeli Arginine Kinase (TrAK) revealed two highly similar copies of the AK gene in this taxon, and a distinct expression profile and activity between replicative and infective forms. Although TrAK expression seems stable during epimastigotes growth, the enzymatic activity increases during the exponential growth phase and decreases from the stationary phase onwards. No differences were observed in activity or expression levels of TrAK during in vitro differentiation from epimastigotes to infective forms, and no detectable AK expression was observed for blood trypomastigotes. Overexpression of TrAK by T. rangeli showed no effects on the in vitro growth pattern, differentiation to infective forms, or infectivity to mice and triatomines. Although differences in TrAK expression and activity were observed among T. rangeli strains from distinct genetic lineages, our results indicate an up-regulation during parasite replication and putative post-translational myristoylation of this enzyme. We conclude that up-regulation of TrAK activity in epimastigotes appears to improve proliferation fitness, while reduced TrAK expression in blood trypomastigotes may be related to short-term and subpatent parasitemia in mammalian hosts.
Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respirator... more Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) from October 2019 until March 2020. Twenty five ml of sewage samples were clarified and viruses concentrated using a glycine buffer method coupled with polyethylene glycol precipitation, and viral RNA extracted using a commercial kit. SARS-CoV-2 RNA was detected by RT-qPCR using oligonucleotides targeting N1, S and two RdRp regions. The results of all positive samples were further confirmed by a different RT-qPCR system in an independent laboratory. S and RdRp amplicons were sequenced to confirm identity with SARS-CoV-2. Genome sequencing was performed using two strategies; a sequence-independent single-primer amplification (SISPA) approach, and by direct metagenomics using Illumina's NGS. SARS-CoV-2 RNA was detected on 27th November 2019 (5.49 ± 0.02 log10 SARS-CoV-2 genome copies (GC) L−1), detection being confirmed by an independent laboratory and genome sequencing analysis. The samples in the subsequent three events were positive by all RT-qPCR assays; these positive results were also confirmed by an independent laboratory. The average load was 5.83 ± 0.12 log10 SARS-CoV-2 GC L−1, ranging from 5.49 ± 0.02 log10 GC L−1 (27th November 2019) to 6.68 ± 0.02 log10 GC L−1 (4th March 2020). Our findings demonstrate that SARS-CoV-2 was likely circulating undetected in the community in Brazil since November 2019, earlier than the first reported case in the Americas (21st January 2020).
We analysed human sewage located in Florianópolis (Santa Catalina, Brazil) from late October unti... more We analysed human sewage located in Florianópolis (Santa Catalina, Brazil) from late October until the Brazil lockdown on early March. We detected SARS-CoV-2 in two samples collected independently on 27th November 2019 (5.49±0.02 log genome copies/L). Subsequent samplings were positive until 4th March 2020 (coinciding with the first COVID-19 case reported in Santa Catalina), with a SARS-CoV-2 RNA increase of one log (6.68±0.02 log genome copies/L). Our results show that SARS-CoV-2 has been circulating in Brazil since late November 2019, much earlier than the first reported case in the Americas (21st January 2020, USA).
Assignment of gene function has been a crucial, laborious, and time-consuming step in genomics. D... more Assignment of gene function has been a crucial, laborious, and time-consuming step in genomics. Due to a variety of sequencing platforms that generates increasing amounts of data, manual annotation is no longer feasible. Thus, the need for an integrated, automated pipeline allowing the use of experimental data towards validation of in silico prediction of gene function is of utmost relevance. Here, we present a computational workflow named AnnotaPipeline that integrates distinct software and data types on a proteogenomic approach to annotate and validate predicted features in genomic sequences. Based on FASTA (i) nucleotide or (ii) protein sequences or (iii) structural annotation files (GFF3), users can input FASTQ RNA-seq data, MS/MS data from mzXML or similar formats, as the pipeline uses both transcriptomic and proteomic information to corroborate annotations and validate gene prediction, providing transcription and expression evidence for functional annotation. Reannotation of t...
COVID-19 has assumed significant and lasting proportions worldwide. Following initial cases in th... more COVID-19 has assumed significant and lasting proportions worldwide. Following initial cases in the Western mesoregion, the State of Santa Catarina (SC), southern Brazil, was heavily affected as a whole by the pandemic in early 2021. This study aimed to evaluate the dynamics of the SARS-CoV-2 virus spreading patterns in the SC state through March 2020 to April 2021 using genomic surveillance. During this period, 23 distinct variants, including two VOCs (Beta and Gamma) were identified, among which, the Gamma and related lineages were predominant in the second pandemic wave within SC. However, a regionalization of P.1-like-II in the Western region was observed, concomitant to the increase in cases, mortality, and case fatality rate (CFR) index. This is the first evidence of the regionalization of the SARS-CoV-2 transmission in the and highlight the importance of tracking variants, dispersion and their impact of SARS-CoV-2 on the public health system in Brazilian states.
Trypanosoma rangeli e um parasito nao patogenico que infecta mamiferos nas Americas, incluindo hu... more Trypanosoma rangeli e um parasito nao patogenico que infecta mamiferos nas Americas, incluindo humanos. Em razao da sua semelhanca morfologica e antigenica com Trypanosoma cruzi , agente etiologico da Doenca de Chagas, a coexistencia desses parasitas em humanos pode levar a erros no diagnostico da doenca. Em infeccoes causadas por T. cruzi , o isotipo IgG se apresenta em elevados titulos, porem, a variacao desse isotipo em T. rangeli e desconhecido . Avaliou-se, neste trabalho, a influencia dos anticorpos da classe IgG na sororeatividade cruzada desses parasitos em camundongos durante a infeccao experimental. Parasitos foram cultivados in vitro ( T. rangeli Choachi e T. cruzi Y) utilizando meio LIT e as formas tripomastigotas foram obtidas em cultura com meio DMEM e culturas envelhecidas, respectivamente. Aproximadamente 10 3 formas tripomastigotas foram inoculadas por via intraperitoneal e os antissoros obtidos por meio de coletas durante 64 dias. Mediante o immunoblotting foi obse...
<p>The blue lines represent regions of homology between the contigs. Annotated genes and ot... more <p>The blue lines represent regions of homology between the contigs. Annotated genes and other sequence characteristics are indicated by colored boxes. Arrows indicate sense transcription. <b>A</b>. Comparison between Scaffold Tr 61 (4,000–53,457 nt) and TcChr27-P (794,000–850,241 nt). <b>B</b>. Comparison between Scaffold Tr 115 (136,482–164,482 nt) and TcChr33-S (975,000–1,041,172 nt). Contig ends were oriented in the 5′ to 3′ direction according to the TriTrypDB assemblies of <i>T. cruzi</i> scaffolds. The accession numbers of the annotated sequences in the <i>T. cruzi</i> scaffolds (TriTrypDB) are displayed below the sequences.</p
Flebotomineos sao dipteros com habitos hematofagos, vetores de agentes etiologicos de leishmanios... more Flebotomineos sao dipteros com habitos hematofagos, vetores de agentes etiologicos de leishmanioses. O estudo objetivou identificar as especies de flebotomineos em municipios do oeste de Santa Catarina e verificar a possivel infeccao destas por Leishmania infantum . As coletas foram realizadas em sete pontos em: Joacaba (1-27° 9'43.77"S; 51°31'36.54"O, 2-27° 9'31.59"S; 51°30'45.35"O, 3-27°10'6.41"S; 51°30'14.04"O), Herval d’Oeste (4-27°10'14.21"S; 51°29'23.21"O), Lacerdopolis (5-27°16'24.23"S; 51°33'35.92"O), Herval Velho (6-27°18'14.31"S; 51°28'21.48"O) e Ouro (7-27°21'9.20"S; 51°41'24.11"O), com auxilio de armadilhas luminosas do tipo Falcao, entre 09/2015 e 03/2016. Os flebotomineos coletados foram preparados e identificados. As femeas, exceto Brumptomyia , tiveram o abdomen e a cabeca separados para a identificacao e o restante do corpo preparado para rea...
Clostridioides difficile BI/NAP1/ribotype 027 is an epidemic hypervirulent strain found worldwide... more Clostridioides difficile BI/NAP1/ribotype 027 is an epidemic hypervirulent strain found worldwide, including in Latin America. We examined the genomes and exoproteomes of two multilocus sequence type (MLST) clade 2 C. difficile strains considered hypervirulent: ICC-45 (ribotype SLO231/UK[CE]821), isolated in Brazil, and NAP1/027/ST01 (LIBA5756), isolated during a 2010 outbreak in Costa Rica. C. difficile isolates were cultured and extracellular proteins were analyzed using high-performance liquid chromatography-tandem mass spectrometry. Genomic analysis revealed that these isolates shared most of the gene composition. Only 83 and 290 NAP1/027 genes were considered singletons in ICC-45 and NAP1/027, respectively. Exoproteome analysis revealed 197 proteins, of which 192 were similar in both strains. Only five proteins were exclusive to the ICC-45 strain. These proteins were involved with catalytic and binding functions and indirectly interacted with proteins related to pathogenicity. ...
O Trypanosoma rangeli e um parasita hemoflagelado, muito similar geneticamente e antigenicamente ... more O Trypanosoma rangeli e um parasita hemoflagelado, muito similar geneticamente e antigenicamente com o Trypanosoma cruzi, agente etiologico da Doenca de Chagas. Porem, se tem conhecimento de varios genes exclusivos de cada especie, e no T. rangeli, alguns destes genes codificam proteinas hipoteticas, que ainda nao foram caracterizadas. Neste contexto, o objetivo deste estudo foi expressar uma proteina hipotetica de superficie do T. rangeli, para posterior avaliacao desta em diagnosticos sorologicos especificos. Primeiramente foi realizada busca no genoma para se obter uma proteina hipotetica especifica do T. rangeli, sendo selecionada a proteina hipotetica KC544871. Com posse da sequencia de nucleotideos, um par de iniciadores especificos para amplificar a sequencia que codifica a proteina de interesse a ser estudada foi construido. A amplificacao deste gene foi realizada atraves de PCR, gerando o produto esperado de 795pb. Apos foi realizado o processo de clonagem em vetor pGEM T-e...
We aimed to find out the sand fly species in five municipalities along the Peixe River in the wes... more We aimed to find out the sand fly species in five municipalities along the Peixe River in the west of Santa Catarina state, Brazil, and verify the presence of females naturally infected with Leishmania ( Leishmania ) infantum Nicolle . Phlebotomine were collected using a Falcao light trap and identified through morphology. A total of 149 specimens of 10 phlebotomine species were identified, four of which were already reported with natural Leishmania spp. infection. However, using amplification of L . ( L .) infantum kDNA through multiplex polymerase chain reaction (PCR) assay, no natural infection was observed in these specimens. The region is non-endemic for leishmaniasis and the low amount of collected specimens may have contributed to the negative results. However, the simultaneous presence of phlebotomine species reported in the literature as naturally infected with Leishmania spp. and a recent report of dogs naturally infected with L . ( L .) infantum demonstrate the need to ...
Diversas areas naturais sao rapidamente transformadas, provocando um desequilibrio ambiental, lev... more Diversas areas naturais sao rapidamente transformadas, provocando um desequilibrio ambiental, levando muitos animais silvestres a viverem e se adaptarem as condicoes urbanas. Com abundância de alimentos, ausencia de predadores e presenca de abrigo, individuos de Hydrochoerus hydrochaeris popularmente conhecidos como capivara, estao cada vez mais presentes nestes ambientes. Sendo assim, adquirem parasitas mais facilmente, podendo vir a transmitir patogenias ao homem, se tornando um problema a saude publica. O objetivo desse estudo e avaliar a presenca de parasitas em fezes de H. hydrochaeris ocorrentes no municipio de Joacaba nas margens do Rio do Peixe – SC. Os pontos de coletas foram divididos em: (P1) e (P4) sendo areas residenciais, (P2) area hospitalar e (P3) e (P5) areas comerciais. No periodo de janeiro a maio foram coletadas duas amostras de cada ponto, totalizando 50, nas quais 47 (94%) apresentaram resultados positivos para um ou mais parasitas. As analises das amostras for...
O Trypanosoma cruzi, agente causador da doenca de Chagas, possui em comum com o Trypanosoma range... more O Trypanosoma cruzi, agente causador da doenca de Chagas, possui em comum com o Trypanosoma rangeli, causador da rangeliose, antigenos que comprometem o diagnostico sorologico diferencial da doenca de Chagas, podendo haver resultados falsos positivos nos exames sorologicos para esta doenca. Deste modo, o presente estudo avaliou um peptideo TKPPPKEGEEYG da proteina GP63 de T. rangeli, como possivel marcador diferencial para o diagnostico sorologico desta doenca. Antissoros policlonais anti-Tc e anti-Tr foram obtidos atraves da infeccao experimental de camundongos Swiss e antissoros anti-peptideos pela imunizacao de camundongos Swiss com 50 μg de cada peptideo. Os ensaios de ELISA foram realizados em placas sensibilizadas com 10 μg/poco do peptideo. Ja os ensaios de imunoblotting foram realizados com 25μg de proteinas soluveis de formas epimastigotas destes parasitos. Os antissoros foram avaliados nas diluicoes de 1:50 a 1:400. Ensaios de ELISA utilizando o peptideo como antigeno resu...
Canine visceral leishmaniasis (CVL) caused by Leishmania (Leishmania) infantum is transmitted by ... more Canine visceral leishmaniasis (CVL) caused by Leishmania (Leishmania) infantum is transmitted by phlebotomine sandflies and a major zoonotic disease in Brazil. Due to the southward expansion of the disease within the country and the central role of dogs as urban reservoirs of the parasite, we have investigated the occurrence of CVL in two municipalities Erval Velho and Herval d’Oeste in the Midwest region of Santa Catarina state. Peripheral blood samples from 126 dogs were collected in both cities and tested for anti-L. infantum antibodies by indirect enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence reaction (IIF) and for the presence of parasite DNA by polymerase chain reaction (PCR) in peripheral blood. From examined dogs, 35.71% (45/126) were positive for at least one of the three tests and two (1.6%) were positive in all performed tests. Twelve dogs (9.5%) were positive for both ELISA and IIF, while 21 dogs were exclusively positive for ELISA (16.7%), an...
Trypanosoma rangeli is a non-virulent hemoflagellate parasite infecting humans, wild and domestic... more Trypanosoma rangeli is a non-virulent hemoflagellate parasite infecting humans, wild and domestic mammals in Central and Latin America. The share of genotypic, phenotypic, and biological similarities with the virulent, human-infective T. cruzi and T. brucei, allows comparative studies on mechanisms of pathogenesis. In this study, investigation of the T. rangeli Arginine Kinase (TrAK) revealed two highly similar copies of the AK gene in this taxon, and a distinct expression profile and activity between replicative and infective forms. Although TrAK expression seems stable during epimastigotes growth, the enzymatic activity increases during the exponential growth phase and decreases from the stationary phase onwards. No differences were observed in activity or expression levels of TrAK during in vitro differentiation from epimastigotes to infective forms, and no detectable AK expression was observed for blood trypomastigotes. Overexpression of TrAK by T. rangeli showed no effects on the in vitro growth pattern, differentiation to infective forms, or infectivity to mice and triatomines. Although differences in TrAK expression and activity were observed among T. rangeli strains from distinct genetic lineages, our results indicate an up-regulation during parasite replication and putative post-translational myristoylation of this enzyme. We conclude that up-regulation of TrAK activity in epimastigotes appears to improve proliferation fitness, while reduced TrAK expression in blood trypomastigotes may be related to short-term and subpatent parasitemia in mammalian hosts.
Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respirator... more Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) from October 2019 until March 2020. Twenty five ml of sewage samples were clarified and viruses concentrated using a glycine buffer method coupled with polyethylene glycol precipitation, and viral RNA extracted using a commercial kit. SARS-CoV-2 RNA was detected by RT-qPCR using oligonucleotides targeting N1, S and two RdRp regions. The results of all positive samples were further confirmed by a different RT-qPCR system in an independent laboratory. S and RdRp amplicons were sequenced to confirm identity with SARS-CoV-2. Genome sequencing was performed using two strategies; a sequence-independent single-primer amplification (SISPA) approach, and by direct metagenomics using Illumina's NGS. SARS-CoV-2 RNA was detected on 27th November 2019 (5.49 ± 0.02 log10 SARS-CoV-2 genome copies (GC) L−1), detection being confirmed by an independent laboratory and genome sequencing analysis. The samples in the subsequent three events were positive by all RT-qPCR assays; these positive results were also confirmed by an independent laboratory. The average load was 5.83 ± 0.12 log10 SARS-CoV-2 GC L−1, ranging from 5.49 ± 0.02 log10 GC L−1 (27th November 2019) to 6.68 ± 0.02 log10 GC L−1 (4th March 2020). Our findings demonstrate that SARS-CoV-2 was likely circulating undetected in the community in Brazil since November 2019, earlier than the first reported case in the Americas (21st January 2020).
We analysed human sewage located in Florianópolis (Santa Catalina, Brazil) from late October unti... more We analysed human sewage located in Florianópolis (Santa Catalina, Brazil) from late October until the Brazil lockdown on early March. We detected SARS-CoV-2 in two samples collected independently on 27th November 2019 (5.49±0.02 log genome copies/L). Subsequent samplings were positive until 4th March 2020 (coinciding with the first COVID-19 case reported in Santa Catalina), with a SARS-CoV-2 RNA increase of one log (6.68±0.02 log genome copies/L). Our results show that SARS-CoV-2 has been circulating in Brazil since late November 2019, much earlier than the first reported case in the Americas (21st January 2020, USA).
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