RESUMO
A phage-display library was generated using a Bus thalamus scorpion toxin (BTK-2) as a peptide scaffold. BTK-2 belongs to the disulfide-rich family of proteins with pronounced structural stability due to the presence of three disulfide bridges that connects antiparallel beta-sheets and one alpha helix. Using BTK-2 as a phage display scaffold, we introduced mutations in five residues located in the alpha-helix and two residues located in the smaller loop, keeping intact the disulfide bridges to create a peptide phage-displayed library with disulfide-rich family properties. The library was subjected to in vivo and in vitro phage display selections against Trypanosoma evansi, the etiological agent of "Surra", a disease that affects a wide range of mammals. The development of T. evansi specific biomarkers is essential to improve diagnostic methods and epidemiological studies leading to a more accurate clinical decision for the treatment of this disease of economic impact for commercial livestock production. In this study, we identified two disulfide-rich peptides targeting T. evansi parasites. Further specificity studies are necessary to investigate the potential of selected peptides as new biomarkers to aid diagnostic and treatment procedures of T. evansi infections.
Assuntos
Dissulfetos , Peptídeos , Trypanosoma/química , Tripanossomíase/diagnóstico , Tripanossomíase/terapia , Sequência de Aminoácidos , Animais , Biomarcadores , Clonagem Molecular , Dissulfetos/química , Camundongos , Camundongos Endogâmicos BALB C , Mutagênese , Oligonucleotídeos/química , Biblioteca de Peptídeos , Peptídeos/química , Peptídeos/genética , Venenos de Escorpião/química , Venenos de Escorpião/genéticaRESUMO
Abstract Trypanosomiasis caused by Trypanosoma evansi can seriously affect both domestic and wild animals. This article reports on an outbreak of canine trypanosomiasis on a farm in the Pantanal region of Brazil. The farm had 38 dogs, 20 of which died before receiving veterinary care. The remaining 18 dogs were underwent anamnesisn, clinical examination, hematological and biochemical evaluations. Blood smears and PCR analysis were performed for the diagnosis. The treatment protocols used according to the clinical recovery or parasitological cure of the dogs, using diminazene diaceturate, isometamidium chloride or quinapyramine sulfate. Post-treatment parasitological evaluation was performed by the microhematocrit technique. 7/18 dogs were PCR positive for T. evansi (confirmed by sequencing). There was clinical findings, which were consistent with both the acute and chronic stages of the disease in dogs. The infected dogs all exhibited at least one clinical sign of the disease. The hematological findings were compatible with trypanosomiasis, highlighting the hypochromic microcytic anemia as the main outcome. No treatment protocol was fully effective and the prolonged use of diminazene diaceturate caused the death of an animal. The trypanosomiasis can cause high rates of morbidity and mortality in dogs and difficulty in establishment an effective and safe therapeutic protocol.
Resumo A tripanossomíase causada por Trypanosoma evansi pode acometer gravemente os animais domésticos e selvagens. Este artigo relata um surto de tripanossomíase canina em uma fazenda na região do Pantanal, Brasil. Na fazenda havia 38 cães, 20 dos quais morreram antes de receber cuidados veterinários. Os 18 cães restantes foram submetidos a anamnese, exame clínico, avaliação hematológica e bioquímica. Esfregaços de sangue e análise da PCR foram realizados para o diagnóstico. Os protocolos de tratamento foram utilizados de acordo com a recuperação clínica ou cura parasitológica dos cães, utilizando diaceturato de diminazeno, cloreto de isometamídio ou sulfato de quinapiramina. A avaliação parasitológica pós-tratamento foi realizada pela técnica de microhematócrito. 7/18 cães foram PCR positivos para T. evansi (confirmado por sequenciamento). Os achados clínicos encontrados, foram consistentes com os estágios agudo e crônico da doença em cães. Todos os cães infectados exibiram pelo menos um sinal clínico da doença. Os achados hematológicos foram compatíveis com a tripanossomíase, destacando a anemia microcítica hipocrômica como principal consequência. Nenhum protocolo de tratamento foi totalmente eficaz e o uso prolongado de diaceturato de diminazeno causou a morte de um animal. A tripanossomíase pode causar altas taxas de morbidade e mortalidade em cães e dificultar o estabelecimento de um protocolo terapêutico eficaz e seguro.
Assuntos
Humanos , Masculino , Feminino , Cães , Fenantridinas/uso terapêutico , Compostos de Quinolínio/uso terapêutico , Tripanossomíase/diagnóstico , Diminazena/análogos & derivados , Doenças do Cão/diagnóstico , Tripanossomíase/terapia , Tripanossomíase/epidemiologia , Brasil/epidemiologia , Reação em Cadeia da Polimerase/veterinária , Surtos de Doenças , Diminazena/uso terapêutico , Doenças do Cão/tratamento farmacológico , Doenças do Cão/epidemiologiaRESUMO
O objetivo deste estudo foi comparar o efeito do plasma imune in natura e congelado de coelhos com infecção aguda no controle profilático e curativo do Trypanosoma evansi em ratos parasitados. Foram utilizados, um coelho e 30 ratos adultos. O coelho foi infectado com 0,5 ml de sangue contendo tripomastigotas. Ao apresentar o parasita na corrente sanguínea, foi coletado o sangue e deste obtido o plasma, o qual foi armazenado em tubos de criopreservação e congelado, "plasma congelado" e a outra metade foi mantida a 37°C por 30 minutos, "plasma in natura". Os ratos foram separados em seis grupos homogêneos, sendo todos infectados intraperitonialmente com 4,5 x 104 tripomastigotas de um isolado de T. evansi. Foram administrados 0,8ml de plasma nos tratamentos profilático (grupos A e B), dois dias antes da inoculação do parasito, e curativo (grupos C e D), dois dias após inoculação do T. evansi. O grupo E recebeu apenas solução fisiológica, grupo controle. O plasma congelado foi administrado nos grupos A e C e o plasma "in natura" nos grupos B e D. Os efeitos do plasma de coelhos com infecção aguda foi avaliado através do período pré-patente, sobrevida e cura dos ratos. Não houve diferença entre os períodos pré-patente. Foi observada uma longevidade superior dos ratos do grupo B que foi tratado com plasma "in natura" profilaticamente. Nenhum animal obteve a cura da infecção, porém podemos concluir que a terapêutica com plasma imune de coelhos aumenta a sobrevida de ratos infectados com T. evansi.
This study was aimed to compare the effect of fresh and frozen immune plasma of rabbits with acute infection in the prophylactic and curative control of Trypanosoma evansi-infected rats. Thirty adult rats and a rabbit were used. The rabbit was infected with 0.5ml of blood containing trypomastigotes. Showing the parasite in the bloodstream, plasma was obtained and divided in frozen and fresh plasma. Frozen plasma was stored in cryopreservation tubes and frozen. Fresh plasma was kept at 37°C for 30 minutes. The rats were separated into six equal groups, all infected intraperitoneally with 4.5 x 104 trypomastigotes of the T. evansi strain. Each rat from prophylatic (A and B) and curative(C and D) groups received 0.8ml of plasma. Animals from group E(control group) received only saline. Frozen plasma was administered in groups A and C and the fresh plasma in groups B and D. The effects of rabbits plasma with acute infection was estimated by evaluation of the prepatency period, longevity and curative effects of rats. No difference was observed in the prepatency period. The longevity of the animals treated prophylactically with fresh plasma (group B) was superior than the other groups. No curative effect was observed in the animals, but based upon the results it is concluded that the treatment with immune plasma of rabbits increases the longevity ofT. evansi-infected rats.
Assuntos
Animais , Coelhos , Ratos , Plasma/imunologia , Tripanossomíase/terapia , Parasitemia/veterinária , Trypanosoma , Prevenção de DoençasRESUMO
Around 1900 Laveran and Mesnil discovered that African trypanosomes do not survive in the blood of some primates and humans. The nature of the trypanolytic factor present in these sera has been the focus of a long-standing debate between different groups. The aim of this study was to investigate the susceptibility of T. evansi isolates to therapy using human blood and plasma in experimentally infected mice. Forty-eight 2-month-old female mice (Mus musculus) were divided into six groups of eight animals per group (A, B, C, D, E and F). Plasma was obtained after blood collection in order to perform therapy. Animals from group A (positive control) were inoculated with T. evansi and treated with 0.2mL of saline solution. Animals from groups B and C were infected with the flagellate and received a curative treatment with 0.2mL of human blood (group B) and 0.2mL of human plasma (group C), 24h after infection. Animals from groups D and E received a prophylactic treatment with 0.2mL of human blood and 0.2mL of human plasma, respectively, 24h prior to the infection. Animals from group F (negative control) were not infected and received 0.2mL of saline solution. The four treatments (B, C, D and E) increased animals longevity when compared to group A. Prepatency period was longer in groups D (15 days) and E (37.7 days) under prophylactic immunotherapy. Moreover, no parasites were found in most of the animals 60 days post-inoculation (PI). Besides the longer longevity, treatments were capable of curing 50% of mice of group B, 37.5% of group C, 37.5% of group D and 25% of the animals from group E.
Assuntos
Sangue , Plasma , Trypanosoma/fisiologia , Tripanossomíase/parasitologia , Tripanossomíase/terapia , Animais , Sangue/parasitologia , Feminino , Humanos , Camundongos , Análise de Sobrevida , Tripanossomicidas/administração & dosagem , Tripanossomíase/mortalidadeRESUMO
INTRODUCTION: Almost three out of every four people in the world who suffer a fatal stroke live in developing countries. A number of different tropical diseases may appear in Europe in the coming years as a consequence of the demographic change that is being brought about by migratory flows. We review the main infectious causes of strokes in the tropics. DEVELOPMENT: There are estimated to be 500 million cases of malaria every year. Cerebral malaria can cause cerebral oedema, diffuse or focal compromise of the subcortical white matter and cortical, cerebellar and pontine infarctions. Chagas disease is an independent risk factor for stroke in South America. At least 20 million people have the chronic form of Chagas disease. The main prognostic factors for Chagas-related stroke are the presence of apical aneurysms, arrhythmia and heart failure. Vascular complications of neurocysticercosis include transient ischemic attacks, ischemic strokes due to angiitis and intracranial haemorrhages. The frequency of cerebral infarction associated with neurocysticercosis varies between 2% and 12%. Gnathostomiasis is a cause of subarachnoid haemorrhage in south-east Asia. Other less common causes of stroke are viral haemorrhagic fevers due to arenavirus and flavivirus. CONCLUSIONS: Several diseases that are endemic in the tropics can be responsible for up to 10% of the cases of strokes in adults.