Modelos preditivos como ferramentas para estimar a quantidade de protozoários parasitos em peixes

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias, Programa de Pós-Graduação em Aquicultura, Florianópolis, 2011Protozoários parasitos são causadores de mortalidade em peixes cultivados, porém existe um número muito pequeno de trabalhos com análises quantitativas sobre o assunto no Brasil. Uma gama de métodos para quantificar parasitos é conhecida, entretanto eles geralmente requerem equipamentos caros e mão de obra especializada. Neste estudo foi desenvolvido um modelo preditivo e, a partir dele, um software, com o objetivo de auxiliar na quantificação de protozoários parasitos nas brânquias de peixe, tornando-a mais precisa e confiável, levando em consideração a realidade brasileira. Tilápias do Nilo (n=129) foram coletadas e tiveram um total de oito variáveis aferidas e testadas no processo de desenvolvimento do modelo preditivo. Os protocolos de coleta e aferição das variáveis, juntamente com a modelagem por meio de MARSplines, provaram ser eficientes e resultaram em um modelo preditivo bastante preciso e robusto. Esses resultados subsidiaram o desenvolvimento de um software, batizado de ProtoCount, que é capaz de estimar a quantidade total de protozoários nas brânquias, a partir dos valores de peso e comprimento do peixe e da média de protozoários observados em amostras de 1,5 ml, retiradas de 200 ml de solução formalina contendo raspado das brânquias do peixe amostrado e contadas em câmara de McMaster. ProtoCount demonstrou ser prático e confiável, assim, espera-se que sua utilização possa contribuir com a expansão, padronização, precisão e confiabilidade das análises quantitativas feitas por laboratórios nacionais de parasitologia de peixes.Protozoan parasites are a major cause of cultured fish mortality, but there are surprisingly few data with quantitative measurements in Brazil. There are few reliable methods for quantifying parasite protozoa, but either they require expensive equipments or highly specialized professionals. This study has developed a predictive model and a software, with the objective of providing more precise and reliable quantifications of protozoan parasites on fish gills more, considering Brazilian reality. Nile tilapia (n = 129) were collected and a total of eight variables were measured and tested for the predictive model development. Colleting and measurement procedures, along with MARSplines modeling, were efficient and resulted in a very precise and robust predicting model. Resulting data helped in developing a software, called ProtoCount, that manages to estimate the total protozoa burned on fish gills, based on fish#s weight, length and mean protozoa count on a 1,5 ml sample. Each 1,5 ml corresponds to 10 homogenized samples, taken from 200 ml formalin solution, containing scraped gills from sample fish, and counted using McMaster#s chamber. ProtoCount proved itself to be simples and reliable, therefore capable of increasing the number, standardization, precision and reliability of quantitative analysis performed in national fish parasitology laboratories

Induced aerocystitis and hemato-immunological parameters in Nile tilapia fed supplemented diet with essential oil of Lippia alba

O presente estudo avaliou a suplementação dietária com óleo essencial de Lippia alba sobre os parâmetros hemato-imunológicos em tilápias do Nilo (Oreochromis niloticus) submetidas à inflamação aguda induzida por carragenina na bexiga natatória. Pelo período de 45 dias, 96 peixes foram divididos em quarto tratamentos em triplicata: a) peixes suplementados com óleo esencial de L. alba (4 mL kg-1 de ração) injetados com carragenina; b) peixes suplementados com álcool de cereais injetados com carragenina; peixes não suplementados com óleo essencial injetados com carragenina; c) peixes não suplementados não injetados. Os níveis de cortisol, eritrograma, leucograma e o infiltrado inflamatório foram analisados seis horas após o estímulo inflamatório. Peixes injetados com carragenina apresentaram reação inflamatória aguda na bexiga natatória caracterizada por maior infiltrado de neutrófilos e monócitos. O número de neutrófilos circulantes foi significativamente maior nos peixes suplementados com L. alba quando comparado aos outros tratamentos. Não houve diferença nos níveis de cortisol. Para a dose, o tempo e a forma de administração testada, a suplementação com óleo essencil de L. alba não apresentou atividade anti-inflamatória. Por outro lado, foi constatada influência da suplementação dietária no número de neutrófilos após a aerocistite enfatizando a sua característica imunomoduladora.The present study evaluated the dietary supplementation with essential oil of Lippia alba on the hemato-immunological parameters of Nile tilapia (Oreochromis niloticus) submitted to acute inflammation induced by carrageenin injection in the swim bladder. For a period of 45 days, 96 fish were divided in four treatments in triplicate, as follows: fish fed supplemented diet with essential oil of L. alba (4 mL kg-1 dry ration) injected with carrageenin; fish fed supplemented diet with cereal alcohol injected with carrageenin; fish fed unsupplemented diet with essential oil injected with carrageenin; fish fed unsupplemented diet and noninjected. Cortisol levels, erythrogram, leukogram and the inflammatory infiltrate were analyzed 6 h after inflammatory stimulus. Carrageenin-injected fish showed acute inflammatory reaction in the swim bladder characterized by higher infiltrate of neutrophils and monocytes. The circulating neutrophils number was significantly higher in fish fed L. alba when compared to other treatments. No difference in cortisol levels was found. For dose, time and administration form tested, supplementation with essential oil of L. alba did not present anti-inflammatory activity. On the other hand, influence of dietary supplementation was observed on the neutrophils number after induced aerocystitis highlighting its immunomodulatory characteristic

MICRO HÁBITAT BRANQUIAL DE MONOGENEAY COPEPODITOS DE LERNAEACYPRINACEA EN LAS BRANQUIAS DE CUATRO PECES BRASILEÑOS DE AGUA DULCE

This study evaluated the ectoparasite distribution on the gill archs of pond-reared pacu (Piaractus mesopotamicusHolmberg, 1887), hybrid patinga (P. mesopotamicusfemale x P. brachypomus(Cuvier, 1817) male), hybrid tambacu (Colossoma macropomum(Cuvier, 1816) female x P. mesopotamicus male) and hybrid surubim (Pseudoplatystoma reticulatumfemale x P. corruscansSpix & Agassiz, 1829 male). Atotal of 300 fish were examined from fish farms in West-Central Brazil for parasitological assessment of the gill arches I, II, III and IV, from the outer to inner arch, respectively. Only the monogenean Mymarothecium boegeriCohen & Kohn, 2005 and Anacanthorus penilabiatusBoeger, Husak & Martins, 1995 from the hybrid patinga showed the greatest (p<0.05) mean intensities on the gill arch I when compared to gill arch IV. Microhabitat preference was not observed in the other fish examined. Copepodids of Lernaea cyprinaceaLinnaeus 1758 showed no microhabitat preference. This study highlighted the fact that under culture conditions, homogeneous distribution of parasites on the gills may occur.Este estudio evaluó la distribución de ectoparásitos en los arcos branquiales de pacu (Piaractus mesopotamicus), hibrido patinga (P. mesopotamicus hembra x P. brachypomus macho), híbrido tambacu (Colossoma macropomumhembra x P. mesopotamicus macho) e híbrido surubim (Pseudoplastystoma reticulatumhembra x P. corruscansmacho) cultivados en viveros. Un total de 300 peces fueran examinados en piscifactorías del Centro Oeste de Brasil para un levantamiento parasitológico en los arcos branquiales denominados I, II, III y IV, del arco más externo al más interno, respectivamente. Solamente los monogeneos Mymarothecium viatorumy Anacanthorus penilabiatus, parásitos del híbrido patinga, presentaran mayores (p<0,05) intensidades promedios en el I arco cuando es comparado con el IVarco branquial. Aun así, ninguna preferencia por micro-hábitat por los parásitos en los otros peces fuera observado. Copepoditos de Lernaea cyprinaceano presentan preferencia por micro-hábitat. Este estudio muestra que en condiciones de cautiverio puede ocurrir una distribución homogénea de los parásitos en las branquias

Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications

Visceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy and Trypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected with Leishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n 31) compared to those from vaccinated dogs (n 21), experimentally infected dogs with cross-reactive parasites (n 23), and healthy controls (n 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no crossreactivity with T. cruzi- or Ehrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes of L. infantum antigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programsThis work was supported by grants from the Pró-Reitoria de Pesquisa of UFMG (supported 03/2013), the Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (INCT Nano-Biofar), Rede Nanobiotec/Brasil-UFU (CAPES), PRONEX-FAPEMIG (APQ-01019- 09), FAPEMIG (APQ-00496-11 and APQ-00819-12), and CNPq (APQ- 472090/2011-9 and APQ-482976/2012-8). E.A.F.C. and L.R.G. are recipients of grants from CNPq. M.A.C.-F. is the recipient of a grant from FAPEMIG/CAPE

Erratum to: The study of cardiovascular risk in adolescents – ERICA: rationale, design and sample characteristics of a national survey examining cardiovascular risk factor profile in Brazilian adolescents

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A list of land plants of Parque Nacional do Caparaó, Brazil, highlights the presence of sampling gaps within this protected area

Brazilian protected areas are essential for plant conservation in the Atlantic Forest domain, one of the 36 global biodiversity hotspots. A major challenge for improving conservation actions is to know the plant richness, protected by these areas. Online databases offer an accessible way to build plant species lists and to provide relevant information about biodiversity. A list of land plants of “Parque Nacional do Caparaó” (PNC) was previously built using online databases and published on the website "Catálogo de Plantas das Unidades de Conservação do Brasil." Here, we provide and discuss additional information about plant species richness, endemism and conservation in the PNC that could not be included in the List. We documented 1,791 species of land plants as occurring in PNC, of which 63 are cited as threatened (CR, EN or VU) by the Brazilian National Red List, seven as data deficient (DD) and five as priorities for conservation. Fifity-one species were possible new ocurrences for ES and MG states