Detection and molecular analysis of Toxoplasma gondii and Neospora caninum from dogs with neurological disorders

Introduction: Toxoplasma gondii and Neospora caninum are related Apicomplexa parasites responsible for systemic diseases in many species of animals, including dogs. Methods: This study aimed to determine the occurrence of T. gondii and N. caninum infections in 50 dogs with neurological signs that were admitted to the Veterinary Hospital of Universidade Estadual Paulista, City of Botucatu, Brazil. All animals were screened for antibodies using an immunofluorescent antibody test for both parasites. Tissues of positive animals were bioassayed in mice (T. gondii) and gerbils (N. caninum), and DNA was analyzed using the polymerase chain reaction (PCR). Positive samples for T. gondii by PCR were typed using restriction fragment length polymorphism-PCR for 11 markers: SAG1, SAG2 (5'-3'-SAG2 and alt.SAG2), SAG3, Btub, GRA6, L358, c22-8, c29-6, PK1 and Apico, and CS3 marker for virulence analysis. Results: Specific antibodies were detected in 11/50 (22%; 95% confidence interval (CI95%), 12.8-35.3%) animals for T. gondii and 7/50 (14%; CI95%, 7.02-26.3%) for N. caninum. In the bioassay and PCR, 7/11 (63.6%; CI95%, 34.9-84.8%) samples were positive for T. gondii and 3/7 (42.9%; CI95% I, 15.7-75.5%) samples were positive for N. caninum. Three different genotypes were identified, but only 1 was unique. Conclusions: These data confirm the presence of T. gondii and N. caninum in dogs from Brazil, indicating the importance of this host as a sentinel of T. gondii for human beings, and the genotypic variation of this parasite in Brazil.

Detection of Toxoplasma gondii in the milk of experimentally infected Wistar female rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genotypic characterization of Toxoplasma gondii in sheep from Brazilian slaughterhouses: New atypical genotypes and the clonal type II strain identified

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chlamydophila psittaci and Toxoplasma gondii infection in pigeons (Columba livia) from São Paulo State, Brazil

Pigeons (Columba livia) cohabit with humans in urban and rural areas, representing a public health problem since microorganisms are transmitted through the inhalation of dust from their dry feces (chlamydiosis) and through ingestion of their undercooked or poorly refrigerated meat (toxoplasmosis). This study aimed to evaluate the presence of Chlamydophila psittaci and Toxoplasma gondii in pigeons from four cities in São Paulo State, Brazil. C psittaci was evaluated through hemi-nested polymerase chain reaction (hnPCR) using cloacal and tracheal swabs, whereas T. gondii specific antibodies were assessed by means of modified agglutination test (MAT), mouse brain and muscle bioassay, and polymerase chain reaction (PCR). To confirm the infection in mice, T. gondii antibodies were assayed by using indirect fluorescent antibody test (IFAT). Considering C. psittaci, 40/238 (16.8%; 95%CI 12.6-22.1%) samples were positive according to hnPCR, especially for the cities of São Paulo (42.5%) and Bauru (35%). As regards T. gondii, 12/238 (5%; 95%CI 2.9-8.6%) serum samples were positive according to MAT. of these, five samples had titer equal to 1:8; six samples, 1:16; and one sample, 1:32. Bioassay, IFAT and PCR were negative for mouse toxoplasmosis. The absence of T. gondii antibodies suggests that pigeons may be infected with a low concentration of the agent, not detected by the antigen test. Thus, C. psittaci represents an actual problem concerning bird health. (C) 2010 Elsevier B.V. All rights reserved.

Recrudescence of Toxoplasma gondii infection in chronically infected rats (Rattus norvegicus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation and multilocus genotyping of Toxoplasma gondii in seronegative rodents in Brazil

Synanthropic rodents, mainly rats and mice, become ecologically associated with men due to changes in their ecosystems caused by human activities. These animals may take part in the epidemiological cycles of several diseases, including toxoplasmosis. The presence of serum antibodies to Toxoplasma gondii in 43 rodents captured in the urban area of Umuarama, PR, Brazil, was verified by modified agglutination test (MAT). Brain and heart samples were also collected and bioassayed in mice for the isolation of the parasite. Isolated samples were analyzed by 12 multilocus genotyping. Although all rodents were seronegative, the parasite was isolated in one mouse (Mus musculus) and one rat (Rattus rattus). Genotyping showed that these samples were similar to those previously isolated from cats in the state of Parana, Brazil. (C) 2010 Elsevier B.V. All rights reserved.

Parâmetros de fermentação ruminal em bovinos alimentados com grãos de milho ou sorgo de alta umidade ensilados

Os parâmetros de fermentação ruminal de dietas contendo silagem de sorgo úmido em substituição à de milho úmido foram estudados em 12 fêmeas bovinas, com peso médio de 584 kg. O delineamento foi inteiramente casualizado com três tratamentos: substituição do milho úmido pelo sorgo úmido ensilado, nos níveis de 0, 50 e 100%. As dietas continham grão úmido de milho ou de sorgo ensilados, soja extrusada, uréia, feno de aveia (Avena sativa sp.), suplemento mineral e monensina. Adicionalmente, foi avaliada a degradabilidade in situ da matéria seca e da fibra em detergente neutro do feno de aveia. Não houve diferença sobre produção total de ácidos graxos voláteis (AGVs) no rúmen, porcentagem molar dos ácidos acético, propiônico e butírico, relação acético/propiônico, pH ruminal, concentração de N-NH3 no rúmen, fluxo e volume de líquidos do rúmen, nos diferentes tratamentos. A degradabilidade da matéria seca e da fibra em detergente neutro do feno não apresentou diferenças. Não se constatou melhora nos parâmetros de fermentação ruminal com a associação dos grãos.

Desempenho de novilhos de corte alimentados com dietas contendo quantidades crescentes de concentrado associado à forragem de aveia (Avena strigosa) tratada com uréia

Este trabalho teve por objetivo avaliar o efeito dos níveis 40, 55 e 70% de concentrado associados a um volumoso de baixa qualidade sobre o consumo e ganho de peso de 16 novilhos mestiços, castrados, com 10 meses de idade e peso vivo inicial médio de 312kg, em delineamento inteiramente ao acaso, com três repetições, durante 63 dias. O volumoso usado foi a aveia (Avena strigosa), tratada com uréia e o concentrado era constituído de milho, farelo de arroz, farelo de soja e minerais. Todas as dietas foram ajustadas para conter 12% de proteína bruta. Houve resposta positiva e linear à proporção de concentrado na dieta, para as variáveis ganho de peso diário e consumo de matéria seca expresso em kg/animal/dia, em percentagem do peso vivo e em g por kg de peso metabólico em função da proporção de concentrado na dieta. Houve resposta linear negativa para conversão alimentar. O consumo de matéria seca e o ganho de peso diário aumentaram na medida em que se elevou a proporção de concentrado na dieta. O volumoso de baixa qualidade empregado possivelmente limitou o consumo e conseqüentemente o aporte de nutrientes.

Type C botulism in swine fed on restaurant waste

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Desenvolvimento da aveia branca e disponibilidade de fósforo em razão da aplicação de silicato de potássio

A aplicação de silicato de potássio pode aumentar a disponibilidade de fósforo no solo, além de influenciar no crescimento da aveia branca. Objetivou-se neste trabalho avaliar o efeito da aplicação de doses de silicato de potássio no solo no desenvolvimento da aveia branca e na disponibilidade de fósforo. O delineamento experimental foi inteiramente ao acaso, em esquema fatorial 2 x 4, com seis repetições. Os tratamentos foram 20 e 200 mg dm-3 de P2O5 combinados com 0, 150, 300 e 450 mg dm-3 de Si, aplicados por ocasião da semeadura. Após a colheita, determinaram-se a matéria seca de folhas + colmos, das panículas e o total da parte aérea. A palha das panículas foi separada para avaliação dos teores de P e Si nas folhas + colmos, grãos e palha das panículas. As concentrações de Si e P no solo foram avaliadas após colheita das plantas. A aplicação de silicato aumenta os teores de Si no solo e influencia na maior disponibilidade de fósforo. A matéria seca de folhas + colmos é incrementada pelo silicato quando os teores iniciais de P são baixos. Mesmo com a dose elevada de P, a aplicação de silicato aumenta a disponibilidade de P no solo e a absorção pelas plantas. Os teores de Si na palha das panículas, folhas + colmos e grãos são influenciados pela maior disponibilidade do elemento no solo.

Studies on anti-allergic constituents in the leaves and stems of Anchietia salutaris var. martiana (Violaceae)

The anti-allergic active fractionation of hexane extracts of the leaves and stems of Anchietia salutaris,ar. martiana (family Violaceae) nas performed by monitoring their activities with an in vitro bioassay system measuring the inhibitory effects on induced histamine release from guinea pig lung cells. Three known pentacyclic triterpenes (friedelin, alpha-amyrin, beta-amyrin) were isolated, but these compounds were inactive. Aliphatic hydrocarbons and methyl esters of fatty acids (palmitic, oleic, linoleic, linolenic acids) were detected in active fractions. All compounds isolated were detected for the first time in this medicinal plant.

Study of Toxoplasma infection in Brazilian wild mammals: Serological evidence in Dasypus novemcinctus Linnaeus, 1758 and Euphractus sexcinctus Wagler, 1830

Toxoplasmosis is a worldwide distributed zoonosis that affects man and most warm-blooded animals, with a great economic impact in animal and public health. Serum samples from nine 9-banded armadillos, three 6-banded armadillos, three coatimundis, two opossums and one nutria were submitted for anti-Toxoplasma gondii antibody detection by means of a modified direct agglutination method. Encephalic tissue of three 6-banded armadillos, one 9-banded armadillo, one coatimundi and one nutria were digested in acid pepsin solution and inoculated into Swiss mice for parasite isolation. Only one serum sample from a nine-banded armadillo and two from six-banded armadillos reacted producing titers equal to 256, 512 and 512, respectively. T gondii was isolated in two 6-banded armadillos, one of which was not positive in the serological test. (c) 2005 Elsevier B.V. All rights reserved.

Risk assessment of cadmium toxicity on hepatic and renal tissues of rats

Pollution and industrial practices result in concentrations of metals and other environmental agents that are related to environmental toxicity. A rat bioassay was utilized for the identification of toxic effects of cadmium intake. This demonstrated increased total urinary proteins and increased kidney weights in rats exposed to CdCl2, for 7 days, in drinking water (100 mg/L). Serum creatinine, total and direct bilirubin concentrations and alanine transaminase activity were increased in Cd-exposed rats, indicating renal and hepatic toxicity. It was also observed that lipoperoxide concentrations were increased, while Cu-Zn superoxide dismutase activity was decreased in rats treated with cadmium. This indicated that the renal and hepatic toxicity induced by cadmium involved superoxide radicals. (C) 1998 Academic Press.

Mass spectrometric characterization of two novel inflammatory peptides from the venom of the social wasp Polybia paulista

The social wasp P. paulista is relatively common in southeast Brazil causing many medically important stinging incidents. The seriousness of these incidents is dependent on the amount of venom inoculated by the wasps into the victims, and the characteristic envenomation symptoms are strongly dependent on the types of peptides present in the venom. In order to identify some of these naturally occurring peptides available in very low amounts, an analytical protocol was developed that uses a combination of reversed-phase and normal-phase high-performance liquid chromatography (HPLC) of wasp venom for peptide purification, with matrix-assisted laser desorption/ionization time-of-flight post-source decay mass spectrometry (MALDI-Tof-PSD-MS) and low-energy collision-induced dissociation (CID) in a quadrupole time-of-flight tandem mass spectrometry (QTof-MS/MS) instrument for peptide sequencing at the sub-picomole level. The distinction between Leu and Ile was achieved both by observing d-type fragment ions obtained under CID conditions and by comparison of retention times of the natural peptides and their synthetic counterparts (with different combinations of I and/or L at N- and C-terminal positions). To distinguish the isobaric residues K and Q, acetylation of peptides was followed by Q-Tof-MS analysis. The primary sequences obtained were INWLKLGKMVIDAL-NH2 (MW 1611.98Da) and IDWLKLGKMVMDVL-NH2 (MW 1658.98Da). Micro-scale bioassay protocols characterized both peptides as presenting potent hemolytic action, mast cell degranulation, and chemotaxis of poly-morphonucleated leukocyte (PMNL) cells. The primary sequences and the bioassay results suggest that these toxins constitute members of a new sub-class of mastoparan toxins, directly involved in the occurrence of inflammatory processes after wasp stinging. Copyright (C) 2004 John Wiley Sons, Ltd.

Morphological alterations induced by boric acid and fipronil in the midgut of worker honeybee (Apis mellifera L.) larvae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)