First identification of Sarcocystis tenella (Railliet, 1886) Moule, 1886 (Protozoa: Apicomplexa) by PCR in naturally infected sheep from Brazil

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

Aspectos ultraestruturais do processo de divisão do Toxoplasma gondii

Neste trabalho é feita uma revisão sobre alguns aspectos biológicos do Toxoplasma gondii, principalmente sobre a ultraestrutura da forma interfásica e as modificações ultraestruturais que ocorrem no parasito durante o seu processo de divisão. Considera-se inicialmente o processo de divisão binária admitindo-se, porém, a possibilidade de que as imagens interpretadas como senão de divisão binária representem estágios da divisão por endodiogenia. Quanto à endodiogenia descrevem-se as alterações que ocorrem na "parasito mãe" durante o processo de formação dos dois "parasitos filhos". Este processo é semelhante no Toxoplasma gondii, Besnoitia jellisoni, Sarcocystis tenella e Frenkelia. Discute-se a possibilidade da formação de mais de dois "parasitos filhos" por um processo de endopoligenia, bem como o processo de esquizogonia. Os resultados mais recentes mostram que não existe esquizogonia nas formas vsgetativas do Toxoplasma gondii, senão que as imagens interpretadas como tal, ao microscópio ótico, são o resultado de endodiogenias sucessivas em que os endozoitas formados permanecem ligados entre si pela região posterior. A esquizogonia é, no entanto, encontrada nas formas que se desenvolvem no interior de células epiteliais do intestino do gato, que é o hospedeiro definitivo do Toxoplasma gondii. Discute-se o conceito de esquizogonia, comparando-o em três protozoários: Eimeria bovis, E. callospermophili e Plasmodium juxtanucleare, que apresentam diferenças entre si quanto ao processo de iniciação da individualização dos "parasitos filhos". Refere-se à recente hipótese que considera a endodiogenia como o processo fundamental de divisão dos esporozoárlos, ocorrendo na fase final da esquizogonia. Finalmente é acentuado o papel que a microscopia eletrônica aliada às modernas técnicas de citoquímica e imunocitoquimica poderá desempenhar no sentido de um melhor conhecimento da biologia do Toxoplasma gondii e da fisiopatogenia da Toxoplasmose.

PREVALENCE OF ANTIBODIES TO TRYPANOSOMA CRUZI, LEISHMANIA INFANTUM, ENCEPHALITOZOON CUNICULI, SARCOCYSTIS NEURONA, AND NEOSPORA CANINUM IN CAPYBARA, HYDROCHOERUS HYDROCHAERIS, FROM SAO PAULO STATE, BRAZIL

Little is known about the importance of capybara. Hydrochoerus hydrochaeris, as reservoirs for parasites of zoonotic or veterinary importance. Sera from 63 capybaras, from 6 counties in the state of Sao Paulo, Brazil, were examined for antibodies to Trypanosoma cruel, Leishmania infantum, Encephalitozoon cuniculi. Sarcacystis neurona, and Neospora caninum using an indirect immunofluorescent antibody test. Five (8%) of the 63 capybaras had antibodies to T cruzi epimastigotes. None of the samples from capybara reacted positively with L. infantum promastigotes or with spores of E. cuniculi. Two (3%) of the serum samples were positive for antibodies to S. neurona merozoites, and 2 (3%) of the serum samples were positive for antibodies to N. caninum tachyzoites. A serum sample from 1 capybara was positive for antibodies to both T cruzi and N. caninum. None of the remaining 62 samples reacted with more than 1 parasite.

Trypanocidal, leislimanicidal and antifungal potential from marine red alga Bostrychia tenella J. Agardli (Rhodomelaceae, Ceraniiales)

Specimens of the red alga Bostrychia tenella J Agardh (Rhodomelaceae, Ceramiales) were collected from the Sao Paulo coast and submitted to loom temperature solvent extraction The resulting extract was fractionated by partitioning with organic solvent The n-hexane (BT-H) and dichloromethane (BT-D) fractions showed antiprotozoal potential in biological tests with Trypanosoma cruzi and Leishmania amazonensis and presented high activity in an antifungal assay with the phytopathogenic fungi Cladosporium cladosporioides and Cladosporium sphaerospermum Chromatography methods were used to generate subfractions from BT-H (H01 to H11) and from BT-D (D01 to 019) The subtractions were analyzed by gas chromatography-mass spectrometry (GC/MS). and the substances were identified by retention index (Kovats) and by comparison to databases of commercial mass spectra The volatile compounds found in marine algae were identified as fatty acids, low molecular mass hydrocarbons, esters and steroids, some of these have been previously described in the literature based on other biological activities Moreover, uncommon substances. such as neophytadiene were also identified In a trypanocidal assay, fractions BT-H and BT-D showed IC(50) values of 168 and 19 1 mu g/mL. respectively, and were mote active than the gentian violet standard (31 mu g/ml.); subfractions H02. H03, D01 and D02 were active against L amasonensis, exhibiting IC(50) values of 1 S. 2 7, 4 4. and 4 3 mu g/mL., respectively (standard amphotericin B IC(50) = 13 mu g/mL.) All fractions showed antifungal potential this work reports the biological activity and identification of compounds by GC/MS for the marine red alga B tenella for the first time (C) 2010 Elsevier B V All lights reserved

Bioactive Chemical Constituents and Comparative Antimicrobial Activity of Callus Culture and Adult Plant Extracts from Alternanthera tenella

Crude extracts of a callus culture (two culture media) and adult plants (two collections) from Alternanthera tenella Colla (Amaranthaceae) were evaluated for their antibacterial and antifungal activity, in order to investigate the maintenance of antimicrobial activity of the extracts obtained from plants in vivo and in vitro. The antibacterial and antifungal activity was determined against thirty strains of microorganisms including Gram-positive and Gram-negative bacteria, yeasts and dermatophytes. Ethanolic and hexanic extracts of adult plants collected during the same period of the years 1997 and 2002 [Ribeirao Preto (SP), collections 1 and 2] and obtained from plant cell callus culture in two different hormonal media (AtT43 and AtT11) inhibited the growth of bacteria, yeasts and dermatophytes with inhibition halos between 6 and 20 mm. For the crude extracts of adult plants bioassay-guided fractionation, purification, and isolation were performed by chromatographic methods, and the structures of the isolated compounds were established by analysis of chemical and spectral evidences (UV, IR, NMR and ES-MS). Steroids, saponins and flavonoids (aglycones and C-glycosides) were isolated. The minimum inhibitory concentration (MIC) of the isolated compounds varied from 50 to 500 mu g/mL.

Bioreduction of Acetophenone Derivatives by Red Marine Algae Bostrychia radicans and B. tenella, and Marine Bacteria Associated

The biocatalytic reduction of acetophenone derivatives was exploited by using algal biomass from Bostrychia radicans and B. tenella producing exclusively (S)-2-phenylethanols with high enantiomeric excess (> 99% ee). Bacterial populations associated with algal biomass were identified as the Bacillus genus. This report deals with the first investigations involving the use of marine bacteria associated with B. radicans and B. tenella marine algae for the biocatalytic reduction of acetophenone derivatives.

Occurrence of Sarcocystis falcatula in Captive Psittacine Birds in Brazil

Thirty-eight captive psittacine birds housed in a bird park in Foz do Iguacu, Parana, Brazil, died within a 15-month period as a result of infection with Sarcocystis falcatula. Although fatalities affected 16 species of psittacine birds, mortality was highest in Old World species, which were most susceptible to the pulmonary form of sarcocystosis. Along with the pathologic findings of this disease outbreak, a review of the pathophysiology of sarcosporidiosis is presented.

Evaluation of immunomodulatory and anti-inflammatory effects and phytochemical screening of Alternanthera tenella Colla (Amaranthaceae) aqueous extracts

Alternanthera tenella Colla extracts are used in Brazilian traditional folk medicine to treat a variety of infectious diseases as well as inflammation and fever. In this work, the immunomodulatory, anti-inflammatory and potential toxic effects of cold (CAE) and hot (HAE) aqueous extracts of A. tenella were investigated in vivo. In addition, we analyzed the phytochemical properties of both extracts. BALB/c mice were immunized in vivo with sheep red blood cells and concomitantly inoculated intraperitoneally (i.p.) with each extract (50, 100 or 200 mg/kg). Specific antibody-producing cells were enumerated using plaque-forming cell assays (PFC) and anti-SRBC IgG and IgM serum levels were measured via enzyme-linked immunosorbent assay. Body and lymphoid organ weights were determined after treatments in order to evaluate toxic effects. Carrageenan-induced paw edema was employed to investigate anti-inflammatory activity in mice inoculated i.p. with CAE or HAE (200 or 400 mg/kg). Phytochemical screening was performed using spectrometric and chromatographic approaches and revealed that CAE possessed higher tannin and flavonoid levels than HAE. PFC numbers were increased after treatment with CAE (100 mg/kg) four days after immunization, as were the serum antibody titers after four and seven days, suggesting immunostimulatory activity through modulation of B lymphocyte functions. Body and organ weights did not show major changes, suggesting that extracts administered to mice did not induce significant toxicity. Both extracts had significant anti-inflammatory activity in the paw edema assay. These results suggested that aqueous extracts from A. tenella contained several chemical compounds that possess positive and/or negative modulator effects on the immune system, which appeared to correlate with tannin and flavonoid levels in those extracts. In summary, these studies provide important insight into the biological activities of A. tenella.

4-(hidroximetil)-benzenossulfonato de potássio: metabólito inédito isolado da alga marinha Bostrychia tenella (Rhodomelaceae, ceramiales)

Chemical investigation of the dichloromethane/methanol extract of the marine alga Bostrychia tenella has led to the isolation of two aromatic compounds, the new sulfate metabolite potassium 4-(hydroxymethyl)-benzenosulfonate (1) and the compound 1-methoxyphenethyl alcohol (2), described previously as a synthetic product. Their structures were determined by spectroscopic methods including NMR, MS, IR and by comparison with literature data.

Aborto ovino associado com infecção por Sarcocystis sp

Infecções por protozoários têm distribuição mundial e podem causar aborto, nascimentos prematuros e ou morte fetal em diversas espécies animais. Em julho de 2004, oito ovinos Corriedale apresentaram problemas reprodutivos caracterizados por aborto e natimortalidade no terço final da gestação. Dessas oito perdas, um natimorto macho foi enviado ao Setor de Patologia Veterinária para necropsia. Alterações macroscópicas não foram observadas durante a necropsia. Lesões histológicas foram observadas principalmente no cérebro e coração e se caracterizaram por encefalite não-supurativa multifocal acentuada associada à presença de protozoários no interior de células endoteliais e vasos sanguíneos e miocardite não-supurativa focal leve. Alguns desses organismos apresentaram formato de roseta. O teste de imunoistoquímica anti-Toxoplasma gondii foi negativo, mas houve reação cruzada com anticorpo anti-Neospora caninum. O exame de imunofluorescência direta para Leptospira sp. foi negativo. A bacteriologia aeróbica e micro-aeróbica não revelou crescimento significativo. Esses achados foram compatíveis com o diagnóstico de Sarcocystis sp.

Sarcocystis cruzi (Apicomplexa: Sarcocystidae) no cachorro-do-mato (Cerdocyon thous)

Esporocistos de Sarcocystis foram identificados nas amostras fecais de um cachorro-do-mato. Eles foram dados por via oral para um bezerro em aleitamento, sendo observados cistos com morfologia compatível com os de Sarcocystis cruzi na musculatura cardíaca e esquelética, três meses após a infecção. Musculatura cardíaca deste bezerro foi dada para um segundo cão doméstico livre de coccídios, que eliminou esporocistos compatíveis com os de Sarcocystis em suas fezes, tendo com períodos pré-patente e patente 11 e 12 dias após a infecção respectivamente. Para comparar a morfologia dos esporocistos e cistos, um segundo cão, também livre de coccídios, foi alimentado com musculatura cardíaca de um bovino infectando naturalmente e positivo para cistos de S. cruzi. Esporocistos compatíveis com os eliminados pelo primeiro cão foram encontrados nas fezes. Apesar dos esporocistos eliminados pelo cachorro-do-mato serem significativamente diferentes dos eliminados pelos cães infectados experimentalmente, pode se considerar com base na morfologia dos esporocistos, cistos e na transmissão biológica que a espécie encontrada nas fezes do cachorro-do-mato é Sarcocystis cruzi.

Anticoccidial effects of acetic acid on performance and pathogenic parameters in broiler chickens challenged with Eimeria tenella

The objective of the present study was to evaluate the anticoccidial effect of the different concentrations of the acetic acid in the broiler chickens in comparison with the amprolium anticoccidial. A total of 198 chicks were placed 11 per pen with three pens per treatment. The different concentrations (1%, 2% and 3%) of acetic acid and amproilum (at the dose rate of 125ppm) were given to the experimental groups in drinking water from 10-19th days of age. One group was kept as infected non medicated control and one as non infected non medicated control. All the groups were inoculated orally with 75,000 sporulated oocysts at the 12th day of age except non infected non medicated control. Anticoccidial effect was evaluated on the basis of performance (weight gain, feed conversion ratio) and pathogenic (oocyst score, lesion score and mortality %age) parameters. Among acetic acid medicated groups, the maximum anticoccidial effect was seen in the group medicated with 3% acetic acid followed by 2% and 1% acetic acid medicated groups. Amprolium and 3% acetic acid were almost equivalent in suppressing the negative performance and pathogenic effects associated with coccidiosis (Eimeria tenella) challenge. In summary, acetic acid has the potential to be used as alternative to chemotherapeutic drugs for Eimeria tenella control. Concentration-dependent anticoccidial effect of acetic acid suggests that further studies should be carried out to determine the possible maximum safe levels of acetic acid with least toxic effects to be used as anticoccidial.

Anticoccidial activity of hydrochloric acid (HCl) against Eimeria tenella in broiler chickens

The present study was planned to evaluate the anticoccidial activity of the different concentrations of the HCl against Eimeria tenella infection in broiler chickens in comparison with the amprolium anticoccidial. For this purpose, a total of 198 chicks were placed 11 per pen with three pens per treatment. The different concentrations of HCl (1000ppm, 2000ppm and 3000ppm) and amproilum (at the dose rate of 125ppm) were given to the experimental groups in drinking water from 10 to 19th days of age. One group was kept as infected non medicated control and one as non infected non medicated control. At the 12th day of age, all the groups were inoculated orally with 75,000 sporulated oocysts except non infected non medicated control. Anticoccidial activity was evaluated on the basis of performance (weight gain, feed conversion ratio) and pathogenic (oocyst score, lesion score and mortality %age) parameters. Among HCl medicated groups, the maximum anticoccidial effect was seen in the group medicated with 1000ppm HCl followed by 2000ppm and 3000ppm HCl medicated groups. Amprolium and 1000ppm HCl were almost equivalent in suppressing the negative performance and pathogenic effects associated with coccidiosis (Eimeria tenella) challenge. In summary, the lower doses of HCl have the potential to be used as alternative to chemotherapeutic drugs for Eimeria tenella control. It is therefore suggested that further studies should be carried out to determine the possible minimum safe levels of HCl with least toxic effects to be used as anticoccidial.

Estudos anatômicos de folhas de espécies de plantas daninhas de grande ocorrência no Brasil: IV - Amaranthus deflexus, Amaranthus spinosus, Alternanthera tenella e Euphorbia heterophylla

Neste trabalho é apresentado um estudo da anatomia das folhas de espécies de plantas daninhas de grande ocorrência no Brasil: Amaranthus deflexus, Amaranthus spinosus, Alternanthera tenella e Euphorbia heterophylla, visando melhor compreensão das barreiras que cada espécie impõe à penetração dos herbicidas e outros compostos utilizados em aplicações foliares. As folhas completamente expandidas do terceiro ao quinto nó foram coletadas de plantas de ocorrência espontânea no campo. Das folhas de cada espécie foram obtidas três amostras da região mediana, com aproximadamente 1 cm², as quais foram utilizadas em estudos da estrutura, clarificação e em observações em microscópio eletrônico de varredura. Todas as espécies avaliadas são anfiestomáticas. As principais barreiras foliares potenciais à penetração de herbicidas observadas nas plantas daninhas A. deflexus e A. spinosus foram, respectivamente, grande espessura da cutícula da face adaxial e da cutícula das duas faces. Já em relação a A. tenella, grande espessura da cutícula das duas faces, elevado teor de cera epicuticular e alta densidade tricomática foram os principais obstáculos potenciais detectados. E. heterophylla apresentou como possíveis principais barreiras foliares à penetração de agroquímicos o alto teor de cera epicuticular, a elevada densidade de laticíferos e a grande espessura da cutícula da face adaxial.

Profundidade de semeadura afetando a emergência de plântulas de Alternanthera tenella

A compreensão a respeito de informações básicas sobre a biologia de plantas daninhas pode contribuir significativamente na construção de estratégias mais adequadas para seu manejo, além de possibilitar o desenvolvimento de ferramentas de controle não-químico. O objetivo deste trabalho foi avaliar o efeito da profundidade de semeadura e da cobertura do solo sobre a emergência de Alternanthera tenella. Os tratamentos foram realizados em esquema fatorial 7x2, considerando-se sete níveis de profundidade (0, 1, 2, 3, 4, 5 e 10 cm) e duas situações de cobertura de solo (com e sem palha). A palha utilizada para cobertura do solo foi de aveia-preta (Avena strigosa), em quantidade equivalente a 2 t ha-1. A semeadura foi feita em colunas de PVC, colocando-se 50 sementes por coluna. A emergência das plântulas foi anotada diariamente até 28 dias após a semeadura. Ao final desse período, calculou-se a porcentagem total de emergência para cada tratamento e o índice de velocidade de emergência (IVE). A emergência das plântulas de A. tenella é influenciada pela profundidade do solo na qual as sementes se encontram. Na ausência e na presença de palha, a emergência é reduzida a partir de 4 e 5 cm de profundidade, respectivamente, e não há emergência a 10 cm de profundidade. Sementes posicionadas na superfície do solo apresentam velocidade de emergência igual ou superior à daquelas colocadas nas demais profundidades, independentemente da presença de palha na superfície do solo.