ABSTRACT Leptospirosis and toxoplasmosis are zoonoses of occupational character that are spread worldwide, mainly in tropical and socioeconomically vulnerable areas. Both diseases have vertebrate hosts, which spread the infectious agent to the environment and are potentially infectious to humans and can progress to fatal cases. The present study reinforces the importance of One Health for the control and prophylaxis of leptospirosis and toxoplasmosis using sentinel animals in areas of socioeconomic vulnerability based on a serological study. A total of 336 blood samples were obtained from the canine population of the urban district, which has high rates of poverty and sanitation. The results were analyzed based on seroreactivity animals and georeferencing by kernel spatial mapping. A total of 43.3% (52/120) of the animals were seroreagents for anti-Leptospira spp., with a predominance of Canicola (35/52 = 67.3%) and Pyrogenes (5/52 = 9.6%), while 52.97% (178/336) of the dogs were seroreactive for anti-Toxoplasma gondii. The presence of seroreagent dogs for both zoonosis alert to the inadequate hygienic-sanitary conditions of the studied region, and highlight the importance of a survey of the social, economic and environmental structure to identify factors related to the links in the epidemiological chain of pathogen transmission and necessary corrective actions.

RESUMO Leptospirose e toxoplasmose são zoonoses de caráter ocupacional disseminadas mundialmente, principalmente em áreas tropicais e socioeconomicamente vulneráveis. Ambas as doenças têm hospedeiros vertebrados, que espalham o agente infeccioso para o meio ambiente e são potencialmente infecciosos para o homem, as quais podem evoluir para casos fatais. O presente estudo reforça a importância da saúde única para o controle e a profilaxia da leptospirose e da toxoplasmose, mediante a utilização de animais sentinela em áreas de vulnerabilidade socioeconômica, com base em estudo sorológico. Um total de 336 amostras de sangue foi obtido da população canina de um distrito urbano que apresenta altos índices de pobreza e ausência de saneamento básico. Os resultados foram analisados com base em animais sororreagentes e georreferenciados por mapeamento espacial de Kernel. Um total de 43,3% (52/120) dos animais foi sororreagente para anti-Leptospira spp., com predominância de Canicola (35/52 = 67,3%) e Pirogenes (5/52 = 9,6%), enquanto 52,97% (178/336) dos cães foram sororreagentes para anti-Toxoplasma gondii. A presença de cães sororreagentes para ambas as doenças alerta para as condições higiênico-sanitárias inadequadas da região estudada e evidencia a importância de um levantamento da estrutura social, econômica e ambiental para identificar fatores relacionados aos elos da cadeia epidemiológica de transmissão do patógeno e ações corretivas necessárias.

RESUMO Neste artigo é proposto um modelo de programação por metas estendido aplicado ao planejamento de radioterapia, em que foi encontrada a melhor combinação de pesos para as metas a serem atingidas. O modelo foi aplicado a um caso real de câncer de próstata e resolvido pelo software CPLEX, em que foi utilizado o Método de Pontos Interiores Barreira Logarítmica como método de resolução.

ABSTRACT In this paper an extended goal programming model applied to radiotherapy planning is proposed, in which the best weight combination was found to each goal. The model was applied to a real prostate cancer case and solved by the software CPLEX, where Logarithmic Barrier Interior Point Method was used.

A bactéria Streptococcus agalactiae é um potente agente causador de surtos por doenças bacterianas em peixes. O estresse provocado pelo manejo zootécnico e pela má qualidade ambiental torna a tilápia susceptível às infecções por essa bactéria. O objetivo do presente trabalho foi avaliar a resistência de tilápias-do-nilo imunizadas com soro hiperimune anti-S. agalactiae, posteriormente desafiadas com cepa homóloga da mesma bactéria. Após determinação da DL 50 de S. agalactiae, 36 tilápias foram distribuídas em quatro aquários, dois para o grupo controle e dois para inoculação celomática para produção de anticorpos anti-S. agalactiae. No 21° e 28° dias, foi coletado sangue para obtenção de soro hiperimune utilizado na transferência passiva. Em seguida, 30 tilápias foram distribuídas em três aquários e submetidas a três tratamentos: GI: controle; GII: imunizadas com o soro inativado; GIII: imunizadas com soro ativo. Após 48 horas e sete, 14, 21, 28 e 35 dias, foram realizadas coletas de sangue para titulação de anticorpos anti-S. agalactiae utilizando-se o teste de aglutinação direta. Para avaliar a taxa de sobrevivência, outras 30 tilápias foram distribuídas em três aquários e submetidas a três tratamentos (GI: controle; GII: imunizadas com soro inativado; GIII: imunizadas com soro ativo). Após 48 horas da imunização, as tilápias foram desafiadas via celomática com 100µL de S. agalactiae e avaliadas duas vezes ao dia, pelo período de 35 dias. Os resultados dos títulos séricos de anticorpos foram detectados pela aglutinação direta até o 21° dia pós-transferência passiva, e, no mesmo período, houve proteção de 80% entre os grupos imunizados com soro inativado e soro ativo contendo anticorpos anti-S. agalactiae. Ao final, os grupos soro inativado e soro ativo apresentaram 60 e 80% de proteção, respectivamente, enquanto no grupo controle 100% dos peixes adoeceram, apresentando sinais graves da infecção, e foram eutanasiados. Não houve diferença estatística significativa na taxa de proteção entre os grupos imunizados.

The Streptococcus agalactiae bacteria is a potent agent which causes outbreaks of bacterial diseases in fish. The stress caused by management and poor environmental quality makes tilapia susceptible to infections, including by bacterium. The aim of this study was to evaluate the resistance of the Nile tilapia immunized with hyperimmune serum against S. agalactiae subsequently challenged with homologous strain of the same bacteria. After determining the DL 50 of S. agalactiae, 36 tilapias were distributed in 4 aquariums, 2 for the control group and 2 for the group via coelomic, inoculated with the DL 50 for anti-S. agalactiae antibodies production. On the 21st and 28th day blood was collected for the obtainment of hiperimmune serum used in passive transference. Then, 30 tilapias were distributed in 3 aquariums submitted to 3 treatments (GI: control; GII: immunized with inactivated-serum; GIII: immunized with non-inactivated serum). After 48 hours, 7, 14, 21, 28 and 35 days blood collections were done for anti-S. agalactiae antibodies titration using the direct agglutination test. For survival evaluation 30 tilapias were distributed in 3 aquariums and submitted to 3 treatments (GI: control; GII: immunized with inactivated-serum; GIII: immunized with non-inactivated serum). After 48 hours of inoculation tilapias were challenged with 100µL of S. agalactiae and monitored twice a day for 35 days. The results showed that titers of antibodies were detected by direct agglutination until the 21st day after passive transference and during the same period the protection between the groups immunized with inactivated serum and non-inactivated serum contending anti-S. agalactiae antibodies was of 80% after challenge with S. agalactiae. In the end, the serum-inactivated serum and active groups had 60 and 80% protection, respectively, while 100% of the fish in the control group developed severe signs of infection and were euthanized. There was no statistical difference in the survival rate between the immunized groups.

RESUMO O objetivo deste trabalho foi determinar a prevalência de anticorpos contra Leptospira spp. em bovinos, caninos, equinos, ovinos e suínos, oriundos de 40 propriedades localizadas na área rural do Município de Jaguapitã, Estado do Paraná. Foram colhidas amostras de sangue de 370 bovinos, 161 equinos, 70 ovinos, 230 suínos e 97 caninos. O número de animais testados em cada propriedade, assim como o número de propriedades, foi determinado utilizando-se o programa Epi-info versão 6. As amostras de soros obtidas foram submetidas à prova de soroaglutinação microscópica (SAM) com 22 sorovares de Leptospira spp. Das 40 propriedades rurais pesquisadas, 38 (95,00%) tiveram pelo menos um animal sororeagente na SAM e dos 928 animais estudados, 316 (34,08%) apresentaram títulos ? 100. A prevalência observada na espécie bovina foi de 42,43%, com 87,18% das propriedades apresentando pelo menos um animal sororeagente. As prevalências de animais e propriedades reagentes para as demais espécies foram, respectivamente, 48,44% e 87,18% para equinos; 38,57% e 100% para ovinos; 18,70% e 28,00% para suínos; 11,34% e 31,25% para cães. O sorovar mais provável encontrado em bovinos foi Hardjo, em equinos Castellonis e Sentot, em ovinos, suínos e cães Icterohaemorrhagiae. Os resultados obtidos neste trabalho demonstram que as cinco espécies animais estudadas na área rural do Município de Jaguapitã tiveram contato com vários sorovares de Leptospira spp. Além disso, os resultados sorológicos sugerem uma possível transmissão do micro-organismo entre espécies animais, provavelmente em decorrência da exposição às mesmas fontes de infecção entre os animais estudados.

ABSTRACT The objective of this work was to determine the prevalence of antibodies against Leptospira spp. in cattle, dogs, horses, sheep and swine from 40 properties located in the rural area of Jaguapitã, state of Paraná, Brazil. Blood samples were taken from 370 cattle, 97 dogs, 161 horses, 70 sheep and 230 swine. The number of animals tested on each property, as well as the number of properties was determined using the program Epi-info version 6. Samples of serum were submitted to microscopic agglutination test (MAT) with 22 Leptospira spp. serovars. From the 40 rural properties investigated, 38 (95.00%) had at least one positive animal according to SAM, and from 928 studied animals, 316 (34.08%) presented titers ? 100. The prevalence observed in the bovine species was 42.43%, with 87.18% of the properties presenting at least one positive animal. The prevalence of animals and properties reactive for the other species were, respectively: 48.44% and 87.18% for horses; 38.57% and 100% for sheep; 18.70% and 28.00% for swine; 11.34% and 31.25% for dogs. The most frequent serovar in bovines was Hardjo, in horses Castellonis and Sentot, in sheep, swine and dogs Icterohaemorrhagiae. The results obtained in this study demonstrated that the 5 studied animal species in the rural area of Jaguapitã had contact with several Leptospira spp. serovars. Moreover, serological results suggest a possible transmission of Leptospira spp. between animal species, probably because of exposure to the same sources of infection among the animals studied.

Realizou-se um estudo para caracterizar a situação epidemiológica da brucelose bovina no Estado do Paraná. O Estado foi estratificado em sete circuitos produtores ou regiões. Em cada circuito foram amostradas aleatoriamente cerca de 300 propriedades e, dentro dessas, foi escolhido de forma aleatória um número pré-estabelecido de animais, dos quais foi obtida uma amostra de sangue. No total, foram amostrados 14.857 animais, provenientes de 2.098 propriedades. Em cada propriedade amostrada foi aplicado um questionário epidemiológico para verificar o tipo de exploração e as práticas zootécnicas e sanitárias que poderiam estar associadas ao risco de infecção pela doença. O protocolo de testes utilizado foi o da triagem com o teste do antígeno acidificado tamponado e o reteste dos positivos com o teste do 2-mercaptoetanol. O rebanho foi considerado positivo se pelo menos um animal foi reagente às duas provas sorológicas. Para o Estado, os resultados de prevalências de focos e de animais infectados foram, respectivamente, de 4,0% [3,2-4,8%] e 1,7% [1,1-2,4%]. Para os circuitos, a prevalência de focos e a de animais foram, respectivamente: circuito 1, 14,7% [10,9-19,2%] e 2,8% [1,2-4,4%]; circuito 2, 8,8% [5,9-12,6%] e 2,4% [1,0-3,8%]; circuito 3, 3,4% [1,6-6,1%] e 0,85% [0,21-1,5%]; circuito 4, 2,3% [0,94-4,8%] e 0,83% [0,02-1,6%]; circuito 5, 2,3% [0,94-4,7%] e 1,6% [0,06-3,3%]; circuito 6, 0,3% [0-1,9%] e 0,09% [0-0,27%]; circuito 7, 1,0% [0,21-2,9%] e 2,2% [0-6,0%]. Os fatores de risco (odds ratio, OR) associados à condição de foco foram: compra de reprodutores (OR= 2,20 [1,42-3,40]) e prática de aluguel de pasto (OR= 2,45 [1,54-3,90]).

A study to characterize the epidemiological situation of bovine brucellosis in the State of Paraná was carried out. In this study, the State was divided into seven regions. Three hundred herds were randomly sampled in each region and a pre-established number of animals were sampled in each of these herds. A total of 1,4857 serum samples from 2,098 herds were collected. In each herd, it was applied an epidemiological questionnaire regarding herd features and also husbandry and sanitary practices that could be associated with risk of infection. The serum samples were screened for antibodies against Brucella spp. by the Rose-Bengal test (RBT), and all RBT-positive sera were re-tested by the 2-mercaptoethanol test (2-ME). A herd was considered positive if at least one animal was positive on both RBT and 2-ME tests. The prevalences of infected herds and infected animals in Paraná State were 4.0% [3.2-4.8%] and 1.7% [1.1-2.4%], respectively. In the regions, the prevalences of infected herds and animals were: region 1, 14.7% [10.9-19.2%] and 2.8% [1.2-4.4%]; region 2, 8.8% [5.9-12.6%] and 2.4% [1.0-3.8%]; region 3, 3.4% [1.6-6.1%] and 0.85% [0.21-1.5%]; region 4, 2.3% [0.94-4.8%] and 0.83% [0.02-1.6%]; region 5, 2.3% [0.94-4.7%] and 1.7% [0.06-3.3%]; region 6, 0.34% [0-1.9%] and 0.09% [0-0.27%]; and region 7, 1.0% [0.21-2.9%] and 2.2% [0-6,0%]. The risck factors (odds ratio, OR) associated with the presence of the infection were: purchase of animals for breeding (OR= 2.20 [1.42-3.40]) and pasture rental practice (OR=2.45 [1.54-3.90]).

Trinta e cinco vacas de rebanhos leiteiros da região Norte do estado do Paraná, com histórico de abortamento, foram pesquisadas sorologicamente para verificar a presença de anticorpos contra Neospora caninum, Toxoplasma gondii, Leptospira spp., Brucella abortus, BHV-1 e BVDV. Vinte e uma vacas apresentaram títulos sorológicos compatíveis com infecção. Todas elas, soropositivas para N. caninum, foram também soropositivas para outros agentes infecciosos, sugerindo a possibilidade de associação desses agentes nos problemas reprodutivos de bovinos, no estado do Paraná.

RESUMO O objetivo deste trabalho foi descrever o quadro clínico de um cão com suspeita de leptospirose e os resultados dos exames específicos e complementares para leptospirose. A anamnese revelou que o animal apresentou episódios de vômito e dificuldade em apreender alimentos sólidos e líquidos. No exame clínico foi constatada desidratação de 5-7%, sensibilidade abdominal, hematoemese, melena, diarréia, necrose na língua, sialorréia e dificuldade de deglutição. Todos os exames considerados específicos, bacteriológico, direto da urina em microscópio de campo escuro, soroaglutinação microscópica e imunoperoxidase indireta do rim apresentaram resultados positivos. A estirpe de Leptospira isolada foi identificada como sorovar Canicola. Os exames complementares revelaram leucocitose com neutrofilia, hipocalcemia, hipoalbuminemia e valores séricos de uréia, creatinina e fosfatos acima dos valores normais enquanto a urinálise revelou proteinúria, leucocitúria e presença de células renais. As alterações renais encontradas no HE foram nefrite intersticial crônica e glomerulite com espessamento da membrana basal. Os sinais clínicos observados e os resultados dos exames laboratoriais indicaram um quadro predominantemente renal, compatível com a infecção pelo sorovar Canicola.

ABSTRACT The purpose of this study was to describe the clinical features of a dog with leptospirosis suspicion, and results of specific and complementary laboratorial tests for leptospirosis. Anamnesis revealed that the animal presented vomiting, difficulty to apprehend both solid and liquid food. In the clinical examination, there was verified dehydration from 5 to 7%, hematoemesis, melena, diarrhea, tong necrosis, sialorrhea and difficulty swallowing. All the specific tests, bacteriologic, direct examination of urine in dark field microscopy, microscopic agglutination test and indirect immunoperoxidase, were positive. The Leptospira strain isolated was identified as serovar Canicola. The complementary tests revealed leukocytosis with neutrophilia, hypocalcemia, hypalbuminemia and blood values of urea, creatinine and phosphates over the normal values while urinalysis showed proteinuria, leukocyturia and presence of kidney cells. Kidney lesions found in HE were chronic interstitial nephritis and glomerulitis with thickened basement membrane. The clinical signs and the results of laboratorial tests found in this study indicated to predominantly to renal feature, consistent with an infection by serovar Canicola.

Ipomoea imperati (Convolvulaceae) lives on the sandy shores of the Brazilian coast and in other areas of the world. The anti-inflammatory activity of a methanol-water extract of the leaves of I. imperati was investigated in experimental models of acute and subchronic inflammation. Topical application of the extract (10 mg/ear) inhibited mouse ear edema induced by croton oil (89.0 ± 1.3% by the lipid fraction with an IC50 of 3.97 mg/ear and 57.0 ± 1.3% by the aqueous fraction with an IC50 of 3.5 mg/ear) and arachidonic acid (42.0 ± 2.0% with an IC50 of 4.98 mg/ear and 31.0 ± 2.0% with an IC50 of 4.72 mg/ear). Phospholipase A2, purified from Apis mellifera bee venom, was also inhibited by the extract (5.0 mg/ml lipid and aqueous fraction) in vitro in a dose-dependent manner (85% by the lipid fraction with an IC50 of 3.22 mg/ml and 25% by the aqueous fraction with an IC50 of 3.43 mg/ml). The methanol-water extract of I. imperati (1000 mg/kg) administered by the oral route also inhibited the formation of cotton pellet-induced granulomas (73.2 ± 1.2% by the lipid fraction and 56.14 ± 2.7% by the aqueous fraction) and did not cause gastric mucosal lesions. I. imperati extracts (10 mg/ml) also inhibited in a dose-dependent manner the muscle contractions of guinea pig ileum induced by acetylcholine and histamine (IC50 of 1.60 mg/ml for the lipid fraction and 4.12 mg/ml for the aqueous fraction). These results suggest the use of I. imperati as an anti-inflammatory and antispasmodic agent in traditional medicine.

Trimethylsulfonium, a compound present in the midgut gland of the sea hare Aplysia brasiliana, negatively modulates vagal response, indicating a probable ability to inhibit cholinergic responses. In the present study, the pharmacological profile of trimethylsulfonium was characterized on muscarinic and nicotinic acetylcholine receptors. In rat jejunum the contractile response induced by trimethylsulfonium (pD2 = 2.46 ± 0.12 and maximal response = 2.14 ± 0.32 g) was not antagonized competitively by atropine. The maximal response (Emax) to trimethylsulfonium was diminished in the presence of increasing doses of atropine (P<0.05), suggesting that trimethylsulfonium-induced contraction was not related to muscarinic stimulation, but might be caused by acetylcholine release due to presynaptic stimulation. Trimethylsulfonium displaced [³H]-quinuclidinyl benzilate from rat cortex membranes with a low affinity (Ki = 0.5 mM). Furthermore, it caused contraction of frog rectus abdominis muscles (pD2 = 2.70 ± 0.06 and Emax = 4.16 ± 0.9 g), which was competitively antagonized by d-tubocurarine (1, 3 or 10 µM) with a pA2 of 5.79, suggesting a positive interaction with nicotinic receptors. In fact, trimethylsulfonium displaced [³H]-nicotine from rat diaphragm muscle membranes with a Ki of 27.1 µM. These results suggest that trimethylsulfonium acts as an agonist on nicotinic receptors, and thus contracts frog skeletal rectus abdominis muscle and rat jejunum smooth muscle via stimulation of postjunctional and neuronal prejunctional nicotinic cholinoreceptors, respectively.

Sea anemones are a rich source of biologically active substances. In crayfish muscle fibers, Bunodosoma cangicum whole venom selectively blocks the I K(Ca) currents. In the present study, we report for the first time powerful hemolytic and neuroactive effects present in two different fractions obtained by gel-filtration chromatography from whole venom of B. cangicum. A cytolytic fraction (Bcg-2) with components of molecular mass ranging from 8 to 18 kDa elicited hemolysis of mouse erythrocytes with an EC50 = 14 µg/ml and a maximum dose of 22 µg/ml. The effects of the neuroactive fraction, Bcg-3 (2 to 5 kDa), were studied on isolated crab nerves. This fraction prolonged the compound action potentials by increasing their duration and rise time in a dose-dependent manner. This effect was evident after the washout of the preparation, suggesting the existence of a reversible substance that was initially masking the effects of an irreversible one. In order to elucidate the target of Bcg-3 action, the fraction was applied to a tetraethylammonium-pretreated preparation. An additional increase in action potential duration was observed, suggesting a blockade of a different population of K+ channels or of tetraethylammonium-insensitive channels. Also, tetrodotoxin could not block the action potentials in a Bcg-3-pretreated preparation, suggesting a possible interaction of Bcg-3 with Na+ channels. The present data suggest that B. cangicum venom contains at least two bioactive fractions whose activity on cell membranes seems to differ from the I K(Ca) blockade described previously.