Reproductive alterations of Biomphalaria glabrata (Say, 1818) infected with Angiostrongylus cantonensis (Chen, 1935) and exposed to Euphorbia milii var. hislopii latex.

The natural phytochemical latex of Euphorbia milii var. hislopii is one of the most promising natural molluscicides for the control of Biomphalaria glabrata, and has been widely studied under laboratory conditions for selective control of schistosomiasis transmission. However, the effect of this product on B. glabrata infected by other helminths had not yet been investigated. The present study reports evaluation of the effect of E. milii var. hislopii latex on the survival and reproductive activity of B. glabrata infected by Angiostrongylus cantonensis. For this purpose, the following groups were formed: control (C), exposed (E), infected (I) and infected and exposed for different time intervals (1 day - I+E-1D, 7 days - I+E-7D, 14 days - I+E-14D, 21 days - I+E-21D and 28 days - I+E-28D). The experimental infection was performed with L1 larvae of A. cantonensis and exposure to 0.08 mg/L (LC50) of E. milii latex for a period of 24 hours. We analyzed the effects of E. milii latex on the survival of snails during four weeks, reproductive parameters and possible histophysiopathological changes in the gonad and albumen gland of the snails. Regarding survival, there was reduction of more than 50% in the groups exposed to latex (E and I + E) compared to the control group. As for the number of ovigerous masses, eggs, and average number of hatched snails, significant increases were observed in the I+E-1D group at the 4th week compared to the control group and the other weeks in the same group. Angiostrongylus cantonensis larvae were observed in the gonad and albumen gland from day 21 and 28 of infection in groups I and I+E, respectively, with granuloma-like formation. At these observation periods and in these groups, an increase in galactogen was observed in the albumen gland, which influenced egg laying, suggesting the existence of a fecundity compensation mechanism phenomenon. It was possible to conclude that both stressors - A. cantonensis infection and exposure to E. milii latex - directly influenced the survival and reproductive parameters of B. glabrata.

Reproductive alterations of Biomphalaria glabrata (Say, 1818) infected with Angiostrongylus cantonensis (Chen, 1935) and exposed to Euphorbia milii var. hislopii latex / Alterações na atividade reprodutiva de Biomphalaria glabrata (Say, 1818) infectada com Angiostrongylus cantonensis (Chen, 1935) e exposta ao látex de Euphorbia milii var. hislopii

The natural phytochemical latex of Euphorbia milii var. hislopii is one of the most promising natural molluscicides for the control of Biomphalaria glabrata, and has been widely studied under laboratory conditions for selective control of schistosomiasis transmission. However, the effect of this product on B. glabrata infected by other helminths had not yet been investigated. The present study reports evaluation of the effect of E. milii var. hislopii latex on the survival and reproductive activity of B. glabrata infected by Angiostrongylus cantonensis. For this purpose, the following groups were formed: control (C), exposed (E), infected (I) and infected and exposed for different time intervals (1 day - I+E-1D, 7 days - I+E-7D, 14 days - I+E-14D, 21 days - I+E-21D and 28 days - I+E-28D). The experimental infection was performed with L1 larvae of A. cantonensis and exposure to 0.08 mg/L (LC50) of E. milii latex for a period of 24 hours. We analyzed the effects of E. milii latex on the survival of snails during four weeks, reproductive parameters and possible histophysiopathological changes in the gonad and albumen gland of the snails. Regarding survival, there was reduction of more than 50% in the groups exposed to latex (E and I + E) compared to the control group. As for the number of ovigerous masses, eggs, and average number of hatched snails, significant increases were observed in the I+E-1D group at the 4th week compared to the control group and the other weeks in the same group. Angiostrongylus cantonensis larvae were observed in the gonad and albumen gland from day 21 and 28 of infection in groups I and I+E, respectively, with granuloma-like formation. At these observation periods and in these groups, an increase in galactogen was observed in the albumen gland, which influenced egg laying, suggesting the existence of a fecundity compensation mechanism phenomenon. It was possible to conclude that both stressors - A. cantonensis infection and exposure to E. milii latex - directly influenced the survival and reproductive parameters of B. glabrata.

O fitoquímico natural látex de Euphorbia milii var. hislopii é um dos mais promissores moluscicidas naturais para o controle de Biomphalaria glabrata, tendo sido amplamente estudado em condições laboratoriais como controle seletivo da transmissão da esquistossomose. No entanto, o efeito deste produto em B. glabrata infectada por outro helminto, ainda não tinha sido investigado. O presente trabalho tem por objetivo avaliar o efeito do látex de E. milii var. hislopii na sobrevivência e atividade reprodutiva de B. glabrata infectada por Angiostrongylus cantonensis. Para isso, foram formados os seguintes grupos: controle (C), exposto (E), infectado (I) e infectado e exposto em diferentes intervalos de tempo de infecção (1 dia - I+E-1D, 7 dias - I+E-7D, 14 dias - I+E-14D, 21 dias - I+E-21D e 28 dias - I+E-28D). A infecção experimental foi realizada com larvas L1 de A. cantonensis e a exposição com 0,08 mg/L (LC50) do látex de E. milii por um período de 24 horas. Foram analisados os efeitos do látex de E. milii quanto à sobrevivência dos moluscos ao longo de quatro semanas, aos parâmetros reprodutivos e as possíveis alterações histofisiopatológicas na gônada e na glândula de albúmen dos moluscos analisados. Em relação à sobrevivência foi possível observar redução de mais de 50% nos grupos expostos ao látex (E e I + E) em comparação ao grupo controle. Quanto ao número de massas ovígeras, ovos e número médio de moluscos eclodidos pode-se observar um aumento significativo no grupo I+E-1D na 4ª semana em comparação ao grupo controle e as demais semanas no mesmo grupo. Larvas de A. cantonensis foram observadas na glândula e na glândula de albúmen, a partir dos 21º dias e 28º dia de infecção nos grupos I e I+E, respectivamente, com a formação de granuloma-like. Nesses períodos de observação e nesses grupos pode-se observar na glândula de albúmen, um aumento do conteúdo de galactogênio, o que influenciou a postura dos ovos, sugerindo um fenômeno de compensação da fecundidade. Foi possível concluir que ambos agentes estressores, infecção por A. cantonensis e exposição ao látex de E. milii influenciou diretamente a sobrevivência e os parâmetros reprodutivos de B. glabrata.

Evaluation of the reproductive profile of Subulina octona (Gastropoda, Subulinidae) experimentally infected by Paratanaisia bragai (Digenea, Eucotylidae) / Avaliação do perfil reprodutivo de Subulina octona (Gastropoda, Subulinidae) infectada experimentalmente por Paratanaisia bragai (Digenea, Eucotylidae)

Snails infected by trematodes may increase or decrease their reproductive activity in response to the presence of infection. Our aim was to verify the reproductive alterations in Subulina octona after infection by Paratanaisia bragai. The infected snails were individually exposed for 24 hours to 20 parasite eggs and four groups were formed (10, 20, 30 and 40 d.p.i.- days after infection), along with control groups. Every 10 days, the number of eggs in the reproductive tract, number of eggs hatched, galactogen content and histopathological changes were evaluated. The reproductive in the control and infected snails presented an alternating pattern, where periods of high production of eggs and newly hatched were followed by periods of low production. However, in relation to the amount of galactogen, both control and infected groups followed the same pattern of variation. In the histology, we observed the presence of male and female gametes with marked reduction in the number of oocytes. The results indicate that the intra-snail development of the parasite affects the reproductive biology of the host.(AU)

Moluscos infectados por trematódeos podem aumentar ou diminuir sua atividade reprodutiva em resposta à presença da infecção. Nosso objetivo foi verificar as alterações reprodutivas de Subulina octona após a infecção por Paratanaisia bragai. Os moluscos infectados foram individualmente expostos durante 24 horas a 20 ovos do parasito e quatro grupos foram formados (10, 20, 30 e 40 d.p.i. dias pós infecção), com respectivos grupos controle. A cada 10 dias, o número de ovos no trato reprodutivo, o número de ovos eclodidos, o conteúdo de galactogênio e alterações histopatológicas eram avaliadas. O padrão reprodutivo nos moluscos controle e infectados evidenciou um comportamento alternante, nos quais períodos de alta produção de ovos e filhotes foram seguidos por períodos de baixa produção. No entanto, em relação ao conteúdo de galactogênio, moluscos controle e infectados seguiram o mesmo padrão de variação. Na histologia, observamos a presença de gametas masculinos e femininos com acentuada redução no número de óocitos. Os resultados indicam que o desenvolvimento intramolusco do parasito afeta a biologia reprodutiva do hospedeiro.(AU)

Evaluation of the reproductive profile of Subulina octona (Gastropoda, Subulinidae) experimentally infected by Paratanaisia bragai (Digenea, Eucotylidae) / Avaliação do perfil reprodutivo de Subulina octona (Gastropoda, Subulinidae) infectada experimentalmente por Paratanaisia bragai (Digenea, Eucotylidae)

Abstract Snails infected by trematodes may increase or decrease their reproductive activity in response to the presence of infection. Our aim was to verify the reproductive alterations in Subulina octona after infection by Paratanaisia bragai. The infected snails were individually exposed for 24 hours to 20 parasite eggs and four groups were formed (10, 20, 30 and 40 d.p.i.- days after infection), along with control groups. Every 10 days, the number of eggs in the reproductive tract, number of eggs hatched, galactogen content and histopathological changes were evaluated. The reproductive in the control and infected snails presented an alternating pattern, where periods of high production of eggs and newly hatched were followed by periods of low production. However, in relation to the amount of galactogen, both control and infected groups followed the same pattern of variation. In the histology, we observed the presence of male and female gametes with marked reduction in the number of oocytes. The results indicate that the intra-snail development of the parasite affects the reproductive biology of the host.

Resumo Moluscos infectados por trematódeos podem aumentar ou diminuir sua atividade reprodutiva em resposta à presença da infecção. Nosso objetivo foi verificar as alterações reprodutivas de Subulina octona após a infecção por Paratanaisia bragai. Os moluscos infectados foram individualmente expostos durante 24 horas a 20 ovos do parasito e quatro grupos foram formados (10, 20, 30 e 40 d.p.i. - dias pós infecção), com respectivos grupos controle. A cada 10 dias, o número de ovos no trato reprodutivo, o número de ovos eclodidos, o conteúdo de galactogênio e alterações histopatológicas eram avaliadas. O padrão reprodutivo nos moluscos controle e infectados evidenciou um comportamento alternante, nos quais períodos de alta produção de ovos e filhotes foram seguidos por períodos de baixa produção. No entanto, em relação ao conteúdo de galactogênio, moluscos controle e infectados seguiram o mesmo padrão de variação. Na histologia, observamos a presença de gametas masculinos e femininos com acentuada redução no número de óocitos. Os resultados indicam que o desenvolvimento intramolusco do parasito afeta a biologia reprodutiva do hospedeiro.

Evaluation of the reproductive profile of Subulina octona (Gastropoda, Subulinidae) experimentally infected by Paratanaisia bragai (Digenea, Eucotylidae).

Snails infected by trematodes may increase or decrease their reproductive activity in response to the presence of infection. Our aim was to verify the reproductive alterations in Subulina octona after infection by Paratanaisia bragai. The infected snails were individually exposed for 24 hours to 20 parasite eggs and four groups were formed (10, 20, 30 and 40 d.p.i.- days after infection), along with control groups. Every 10 days, the number of eggs in the reproductive tract, number of eggs hatched, galactogen content and histopathological changes were evaluated. The reproductive in the control and infected snails presented an alternating pattern, where periods of high production of eggs and newly hatched were followed by periods of low production. However, in relation to the amount of galactogen, both control and infected groups followed the same pattern of variation. In the histology, we observed the presence of male and female gametes with marked reduction in the number of oocytes. The results indicate that the intra-snail development of the parasite affects the reproductive biology of the host.

Health economic assessment of a shift to active transport.

Active transportation (walking or cycling) as a substitute for car trips still represents a small percentage of all daily travels in many European cities. This study aimed to estimate the health and economic co-benefits for the adult population of modal shift from driving to active travel in urban environments. Three scenarios were modelled for the case study, the city of Porto, Portugal, by comparing travel patterns of 2013 to hypothetical scenarios of modal shifts from driving to active transport, namely: i) SC1 - conservative scenario, with a change of 5% from driving to cycling and 10% from driving to walking; ii) SC2 - moderate scenario, with a shift of 10% and 15%, respectively; and iii) SC3 - optimistic scenario, with a shift of 15% and 20%, respectively. The mortality risk reduction for five health outcomes (colon and breast cancers, diabetes, ischemic heart disease, cerebrovascular disease) was assessed, including an estimation of traffic injury and air pollution exposure risks. Results were presented in Disability-Adjusted Life Years (DALYs) avoided. Economic valuation for each scenario was performed using a Willingness-to-Pay approach for morbimortality and a Cost of Illness approach for 2013 hospitalizations and work absenteeism. Significant health benefits were found in all modelled scenarios, ranging from 1657 (16%) to 2881 (28%) DALYs avoided. Total costs averted ranged from €3894 to €6769 million through the scenarios. Cardio and cerebrovascular diseases mortality presented the largest benefit, accounting for about 3/4 of all avoidable DALYs in all scenarios. Reductions in CO2 and PM10 emissions were calculated, showing a decrease from 31.6 to 73.7 kt of CO2 and 7 to 16 t for PM10, respectively. A modal shift towards active transportation could lead to significant health and economic benefits, indicating that the evaluation of health impacts should be included in the analysis of active transport interventions.

Evaluation of the reproductive profile of Subulina octona (Gastropoda, Subulinidae) experimentally infected by Paratanaisia bragai (Digenea, Eucotylidae)

Abstract Snails infected by trematodes may increase or decrease their reproductive activity in response to the presence of infection. Our aim was to verify the reproductive alterations in Subulina octona after infection by Paratanaisia bragai. The infected snails were individually exposed for 24 hours to 20 parasite eggs and four groups were formed (10, 20, 30 and 40 d.p.i.- days after infection), along with control groups. Every 10 days, the number of eggs in the reproductive tract, number of eggs hatched, galactogen content and histopathological changes were evaluated. The reproductive in the control and infected snails presented an alternating pattern, where periods of high production of eggs and newly hatched were followed by periods of low production. However, in relation to the amount of galactogen, both control and infected groups followed the same pattern of variation. In the histology, we observed the presence of male and female gametes with marked reduction in the number of oocytes. The results indicate that the intra-snail development of the parasite affects the reproductive biology of the host.

Resumo Moluscos infectados por trematódeos podem aumentar ou diminuir sua atividade reprodutiva em resposta à presença da infecção. Nosso objetivo foi verificar as alterações reprodutivas de Subulina octona após a infecção por Paratanaisia bragai. Os moluscos infectados foram individualmente expostos durante 24 horas a 20 ovos do parasito e quatro grupos foram formados (10, 20, 30 e 40 d.p.i. dias pós infecção), com respectivos grupos controle. A cada 10 dias, o número de ovos no trato reprodutivo, o número de ovos eclodidos, o conteúdo de galactogênio e alterações histopatológicas eram avaliadas. O padrão reprodutivo nos moluscos controle e infectados evidenciou um comportamento alternante, nos quais períodos de alta produção de ovos e filhotes foram seguidos por períodos de baixa produção. No entanto, em relação ao conteúdo de galactogênio, moluscos controle e infectados seguiram o mesmo padrão de variação. Na histologia, observamos a presença de gametas masculinos e femininos com acentuada redução no número de óocitos. Os resultados indicam que o desenvolvimento intramolusco do parasito afeta a biologia reprodutiva do hospedeiro.

First record of Bulimulus tenuissimus (Mollusca) as potential experimental intermediate host of Angiostrongylus cantonensis (Nematoda) / Primeiro registro de Bulimulus tenuissimus (Mollusca) como hospedeiro intermediário de Angiostrongylus cantonensis (Nematoda)

Abstract Snails are essential to complete the life cycle of the metastrongylid nematode Angiostrongylus cantonensis, the causative agent of infections in domestic and wild animals, mainly rodents, and also of neural angiostrongyliasis or eosinophilic meningitis in humans. There are many reports of mollusks that can act as intermediate hosts of this parasite, especially freshwater snails and the African giant Achatina fulica. The terrestrial gastropod Bulimulus tenuissimus is widely distributed in Brazil and other species of the same genus occur in Brazil and other countries, overlapping regions in which there are reports of the occurrence of A. cantonensis and angiostrongyliasis. In spite of this, there are no records in the literature of this species performing the role of intermediate host to A. cantonensis. The present study analyzed the experimental infection with first-stage larvae of A. cantonensis, under laboratory conditions, of B. tenuissimus, by using histology and electron microscopy techniques. Three weeks after exposure to L1 larvae, it was possible to recover L3 larvae in small numbers from the infected snails. Developing larvae were observed in the cephalopedal mass (foot), ovotestis, and mantle tissues, being located inside a granulomatous structure composed of hemocyte infiltration, but there was no calcium or collagen deposition in these structures in significant amounts. In the third week post exposure, it was possible observe a sheath around the developing larvae. The infected snails presented reduction in the fibrous muscular tissue in the foot region, loss of the acinar organization in the digestive gland, with increase of amorphous material inside the acini and loss of epithelial pattern of nuclear organization in the acinar cells. However, the ovotestis seemed unaffected by the infection, since there was a large number of developing oocytes and spermatozoa in different stages of formation. The digestion of infected snails allows us the third-stage recovery rate of 17.25%, at 14 days post exposure to the L1. These L3 recovered from B. tenuissimus were used to infect rats experimentally, and 43 days post infection first-stage (L1) larvae of A. cantonensis were recovered from fresh feces. The results presented constituted the first report of the role of B. tenuissimus as an experimental intermediate host to A. cantonensis and shed some light on a possible problem, since the overlapping distribution of B. tenuissimus and A. cantonensis in Brazil and other countries where different species of Bulimulus occur enables the establishment and maintenance of the life cycle of this parasite in nature, with wild rodents as reservoirs, acting as a source of infection to humans, causing neural angiostrongyliasis.

Resumo Os moluscos são um requisito essencial para a conclusão do ciclo de vida pelo nematoide metastrogilídeo Angiostrongylus cantonensis, o agente causador de infecções em animais domésticos e selvagens, principalmente roedores, e também de angiostrongilíase neural ou meningite eosinofílica em humanos. Há muitos relatos de moluscos que podem atuar como hospedeiro para este parasito, sendo o foco dado aos moluscos de água doce e no gigante africano Achatina fulica. O gastrópode terrestre Bulimulus tenuissimus é amplamente distribuído no território brasileiro e há outras espécies do mesmo gênero que ocorrem no Brasil e outros países, sobrepondo-se às regiões em que há relatos à ocorrência de A. cantonensis e angiostornigilíase. Apesar disso, não há registro na literatura, acerca desta espécie como hospedeiro intermediário para A. cantonensis. O presente estudo teve como objetivo verificar a possibilidade de infectar experimentalmente, utilizando larvas L1 de A. cantonensis, em condições laboratoriais, o molusco B. tenuissimus, utilizando técnicas de histologia e microscopia eletrônica. Três semanas após a exposição às larvas L1, foi possível recuperar larvas L3 dos moluscos infectados, em pequena quantidade. As larvas em desenvolvimento foram observadas na massa cefalopediosa (pé), ovotestis e nos tecidos do manto, sendo localizadas dentro de uma estrutura granulomatosa constituída por infiltração hemocitária, mas não houve deposição de cálcio ou colágeno nessas estruturas em quantidade significativa. Na terceira semana pós exposição, foi possível observar uma bainha ao redor das larvas em desenvolvimento. Os caracóis infectados apresentaram redução no tecido muscular fibroso na região do pé, perda da organização acinar na glândula digestiva, com aumento de material amorfo dentro dos ácinos e perda do padrão epitelial da organização nuclear nas células acinares. No entanto, o ovotestis, pareceu não ser afetado pela infecção, uma vez que houve um grande número de oócitos em desenvolvimento e espermatozóides em diferentes estágios de formação. A digestão dos moluscos infectados nos permitiu a recuperação de larvas de terceiro estágio (17,25%), aos 14 dias após a exposição à L1 de A. cantonensis . Estas L3 recuperadas de B. tenuissimus foram utilizados para infectar ratos experimentalmente, e 43 dias após a infecção, as larvas do primeiro estágio (L1) foram recuperadas de fezes frescas. Os resultados apresentados representam o primeiro registro do papel de B. tenuissimus como hospedeiro intermediário experimental de A. cantonensis e trazem alguma luz a um problema, até então silencioso, uma vez que a sobreposição da distribuição de B. tenuissimus e A. cantonensis no Brasil, e outros países, onde as diferentes espécies de Bulimulus ocorrem, torna possível o estabelecimento e manutenção do ciclo de vida deste parasito na natureza, com roedores selvagens como reservatório, agindo como fonte de infecção para humanos e causando a angiostrongilíase neural.

First record of Bulimulus tenuissimus (Mollusca) as potential experimental intermediate host of Angiostrongylus cantonensis (Nematoda).

Snails are essential to complete the life cycle of the metastrongylid nematode Angiostrongylus cantonensis, the causative agent of infections in domestic and wild animals, mainly rodents, and also of neural angiostrongyliasis or eosinophilic meningitis in humans. There are many reports of mollusks that can act as intermediate hosts of this parasite, especially freshwater snails and the African giant Achatina fulica. The terrestrial gastropod Bulimulus tenuissimus is widely distributed in Brazil and other species of the same genus occur in Brazil and other countries, overlapping regions in which there are reports of the occurrence of A. cantonensis and angiostrongyliasis. In spite of this, there are no records in the literature of this species performing the role of intermediate host to A. cantonensis. The present study analyzed the experimental infection with first-stage larvae of A. cantonensis, under laboratory conditions, of B. tenuissimus, by using histology and electron microscopy techniques. Three weeks after exposure to L1 larvae, it was possible to recover L3 larvae in small numbers from the infected snails. Developing larvae were observed in the cephalopedal mass (foot), ovotestis, and mantle tissues, being located inside a granulomatous structure composed of hemocyte infiltration, but there was no calcium or collagen deposition in these structures in significant amounts. In the third week post exposure, it was possible observe a sheath around the developing larvae. The infected snails presented reduction in the fibrous muscular tissue in the foot region, loss of the acinar organization in the digestive gland, with increase of amorphous material inside the acini and loss of epithelial pattern of nuclear organization in the acinar cells. However, the ovotestis seemed unaffected by the infection, since there was a large number of developing oocytes and spermatozoa in different stages of formation. The digestion of infected snails allows us the third-stage recovery rate of 17.25%, at 14 days post exposure to the L1. These L3 recovered from B. tenuissimus were used to infect rats experimentally, and 43 days post infection first-stage (L1) larvae of A. cantonensis were recovered from fresh feces. The results presented constituted the first report of the role of B. tenuissimus as an experimental intermediate host to A. cantonensis and shed some light on a possible problem, since the overlapping distribution of B. tenuissimus and A. cantonensis in Brazil and other countries where different species of Bulimulus occur enables the establishment and maintenance of the life cycle of this parasite in nature, with wild rodents as reservoirs, acting as a source of infection to humans, causing neural angiostrongyliasis.

First record of Bulimulus tenuissimus (Mollusca) as potential experimental intermediate host of Angiostrongylus cantonensis (Nematoda) / Primeiro registro de Bulimulus tenuissimus (Mollusca) como hospedeiro intermediário de Angiostrongylus cantonensis (Nematoda)

Snails are essential to complete the life cycle of the metastrongylid nematode Angiostrongylus cantonensis, the causative agent of infections in domestic and wild animals, mainly rodents, and also of neural angiostrongyliasis or eosinophilic meningitis in humans. There are many reports of mollusks that can act as intermediate hosts of this parasite, especially freshwater snails and the African giant Achatina fulica. The terrestrial gastropod Bulimulus tenuissimus is widely distributed in Brazil and other species of the same genus occur in Brazil and other countries, overlapping regions in which there are reports of the occurrence of A. cantonensis and angiostrongyliasis. In spite of this, there are no records in the literature of this species performing the role of intermediate host to A. cantonensis. The present study analyzed the experimental infection with first-stage larvae of A. cantonensis, under laboratory conditions, of B. tenuissimus, by using histology and electron microscopy techniques. Three weeks after exposure to L1 larvae, it was possible to recover L3 larvae in small numbers from the infected snails. Developing larvae were observed in the cephalopedal mass (foot), ovotestis, and mantle tissues, being located inside a granulomatous structure composed of hemocyte infiltration, but there was no calcium or collagen deposition in these structures in significant amounts. In the third week post exposure, it was possible observe a sheath around the developing larvae. The infected snails presented reduction in the fibrous muscular tissue in the foot region, loss of the acinar organization in the digestive gland, with increase of amorphous material inside the acini and loss of epithelial pattern of nuclear organization in the acinar cells. However, the ovotestis seemed unaffected by the infection, since there was a large number of developing oocytes and spermatozoa in different stages of formation. The digestion of infected snails allows us the third-stage recovery rate of 17.25%, at 14 days post exposure to the L1. These L3 recovered from B. tenuissimus were used to infect rats experimentally, and 43 days post infection first-stage (L1) larvae of A. cantonensis were recovered from fresh feces. The results presented constituted the first report of the role of B. tenuissimus as an experimental intermediate host to A. cantonensis and shed some light on a possible problem, since the overlapping distribution of B. tenuissimus and A. cantonensis in Brazil and other countries where different species of Bulimulus occur enables the establishment and maintenance of the life cycle of this parasite in nature, with wild rodents as reservoirs, acting as a source of infection to humans, causing neural angiostrongyliasis(AU).

Os moluscos são um requisito essencial para a conclusão do ciclo de vida pelo nematoide metastrogilídeo Angiostrongylus cantonensis, o agente causador de infecções em animais domésticos e selvagens, principalmente roedores, e também de angiostrongilíase neural ou meningite eosinofílica em humanos. Há muitos relatos de moluscos que podem atuar como hospedeiro para este parasito, sendo o foco dado aos moluscos de água doce e no gigante africano Achatina fulica. O gastrópode terrestre Bulimulus tenuissimus é amplamente distribuído no território brasileiro e há outras espécies do mesmo gênero que ocorrem no Brasil e outros países, sobrepondo-se às regiões em que há relatos à ocorrência de A. cantonensis e angiostornigilíase. Apesar disso, não há registro na literatura, acerca desta espécie como hospedeiro intermediário para A. cantonensis. O presente estudo teve como objetivo verificar a possibilidade de infectar experimentalmente, utilizando larvas L1 de A. cantonensis, em condições laboratoriais, o molusco B. tenuissimus, utilizando técnicas de histologia e microscopia eletrônica. Três semanas após a exposição às larvas L1, foi possível recuperar larvas L3 dos moluscos infectados, em pequena quantidade. As larvas em desenvolvimento foram observadas na massa cefalopediosa (pé), ovotestis e nos tecidos do manto, sendo localizadas dentro de uma estrutura granulomatosa constituída por infiltração hemocitária, mas não houve deposição de cálcio ou colágeno nessas estruturas em quantidade significativa. Na terceira semana pós exposição, foi possível observar uma bainha ao redor das larvas em desenvolvimento. Os caracóis infectados apresentaram redução no tecido muscular fibroso na região do pé, perda da organização acinar na glândula digestiva, com aumento de material amorfo dentro dos ácinos e perda do padrão epitelial da organização nuclear nas células acinares. No entanto, o ovotestis, pareceu não ser afetado pela infecção, uma vez que houve um grande número de oócitos em desenvolvimento e espermatozóides em diferentes estágios de formação. A digestão dos moluscos infectados nos permitiu a recuperação de larvas de terceiro estágio (17,25%), aos 14 dias após a exposição à L1 de A. cantonensis . Estas L3 recuperadas de B. tenuissimus foram utilizados para infectar ratos experimentalmente, e 43 dias após a infecção, as larvas do primeiro estágio (L1) foram recuperadas de fezes frescas. Os resultados apresentados representam o primeiro registro do papel de B. tenuissimus como hospedeiro intermediário experimental de A. cantonensis e trazem alguma luz a um problema, até então silencioso, uma vez que a sobreposição da distribuição de B. tenuissimus e A. cantonensis no Brasil, e outros países, onde as diferentes espécies de Bulimulus ocorrem, torna possível o estabelecimento e manutenção do ciclo de vida deste parasito na natureza, com roedores selvagens como reservatório, agindo como fonte de infecção para humanos e causando a angiostrongilíase neural.(AU)

First record of Bulimulus tenuissimus (Mollusca) as potential experimental intermediate host of Angiostrongylus cantonensis (Nematoda)

Abstract Snails are essential to complete the life cycle of the metastrongylid nematode Angiostrongylus cantonensis, the causative agent of infections in domestic and wild animals, mainly rodents, and also of neural angiostrongyliasis or eosinophilic meningitis in humans. There are many reports of mollusks that can act as intermediate hosts of this parasite, especially freshwater snails and the African giant Achatina fulica. The terrestrial gastropod Bulimulus tenuissimus is widely distributed in Brazil and other species of the same genus occur in Brazil and other countries, overlapping regions in which there are reports of the occurrence of A. cantonensis and angiostrongyliasis. In spite of this, there are no records in the literature of this species performing the role of intermediate host to A. cantonensis. The present study analyzed the experimental infection with first-stage larvae of A. cantonensis, under laboratory conditions, of B. tenuissimus, by using histology and electron microscopy techniques. Three weeks after exposure to L1 larvae, it was possible to recover L3 larvae in small numbers from the infected snails. Developing larvae were observed in the cephalopedal mass (foot), ovotestis, and mantle tissues, being located inside a granulomatous structure composed of hemocyte infiltration, but there was no calcium or collagen deposition in these structures in significant amounts. In the third week post exposure, it was possible observe a sheath around the developing larvae. The infected snails presented reduction in the fibrous muscular tissue in the foot region, loss of the acinar organization in the digestive gland, with increase of amorphous material inside the acini and loss of epithelial pattern of nuclear organization in the acinar cells. However, the ovotestis seemed unaffected by the infection, since there was a large number of developing oocytes and spermatozoa in different stages of formation. The digestion of infected snails allows us the third-stage recovery rate of 17.25%, at 14 days post exposure to the L1. These L3 recovered from B. tenuissimus were used to infect rats experimentally, and 43 days post infection first-stage (L1) larvae of A. cantonensis were recovered from fresh feces. The results presented constituted the first report of the role of B. tenuissimus as an experimental intermediate host to A. cantonensis and shed some light on a possible problem, since the overlapping distribution of B. tenuissimus and A. cantonensis in Brazil and other countries where different species of Bulimulus occur enables the establishment and maintenance of the life cycle of this parasite in nature, with wild rodents as reservoirs, acting as a source of infection to humans, causing neural angiostrongyliasis.

Resumo Os moluscos são um requisito essencial para a conclusão do ciclo de vida pelo nematoide metastrogilídeo Angiostrongylus cantonensis, o agente causador de infecções em animais domésticos e selvagens, principalmente roedores, e também de angiostrongilíase neural ou meningite eosinofílica em humanos. Há muitos relatos de moluscos que podem atuar como hospedeiro para este parasito, sendo o foco dado aos moluscos de água doce e no gigante africano Achatina fulica. O gastrópode terrestre Bulimulus tenuissimus é amplamente distribuído no território brasileiro e há outras espécies do mesmo gênero que ocorrem no Brasil e outros países, sobrepondo-se às regiões em que há relatos à ocorrência de A. cantonensis e angiostornigilíase. Apesar disso, não há registro na literatura, acerca desta espécie como hospedeiro intermediário para A. cantonensis. O presente estudo teve como objetivo verificar a possibilidade de infectar experimentalmente, utilizando larvas L1 de A. cantonensis, em condições laboratoriais, o molusco B. tenuissimus, utilizando técnicas de histologia e microscopia eletrônica. Três semanas após a exposição às larvas L1, foi possível recuperar larvas L3 dos moluscos infectados, em pequena quantidade. As larvas em desenvolvimento foram observadas na massa cefalopediosa (pé), ovotestis e nos tecidos do manto, sendo localizadas dentro de uma estrutura granulomatosa constituída por infiltração hemocitária, mas não houve deposição de cálcio ou colágeno nessas estruturas em quantidade significativa. Na terceira semana pós exposição, foi possível observar uma bainha ao redor das larvas em desenvolvimento. Os caracóis infectados apresentaram redução no tecido muscular fibroso na região do pé, perda da organização acinar na glândula digestiva, com aumento de material amorfo dentro dos ácinos e perda do padrão epitelial da organização nuclear nas células acinares. No entanto, o ovotestis, pareceu não ser afetado pela infecção, uma vez que houve um grande número de oócitos em desenvolvimento e espermatozóides em diferentes estágios de formação. A digestão dos moluscos infectados nos permitiu a recuperação de larvas de terceiro estágio (17,25%), aos 14 dias após a exposição à L1 de A. cantonensis . Estas L3 recuperadas de B. tenuissimus foram utilizados para infectar ratos experimentalmente, e 43 dias após a infecção, as larvas do primeiro estágio (L1) foram recuperadas de fezes frescas. Os resultados apresentados representam o primeiro registro do papel de B. tenuissimus como hospedeiro intermediário experimental de A. cantonensis e trazem alguma luz a um problema, até então silencioso, uma vez que a sobreposição da distribuição de B. tenuissimus e A. cantonensis no Brasil, e outros países, onde as diferentes espécies de Bulimulus ocorrem, torna possível o estabelecimento e manutenção do ciclo de vida deste parasito na natureza, com roedores selvagens como reservatório, agindo como fonte de infecção para humanos e causando a angiostrongilíase neural.

First record of Bulimulus tenuissimus (Mollusca) as potential experimental intermediate host of Angiostrongylus cantonensis (Nematoda)

Abstract Snails are essential to complete the life cycle of the metastrongylid nematode Angiostrongylus cantonensis, the causative agent of infections in domestic and wild animals, mainly rodents, and also of neural angiostrongyliasis or eosinophilic meningitis in humans. There are many reports of mollusks that can act as intermediate hosts of this parasite, especially freshwater snails and the African giant Achatina fulica. The terrestrial gastropod Bulimulus tenuissimus is widely distributed in Brazil and other species of the same genus occur in Brazil and other countries, overlapping regions in which there are reports of the occurrence of A. cantonensis and angiostrongyliasis. In spite of this, there are no records in the literature of this species performing the role of intermediate host to A. cantonensis. The present study analyzed the experimental infection with first-stage larvae of A. cantonensis, under laboratory conditions, of B. tenuissimus, by using histology and electron microscopy techniques. Three weeks after exposure to L1 larvae, it was possible to recover L3 larvae in small numbers from the infected snails. Developing larvae were observed in the cephalopedal mass (foot), ovotestis, and mantle tissues, being located inside a granulomatous structure composed of hemocyte infiltration, but there was no calcium or collagen deposition in these structures in significant amounts. In the third week post exposure, it was possible observe a sheath around the developing larvae. The infected snails presented reduction in the fibrous muscular tissue in the foot region, loss of the acinar organization in the digestive gland, with increase of amorphous material inside the acini and loss of epithelial pattern of nuclear organization in the acinar cells. However, the ovotestis seemed unaffected by the infection, since there was a large number of developing oocytes and spermatozoa in different stages of formation. The digestion of infected snails allows us the third-stage recovery rate of 17.25%, at 14 days post exposure to the L1. These L3 recovered from B. tenuissimus were used to infect rats experimentally, and 43 days post infection first-stage (L1) larvae of A. cantonensis were recovered from fresh feces. The results presented constituted the first report of the role of B. tenuissimus as an experimental intermediate host to A. cantonensis and shed some light on a possible problem, since the overlapping distribution of B. tenuissimus and A. cantonensis in Brazil and other countries where different species of Bulimulus occur enables the establishment and maintenance of the life cycle of this parasite in nature, with wild rodents as reservoirs, acting as a source of infection to humans, causing neural angiostrongyliasis.

Resumo Os moluscos são um requisito essencial para a conclusão do ciclo de vida pelo nematoide metastrogilídeo Angiostrongylus cantonensis, o agente causador de infecções em animais domésticos e selvagens, principalmente roedores, e também de angiostrongilíase neural ou meningite eosinofílica em humanos. Há muitos relatos de moluscos que podem atuar como hospedeiro para este parasito, sendo o foco dado aos moluscos de água doce e no gigante africano Achatina fulica. O gastrópode terrestre Bulimulus tenuissimus é amplamente distribuído no território brasileiro e há outras espécies do mesmo gênero que ocorrem no Brasil e outros países, sobrepondo-se às regiões em que há relatos à ocorrência de A. cantonensis e angiostornigilíase. Apesar disso, não há registro na literatura, acerca desta espécie como hospedeiro intermediário para A. cantonensis. O presente estudo teve como objetivo verificar a possibilidade de infectar experimentalmente, utilizando larvas L1 de A. cantonensis, em condições laboratoriais, o molusco B. tenuissimus, utilizando técnicas de histologia e microscopia eletrônica. Três semanas após a exposição às larvas L1, foi possível recuperar larvas L3 dos moluscos infectados, em pequena quantidade. As larvas em desenvolvimento foram observadas na massa cefalopediosa (pé), ovotestis e nos tecidos do manto, sendo localizadas dentro de uma estrutura granulomatosa constituída por infiltração hemocitária, mas não houve deposição de cálcio ou colágeno nessas estruturas em quantidade significativa. Na terceira semana pós exposição, foi possível observar uma bainha ao redor das larvas em desenvolvimento. Os caracóis infectados apresentaram redução no tecido muscular fibroso na região do pé, perda da organização acinar na glândula digestiva, com aumento de material amorfo dentro dos ácinos e perda do padrão epitelial da organização nuclear nas células acinares. No entanto, o ovotestis, pareceu não ser afetado pela infecção, uma vez que houve um grande número de oócitos em desenvolvimento e espermatozóides em diferentes estágios de formação. A digestão dos moluscos infectados nos permitiu a recuperação de larvas de terceiro estágio (17,25%), aos 14 dias após a exposição à L1 de A. cantonensis . Estas L3 recuperadas de B. tenuissimus foram utilizados para infectar ratos experimentalmente, e 43 dias após a infecção, as larvas do primeiro estágio (L1) foram recuperadas de fezes frescas. Os resultados apresentados representam o primeiro registro do papel de B. tenuissimus como hospedeiro intermediário experimental de A. cantonensis e trazem alguma luz a um problema, até então silencioso, uma vez que a sobreposição da distribuição de B. tenuissimus e A. cantonensis no Brasil, e outros países, onde as diferentes espécies de Bulimulus ocorrem, torna possível o estabelecimento e manutenção do ciclo de vida deste parasito na natureza, com roedores selvagens como reservatório, agindo como fonte de infecção para humanos e causando a angiostrongilíase neural.

Evaluation of the reproductive profile of Subulina octona (Gastropoda, Subulinidae) experimentally infected by Paratanaisia bragai (Digenea, Eucotylidae)

Abstract Snails infected by trematodes may increase or decrease their reproductive activity in response to the presence of infection. Our aim was to verify the reproductive alterations in Subulina octona after infection by Paratanaisia bragai. The infected snails were individually exposed for 24 hours to 20 parasite eggs and four groups were formed (10, 20, 30 and 40 d.p.i.- days after infection), along with control groups. Every 10 days, the number of eggs in the reproductive tract, number of eggs hatched, galactogen content and histopathological changes were evaluated. The reproductive in the control and infected snails presented an alternating pattern, where periods of high production of eggs and newly hatched were followed by periods of low production. However, in relation to the amount of galactogen, both control and infected groups followed the same pattern of variation. In the histology, we observed the presence of male and female gametes with marked reduction in the number of oocytes. The results indicate that the intra-snail development of the parasite affects the reproductive biology of the host.

Resumo Moluscos infectados por trematódeos podem aumentar ou diminuir sua atividade reprodutiva em resposta à presença da infecção. Nosso objetivo foi verificar as alterações reprodutivas de Subulina octona após a infecção por Paratanaisia bragai. Os moluscos infectados foram individualmente expostos durante 24 horas a 20 ovos do parasito e quatro grupos foram formados (10, 20, 30 e 40 d.p.i. dias pós infecção), com respectivos grupos controle. A cada 10 dias, o número de ovos no trato reprodutivo, o número de ovos eclodidos, o conteúdo de galactogênio e alterações histopatológicas eram avaliadas. O padrão reprodutivo nos moluscos controle e infectados evidenciou um comportamento alternante, nos quais períodos de alta produção de ovos e filhotes foram seguidos por períodos de baixa produção. No entanto, em relação ao conteúdo de galactogênio, moluscos controle e infectados seguiram o mesmo padrão de variação. Na histologia, observamos a presença de gametas masculinos e femininos com acentuada redução no número de óocitos. Os resultados indicam que o desenvolvimento intramolusco do parasito afeta a biologia reprodutiva do hospedeiro.

Variation of surface ozone in Campo Grande, Brazil: meteorological effect analysis and prediction.

The effect of meteorological variables on surface ozone (O3) concentrations was analysed based on temporal variation of linear correlation and artificial neural network (ANN) models defined by genetic algorithms (GAs). ANN models were also used to predict the daily average concentration of this air pollutant in Campo Grande, Brazil. Three methodologies were applied using GAs, two of them considering threshold models. In these models, the variables selected to define different regimes were daily average O3 concentration, relative humidity and solar radiation. The threshold model that considers two O3 regimes was the one that correctly describes the effect of important meteorological variables in O3 behaviour, presenting also a good predictive performance. Solar radiation, relative humidity and rainfall were considered significant for both O3 regimes; however, wind speed (dispersion effect) was only significant for high concentrations. According to this model, high O3 concentrations corresponded to high solar radiation, low relative humidity and wind speed. This model showed to be a powerful tool to interpret the O3 behaviour, being useful to define policy strategies for human health protection regarding air pollution.

Short-term effects of air pollution on respiratory morbidity at Rio de Janeiro--Part II: health assessment.

The effects of air pollution on health have been studied worldwide. Given that air pollution triggers oxidative stress and inflammation, it is plausible that high levels of air pollutants cause higher number of hospitalisations. This study aimed to assess the impact of air pollution on the emergency hospitalisation for respiratory disease in Rio de Janeiro, Brazil. The study was divided in two parts: Part I specifically addressing the air pollution assessment and Part II addressing the health assessment. Accordingly, this Part II aimed to estimate the association between the concentrations of PM10, SO2 and CO observed in Rio de Janeiro and the number of emergency hospitalisations at a central hospital due to respiratory diseases. The pollutant concentrations were measured at two different sites in Rio de Janeiro, but the excess relative risks were calculated based on the concentrations observed at one of the sites, where limits were generally exceeded more frequently, between September 2000 and December 2005. A time series analysis was performed using the number of hospitalisations, divided in three categories (children until 1 year old, children aged between 1 and 5 years old and elderly with 65 years old or more) as independent variable, the concentrations of pollutants as dependent variables and temperature, relative humidity, long term trend, and seasonality as confounders. Data were analysed using generalised additive models with smoothing for some of the dependent variables. Results showed an excess risk of hospitalisation for respiratory disease higher than 2% per 10 µg m⁻³ increase in PM10 concentrations for children under 5 years old, of 2% per 10 µg m⁻³ increase in SO2 for elderly above 65 years old and around 0.1% per 10 µg m⁻³ increase in CO for children under 1 year and elderly. Other studies have found associations that are in agreement with the results achieved in this study. The study suggests that the ambient levels of air pollutants experienced in Rio de Janeiro between 2000 and 2005 were linked to the number of hospitalisations for respiratory diseases among children and elderly.

Short-term effects of air pollution on respiratory morbidity at Rio de Janeiro--PART I: Air pollution assessment.

Exposure to air pollution has been related with the most varied adverse health outcomes. This study aims to assess the impact of air pollution on the emergency hospitalization for respiratory disease in Rio de Janeiro, Brazil. The study was divided in two parts: Part I specifically addressing the air pollution assessment and Part II addressing the health assessment. Accordingly, this Part I aims to: i) evaluate the concentrations of PM(10), SO(2) and CO at two sites in Rio de Janeiro and compare them; ii) analyse the concentrations observed according to the national and international standards; and iii) analyse the air pollutants behaviour, namely, annually, seasonally, daily and considering weekdays/weekends variations. The pollutant concentrations were measured at two different sites in Rio de Janeiro and the analysis was performed for the period between September 2000 and December 2005. Results showed that PM(10) concentrations in Rio de Janeiro exceeded the daily and annual standards imposed by the European Union, the Brazilian legislation and WHO guidelines. Regarding SO(2) and CO, concentrations were, generally, below both European and Brazilian standards. Nevertheless, considering WHO guidelines, SO(2) threshold for daily concentrations (20 µg m(-3)) was exceeded around 150 times. Behaviour assessment showed that the influence of traffic is a major factor affecting the air pollution in Rio de Janeiro. Considering the results achieved and the proven health effects of air pollution, strategies should be defined for its reduction, particularly concerning particulate matter, and consequently contribute to the protection of public health.