Infectivity of Chordodes nobilii larvae (Gordiida: Nematomorpha) / Infectividad de las larvas de Chordodes nobilii (Gordiida: Nematomorpha)

AbstractThe gordiids are freshwater representatives of the parasite phylum Nematomorpha that function as a link between aquatic and terrestrial ecosystems. In recent years, different ecotoxicologic studies have been made with the South-American gordiid species, Chordodes nobilii, that have demonstrated the capacity of this group to act as a bioindicator of contamination. Despite the Gordiida's ecologic relevance, further studies are still needed to elucidate different aspects of the biology of the class, and among those, the infective capacity, a parameter that can be evaluated by the infection index mean abundance (IIMA). A knowledge of the intrinsic variability in the infective capacity of C. nobilii would warrant priority in order to establish, the range of acceptable responses for normal or standard conditions in the laboratory, and, to compare the criteria among different assays. The objective of this study was to establish a baseline value for the infective capacity for C. nobilii larvae, under controlled laboratory conditions, by employing the IIMA as the evaluation parameter. To that end, we analyzed the infective capacity of C. nobilii larvae that had hatched from different strings of eggs laid in the laboratory by a total of 12 females. The C. nobilii adults were collected from streams within the Argentina Sauce Grande basin, between 2006 and 2009. Once in the laboratory, after mating, the females were placed in individual containers for oviposition. The egg strings obtained from each female were cut in 3 mm long segments; and when free larvae were observed, the segments (N= 90) were placed together with 30 Aedes aegypti larvae for evaluation of the gordiids' infective capacity. After 72 h, the mosquito larvae were observed by microscopy in order to quantify the C. nobilii larvae in body cavities. The IIMAs were calculated as the total number of C. nobilii larvae present divided by total number of A. aegypti larvae examined. For analysis of the IIMAs obtained, the data were grouped according to the female who made the original ovoposition. Our results enabled the corroboration of an ample range of responses in the infective capacity of this species, a characteristic that would normally be linked to the progenitors originating the hatch. Because this relationship prevents the establishment of a baseline for making comparisons among assays with gordiids, through the IIMA as a response parameter, we recommend expressing the IIMA values in each assay relative to their respective controls. These findings also provide evidence for the greater success in infections by certain members of the progeny over others. Finally, on the basis of the results obtained from this study, we stress the relevance of the use of the IIMA as a decisive aspect to be considered in different studies on the biology of Gordiida. Rev. Biol. Trop. 65 (1): 1-8. Epub 2017 March 01.

ResumenLos gordiidos son representantes dulceacuícolas del Phylum parásito Nematomorpha que actúan como un enlace entre ecosistemas acuáticos y terrestres. En años recientes, diferentes estudios ecotoxicológicos se han desarrollado con una especie sudamericana de gordiido, C. nobilii, que ha demostrado la capacidad de este grupo de actuar como bioindicador de contaminación. A pesar de su evidente importancia ecológica, aún se necesitan realizar estudios para dilucidar distintos aspectos de su biología, entre estos, la capacidad infectiva, un parámetro que puede evaluarse utilizando el Índice de Infección Abundancia Media (IIMA). El conocimiento de la variabilidad intrínseca en la capacidad infectiva de C. nobilii merece prioridad con el objeto de establecer el ámbito de respuesta aceptable para condiciones normales o estándar en el laboratorio, y que permita comparar los resultados entre distintos ensayos. El objetivo de este estudio es establecer la línea de base de la capacidad infectiva del gordiido C. nobilii en condiciones controladas de laboratorio, empleando el IIMA como parámetro de evaluación. Con este fin, se analizó la capacidad infectiva de larvas de C. nobilii que eclosionaron de diferentes cordones de huevos depositados por un total de 12 hembras, mantenidas en laboratorio. Los adultos de C. nobilii fueron recolectados de arroyos de la cuenca argentina Sauce Grande, entre 2006 y 2009. Una vez en el laboratorio, después de la cópula, las hembras se ubicaron en recipientes individuales a la espera de la oviposición. Los cordones de huevos obtenidos de cada hembra se cortaron en segmentos de 3 mm de longitud; y cuando las larvas libres fueron observadas al microscopio, los segmentos (N= 90) fueron ubicados junto con 30 larvas de Aedes aegypti para evaluar la capacidad infectiva del gordiido. Después de 72 h, las larvas del mosquito fueron observadas al microscopio para contabilizar las larvas de C. nobilii en las cavidades corporales. El IIMA fue calculado como el número total de larvas de C. nobilii presentes dividido entre el número total de larvas de A. aegypti examinadas. Para el análisis de los IIMAs obtenidos, los datos fueron agrupados de acuerdo a la hembra que hizo la oviposición. Nuestros resultados permiten corroborar un amplio rango de respuesta en la capacidad infectiva de esta especie, que estaría vinculada al origen de la camada. Debido a que no se pudo establecer una línea de base para realizar comparaciones entre estudios en los gordiidos utilizando el IIMA como parámetro de respuesta, se aconseja relativizar los valores de los IIMAs a sus respectivos controles. Estos resultados también pusieron en evidencia la ventaja en el éxito de infección de algunas progenies sobre las restantes. Finalmente, con base en los resultados obtenidos a partir de este estudio se plantea la importancia del uso del IIMA como punto final a considerar en distintos estudios sobre la biología de los gordiida.

Infectivity of Chordodes nobilii larvae (Gordiida: Nematomorpha).

The gordiids are freshwater representatives of the parasite phylum Nematomorpha that function as a link between aquatic and terrestrial ecosystems. In recent years, different ecotoxicologic studies have been made with the South-American gordiid species, Chordodes nobilii, that have demonstrated the capacity of this group to act as a bioindicator of contamination. Despite the Gordiida's ecologic relevance, further studies are still needed to elucidate different aspects of the biology of the class, and among those, the infective capacity, a parameter that can be evaluated by the infection index mean abundance (IIMA). A knowledge of the intrinsic variability in the infective capacity of C. nobilii would warrant priority in order to establish, the range of acceptable responses for normal or standard conditions in the laboratory, and, to compare the criteria among different assays. The objective of this study was to establish a baseline value for the infective capacity for C. nobilii larvae, under controlled laboratory conditions, by employing the IIMA as the evaluation parameter. To that end, we analyzed the infective capacity of C. nobilii larvae that had hatched from different strings of eggs laid in the laboratory by a total of 12 females. The C. nobilii adults were collected from streams within the Argentina Sauce Grande basin, between 2006 and 2009. Once in the laboratory, after mating, the females were placed in individual containers for oviposition. The egg strings obtained from each female were cut in 3 mm long segments; and when free larvae were observed, the segments (N= 90) were placed together with 30 Aedes aegypti larvae for evaluation of the gordiids' infective capacity. After 72 h, the mosquito larvae were observed by microscopy in order to quantify the C. nobilii larvae in body cavities. The IIMAs were calculated as the total number of C. nobilii larvae present divided by total number of A. aegypti larvae examined. For analysis of the IIMAs obtained, the data were grouped according to the female who made the original ovoposition. Our results enabled the corroboration of an ample range of responses in the infective capacity of this species, a characteristic that would normally be linked to the progenitors originating the hatch. Because this relationship prevents the establishment of a baseline for making comparisons among assays with gordiids, through the IIMA as a response parameter, we recommend expressing the IIMA values in each assay relative to their respective controls. These findings also provide evidence for the greater success in infections by certain members of the progeny over others. Finally, on the basis of the results obtained from this study, we stress the relevance of the use of the IIMA as a decisive aspect to be considered in different studies on the biology of Gordiida.

Effect of temperature changes on the cercarial-shedding rate of two trematodes / Efecto de cambios en la temperatura sobre la emergencia de cercarías de dos trematodes

Considerando que la temperatura es un factor clave en la emergencia de cercarias, el objetivo de este trabajo fue evaluar el efecto de ésta variable sobre la emergencia de dos tipos morfológicos de cercarias. Los bioensayos se realizaron con caracoles naturalmente infectados con cercarias de los tipos monostoma o pleurolofocerca, debido a que estos dos tipos fueron los más prevalentes en el ambiente en estudio. Los caracoles fueron ubicados en cápsulas plásticas de 6 ml conteniendo 3 ml de agua reconstituida, y entonces fueron secuencialmente expuestos por 24 h con un ciclo 14:10 h luz:oscuridad a 20, 25 y 30 °C. Los datos fueron analizados a un nivel de significación de p<0.05 con el programa InfoStat. Los resultados mostraron que la temperatura óptima de la emergencia de cercarias estuvo en el rango de 25 °C, y disminuyó significativamente a 30 °C. Se observó también que no existía diferencia significativa en el número de cercarias emergentes entre los dos tipos morfológicos.(AU)

Since temperature is a key condition in the initiation of cercarial emergence, the objective of the work reported here was to evaluate the effect of that variable on the shedding of two morphologic and taxonomic types of cercariae. Bioassays were accordingly performed with snails naturally infected with either monostome or pleurolophocercous cercariae because those two cercarial types were found to be the most prevalent in the environment under study. The snails were placed in 6-ml circular plastic dishes with soft plastic stoppers containing 3 ml of reconstituted water and then sequentially exposed for 24 h to a 14:10-h light:dark cycle at 20, 25, or 30 °C. The data were analyzed at a significance level of p <0.05 by means of the InfoStat program. The optimum temperature for cercarial emergence was typically in the range of 25 °C and decreased significantly at 30 °C. We observed no significant difference in the number of cercariae emerging between the two morphologic types.(AU)

Effect of temperature changes on the cercarial-shedding rate of two trematodes / Efecto de cambios en la temperatura sobre la emergencia de cercarías de dos trematodes

Considerando que la temperatura es un factor clave en la emergencia de cercarias, el objetivo de este trabajo fue evaluar el efecto de ésta variable sobre la emergencia de dos tipos morfológicos de cercarias. Los bioensayos se realizaron con caracoles naturalmente infectados con cercarias de los tipos monostoma o pleurolofocerca, debido a que estos dos tipos fueron los más prevalentes en el ambiente en estudio. Los caracoles fueron ubicados en cápsulas plásticas de 6 ml conteniendo 3 ml de agua reconstituida, y entonces fueron secuencialmente expuestos por 24 h con un ciclo 14:10 h luz:oscuridad a 20, 25 y 30 °C. Los datos fueron analizados a un nivel de significación de p<0.05 con el programa InfoStat. Los resultados mostraron que la temperatura óptima de la emergencia de cercarias estuvo en el rango de 25 °C, y disminuyó significativamente a 30 °C. Se observó también que no existía diferencia significativa en el número de cercarias emergentes entre los dos tipos morfológicos.

Since temperature is a key condition in the initiation of cercarial emergence, the objective of the work reported here was to evaluate the effect of that variable on the shedding of two morphologic and taxonomic types of cercariae. Bioassays were accordingly performed with snails naturally infected with either monostome or pleurolophocercous cercariae because those two cercarial types were found to be the most prevalent in the environment under study. The snails were placed in 6-ml circular plastic dishes with soft plastic stoppers containing 3 ml of reconstituted water and then sequentially exposed for 24 h to a 14:10-h light:dark cycle at 20, 25, or 30 °C. The data were analyzed at a significance level of p <0.05 by means of the InfoStat program. The optimum temperature for cercarial emergence was typically in the range of 25 °C and decreased significantly at 30 °C. We observed no significant difference in the number of cercariae emerging between the two morphologic types.

Effect of temperature changes on the cercarial-shedding rate of two trematodes

RESUMEN Considerando que la temperatura es un factor clave en la emergencia de cercarias, el objetivo de este trabajo fue evaluar el efecto de ésta variable sobre la emergencia de dos tipos morfológicos de cercarias. Los bioensayos se realizaron con caracoles naturalmente infectados con cercarias de los tipos monostoma o pleurolofocerca, debido a que estos dos tipos fueron los más prevalentes en el ambiente en estudio. Los caracoles fueron ubicados en cápsulas plásticas de 6 ml conteniendo 3 ml de agua reconstituida, y entonces fueron secuencialmente expuestos por 24 h con un ciclo 14:10 h luz:oscuridad a 20, 25 y 30 °C. Los datos fueron analizados a un nivel de significación de p 0.05 con el programa InfoStat. Los resultados mostraron que la temperatura óptima de la emergencia de cercarias estuvo en el rango de 25 °C, y disminuyó significativamente a 30 °C. Se observó también que no existía diferencia significativa en el número de cercarias emergentes entre los dos tipos morfológicos.

ABSTRACT Since temperature is a key condition in the initiation of cercarial emergence, the objective of the work reported here was to evaluate the effect of that variable on the shedding of two morphologic and taxonomic types of cercariae. Bioassays were accordingly performed with snails naturally infected with either monostome or pleurolophocercous cercariae because those two cercarial types were found to be the most prevalent in the environment under study. The snails were placed in 6-ml circular plastic dishes with soft plastic stoppers containing 3 ml of reconstituted water and then sequentially exposed for 24 h to a 14:10-h light:dark cycle at 20, 25, or 30 °C. The data were analyzed at a significance level of p 0.05 by means of the InfoStat program. The optimum temperature for cercarial emergence was typically in the range of 25 °C and decreased significantly at 30 °C. We observed no significant difference in the number of cercariae emerging between the two morphologic types.

Validation test with embryonic and larval stages of Chordodes nobilii (Gordiida, Nematomorpha): sensitivity to three reference toxicants.

Chordodes nobilii is a parasite whose pre- and postparasitic stages are found in different types of freshwater bodies. Due to the peculiarities of its life cycle, it acts as a link between freshwater bodies and terrestrial ecosystems. There is little toxicological information on the group Gordiida. It is only known that embryos and larvae of C. nobilii are sensitive to glyphosate and malathion at relevant concentrations in the environment. On this basis, the aims of this study were to characterize the sensitivity of the pre-parasitic stages of C. nobilii to three reference toxicants: sodium dodecyl sulfate (SDS), cadmium chloride and potassium dichromate (Cr(6+)), and to validate a previous experimental protocol for ecotoxicological risk assessment. The protocol involved acute exposure of early embryonic stages and larvae to the three toxicants for 96 h and 48 h, respectively. Embryo development was inhibited only by Cr(6+) which presented a IC(50) of 0.71 mg Cr(6+)L(-1). The development of the eggs exposed to SDS and those exposed to cadmium chloride showed no differences as compared to that of controls. However, the infective capacity of larvae derived from the eggs exposed to the three toxicants was lower than that of controls. Larval survival was affected even at the lowest concentration of the three toxicants assayed. In relation to other freshwater organisms, C. nobilii can be characterized as an organism medium to highly sensitive to the toxicants tested.

Sensitivity of preparasitic stages of Chordodes nobilii (Gordiida, Nematomorpha) to malathion.

The largest part of the life cycle of Gordiida, known as horsehair worms, occurs in aquatic environments usually affected by agricultural activities. The free-living adults reproduce in freshwater environments, where preparasitic larvae undergo development. Since malathion is an insecticide used in the distribution area of Chordodes nobilii, the aim of this study is to evaluate the effect of malathion concentrations which might be expected in the environment on preparasitic stages of this species. The embryonic development and the viability of larvae were analyzed after a short-term exposure to malathion concentrations ranging between 36 and 220 microg a.i./l. Embryo development was inhibited at 220 microg a.i./l and the infective capacity of larvae derived from malathion-exposed eggs was significantly decreased starting from the lowest concentration. Larvae developed from malathion-exposed eggs exhibited malformations. Directly exposed larvae also showed decreased infectivity since the lowest assayed concentration. Our results indicate that a short-term exposure to malathion at levels potentially present in the surface water in environments inhabited by Chordodes nobilii affects significantly its preparasitic stages and the infective capacity of parasitic larvae.

Toxicity of the herbicide glyphosate to Chordodes nobilii (Gordiida, Nematomorpha).

Nematomorpha (horsehair worms) is a poorly known group of worm-like animals similar to nematodes. Adults are free-living and reproduction takes place in freshwater environments, where preparasitic larvae undergo development. All species have a parasitic juvenil stage and infection may result in the host's death, insects being the most frequent host. Most of the life cycle occurs in freshwater environments, which are often contaminated by different pollutants. Based on the lack of information on the toxicity of herbicides to horsehair worms, the objective of this study is to evaluate the effect of different concentrations of glyphosate (technical grade and formulated product) on Chordodes nobilii (Gordiida, Nematomorpha). Bioassays were performed with embryos and larvae (preparasitic stages), and adults (postparasitic stage). Test organisms were exposed for a short period of time to concentrations ranging between 0.1 and 8 mga.e.l(-1) of glyphosate (technical and formulated). Although embryo development was not inhibited, there was a significant decrease in the infective capacity of larvae derived from eggs that had been exposed to >or= 0.1mg/l. Similar results were obtained for directly exposed larvae. No differences in toxicity were detected between the active ingredient and formulated product. Adult exposed for 96 h to 1.76 mgl(-1) formulated Gly shown a mortality of 50%. Results indicate that C. nobilii is affected at glyphosate concentrations lower than those expected to be found in freshwater environments and those specified in the legislation.