ABSTRACT
Currently, the selection of non-pathogenic microorganisms that lack clinically relevant antimicrobial resistance is crucial to bioaugmentation strategies. Pseudomonas sp. P26 (P26) is an environmental bacterium of interest due to its ability to remove aromatic compounds from petroleum, but its safety characteristics are still unknown. The study aimed to: a) determine P26 sensitivity to antimicrobials, b) investigate the presence of quinolone and ß-lactam resistance genes, c) determine the presence of virulence factors, and d) evaluate the effect of P26 on the viability of Galleria mellonella (an invertebrate animal model). P26 antimicrobial sensitivity was determined in vitro using the Kirby-Bauer agar diffusion method and the VITEK 2 automated system (BioMerieux®). Polymerase Chain Reaction was employed for the investigation of genes associated with quinolone resistance, extended-spectrum ß-lactamases, and carbapenemases. Hemolysin and protease production was determined in human blood agar and skimmed-milk agar, respectively. In the in vivo assay, different doses of P26 were injected into Galleria mellonella larvae and their survival was monitored daily. Control larvae injected with Pseudomonas putida KT2440 (a strain considered as safe) and Pseudomonas aeruginosa PA14 (a pathogenic strain) were included. Pseudomonas sp. P26 was susceptible to most evaluated antimicrobials, except for trimethoprim-sulfamethoxazole. No epidemiologically relevant genes associated with quinolone and ß-lactam resistance were identified. Hemolysin and protease production was only evidenced in the virulent strain (PA14). Furthermore, the results obtained in the in vivo experiment demonstrated that inocula less than 108 CFU/mL of P26 and P. putida KT2440 did not significantly affect larval survival, whereas larvae injected with the lowest dose of the pathogenic strain P. aeruginosa PA14 experienced instant mortality. The results suggest that Pseudomonas sp. P26 is a safe strain for its application in environmental bioremediation processes. Additional studies will be conducted to ensure the safety of this bacterium against other organisms.
Subject(s)
Anti-Infective Agents , Moths , Quinolones , Animals , Humans , Pseudomonas/genetics , Agar/pharmacology , Hemolysin Proteins/pharmacology , Moths/microbiology , Larva , Pseudomonas aeruginosa , Anti-Infective Agents/pharmacology , Peptide Hydrolases , Anti-Bacterial Agents/toxicityABSTRACT
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.
Subject(s)
Animals , Parasite Load/methods , Helminths/pathogenicity , Larva/pathogenicity , Oviposition/physiology , Reference Standards , Sex Factors , Statistics, Nonparametric , Aedes/parasitology , Fresh Water/parasitologyABSTRACT
We characterized the infective and sporulation capacities of microbial consortia of arbuscular mycorrhizal fungi (AMF) native of Buenos Aires province (Argentina) and determined if some soil characteristics and mycorrhizal parameters could allow to select potentially beneficial inocula. Soil samples were selected from seven locations in Buenos Aires province all under agricultural (A) and pristine (P) conditions. The AMF were multiplied and mycorrhizal root colonization of trap plants was observed at 10 weeks of growth. Spore number in field was low; however, after multiplication spore density accounted for 80-1175 spores per 100g of soil. The principal component analysis showed that the P and Fe soil contents are the main modulators of infectivity and sporulation capacity. The mycorrhizal potential was determined in three locations, being high in Pristine Lobería and Agricultural Trenque Lauquen and low in Junín. Agricultural Lobería (AL) and Pristine Lobería (PL) inocula were selected and their efficiency was evaluated under controlled conditions. Even though shoot dry matter increases after inoculation was not significant (p>0.05) mycorrhizal response was greater than 40% for tomato and 25% for corn, particularly after inoculation with inocula from the agricultural management. These results could be associated to the incipient development of mycorrhizae in both species. Additional research should be conducted to further develop our findings in order to determine the factors involved in the selection of efficient inocula.
Subject(s)
Fungi , Microbial Consortia , Mycorrhizae/physiology , Soil Microbiology , Agriculture , ArgentinaABSTRACT
Se evaluó la capacidad micotrófica y de esporulación de consorcios microbianos con hongos micorrícicos arbusculares (HMA) nativos de suelos de la provincia de Buenos Aires (Argentina), y se determinó si las características edáficas y los parámetros micorrícicos podrían permitir seleccionar inóculos potencialmente benéficos. Se seleccionaron muestras de suelo provenientes de 7 localidades, cada una bajo manejo agrícola (A) y prístino (P). Se instalaron plantas trampa y a las 10 semanas de crecimiento se evidenció colonización micorrícica en la raíces. El número de esporas del suelo de campo fue bajo, mientras que en el sustrato donde crecieron las plantas trampa varió entre 80-1175 esporas/100 g. El análisis de componentes principales indicó que los contenidos de P y Fe en el suelo fueron los principales moduladores de la capacidad micotrófica y de esporulación. Se determinó el potencial micorrícico (PM) de muestras provenientes de tres localidades: Lobería, Junín y Trenque Lauquen. Se verificó un PM elevado en las muestras provenientes de Lobería con suelo prístino y en las de Trenque Lauquen bajo manejo agrícola, mientras que fue bajo en las de Junín. Finalmente, se evaluó la eficiencia en condiciones controladas de suelos-inóculo de Lobería bajo manejo agrícola o prístino en plantas de maíz y de tomate. Si bien los incrementos en la materia seca asociados a la inoculación fueron no significativos (p > 0,05), la respuesta micorrícica fue superior al 40 % en tomate y al 25 % en maíz, particularmente con el inóculo proveniente del sitio agrícola. Los bajos incrementos de crecimiento estarían asociados con la incipiente micorrización en ambas especies. Se plantea la necesidad de profundizar los estudios a efectos de determinar los factores involucrados que permitirían seleccionar inóculos eficientes.
We characterized the infective and sporulation capacities of microbial consortia of arbuscular mycorrhizal fungi (AMF) native of Buenos Aires province (Argentina) and determined if some soil characteristics and mycorrhizal parameters could allow to select potentially beneficial inocula. Soil samples were selected from seven locations in Buenos Aires province all under agricultural (A) and pristine (P) conditions. The AMF were multiplied and mycorrhizal root colonization of trap plants was observed at 10 weeks of growth. Spore number in field was low; however, after multiplication spore density accounted for 80-1175 spores per 100 g of soil. The principal component analysis showed that the P and Fe soil contents are the main modulators of infectivity and sporulation capacity. The mycorrhizal potential was determined in three locations, being high in Pristine Lobería and Agricultural Trenque Lauquen and low in Junín. Agricultural Lobería (AL) and Pristine Lobería (PL) inocula were selected and their efficiency was evaluated under controlled conditions. Even though shoot dry matter increases after inoculation was not significant (p > 0.05) mycorrhizal response was greater than 40% for tomato and 25% for corn, particularly after inoculation with inocula from the agricultural management. These results could be associated to the incipient development of mycorrhizae in both species. Additional research should be conducted to further develop our findings in order to determine the factors involved in the selection of efficient inocula.
Subject(s)
Fungi , Microbial Consortia , Mycorrhizae/physiology , Soil Microbiology , Agriculture , ArgentinaABSTRACT
Se evaluó la capacidad micotrófica y de esporulación de consorcios microbianos con hongos micorrícicos arbusculares (HMA) nativos de suelos de la provincia de Buenos Aires (Argentina), y se determinó si las características edáficas y los parámetros micorrícicos podrían permitir seleccionar inóculos potencialmente benéficos. Se seleccionaron muestras de suelo provenientes de 7 localidades, cada una bajo manejo agrícola (A) y prístino (P). Se instalaron plantas trampa y a las 10 semanas de crecimiento se evidenció colonización micorrícica en la raíces. El número de esporas del suelo de campo fue bajo, mientras que en el sustrato donde crecieron las plantas trampa varió entre 80-1175 esporas/100 g. El análisis de componentes principales indicó que los contenidos de P y Fe en el suelo fueron los principales moduladores de la capacidad micotrófica y de esporulación. Se determinó el potencial micorrícico (PM) de muestras provenientes de tres localidades: Lobería, Junín y Trenque Lauquen. Se verificó un PM elevado en las muestras provenientes de Lobería con suelo prístino y en las de Trenque Lauquen bajo manejo agrícola, mientras que fue bajo en las de Junín. Finalmente, se evaluó la eficiencia en condiciones controladas de suelos-inóculo de Lobería bajo manejo agrícola o prístino en plantas de maíz y de tomate. Si bien los incrementos en la materia seca asociados a la inoculación fueron no significativos (p > 0,05), la respuesta micorrícica fue superior al 40 % en tomate y al 25 % en maíz, particularmente con el inóculo proveniente del sitio agrícola. Los bajos incrementos de crecimiento estarían asociados con la incipiente micorrización en ambas especies. Se plantea la necesidad de profundizar los estudios a efectos de determinar los factores involucrados que permitirían seleccionar inóculos eficientes.(AU)
We characterized the infective and sporulation capacities of microbial consortia of arbuscular mycorrhizal fungi (AMF) native of Buenos Aires province (Argentina) and determined if some soil characteristics and mycorrhizal parameters could allow to select potentially beneficial inocula. Soil samples were selected from seven locations in Buenos Aires province all under agricultural (A) and pristine (P) conditions. The AMF were multiplied and mycorrhizal root colonization of trap plants was observed at 10 weeks of growth. Spore number in field was low; however, after multiplication spore density accounted for 80-1175 spores per 100 g of soil. The principal component analysis showed that the P and Fe soil contents are the main modulators of infectivity and sporulation capacity. The mycorrhizal potential was determined in three locations, being high in Pristine Lobería and Agricultural Trenque Lauquen and low in Junín. Agricultural Lobería (AL) and Pristine Lobería (PL) inocula were selected and their efficiency was evaluated under controlled conditions. Even though shoot dry matter increases after inoculation was not significant (p > 0.05) mycorrhizal response was greater than 40% for tomato and 25% for corn, particularly after inoculation with inocula from the agricultural management. These results could be associated to the incipient development of mycorrhizae in both species. Additional research should be conducted to further develop our findings in order to determine the factors involved in the selection of efficient inocula.(AU)
ABSTRACT
Se evaluó la capacidad infectiva de promastigotes de Leishmania amazonensis al ser tratados con una serie de 10 derivados de la tiadiazina. Los parásitos fueron incubados durante 24 h con 1 o 0,1 mg/mL de cada compuesto y posteriormente, se infectaron macrófagos peritoneales de ratones BALB/c en cultivos. Todos los compuestos causaron una reducción de la capacidad infectiva de los parásitos mayor que 50 %. El producto T1O fue el que causó un mayor efecto, disminuyendo la infección en 92,8 % de infección.
The infective capacity of Leishmania amazonensis promastigotes was evaluated after they were treated with a series of 10 thiadiazine derivatives. The parasites were incubated for 24 hours with 1 or 0,1 mg/ml of each compound and then, peritoneal macrophages from BALB/c mice were infected in cultures. All the compounds managed to reduce the infective capacity of parasites by more than 50%. T10 product exhibited the highest effect since it reduced infection by 92,8%.