Prototheca bovis in goats: Experimental mastitis and treatment with polypyrrole.

Prototheca bovis has been associated with several cases of mastitis in cattle but no record of intramammary infections has been reported in goats. This infection does not respond to available treatments and the disposal recommendation of affected animals cause great damage to the dairy industry. Alternatives for dealing with infections caused by Prototheca spp. are required worldwide. In vitro results suggest polypyrrole as promising molecule for combating this alga, because an algaecide effect was observed on tested Prototheca spp. isolates. Thus, this study evaluated goats as an experimental model for intramammary infection by P. bovis and a protocol for treating these animals with an intramammary polypyrrole solution. The possibility of P. bovis promoting an intramammary infection in goats was experimentally proven, demonstrating this species as an important model for studies involving algae mastitis. Furthermore, polypyrrole reduced the counts of Prototheca sp. in the analyzed samples, showing potential to fight this microorganism also in vivo. The results obtained in this study demonstrate the ability of P. bovis to colonize breast tissue in lactating goats and the highly soluble molecule of polypyrrole has potential use for the treatment of protothecosis.

Effect of Paecilomyces lilacinus, Trichoderma harzianum and Trichoderma virens fungal extracts on the hatchability of Ancylostoma eggs / Efecto de los extractos de los hongos Paecilomyces lilacinus, Trichoderma harzianum y Trichoderma virens en la eclosionabilidad de huevos de Ancylostoma

Background. Ancylostoma species have demanded attention due to their zoonotic potential. The use of anthelmintics is the usual method to prevent environmental contamination by Ancylostoma eggs and larvae. Nematophagous fungi have been widely used in their biological control due to the fungus ability to capture and digest free nematode forms. Aims. The aim of this study was to evaluate the effect of four different fungal extracts of Paecilomyces lilacinus (n=2), Trichoderma harzianum (n=1) and Trichoderma virens (n=1) isolates on the hatchability of Ancylostoma eggs. Methods. Fungal extracts consisted of fungal broth culture supernatant without filtration (crude extract) and filtered broth (filtered extract), macerated mycelium (crude macerate), and macerated mycelium submitted to filtration (filtered macerate). The Ancylostoma eggs were obtained from the feces of naturally infected dogs. In vitro assays were performed in five replicates and consisted of four treatments and one control group. Results. The activity of the fungal extracts of each evaluated fungus differed (p<0.05) from those of the control group, showing significant ovicidal activity. The hatching of the eggs suffered reduction percentages of 68.43% and 47.05% with P. lilacinus, and 56.43% with T. harzianum, when crude macerate extract was used. The reduction with the macerate extract of T. virens was slightly lower (52.25%) than that for the filtered macerate (53.64%). Conclusions. The results showed that all extracts were effective in reducing the hatchability of Ancylostoma eggs. The ovicidal effect observed is likely to have been caused by the action of hydrolytic enzymes secreted by the fungi (AU)

Antecedentes. Las especies del género Ancylostoma son de gran importancia debido a su potencial zoonótico. El uso de antihelmínticos es el método habitual en la prevención de la contaminación ambiental por huevos y larvas del género Ancylostoma. Los hongos nematófagos se utilizan ampliamente en el control biológico de aquellos, debido a su capacidad de capturar y digerir nematodos libres. Objetivo. El objetivo del estudio fue evaluar el efecto de cuatro extractos diferentes de hongos (Paecilomyces lilacinus [n=2], Trichoderma harzianum [n=1] y Trichoderma virens [n=1]) en la eclosionabilidad de huevos de especies de Ancylostoma. Métodos. Los extractos de hongos constaban del sobrenadante del cultivo en caldo fúngico sin filtración (extracto crudo) y caldo filtrado (extracto filtrado), micelio macerado (macerado crudo) y micelio macerado sometido a filtración (macerado filtrado). Los huevos de Ancylostoma se obtuvieron a partir de heces de perros infectados de manera natural. Se realizaron cinco repeticiones de los ensayos in vitro con cuatro tratamientos y un grupo control. Resultados. La actividad de los extractos fúngicos de cada hongo evaluado difiere (p<0,05) de la de aquellos del grupo control, lo que demuestra una actividad ovicida significativa. Con el extracto crudo macerado, la reducción de la eclosión mostró porcentajes del 68,43 y el 47,05% en el caso de P. lilacinus y del 56,43% para el caso de T. harzianum. El porcentaje de reducción en el uso del macerado crudo en T. virens fue del 52,25%, algo inferior respecto al macerado filtrado (53,64%). Conclusiones. Los resultados mostraron que todos los extractos fueron eficaces en la reducción de la eclosionabilidad de huevos de Ancylostoma. Es probable que el efecto ovicida observado haya sido causado por la acción de enzimas hidrolíticas secretadas por los hongos (AU)

Trichoderma virens as a biocontrol of Toxocara canis: In vivo evaluation / Trichoderma virens como control biológico de Toxocara canis: evaluación in vivo

Background. Microorganisms have been widely studied as biological control agents of parasites of medical and veterinary importance. Coprophagous arthropods, bacteria and fungi are among the different organisms evaluated as potential biological control agents. Nematophagous fungi capture and digest the free forms of nematodes in the soil. Due to its zoonotic potential, Toxocara canis have been brought to the attention of researchers. Aims. The aim of the present study was to determine whether the administration of embryonated T. canis eggs exposed to the nematophagous fungus Trichoderma virens reduces parasite infection in experimental animals. Methods. Embryonated T. canis eggs were exposed to T. virens mycelium for 15 days at 25°C. Subsequently, 100 fungus-exposed eggs were orally administered to 20 Swiss mice. As a positive control, another 20 mice received 100 embryonated eggs that were not exposed to the fungus. After 48h, the animals were killed, and heart, lungs and liver were harvested for the recovery of larvae. Results. The organs of the animals that received embryonated T. canis eggs exposed to the fungus showed a lower mean larval recovery when compared with the animals that received embryonated eggs without fungus exposure (p<0.05). Conclusions. The exposure of T. canis eggs to T. virens reduces the experimental infection, demonstrating the potential of this nematophagous fungus as a biocontrol agent (AU)

Antecedentes. Algunos microorganismos han sido ampliamente estudiados como agentes de control biológico de parásitos de importancia médica y veterinaria. Los artrópodos coprófagos, las bacterias y los hongos están entre los diferentes organismos que sirven como agentes para el control con potencial biológico. Los hongos nematófagos capturan y digieren las formas libres de nematodos en el suelo. Toxocara canis, debido a su potencial zoonótico, ha captado la atención de los investigadores en estos estudios. Objetivos. El objetivo del presente estudio fue evaluar si la exposición de huevos embrionados de T. canis al hongo nematófago Trichoderma virens reduce la infección parasítica en un modelo experimental animal. Métodos. Los huevos embrionados de T. canis fueron expuestos al micelio de T. virens durante 15días a 25°C. Posteriormente, 100huevos de T. canis expuestos al hongo fueron administrados por vía oral a un grupo de 20ratones Swiss. Como control positivo se usó otro grupo de 20ratones que recibieron 100huevos embrionados no expuestos al hongo. Después de 48h, los animales fueron sacrificados y corazón, pulmones e hígado fueron extraídos para la posterior obtención de larvas. Resultados. El número de larvas obtenidas en los diferentes órganos fue menor en el grupo de animales que fueron infectados con los huevos embrionados de T. canis expuestos al hongo en comparación con el grupo de animales que recibieron huevos embrionados sin la exposición al hongo (p<0,05). Conclusiones. La exposición de los huevos de T. canis a T. virens reduce la infección experimental, lo que demuestra el potencial de este hongo nematófago como agente para el control biológico (AU)

Trichoderma virens as a biocontrol of Toxocara canis: In vivo evaluation.

BACKGROUND: Microorganisms have been widely studied as biological control agents of parasites of medical and veterinary importance. Coprophagous arthropods, bacteria and fungi are among the different organisms evaluated as potential biological control agents. Nematophagous fungi capture and digest the free forms of nematodes in the soil. Due to its zoonotic potential, Toxocara canis have been brought to the attention of researchers. AIMS: The aim of the present study was to determine whether the administration of embryonated T. canis eggs exposed to the nematophagous fungus Trichoderma virens reduces parasite infection in experimental animals. METHODS: Embryonated T. canis eggs were exposed to T. virens mycelium for 15 days at 25°C. Subsequently, 100 fungus-exposed eggs were orally administered to 20 Swiss mice. As a positive control, another 20 mice received 100 embryonated eggs that were not exposed to the fungus. After 48h, the animals were killed, and heart, lungs and liver were harvested for the recovery of larvae. RESULTS: The organs of the animals that received embryonated T. canis eggs exposed to the fungus showed a lower mean larval recovery when compared with the animals that received embryonated eggs without fungus exposure (p<0.05). CONCLUSIONS: The exposure of T. canis eggs to T. virens reduces the experimental infection, demonstrating the potential of this nematophagous fungus as a biocontrol agent.

Effect of Paecilomyces lilacinus, Trichoderma harzianum and Trichoderma virens fungal extracts on the hatchability of Ancylostoma eggs.

BACKGROUND: Ancylostoma species have demanded attention due to their zoonotic potential. The use of anthelmintics is the usual method to prevent environmental contamination by Ancylostoma eggs and larvae. Nematophagous fungi have been widely used in their biological control due to the fungus ability to capture and digest free nematode forms. AIMS: The aim of this study was to evaluate the effect of four different fungal extracts of Paecilomyces lilacinus (n=2), Trichoderma harzianum (n=1) and Trichoderma virens (n=1) isolates on the hatchability of Ancylostoma eggs. METHODS: Fungal extracts consisted of fungal broth culture supernatant without filtration (crude extract) and filtered broth (filtered extract), macerated mycelium (crude macerate), and macerated mycelium submitted to filtration (filtered macerate). The Ancylostoma eggs were obtained from the feces of naturally infected dogs. In vitro assays were performed in five replicates and consisted of four treatments and one control group. RESULTS: The activity of the fungal extracts of each evaluated fungus differed (p<0.05) from those of the control group, showing significant ovicidal activity. The hatching of the eggs suffered reduction percentages of 68.43% and 47.05% with P. lilacinus, and 56.43% with T. harzianum, when crude macerate extract was used. The reduction with the macerate extract of T. virens was slightly lower (52.25%) than that for the filtered macerate (53.64%). CONCLUSIONS: The results showed that all extracts were effective in reducing the hatchability of Ancylostoma eggs. The ovicidal effect observed is likely to have been caused by the action of hydrolytic enzymes secreted by the fungi.

In vitro photodynamic inactivation of Sporothrix schenckii complex species.

Photodynamic therapy has been applied successfully against cutaneous and subcutaneous mycoses. We applied methylene blue as a photosensitizing agent and light emitting diode (InGaAlP) against Sporothrix schenckii complex species in an in vitro assay. The viability of the conidia was determined by counting colony-forming units. Methylene blue in conjunction with laser irradiation was able to inhibit the growth of all tested samples. The in vitro inhibition of Sporothrix spp. isolates by laser light deserves in vivo experimental and clinical studies since it may be a promising treatment for cutaneous and subcutaneous sporotrichosis.

Fungal ovicidal activity on Toxocara canis eggs

Background. Visceral toxocariasis is a parasitic zoonosis caused by Toxocara canis. The prevalence of this parasite in dogs, soil contamination and the resistance of eggs increase human exposure to the disease. Moreover, the difficulties of the control measures justify the need for alternative ones. Aims. The objective of this study was to evaluate the in vitro ovicidal activity of fungi isolated from soils from public places in the city of Pelotas, Rio Grande do Sul, Brazil, on Toxocara canis. Methods. Samples of soil from ten localities were inoculated onto Petri dishes with 2% water–agar (WA) that contained antibiotics, and incubated at 25 °C/21 days. Isolated fungi were tested in vitro for ovicidal activity, with five replicates. One mL of an embryonated Toxocara canis egg suspension (103 eggs) was poured over the fungal cultures after 10 days of growth. At intervals of 7, 14 and 21 days, 100 eggs were removed from each plaque and evaluated by optical microscopy. Results. Acremonium, Aspergillus, Bipolaris, Fusarium, Gliocladium, Mucor and Trichoderma were isolated from the soil. A significant ovicidal type 3 effect was observed in Trichoderma, Fusarium solani complex and Acremonium. Those isolates from the genus Trichoderma showed their ovicidal effect on the 14th day of fungus–egg interaction. The other fungal genera tested showed a type 2 effect. Conclusions. These results suggest that the use of Trichoderma and Fusarium solani complex in biological control of T. canis is promising; however, further studies should be performed (AU)

Fungal ovicidal activity on Toxocara canis eggs.

BACKGROUND: Visceral toxocariasis is a parasitic zoonosis caused by Toxocara canis. The prevalence of this parasite in dogs, soil contamination and the resistance of eggs increase human exposure to the disease. Moreover, the difficulties of the control measures justify the need for alternative ones. AIMS: The objective of this study was to evaluate the in vitro ovicidal activity of fungi isolated from soils from public places in the city of Pelotas, Rio Grande do Sul, Brazil, on Toxocara canis. METHODS: Samples of soil from ten localities were inoculated onto Petri dishes with 2% water-agar (WA) that contained antibiotics, and incubated at 25°C/21 days. Isolated fungi were tested in vitro for ovicidal activity, with five replicates. One mL of an embryonated Toxocara canis egg suspension (10(3) eggs) was poured over the fungal cultures after 10 days of growth. At intervals of 7, 14 and 21 days, 100 eggs were removed from each plaque and evaluated by optical microscopy. RESULTS: Acremonium, Aspergillus, Bipolaris, Fusarium, Gliocladium, Mucor and Trichoderma were isolated from the soil. A significant ovicidal type 3 effect was observed in Trichoderma, Fusarium solani complex and Acremonium. Those isolates from the genus Trichoderma showed their ovicidal effect on the 14th day of fungus-egg interaction. The other fungal genera tested showed a type 2 effect. CONCLUSIONS: These results suggest that the use of Trichoderma and Fusarium solani complex in biological control of T. canis is promising; however, further studies should be performed.

Diversity of yeasts from bovine mastitis in Southern Brazil.

Mastitis is one of the most serious problems in the dairy cattle farms. The great majority of the cases are caused by bacteria, but lately there have been an increasing number of reports about cases of mycotic etiology. The objective of this work was to characterize the yeasts and yeast-like fungi associated with milk of cows with mastitis. Milk samples (n = 248) from a dairy belt situated around the region of Passo Fundo, hinterland of the state of Rio Grande do Sul, Southern Brazil, were analyzed. Aliquots of 0.1 ml of milk were inoculated on yeast extract-malta agar with chloramphenicol. After a period of incubation of 3-5 days at 22-25 degrees C, the counting of the morphologically distinct colonies was performed, as well as the isolation and identification through phenotypical and physiological criteria. It was possible to isolate 68 yeast species from 43 (17.3%) of the samples. The most frequent genera were Candida (37.9%), Pichia (19.1%), Cryptococcus (10.3%) and Rhodotorula (10.3%).

Diversity of yeasts from bovine mastitis in Southern Brazil

La mastitis es uno de los problemas más graves en los rebaños lecheros.La gran mayoría de los casos son causados por bacterias, aunqueúltimamente la literatura está registrando un aumento de los casos deetiología micológica. El objetivo de este trabajo fue caracterizar levaduras yotros hongos levaduriformes aislados de leche de vacas con mastitis.Fueron analizadas 248 muestras de leche del cinturón lechero situadoalrededor de la región de Passo Fundo, interior del estado de Rio Grande doSul, sur de Brasil. Alicuotas de 0,1 ml de leche fueron inoculadas en agarextracto de levadura-malta con cloranfenicol. Después de un período deincubación de 3-5 días a 22-25 ºC fue realizado el recuento de las coloniasdiferenciadas morfológicamente, así como el aislamiento e identificación através de criterios fenotípicos y fisiológicos. En 43 (17,3%) de las muestrasfue posible aislar 68 especies. Los géneros más frecuentes fueron Candida(37,9%), Pichia (19,1%), Cryptococcus (10,3%) y Rhodotorula (10,3%)(AU)

Mastitis is one of the most serious problems in the dairy cattle farms.The great majority of the cases are caused by bacteria, but lately there havebeen an increasing number of reports about cases of mycotic etiology.The objective of this work was to characterize the yeasts and yeast-like fungiassociated with milk of cows with mastitis. Milk samples (n = 248) from a dairybelt situated around the region of Passo Fundo, hinterland of the state of RioGrande do Sul, Southern Brazil, were analyzed. Aliquots of 0.1 ml of milk wereinoculated on yeast extract-malta agar with chloramphenicol. After a period ofincubation of 3-5 days at 22-25 °C, the counting of the morphologicallydistinct colonies was performed, as well as the isolation and identificationthrough phenotypical and physiological criteria. It was possible to isolate68 yeast species from 43 (17.3%) of the samples. The most frequent generawere Candida (37.9%), Pichia (19.1%), Cryptococcus (10.3%) andRhodotorula (10.3%)(AU)