Statistical tools application on dextranase production from Pochonia chlamydosporia (VC4) and its application on dextran removal from sugarcane juice.

The aim of this study was to optimize the dextranase production by fungus Pochonia chlamydosporia (VC4) and evaluate its activity in dextran reduction in sugarcane juice. The effects, over the P. chlamydosporia dextranase production, of different components from the culture medium were analyzed by Plackett-Burman design and central composite design. The response surface was utilized to determine the levels that, among the variables that influence dextranase production, provide higher production of these enzymes. The enzymatic effect on the removal of dextran present in sugarcane juice was also evaluated. It was observed that only NaNO3 and pH showed significant effect (p<0.05) over dextranase production and was determined that the levels which provided higher enzyme production were, respectively, 5 g/L and 5.5. The dextranases produced by fungus P. chlamydosporia reduced by 75% the dextran content of the sugarcane juice once treated for 12 hours, when compared to the control treatment.

Statistical tools application on dextranase production from Pochonia chlamydosporia (VC4) and its application on dextran removal from sugarcane juice

ABSTRACT The aim of this study was to optimize the dextranase production by fungus Pochonia chlamydosporia (VC4) and evaluate its activity in dextran reduction in sugarcane juice. The effects, over the P. chlamydosporia dextranase production, of different components from the culture medium were analyzed by Plackett-Burman design and central composite design. The response surface was utilized to determine the levels that, among the variables that influence dextranase production, provide higher production of these enzymes. The enzymatic effect on the removal of dextran present in sugarcane juice was also evaluated. It was observed that only NaNO3 and pH showed significant effect (p<0.05) over dextranase production and was determined that the levels which provided higher enzyme production were, respectively, 5 g/L and 5.5. The dextranases produced by fungus P. chlamydosporia reduced by 75% the dextran content of the sugarcane juice once treated for 12 hours, when compared to the control treatment.

Occurrence of ticks in dogs in a hospital population in the state of Espírito Santo, Brazil / Ocorrência de carrapatos em cães de população hospitalar no Espírito Santo

Ticks, along with fleas, are considered the main ectoparasites affecting dogs in Brazil. The literature is rich in reports on the occurrence of ticks, which are vectors of diseases in both dogs and humans. The study of the epidemiology of these arthropods provides important data that can help control the infestation in residential areas where dogs are present. Thus, the study of their occurrence in urban areas and veterinary services is very important. The aim of the present study was to determine the occurrence of ticks in dogs in a veterinary hospital population in the state of Espírito Santo, Brazil. Over a one-year period (July 2012 to July 2013), 1483 dogs brought to veterinary services in south-eastern Brazil were examined. Among the dogs examined, 8.97% presented tick infestation, of which 100% were identified as Rhipicephalus sanguineus sensu lato. Given that this species of tick acts as a vector for diseases, it is important that the official health services monitor the occurrence of infestation in dogs and humans, in order to identify diseases transmitted by these ticks in this region.(AU)

Os carrapatos, juntamente com as pulgas, são considerados, no Brasil, como os principais ectoparasitas do cão. A literatura é rica em relatos considerando a ocorrência de carrapatos, que são vetores de doenças para cães e humanos. O estudo da epidemiologia de artrópodes é um dado importante que pode ajudar a controlar a infestação em áreas domiciliadas com a presença de cães. Nesse sentido, o estudo de sua ocorrência em áreas urbanas e ou mesmo em serviços médicos veterinários é importante. O objetivo do presente estudo foi verificar a ocorrência de carrapatos em cães em uma população hospitalar do estado de Espírito Santo, Brasil. Durante um ano (julho de 2012 a julho de 2013), foram examinados 1483 cães atendidos em serviços hospitalares no sudeste do Brasil. Entre os cães examinados, 8,97% apresentaram infestação de carrapatos, sendo 100% deles identificados como Rhipicephalus sanguineus sensu lato. Uma vez que se trata de uma espécie de carrapato que atua como vetor de doenças, é importante que os serviços oficiais de saúde mantenham um monitoramento da ocorrência de infestação tanto em cães como em humanos, a fim de identificar doenças transmitidas por esses carrapatos nesta região.(AU)

Pathological and parasitological characterization of Prosthenorchis elegans in a free-ranging marmoset Callithrix geofroyi from the Brazilian Atlantic Forest / Caracterização patológica e parasitológica de Prosthenorchis elegans em Callithrix geoffroyi primate de vida livre da Mata Atlântica

Prosthenorchis elegans is an acanthocephalan intestinal parasite reported in neotropical primates. Despite parasitism by P. elegans having already been described in wild marmosets in the Brazilian Atlantic Forest, there are no reports of this infection in wild Geoffroy's marmoset (Callithrix geofroyi). The aim of this study is to report one case of P. elegans parasitism in a free-ranging C. geoffroyi from Brazilian Atlantic Forest in Espírito Santo state, and characterize the pathological and parasitological findings of this infection. One Geoffroy's marmoset necropsied at the Vila Velha University's Veterinary Pathology Laboratory presented intense chronic transmural ulcerative enteritis associated with twenty cylindrical helminths present in the jejunum and ileum. We can conclude that parasitism by P. elegans occurs in free-ranging groups of Geoffroy's marmosets. Its infection produced severe intestinal lesions even in free-ranging marmoset and therefore is a threat to this animal's survival in wildlife and can have some impact on primate conservation in the Brazilian Atlantic Forest.(AU)

Prosthenorchis elegans é um acantocéfalo intestinal descrito em primatas neotropicais. Apesar do parasitismo por P. elegans já ter sido descrito de saguis da Mata Atlântica brasileira, não há relatos da infecção em saguis-da-cara-branca (Callithrix geofroyi) de vida livre. O objetivo deste estudo é relatar um caso de parasitismo por P. elegans em um C. geoffroyi de vida livre proveniente da Mata Atlântica brasileira no Estado do Espírito Santo e caracterizar os achados patológicos e parasitológicos dessa infecção. Um sagui-da-cara-branca foi necropsiado no Laboratório de Patologia Veterinária da Universidade de Vila Velha, onde foi observada intensa enterite ulcerativa transmural crônica associada a vinte helmintos cilíndricos presentes no jejuno e íleo. Conclui-se que o parasitismo por P. elegans ocorre em grupos livres de saguis-da-cara-branca e a sua infecção leva a lesões intestinais graves; portanto, este parasita pode prejudicar a sobrevivência deste animal na vida selvagem e pode ter algum impacto na conservação de primatas na Mata Atlântica brasileira.(AU)

Avaliação de parâmetros bioquímicos séricos em Tartarugas-da-Amazônia (Podocnemis expansa) mantidas em cativeiro / Evaluation of serum biochemical parameters in Amazon turtles (Podocnemis expansa) hatchlings kept in captivity

Amostras de sangue de Tartaruga-da-Amazônia mantidas em cativeiro foram analisadas com o propósito de avaliar os constituintes bioquímicos séricos e de relacionar ao que já foi relatado por outros pesquisadores, além de gerar mais informações, tendo em vista a escassez de dados envolvendo a espécie estudada. Foram utilizados 35 animais, com idade entre 3 e 7 meses, os quais foram contidos fisicamente para coleta de sangue em tubo sem anticoagulante. As análises bioquímicas foram processadas através de métodos cinéticos (enzimas) e colorimétricos (demais análises), e os valores médios obtidos foram: cálcio 7,0±1,9mg/dL, fóforo 2,8±1,2mg/dL, ácido úrico 1,8±0,8mg/dL, uréia 10,4±3,9mg/dL, alanina aminotransferase (AST) 75,5±33,6 UI/L, proteína total 1,9±0,8g/dL, triglicerídeos 22,9±9,8mg/dL e colesterol 45,3±19,7mg/dL. Observou-se que os valores dos componentes séricos avaliados assemelham-se aos que foram encontrados por outros pesquisadores em estudos bioquímicos séricos com Podocnemis expansa e outras tartarugas de água doce. Importante ressaltar que, no decorrer deste estudo observou-se a escassez de dados publicados sobre parâmetros sanguíneos de tartarugas do gênero Podocnemis, especialmente a espécie Podocnemis expansa, o que corroborou a alta relevância do desenvolvimento de mais pesquisas que envolvam a questão.(AU)

Blood samples from the giant river turtle kept in captivity were analyzed in order to evaluate the serum biochemical constituents and relate to what has been reported by other researchers, beyond the purpose of generating further information, given the paucity of data involving this species. The turtle eggs were from the state of Tocantins and were obtained from direct sampling in their natural habitat. After being collected the eggs were kept in incubators and then the puppies were kept in tanks. Samples were collected after chemical restraint of these animals by intracardiac puncture or cardiocentesis. The samples were centrifuged to obtain serum. Biochemical analyzes were processed by kinetic and colorimetric methods, as described by the manufacturer, and the values obtained were: calcium 7.0±1.9mg/dL, phosphorus 2.8±1.2mg/dL, uric acid 1.8±0.8mg/dL, BUN 10.4±3.9mg/dL, alanine aminotransferase (AST) 75.5±33.6 IU/L, total protein 1.9±0.8g/dL, triglycerides 22.9±9.8mg/dL and cholesterol 45.3±19.7mg/dL. It was observed that the values of the evaluated blood components are similar to those found by other researchers in serum biochemical studies with samples of Podocnemis expansa. During this study we observed the paucity of published data on blood parameters of Podocnemis genus, especially P. expansa species, which confirmed the high relevance of the development of further research involving this topic.(AU)

Atividade da colinesterase plasmática como biomarcador de impacto ambiental em tartarugas verdes (Chelonia mydas) no litoral do Arquipélago de Fernando de Noronha, Pernambuco / Plasma cholinesterase activity as environmental impact biomarker for green turtles (Chelonia mydas) on the coast of Fernando de Noronha, Pernambuco, Brazil

Biomarcadores podem ser usados de forma preditiva, permitindo que sejam tomadas ações de controle antes que ocorram danos ambientais irreversíveis com consequências ecológicas severas, no entanto, espécies sentinelas são necessárias para avaliação desses marcadores. As tartarugas marinhas são consideradas espécies sentinelas quando acometidas por fibropapilomas, sendo sinalizadora do desequilíbrio ambiental marinho nas suas áreas de ocorrência. Com o objetivo de propor a determinação da atividade da colinesterase plasmática em tartarugas verdes (Chelonia mydas) como biomarcador, procedeu-se a determinação da atividade enzimática em animais saudáveis e em localidade de baixo impacto antrópico (Arquipélago de Fernando de Noronha, Pernambuco, Brasil) para servir como referência para comparação com animais capturados em locais de maior impacto antrópico. Ao todo foram analisadas amostras de plasma heparinizado de 35 animais capturados. Todas as amostras analisadas apresentaram alguma atividade enzimática de colinesterase plasmática. Os valores obtidos de colinesterase variaram de 162 a 379 UI/L, com média e desvio padrão de 216,4 ± 51,4 UI/L. Nos estudos de repetibilidade e reprodutibilidade obtiveram-se coeficientes de variação menor que 5% em todas as análises, portanto a metodologia analítica utilizada se mostrou confiável. A longevidade das tartarugas marinhas da espécie C. mydas, o comportamento alimentar, juntamente com o fato de possuirem atividade enzimática detectável podem indicar essa espécie como bioindicadora de exposição a poluentes que influenciam na atividade da colinesterase plasmática.(AU)

Biomarkers can be used in a predictive manner, permitting actions control to be taken before they occur irreversible damage with severe environmental ecological consequences, however, sentinel species are needed to evaluate these markers. Sea turtles affected by fibropapillomas are considered sentinel species, signaling the marine environmental imbalance in their areas of occurrence. With the objective to use the plasma cholinesterase activity in green turtles (Chelonia mydas) as a biomarker, the measurement of enzyme activity was evaluated in healthy animals and in location of low pollution impact (Fernando de Noronha, Pernambuco, Brazil) to serve as a reference for comparison with animals trapped in places with higher pollution impact. Were analyzed heparinized plasma samples from 35 captured animals. All samples analyzed showed enzymatic activity of plasma cholinesterase. The values obtained ranged between 162-379 IU/L, with mean and standard deviation of 216.4 ± 51.4 IU/L. In studies of repeatability and reproducibility were obtained variance coefficients of less than 5% in all analyzes, so the analytical methodology proved successful. The longevity of marine turtles of the species C. mydas, feeding behavior, along with the fact possess detectable enzyme activity may indicate this species as a bioindicator of exposure to pollutants that influence plasma cholinesterase activity.(AU)

Biological control of infective larvae of Ancylostoma spp. in beach sand / Control biológico de las larvas infectantes de Ancylostoma spp. en arena de playa

Background. Geohelminths are parasites that stand out for their prevalence and wide distribution, depending on the soil for their transmission. Aims. The aim of this work was to evaluate the predatory capacity of the fungal isolate of the genus Duddingtonia (CG768) on third stage larvae (L3) of Ancylostoma spp. in beach sand under laboratory conditions. Methods. In the assay A five treatment groups and 1 control group were formed. The treatment groups contained 5000, 10,000, 15,000, 20,000 or 25,000 chlamydospores of the fungal isolate and 1000 Ancylostoma spp. L3 in pots containing 30 g of sand. The control group (without fungus) contained only 1000 Ancylostoma spp. L3 and distilled water in pots with 30 g of sand. Results. Evidence of predatory activity was observed at the end of 15 days, where we observed the following percentages of reduction of L3: Group 1 (4.5%); Group 2 (24.5%); Group 3 (59.2%); Group 4 (58.8%); Group 5 (63%). However, difference was noted (p < 0.01) only at concentrations 15,000, 20,000 and 25,000 in relation to control group. In the assay B two groups were formed in Petri dishes of 9 cm in diameter containing agar water 2% medium. In the treated group, each Petri dish contained 500 Ancylostoma spp. L3 and 5 g of sand containing the isolate CG 768 at a concentration of 25,000 chlamydospores/g of sand, and the control group (without fungus) contained only 500 L3. At the end of 7 days the non-predation L3 of Petri dishes using the method of Baermann were recovered. Difference (p < 0.01) between groups on reducing the average number of Ancylostoma spp. L3 (percent reduction of 84%) was observed. Conclusions. The results of this study confirm earlier work on the efficiency of the Duddingtonia genus in the control of Ancylostoma spp. infective larvae (AU)

Antecedentes. Los geohelmintos son parásitos que destacan por su prevalencia y amplia distribución, puesto que su transmisión depende del suelo. Objetivos. El objetivo del presente estudio fue evaluar la capacidad predatoria de aislamientos fúngicos del género Duddingtonia (CG768) sobre las larvas de estadio 3 (L3) de Ancylostoma spp. en arena de playa, en condiciones de laboratorio. Métodos. En el ensayo A se formaron 5 grupos de tratamiento y un grupo de control. Los grupos de tratamiento contenían 5000, 10.000, 15.000, 20.000 o 25.000 clamidosporas del aislamiento fúngico y 1000 larvas L3 de Ancylostoma spp. en recipientes con 30 g de arena. Los recipientes del grupo de control (sin clamidosporas) solo contenían 1000 larvas L3 de Ancylostoma spp. y agua destilada con 30 g de arena. Resultados. Al término de 15 días, fue evidente la actividad predatoria, con los porcentajes siguientes de reducción de larvas L3: grupo 1 (4.5%); grupo 2 (24.5%); grupo 3 (59.2%); grupo 4 (58.8%), y grupo 5 (63%). Sin embargo, en relación con el grupo control, solo se identificaron diferencias significativas (p < 0.01) a las concentraciones de 15.000, 20.000 y 25.000. En el ensayo B, en placas de Petri de 9 cm de diámetro, que contenían un medio de agar agua al 2%, se formaron 2 grupos. En el grupo tratado, cada placa de Petri contenía 500 larvas L3 de Ancylostoma spp. y 5 g de arena con el aislamiento CG768 a una concentración de 25.000 clamidosporas/g de arena, y el grupo de control (sin hongo) solo contenía 500 larvas L3. Al cabo de 7 días, utilizando el método de Baermann, a partir de las placas de Petri se obtuvieron larvas L3 no sometidas a predación por el hongo. Entre los grupos se observó una diferencia significativa (p < 0.01) en la reducción del número medio de larvas L3 de Ancylostoma spp. (porcentaje de reducción del 84%). Conclusiones. Los resultados del presente estudio confirman los datos de investigaciones previas sobre la eficiencia del género Duddingtonia en el control de las larvas infectantes de Ancylostoma spp (AU)

Predatory capability of the nematophagous fungus Arthrobotrys robusta preserved in silica gel on infecting larvae of Haemonchus contortus.

Biological control of gastrointestinal nematodiasis in ruminants is an alternative to reduce the number of infective larvae. The fungal isolates predatory activity preservation is a basic requirement for the success of this control type. The aim of this work is to evaluate the predatory capacity of the fungus Arthrobotrys robusta (isolate I-31), preserved on silica gel on infective larvae of Haemonchus contortus under laboratory conditions on 2 % water agar (2 % WA). In this essay, A. robusta storage on silica gel showed successful predatory activity on H. contortus L3 larvae (p < 0.01) compared to the control group. Nematophagous fungi were not observed in the control group during the experiment. There was a significant reduction (p < 0.01) of 73.84 % in the means of H. contortus (L3) recovered from treatment with isolate I-31 compared to the control without fungi. Results indicate that A. robusta (I-31) could survive stored on silica gel for at least 7 years and keep its predatory activity on H. contortus (L3).

Biological control of infective larvae of Ancylostoma spp. in beach sand.

BACKGROUND: Geohelminths are parasites that stand out for their prevalence and wide distribution, depending on the soil for their transmission. AIMS: The aim of this work was to evaluate the predatory capacity of the fungal isolate of the genus Duddingtonia (CG768) on third stage larvae (L3) of Ancylostoma spp. in beach sand under laboratory conditions. METHODS: In the assay A five treatment groups and 1 control group were formed. The treatment groups contained 5000, 10,000, 15,000, 20,000 or 25,000 chlamydospores of the fungal isolate and 1000 Ancylostoma spp. L3 in pots containing 30g of sand. The control group (without fungus) contained only 1000 Ancylostoma spp. L3 and distilled water in pots with 30g of sand. RESULTS: Evidence of predatory activity was observed at the end of 15 days, where we observed the following percentages of reduction of L3: Group 1 (4.5%); Group 2 (24.5%); Group 3 (59.2%); Group 4 (58.8%); Group 5 (63%). However, difference was noted (p<0.01) only at concentrations 15,000, 20,000 and 25,000 in relation to control group. In the assay B two groups were formed in Petri dishes of 9cm in diameter containing agar water 2% medium. In the treated group, each Petri dish contained 500 Ancylostoma spp. L3 and 5g of sand containing the isolate CG 768 at a concentration of 25,000 chlamydospores/g of sand, and the control group (without fungus) contained only 500 L3. At the end of 7 days the non-predation L3 of Petri dishes using the method of Baermann were recovered. Difference (p<0.01) between groups on reducing the average number of Ancylostoma spp. L3 (percent reduction of 84%) was observed. CONCLUSIONS: The results of this study confirm earlier work on the efficiency of the Duddingtonia genus in the control of Ancylostoma spp. infective larvae.

Nematicidal activity of three novel extracellular proteases of the nematophagous fungus Monacrosporium sinense.

Extracellular proteases are an important virulence factor for the nematophagous fungi Monacrosporium. The objective of this study was to optimize, purify, partially characterize, and to evaluate the nematicidal activity of the proteases produced by the nematophagous fungus Monacrosporium sinense (SF53) by solid-state fermentation. Wheat bran was used as substrate for protease production. The variables moisture, pH, incubation time, temperature, glucose, yeast extract, and the number of conidia were tested for their influences on protease production by SF53. To determine the optimal level of the selected variables the central composite design was applied. The crude extract obtained was purified in two steps, an ion exchange chromatography and a gel excision. SDS-PAGE and zymogram were performed for analysis of the purification process. Proteolytic activity was also tested at different pHs and temperatures. In the in vitro assay, the nematicidal activity of the three proteases was evaluated. pH and incubation time showed a significant effect (p<0.05) on production of protease. The highest value of activity was 38.0 (U/ml) under the conditions of pH 5.0 and incubation time of 211 h. SF53 produced three different proteases (Ms1, Ms2, and Ms3) which were directly purified from the zymogram. Ms1, Ms2, and Ms3 showed the following percentage of reduction (p<0.05) on the number of Panagrellus redivivus compared to control after 24 h: 76.8, 68.1, and 92.1%. This is the first report of the use of proteases of the isolate SF53 on a phytonematode, which may be a research tool in future works.

In vitro activity of a serine protease from Monacrosporium thaumasium fungus against first-stage larvae of Angiostrongylus vasorum.

A serine protease from the nematophagous fungus Monacrosporium thaumasium (NF34a) was purified, partially characterized and tested in vitro in control of the first larval stage of Angiostrongylus vasorum. NF34a grew in liquid culture medium, producing its crude extract that was purified by ion exchange chromatography. The fractions with high protease activity were collected in a pool, and elution of proteases was monitored by enzymatic assay and protein content. Purification steps were monitored by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Protease activity was determined under different pH and temperature conditions, and the inhibitor effects of metal ions and phenylmethylsulfonyl fluoride (PMSF) were assessed. In an experimental test, the infection process of NF34a on first-stage larvae of A. vasorum was investigated. A purified serine protease (Mt1) was identified, with an approximate molecular mass of 40 kDa and apparent homogeneity in SDS-PAGE, having optimal activity at pH 7.0 to 8.0 and temperature of 60°C. Mg(2+) and Zn(2+) partially inhibited the activity of Mt1 while PMSF inhibited it completely. Mt1 production was observed when NF34a was grown using first-stage larvae of A. vasorum as the only source of carbon and nitrogen. These results show that the enzyme may have a possible role in the infection process of the larvae. In the in vitro test of applicability against A. vasorum L(1), we observed a reduction in the number of larvae of 23.9% (p < 0.05) in the group treated with Mt1 compared with the control group. However, even this low reduction demonstrates that the Mt1 is important in the infection process.

Survival of Pochonia chlamydosporia in the gastrointestinal tract of experimentally treated dogs.

The predatory capacity of the nematophagous fungus Pochonia chlamydosporia (isolate VC4) after passage through the gastrointestinal tract of dogs was assessed in vivo against Toxocara canis eggs. Twelve dogs previously wormed were divided into two groups of six animals and caged. The treatments consisted of a fungus-treated group (VC4) and a control group without fungus. Each dog of the fungus-treated group received a single 4 g dose of mycelial mass of P. chlamydosporia (VC4). Fecal samples from animals of both groups (treated and control) were collected at five different times (6, 12, 24, 36, and 48 h) after fungal administration, and placed in Petri dishes. Each Petri dish of both groups for each studied time interval received approximately 1000 T. canis eggs. Thirty days after the fecal samples were collected, approximately one hundred eggs were removed from each Petri dish of each studied time interval and evaluated by light microscopy (LM) and scanning electron microscopy (SEM). Microscopy examination of plates inoculated with the fungus showed that the isolate VC4 was able to destroy the T. canis eggs with destruction percentages of 28.6% (6 h), 29.1% (12 h), 32.0% (24 h), 31.7% (36 h), and 37.2% (48 h). These results suggest that P. chlamydosporia can be used as a tool for the biological control of T. canis eggs in feces of contaminated dogs.

Biological control of Fasciola hepatica eggs with the Pochonia chlamydosporia fungus after passing through the cattle gastrointestinal tract.

Fasciolosis is a disease caused by Fasciola hepatica responsible for causing significant losses in livestock. This study aimed to evaluate the Pochonia chlamydosporia fungus (isolate VC1) on F. hepatica eggs after passing through the cattle gastrointestinal tract. For this evaluation, 1 g pellet was given in sodium alginate matrix per kilogram live weight containing 25% of fungal mycelium from isolate VC1 per animal. Twelve animals were used, six treated and six untreated (control). Some stool samples were collected from the groups of treated and control animals, at the times of 12, 18, 24, 48, 72, and 96 h after the pellets' administration. Then, from each stool sample of treated and control groups, 2 g was placed in a Petri dish of 9 cm in diameter, containing 2% water-agar and 1,000 eggs of F. hepatica. It was observed that the fungus was effective in preying upon the eggs in the samples recovered at all of the schedules starting at 12 h. Furthermore, differences were observed (p < 0.01) in the destruction of eggs in the Petri dishes in the treated group compared with the control group. The ovicidal effect was observed after 7 days of interaction. The ovicidal P. chlamydosporia fungus was effective in destroying F. hepatica eggs; therefore, it is suggested that this fungus could be employed as agent for the control of helminth eggs.

Ovicidal activity of seven Pochonia chlamydosporia fungal isolates on Ascaris suum eggs.

The ovicidal effect of the nematophagous fungus Pochonia chlamydosporia on eggs of Ascaris suum was tested under laboratory conditions. A. suum eggs were plated on 2% water-agar with seven fungal isolates (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) and control without fungus. After 5, 7, 10, 14, 15 and 21 days of incubation, approximately 100 eggs were removed from the plates and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo showing hyphal penetration and internal egg colonization. The isolates effectively destroyed A. suum eggs and all types of effects were observed during the experiment. There was no variation in ovicidal capacity (type 3 effect) among the isolates (p>0.05) throughout the experiment. After 21 days, isolate 5 showed the highest percentages of type 3 effect (58.33%). The results indicated that P. chlamydosporia (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) can destroy A. suum eggs and is, therefore, a potential biological control agent of nematodes.

In vitro predatory activity of the fungi Duddingtonia flagrans, Monacrosporium thaumasium, Monacrosporium sinense and Arthrobotrys robusta on Ancylostoma ceylanicum third-stage larvae.

The potential role of companion animals as reservoirs for zoonotic diseases has been recognised as a significant public health problem worldwide. Ancylostoma ceylanicum is the only ancylostomatidae species known for infecting human beings. This article aimed to compare the predatory capacity of predatory fungi isolates Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), Monacrosporium sinense (SF53) and Arthrobotrys robusta (I31) on A. ceylanicum infectious larvae (L(3)) in a 2% water-agar plate. There was no predatory capacity variation among the fungi tested (P>0.05) over the 7-day period experimental assay. When compared to the control (without fungi), there was a significant reduction (P<0.05) of 95.6%, 85.1%, 87.4% and 90.2% on the A. ceylanicum L(3) mean recovered from treatments with isolates AC001, NF34, SF53 and I31, respectively. Regarding linear regression coefficients, negative values were noted for treatments, therefore indicating A. ceylanicum non-predated larvae reduction over 7 days. In this work, all predatory fungi isolates were efficient at capturing and destroying in vitro the A. ceylanicum L(3); therefore being able to be used as biological controllers of such nematode.

In vitro predatory activity of nematophagous fungi and after passing through gastrointestinal tract of equine on infective larvae of Strongyloides westeri.

Three isolates of predator fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), and Arthrobotrys robusta (I-31) were assessed in in vitro test regarding the capacity of prey infective larvae (L(3)) Strongyloides westeri. Compared to control, without fungus, there was a significant decrease (P < 0.01) of 80.4%, 67.9%, and 72.8% in means of infective larvae S. westeri recovered from treatments with isolates AC001, NF34, and I-31, respectively. All tested isolates were efficient in the capture of S. westeri (P > 0.01) in vitro test. Linear regression coefficients of treated and control groups were -0.21 for control, -0.32 for D. flagrans, -0.34 for M. thaumasium, and -0.22 for A. robusta. In the following, isolates AC001 and NF34 were assessed in vivo regarding the capacity of supporting the passage through equine gastrointestinal tract without loss of ability of preying infective larvae S. westeri. Fungal isolates survived the passage and were efficient in preying L(3) since the first 12 h of collection (P < 0.01) in relation to the control group (without fungus). Compared to control, there was a significant decrease (P < 0.01) of 76.4% and 76.7% (12 h), 86.4% and 85.9% (24 h), 88.3% and 87.7% (48 h), and 89.9% and 87.2% (72 h) in means of infective larvae S. westeri recovered from treatments with isolates AC001 and NF34, respectively. Linear regression coefficients of L(3) of recovered S. westeri regarding the collections due to time were 1.93 for control, -3.52 for AC001, and -2.64 for NF34. Fungi D. flagrans and M. thaumasium (NF34) have demonstrated to be promising for use in the biological control of equine parasite S. westeri.

Efficacy of Duddingtonia flagrans and Arthrobotrys robusta in controlling sheep parasitic gastroenteritis.

The aim of this study was to evaluate the efficacy of formulations of sodium alginate matrix (pellets) of the nematode predatory fungi, Duddingtonia flagrans (AC001 isolate) and Arthrobotrys robusta (I-31 isolate), in the biological control of sheep gastrointestinal nematode infections. Thirty young Bergamacia ewes were allocated into three groups: In group 1 (control), the animals received 2 g/10 kg of live weight (l.w.) of pellets without fungus; in group 2, each animal received 2 g/10 kg of l.w. of pellets of D. flagrans (0.2 g of fungus/10 kg l.w.); and in group 3, each animal received 2 g/10 kg of l.w. of pellets of A. robusta (0.2 g of fungus/10 kg l.w.). The animals of each group were kept separately under rotational grazing. Pellets, with or without fungi, were mixed with 1 kg animal food and administered twice a week for 6 months. There was no significant difference in mean live weight and packed cell volume among groups (P > 0.05). Mean nematode fecal egg counts (FEC) did not significantly differ between the control and the remaining groups, except in one or two collections, when FEC was higher in the control group than in group 2 and group 3, respectively. The group that received A. robusta pellets needed less salvage anthelmintic treatments. Haemonchus contortus was the predominant species recovered from tracer lambs. The nematophagous fungi, D. flagrans and A. robusta, did not provide satisfactory results in the prophylaxis of parasitic gastroenteritis in sheep, under the conditions of the present study.

Predatory activity of the nematophagous fungus Duddingtonia flagrans on horse cyathostomin infective larvae.

This work was performed to determine the predatory capacity in vitro of the nematophagous fungus Duddingtonia flagrans (isolate AC001) on cyathostomin infective larvae of horse (L(3)). The experimental assay was carried out on plates with 2% water-agar (2% WA). In the treated group, each plate contained 1.000 L(3) and 1.000 conidia of the fungus. The control group without fungus only contained 1.000 L(3) in the plates. Ten random fields (4 mm diameter) were examined per plate of treated and control groups, every 24 h for seven days under an optical microscope (10x and 40x objective lens) for non-predated L(3) counts. After 7 days, the non-predated L(3) were recovered from the Petri dishes using the Baermann method. The interaction there was a significant reduction (p < 0.01) of 93.64% in the cyathostomin L(3) recovered. The results showed that the D. flagrans is a potential candidate to the biological control of horse cyathostomin L(3).

Predatory activity of Pochonia chlamydosporia fungus on Toxocara (syn. Neoascaris) vitulorum eggs.

Toxocara (Neoascaris) vitulorum is a gastrointestinal nematode parasite of young ruminants, responsible for high mortality rates in parasitized cattle and buffalo calves. The objective of this work was to compare the predatory capacity under laboratory conditions of four fungal isolates of the nematophagous fungus Pochonia chlamydosporia (VC1, VC4, VC5 and VC12) on T. vitulorum eggs in 2% water-agar (2% WA). T. vitulorum eggs were plated on 2% WA Petri dishes which contained cultured fungal isolates and control plates without fungi. After 10 and 15 days one hundred eggs were removed and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo in addition to hyphal penetration and internal egg colonization. The fungal isolates were effective in the destruction of T. vitulorum eggs presenting the type 3 effect at 10 and 15 days after contact with the fungus. No nematophagous fungi were observed in the control group during the experiment. There was no variation in the predatory capacity of the fungal isolates (P > 0.01) at the intervals of 10 and 15 days. These results indicate that P. chlamydosporia (VC1, VC4, VC5 and VC12) negatively influenced the development of T. vitulorum eggs and can be considered a potential candidate for the biological control of nematodes.

[In vitro evaluation of nematode predacious fungus Duddingtonia fagrans on cyathostomes infective larvae of equines (Nematoda: Cyathostominae)]. / Avaliação in vitro do fungo predador de nematoides Duddingtonia flagrans sobre larvas infectantes de ciatostomíneos de equinos (Nematoda: Cyathostominae).

The predatory capacity of one isolate of nematode-trapping fungus Duddingtonia flagrans (AC001) on infective larvae of cyathostomes was evaluated in laboratorial conditions in medium water-agar 2% (WA 2%). There was significant reduction (p<0.01) of 93.64% in the average of infective larvae of cyathostomes recovered of medium WA 2% at seven day. These results show that the isolated AC001 could be used in the biological control of cyathostomes of horses.