Multi-enzyme complex of white rot fungi in saccharification of lignocellulosic material

ABSTRACT The multi-enzyme complex (crude extract) of white rot fungi Pleurotus ostreatus, Pleurotus eryngii, Trametes versicolor, Pycnosporus sanguineus and Phanerochaete chrysosporium were characterized, evaluated in the hydrolysis of pretreated pulps of sorghum straw and compared efficiency with commercial enzyme. Most fungi complexes had better hydrolysis rates compared with purified commercial enzyme.

Nematicidal activity of Paecilomyces marquandii proteases on infective larvae of Ancylostoma spp

ABSTRACT The present study aimed to evaluate the action of Paecilomyces marquandii proteases on Ancylostoma spp L3. White halos in the zymogram confirmed the proteolytic action. Difference (p <0.01) between the number of L3 in the differents groups was found, with 41.4% of reduction of Ancylostoma spp. L3 before 24 hours.

Platynosomum fastosum in an asymptomatic cat in the state of Espírito Santo: first report

Platynosomum fastosum is a parasite that affects cats, and among these, domestic cats, especially those originating from areas with tropical and subtropical climates. It is the most common liver parasite of domestic cats and is usually located in the bile ducts and gall bladder, but may also be found in the small intestine and pancreas. Infection occurs through ingestion of geckos containing metacercariae, causing cholangitis in the bile ducts of domestic cats. The aim of this study was to report the occurrence of P. fastosum after necropsy of an asymptomatic domestic cat, with a history of sudden death, received at the Animal Pathology sector of the Ricardo Alexandre Hippler Veterinary Hospital, Espírito Santo, Brazil. In macroscopic examination adult specimens of P. fastosum were observed inside the visibly dilated bile ducts. The dilatation, the presence of parasites and papillary proliferations into the lumen of the ducts were confirmed by histopathological examination. Then, a cholangiohepatitis associated to P. fastosum infection was classified. This is the first report of the occurrence of P. fastosum in a domestic cat in the state of Espírito Santo

Optimization of protease production by the fungus Monacrosporium thaumasium and its action against Angiostrongylus vasorum larvae / Otimização da produção de protease pelo fungo Monacrosporium thaumasium e sua ação contra larvas de Angiostrongylus vasorum

The objectives of this study were to optimize protease production from the nematophagous fungus Monacrosporium thaumasium (NF34a) and evaluate its larvicidal activity and biological stability. An isolate of the nematophagous fungus Monacrosporium thaumasium (NF34a) was used to produce the enzyme. The Plackett-Burman design was used in order to scan which components of the culture medium could have a significant influence on protease production by the fungus NF34a. An in vitro assay was also performed to evaluate the larvicidal activity of NF34a. It was observed that only one component of the culture medium (yeast extract), at the levels studied, had any significant effect (p < 0.05) on protease production. There was a reduction (p < 0.01) in the mean number of larvae recovered from the treated groups, compared with the control groups. The results confirm previous reports on the efficiency of nematophagous fungi for controlling nematode larvae that are potentially zoonotic. Thus, given the importance of biological control, we suggest that further studies should be conducted on the protease produced by the fungus Monacrosporium thaumasium.

O objetivo deste trabalho foi otimizar a produção de proteases do fungo nematófago Monacrosporium thaumasium (NF34a), avaliar sua atividade larvicida e sua estabilidade biológica. Foi utilizado um isolado do fungo nematófago Monacrosporium thaumasium (NF34a) para a produção de enzima. O delineamento Plackett-Burman foi utilizado com o objetivo de se escanear quais componentes do meio de cultura, poderiam ter influência significava na produção de protease pelo fungo NF34a. Foi realizado um ensaio in vitro para avaliar a atividade larvicida de NF34a. Observou-se que apenas um dos componentes do meio de cultura (extrato de levedura), nos níveis avaliados, apresentou um efeito significativo (p < 0,05) sobre a produção de protease. Houve redução (p < 0,01) entre as médias de larvas recuperadas dos grupos tratados em relação às dos grupos controle. Os resultados confirmam trabalhos anteriores da eficiência de fungos nematófagos no controle de larvas de nematóides potencialmente zoonóticos. Assim, devido a importância do controle biológico, os autores sugerem estudos mais aprofundados sobre a protease produzida pelo fungo Monacrosporium thaumasium.

Control of infective larvae of gastrointestinal nematodes in heifers using different isolates of nematophagous fungi / Controle de larvas infectantes de nematóides gastrintestinais de novilhas por diferentes isolados dos fungos nematófagos

The effect of different nematophagous fungi [Duddingtonia flagrans (AC001 and CG722) and Monacrosporium thaumasium (NF34)] with regard to controlling infective larvae (L3) of nematodes after gastrointestinal transit in female cattle (3/4 Holstein × Zebu) was evaluated. A total of 24 pubescent female cattle were used, weighing approximately 320 kg each one. There were three treatment groups, each contained six animals that received 150 g of pellets (0.2 g of mycelium), orally in a single dose, in a sodium alginate matrix containing mycelial mass of the fungus D. flagrans (AC001 or CG722) or M. thaumasium (NF34); and one control group (without fungi). Fecal samples were collected from the animals at intervals of 12, 15, 18, 21, 24, 48, and 72 hours. At the end of 17 days, the L3 not subjected to predation were recovered by means of the Baermann method. The fungal isolates tested were capable of destroying the L3 after gastrointestinal transit. It was observed that within 72 hours, the isolates AC001, CG722, and NF34 showed a higher predatory activity (81.2%, 97.3%, and 98.3%, respectively). The results justify the need for studies in the field, and over longer intervals, in order to observe the efficiency of the fungus D. flagrans, or even M. thaumasium, for environmental control over nematodes in naturally infected cattle.

No presente estudo, foi avaliado o efeito de diferentes fungos nematófagos [Duddingtonia flagrans (AC001 e CG722) eMonacrosporium thaumasium (NF34)] no controle de larvas infectantes (L3) de nematóides após o trânsito gastrointestinal em fêmeas bovinas (3/4 Holandês x Zebu). Um total de 24 fêmeas bovinas pubescentes foram utilizadas, pesando aproximadamente 320 kg cada. Foram utilizados três grupos de tratamento; cada um contendo seis animais que receberam por via oral de 150 g de péletes (0,2 g de micélio), em dose única, em uma matriz de alginato de sódio contendo massa micelial dos fungos D. flagrans (AC001 ou CG722), M. thaumasium (NF34), além de um grupo controle (sem fungo). Amostras de fezes foram colhidas dos animais em intervalos de 12, 15, 18, 21, 24, 48 e 72 horas. No final de 17 dias, as L3 não predadas foram recuperadas pelo método de Baermann. Os isolados de fungos testados foram capazes de destruir as L3 após trânsito gastrointestinal. Observou-se após 72 horas, os isolados AC001, CG722 e NF34 mostraram uma maior atividade predatória (81,2%, 97,3% e 98,3%, respectivamente). Estes resultados justificam a necessidade de estudos a campo e em intervalos mais longos, a fim de observar a eficácia dos fungos D. flagrans ou mesmoM. thaumasium no controle ambiental dos nematóides de bovinos naturalmente infectados.

First report of interaction of nematophagous fungi on Libyostrongylus douglassii (Nematoda: Trichostrongylidae) / Primeiro relato da interação de fungos nematófagos sobre Libyostrongylus douglassii (Nematoda: Trichostrongilidae)

Libyostrongylus douglassii is a gastrointestinal nematode parasite of ostriches that can cause up to 50% mortality in young birds. The objective of this study was to compare the predatory capacity of two isolates of the predatory fungi Duddingtonia flagrans(AC001 and CG722 isolates) and one of Arthrobotrys cladodes (CG719) on infective larvae (L3) of L. douglassii under laboratory conditions, in 2% water-agar medium. The results showed that the fungi tested were effective in preying upon the L3 of L. douglassii (P < 0.05), compared with the control group. However, there was no difference in predatory capacity between the fungi tested (P > 0.05) during the seven days of experimental testing. In comparison with the control, without fungus, there were significant decreases (P < 0.05) of 85.2% (AC001), 81.2% (CG722) and 89.2% (CG719) in the average numbers of L3 of L. douglassii recovered from treatments with the isolates tested. In the present study, the three isolates of the predatory fungi D. flagrans (AC001 and CG722) and A. cladodes (CG719) were efficient at in vitro destruction of the L3 of L. douglassii.

Libyostrongylus douglassii é um nematóide parasito gastrintestinal de avestruzes que pode causar até 50% de mortalidade em aves jovens. O objetivo deste trabalho foi comparar a capacidade predatória de dois isolados de fungos predadores Duddingtonia flagrans (isolados AC001 e CG722) e um Arthrobotrys cladodes (CG719) sobre larvas infectantes (L3) de L. douglassii em condições laboratoriais, em meio ágar­água 2%. Os resultados demonstraram que os fungos testados foram eficientes em predar as L3 de L. douglassii (P < 0,05) em relação ao grupo controle. Contudo, não foi observada nenhuma diferença na capacidade predatória entre os fungos testados (P > 0,05) durante os sete dias do ensaio experimental. Em comparação ao controle, sem fungo, houve uma redução significativa (P < 0,05) de 85,2% (AC001); 81,2% (CG722) e 89,2% (C719) na média de L3 recuperadas nas placas do grupo tratado com os isolados testados. No presente trabalho, os três isolados de fungos predadores D. flagrans (AC001 e CG722) e A. cladodes (CG719) foram eficientes na destruição in vitro das L3 de L. douglassii.

Proteolytic action of the crude extract of Duddingtonia flagrans on cyathostomins (Nematoda: Cyathostominae) in coprocultures / Ação proteolítica do extrato bruto de Duddingtonia flagrans sobre ciatostomíneos (Nematoda: Cyathostominae) em coproculturas

The objective of this study was to examine the action of the crude extract of Duddingtonia flagrans (isolates AC001 and CG722) on infective larvae (L3) of cyathostomins in coprocultures and to confirm its proteolytic activity by means of a zymogram. The following groups were formed in coprocultures: Group 1: 10 mL of crude extract of D. flagrans (AC001); group 2: 10 mL of crude extract of AC001 with 10 mM of Ca2+; group 3: 10 mL of crude extract of D. flagrans (CG722); group 4: 10 mL of crude extract of CG722 with 10 mM of Ca2+; and group 5: control group (distilled water). The third-stage larvae (L3) were obtained after eight days. The crude extract of D. flagrans was effective in reducing the number of L3, with the following percentage reductions: group 1, 49.5%; group 2, 52.5%; group 3, 36.8%; and group 4, 57.7%; in relation to the control group (p > 0.05). The proteolytic activity of the crude extract was confirmed through the zymogram. The results from this study confirmed that the crude extract of the fungus D. flagrans could be used for controlling cyathostomin L3, and suggested that at least one protease of approximately 38 kDa was present.

O objetivo deste trabalho foi estudar a ação do extrato bruto de Duddingtonia flagrans (isolados AC001 e CG722) sobre larvas infectantes (L3) de ciatostomíneos em coproculturas e confirmar a sua atividade proteolítica por meio de um zimograma. Foram formados os seguintes grupos em coproculturas: grupo 1: 10 mL de extrato bruto de D. flagrans (AC001); grupo 2: 10 mL de extrato bruto de AC001 com íons Ca2+ 10 Mm; grupo 3: 10 mL de extrato bruto de D. flagrans (CG722); grupo 4: 10 mL de extrato bruto de CG722 com íons Ca2+ 10 Mm; e grupo 5 como controle (água destilada), obtendo-se as L3 ao final de 8 dias. O extrato bruto de D. flagrans foi eficiente na redução do número de L3 com os seguintes percentuais de redução: grupo 1 (49,5%); grupo 2 (52,5%); grupo 3 (36,8%) e grupo 4 (57,7%) em relação ao grupo controle (p > 0,05). Confirmou-se a atividade proteolítica por meio do zimograma. Os resultados do presente trabalho confirmam a utilização do extrato bruto do fungo D. flagrans no controle de L3 de ciatostomíneos e sugere a presença de pelo menos uma protease de aproximadamente 38 kDa.

In vitro predatory activity of conidia of fungal isolates of the Duddingtonia flagrans on Angiostrongylus vasorum first-stage larvae

INTRODUCTION:Angiostrongylus vasorum is a nematode that parasitizes molluscs, dogs, and even man. METHODS:The objective was to evaluate the predatory activity of the conidia of two fungal isolates of Duddingtonia flagrans (AC001 and CG722) on first-stage larvae (L1) of A. vasorum in laboratory conditions. RESULTS: At the end of the experiment, there were significant reductions (p<0.01) of 74.5% and 63.2%, on average, in the A. vasorum (L1) recovered in the AC001 and CG722 treatment conditions, respectively. CONCLUSIONS: The two isolates of fungi were efficient in the capture and destruction of A. vasorum (L1).

Control of Strongyloides westeri by nematophagous fungi after passage through the gastrointestinal tract of donkeys / Controle de Strongyloides westeri por fungos nematófagos após trânsito gastrintestinal em jumentas

Strongyloides westeri is the most prevalent nematode among equines aged up to four months and causes gastrointestinal disorders. The objective of this study was to observe the control of infective S. westeri larvae (L3) by the nematophagous fungi Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34) after passage through the gastrointestinal tract of female donkeys. Twelve dewormed female donkeys that were kept in stables were used. Two treatment groups each comprising four animals received orally 100 g of pellets made of sodium alginate matrix containing a mycelial mass of either D. flagrans (AC001) or M. thaumasium (NF34). The control group consisted of four animals that received pellets without fungus. Feces samples were then collected from the animal groups at different times (after 12, 24, 48 and 72 hours). These feces were placed in Petri dishes containing 2% water-agar medium and 1000 L3 of S. westeri. AC001 and NF34 isolates showed the ability to destroy the L3, after gastrointestinal transit, thus demonstrating their viability and predatory activity.

O Strongyloides westeri é o nematóide de maior prevalência entre equídeos com idade até quatro meses, causando distúrbios gastrintestinais. O objetivo do presente trabalho foi observar o controle de larvas infectantes (L3) de Strongyloides westeri pelos fungos nematófagos Duddingtonia flagrans (AC001) e Monacrosporium thaumasium (NF34) após trânsito gastrintestinal em jumentas. Foram utilizadas 12 jumentas, estabuladas e previamente vermifugadas. A seguir, dois grupos tratados, contendo cada um 4 animais receberam por via oral 100 g de péletes em matriz de alginato de sódio, contendo massa miceliana dos fungos D. flagrans (AC001) ou M. thaumasium (NF34). O grupo controle foi constituído de 4 animais que receberam péletes sem fungo. A seguir, amostras de fezes dos grupos de animais foram coletadas em distintos intervalos de horas (12, 24, 48 e 72). Essas fezes foram vertidas em placas de Petri contendo meio sólido ágar-água 2% e 1000 L3 de S. westeri. Os isolados AC001 e NF34 apresentaram capacidade de destruir as L3 após o trânsito, demonstrando sua viabilidade e atividade predatória.

Atividade larvicida do extrato bruto enzimático do fungo Duddingtonia flagras sobre larvas de primeiro estádio de Angiostrongylus vasorum / Larvicidal activity of a crude enzyme extract of the fungus Duddingtonia flagrans on first-stage larvae of Angiostrongylus vasorum

INTRODUÇÃO: Angiostrongylus vasorum é um nematóide que parasita cães domésticos e eventualmente o homem. MÉTODOS: O objetivo deste trabalho foi observar a atividade predatória in vitro do extrato bruto enzimático do fungo Duddingtonia flagrans sobre larvas de primeiro estádio A. vasorum em condições laboratoriais no meio ágar-água 2 por cento. RESULTADOS: Ao final do experimento, os percentuais de redução das L1 de A. vasorum observados foram de: 53,5 por cento (24h) e 71,3 por cento (48h) CONCLUSÕES: O extrato bruto enzimático do fungo D. flagrans destruiu in vitro as L1, podendo ser utilizado como controle biológico desse nematóide.

INTRODUCTION: Angiostrongylus vasorum is a nematode parasite of domestic dogs and potentially of humans. METHODS: This study aimed to observe the predatory activity in vitro of a crude enzyme extract of the fungus Duddingtonia flagrans on first-stage larvae of A. vasorum in laboratory conditions on 2 percent water-agar. RESULTS: At the end of the experiment, the percentage reductions observed for A. vasorum L1 were 53.5 percent (24h) and 71.3 percent (48h). CONCLUSIONS: Crude enzyme extract of the fungus D. flagrans destroyed the L1 in vitro and can be used as a biological control for this nematode.

Ação ovicida do extrato bruto enzimático do fungo Pochonia chlamydosporia sobre ovos de Ancylostoma sp / Ovicidal action of a crude enzymatic extract of fungus Pochonia chlamydosporia against Ancylostoma sp eggs

INTRODUÇÃO: Ancylostoma sp é um geo-helminto potencialmente zoonótico. MÉTODOS: O objetivo deste trabalho foi avaliar in vitro a ação do extrato bruto enzimático de Pochonia chlamydosporia (VC4) sobre ovos de Ancylostoma sp, em meio ágar-água 2 por cento e em cultura de fezes. RESULTADOS: Observou-se um percentual de redução na eclosão dos ovos de Ancylostoma sp, de 76,8 por cento na placas de Petri do grupo tratado em relação ao grupo controle. CONCLUSÕES: O extrato bruto enzimático de Pochonia chlamydosporia foi eficiente na redução da eclosão dos ovos de Ancylostoma sp, podendo ser utilizado como controlador biológico desse nematoide.

INTRODUCTION: Ancylostoma sp is a potentially zoonotic geohelminth. METHODS: This study aimed to evaluate in vitro the action of crude enzyme extract of Pochonia chlamydosporia (VC4) on eggs of Ancylostoma sp in 2 percent water-agar and in fecal cultures. RESULTS: The percentage reduction in Ancylostoma sp egg eclosion was 76.8 percent in Petri dishes of the treated group compared to the control group. CONCLUSIONS: The crude enzyme extract of Pochonia chlamydosporia was effective at reducing Ancylostoma sp egg eclosion and can be used as biological control of this nematode.