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The aim of this study was to evaluate the isolated and associated use of the nematophagous fungi Pochonia chlamydosporia (isolate VC4) as an ovicide and Duddingtonia flagrans (isolate AC001) as a predator in the control of Taenia saginata eggs. Viable T. saginata eggs were obtained by dissecting mature proglottids from a specimen. For the experimental assay, four groups were formed in microtubes, as follows: (G1—T. saginata eggs/control); (G2—100 T. saginata eggs + VC4); (G3—100 T. saginata eggs + AC001); (G4—100 T. saginata eggs + VC4 + AC001). All the groups were stored in a B.O.D. incubation chamber at a temperature of 27 °C for 15 days and then the contents of the microtubes were analyzed using an optical microscope with a 40x objective. At the end of the assay the treated groups (G2 to G4) showed ovicidal activity (destruction of eggs) compared to the control group (G1). The highest ovicidal percentage was observed in group G2 (eggs + VC4), with 43.3%. In groups G3 and G4 (combination of fungal isolates), the ovicidal percentages were 25.7% and 25.6%, respectively. The results of this study shed light on a new possibility for the combined use of different species of nematophagous fungi, which could be used in the future for environmental biological control of T. saginata eggs.

Alphitobius diaperinus, popularmente conhecido como cascudinho, atualmente é um dos principais problemas da avicultura mundial. Objetivou-se avaliar o potencial do fungo Pochonia chlamydosporia (VC4) no controle biológico de A. diaperinus em condições laboratoriais. Exemplares adultos do inseto foram coletados em uma granja avícola, no sudeste do Brasil. Em seguida, foi realizado crescimento do fungo P. chlamydosporia em meio ágar quitina (AQ2%). Após 10 dias, foi obtida uma concentração de estruturas fúngicas (conídios/clamidósporos) de 1500/?L. Posteriormente, foram formados dois grupos: grupo tratamento (G1), que consistiu em placas de Petri contendo meio de cultura ágar-água a 2% (AA2%) com 10 insetos vivos + 10µL de solução fúngica (vertida) sobre cada inseto; e o grupo controle (G2), que foi constituído por placas de Petri contendo AA 2% + 10 insetos vivos + 10µL de água destilada vertidos sobre cada inseto. Ambos os grupos foram realizados em triplicata. As placas de ambos os grupos G1 e G2 foram incubadas em BOD a 25±1°C e 80±10% de umidade relativa, durante 7 dias. Ao final do sétimo dia de interação, no grupo G1 foi observada a colonização e morte de 100% de A. diaperinus e visualização de hifas de VC4 sobre os mesmos, sugerindo uma possível atividade entomopatôgenica. No grupo G2 foram observados insetos vivos. Conclui-se que o controle biológico de A. diaperinus pelo fungo P. chlamydosporia (VC4) foi eficaz em condições laboratoriais e mais delineamentos experimentais, em condições a campo, podem vir a colaborar com a sua aplicabilidade.

Nematophagous, or helminthophagous fungi of the genera Duddingtonia, Arthrobotrys, Monacrosporium, Pochonia, Paecilomyces, and Mucor, have been used over the years in in vitro and in vivo experiments to control helminth parasites that are potentially zoonotic. These fungi have shown efficacy against the following helminth genera: Ancylostoma, Toxocara, Enterobius, Strongyloides, Angiostrongylus, Taenia, Fasciola, and Schistosoma. The results obtained from these experiments, together with studies on soil contamination, suggest the viability of their use as a sustainable and effective strategy to reduce environmental contamination by these zoonotic parasites. Therefore, the aim of this review was to address the role of helminthophagous fungi in the biological control of potentially zoonotic helminths. To this end, we describe (1) a brief history of helminthophagous fungi; (2) a discussion of some potentially zoonotic intestinal parasites; (3) the importance of helminthophagous fungi in the control of nematodes, cestodes, and trematodes; and (4) the potential of helminthophagous fungi as a practical and sustainable strategy.

Equids can be infected by Neospora spp., and its infection is associated with neurological and reproductive disorders. This systematic review and meta-analysis aimed to assess the prevalence of anti-Neospora caninum antibodies in horses, donkeys and mules worldwide between 2012 and 2022. Five scientific databases were consulted: MEDLINE, ScienceDirect, PubMed, LILACS and SciELO, using the descriptors “protozoan” or “neospora” AND “horses” or “donkeys” or “mules”. A total of 1909 studies were screened, and 25 studies met the inclusion criteria. The pooled prevalence of Neospora spp. infection in equids, based on a total of 11,076, was 8.34% (95% CI: 4.78–14.17%). Further analysis by species revealed no statistically significant difference in the prevalence of Neospora spp. infection across equids: 8.5% (95% CI 5.03–13.99%) in horses, 7.36% (CI 2.08–22.94%) in donkeys and 6.07% (95% CI 0.71–36.83%) in mules. Regarding diagnostic tests, the prevalence of Neospora spp. infection determined by the indirect fluorescent antibody test (IFAT) was 8.08% (95% CI: 3.74–16.61%), by the enzyme-linked immunosorbent assay (ELISA) was 7.91% (95% CI: 3.71–16.08%), and by the modified agglutination test (MAT) was 20% (95% CI: 15.37–25.60%). This meta-analysis provided valuable and comprehensive insights into the prevalence of Neospora spp. infection in horses, donkeys, and mules using serological detection methods. The results highlight the wide distribution of Neospora spp. infection in equids across all continents, indicating the importance of this infection. Transmission of the parasite can occur both horizontally (directly between animals) and vertically (from mother to offspring), further emphasizing the significance of understanding and managing this infection in equid populations.

ABSTRACT: Rabies is a viral encephalitis that affects mammals, including humans. Rapid and effective laboratory diagnosis of the rabies virus is critical for public health. This study evaluated the operational, technical, and financial viability of the RT-qPCR in replacement of inoculation in mice for diagnosing rabies in the laboratory routine. A total of 316 samples of mammalian brains were analyzed, 121 positives and 195 negatives, previously diagnosed by direct immunofluorescence (DIF) and mouse inoculation test (MIT). The samples were submitted to the duplex TaqMan RT-qPCR technique. The accuracy, sensitivity, and specificity of RT-qPCR were analyzed. We analyzed the costs for performing the RT-qPCR technique and compared it with the cost of MIT. The results showed 99.37% accuracy, 99.17% sensitivity, and 99.49% specificity by RT-qPCR when related to DIF and MIT results, which proved to be a robust and repeatable technique. The minimum time for a positive diagnosis was reduced in RT-qPCR (1 day) if compared to MIT (9.64 days), with a 17.7% reduction in the cost of the molecular technique. The present study demonstrated that the molecular biology technique is an efficient tool to diagnose rabies in the laboratory routine, being able to replace MIT.

O objetivo do presente trabalho foi avaliar diferentes concentrações de conídios do fungo Duddingtonia flagrans (AC001) e hipoclorito de sódio (NaOCl) a 5% (v/v) sob a eclosão de ovos de nematódeos gastrintestinais de equinos. Foram formados cinco grupos experimentais: Grupo 1 (3 x 106 conídios de AC001 + 20 g de fezes), Grupo 2 (6 x 106 conídios de AC001 + 20 g de fezes), Grupo 3 (9 x 106 conídios de AC001 + 20 g de fezes), Grupo 4 (5 mL de hipoclorito de sódio 5% (v/v) + 20 g de fezes) e Grupo 5 (5 mL de água destilada + 20 g de fezes). Em seguida, as coproculturas foram incubadas por 10 dias no escuro 26 ± 1° a 2°C e, posteriormente, procedeu-se a recuperação de larvas infectantes (L3) de nematódeos por meio da técnica de Baermann. Ao final do período experimental, constatou-se a presença absoluta de L3 de nematódeos ciatostomíneos. Os resultados demonstraram que houve redução significativa (p0,01) entre as três concentrações de conídios testadas nos grupos G2 e G3. Conclui-se que o fungo D. flagrans e o hipoclorito de sódio a 5% (v/v) foram eficientes na redução de L3 de nematódeos gastrintestinais nas coproculturas, podendo esse resultado ser utilizado em futuros delineamentos experimentais.

O objetivo deste estudo foi avaliar a ação de uma formulação comercial à base de Duddingtonia flagrans (Bioverm®) sobre os ovos de Ascaris suum e larvas infectantes (L3) de Oesophagostomum spp. e Hyostrongylus rubidus de suínos. Doze suínos machos foram divididos em dois grupos: tratado, que recebeu dose única de 1 g/10 kg de peso vivo (105 clamidósporos de D. flagrans); e controle, que permaneceram sem tratamento. Amostras fecais (100g) foram coletadas individualmente às 0, 12, 24, 36, 48, 60 e 72 horas após os tratamentos. No ensaio A, 2 g de fezes e 1000 ovos de A. suum foram adicionados a placas de Petri, e a predação larval foi avaliada para classificar os efeitos da predação. No ensaio B, 2000 L3 de Oesophagostomum spp. e H. rubidus foram adicionados às coproculturas, incubadas por 10 dias e recuperadas para quantificação das L3 não predadas. No ensaio A, houve apenas efeito tipo 1, com crescimento fúngico em ovos, mas sem ação lítica. No ensaio B, observou-se uma redução de L3 de Oesophagostomum spp. e H. rubidus de 73,9% (p

Objetivo. El objetivo de este estudio fue evaluar la asociación in vitro del hongo Duddingtonia flagrans (AC001) e ivermectina en el control de nematodos gastrointestinales de terneros búfalo. Materiales y métodos. Se formaron cuatro grupos experimentales en microtubos, con cinco réplicas para cada grupo: G1 (nematodos + AC001), G2 (nematodos + ivermectina 1%), G3 (nematodos + AC001 + ivermectina 1%) y G4 (nematodos + agua destilada). Para cada grupo, después de 36 horas de interacción se leyó el contenido de los microtubos mediante microscopía óptica, contabilizando el número de nematodos por grupo. Resultados. Hubo una reducción larvaria significativa de los grupos tratados, con los siguientes porcentajes con relación al G4 (control): G1: 43,7%; G2: 82,3% y G3: 65,7%. También se observó que la asociación in vitro de D. flagrans con ivermectina fue más efectiva en la reducción de L3 en comparación con el uso aislado de este hongo. Conclusiones. Se concluyó que el uso conjunto de D. flagrans con ivermectina puede potenciar la eficacia del control biológico de los nematodos gastrointestinales de los búfalos, previendo su uso en las condiciones naturales de la cría de búfalos.

The objective of this study was to verify the occurrence of Anaplasma marginale, Babesia bovis and Babesia bigemina in dairy cattle from the central mesoregion of the State of Espírito Santo, through direct parasitological tests (blood smears) and serological tests (Enzyme Linked Immunosorbent Assay - ELISA). Blood samples were collected from 159 animals from 6 farms, concentrated in the central mesoregion of the state of Espírito Santo. The blood collected was stored in EDTA tubes, used to perform blood smears, and in tubes without anticoagulant, to obtain serum. The sera obtained were used for the detection of IgG antibodies to A. marginale, B. bovis and B. bigemina. Of the 159 blood smears analyzed, 64 (40.2%) and 11 (6.9%) were positive for A. marginale and Babesia sp., respectively. The ELISA of the 159 samples, 9 (5.7%), 59 (37.1%) and 47 (29.5%) were seropositive for A. marginale, B. bovis and B. bigemina, respectively. The results obtained characterize these farms of the central mesoregion of the state of Espírito Santo, as of enzootic instability for Bovine Parasitic Sadness (TPB) agents, presenting a high risk of occurrence of anaplasmosis and babesiosis outbreaks.

ABSTRACT: Bovine cysticercosis (BCC) is an important disease in Brazil due to its detection in abattoirs, resulting in economic losses for cattle farmers, and a public health corcern. Besides its importance, the knowledge about BCC epidemiology and impact remains not fully described in several areas in this country, requiring more studies. Thus, this study focused on establishing BCC prevalence in the regions and municipalities of the state of Espírito Santo from 2017 to 2019, establishing the associated risk factors, and estimating the economic losses for cattle farmers. A set of 2,330 cases of BCC was detected in the 407,529 bovine inspected by Federal Inspection Service (0.57%; C.I. 95% 0.55 - 0.60%) with a higher detection of unviable cysticercus (66.14%). The beef producers lost at least US$153,000.20 due to BCC through this period. The highest risk for BCC infection occurred in Metropolitana (OR = 9.19), Litoral Sul (OR = 7.77), Caparó (OR = 7.44), and Central Sul (OR = 7.19) mesoregions, respectively. The BCC was detected in animals from 67 municipalities and its prevalence increased in areas with high human population density (OR = 1.58;; p = 0.01). In conclusion, BCC is an important disease for beef production chain in the Brazilian state of Espírito Santo and a cause of economic losses, mainly in some areas, and urgently requires the adoption of prophylactic strategies to reduce the ocurrence of this parasite.

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The fungi Arthrobotrys musiformis (A144 isolate) and Monacrosporium sinense (SF53 isolate) were evaluated for controlling L3 of cyathostomins. In vitro, Petri dishes containing 2% water-agar were divided into groups: 1 (A144 + SF53); 2 (SF53); 3 (A144); 4 (control group). In vivo, 24 animals received pellets containing fungal mycelium. The animals were divided into four groups: 1 (100 g of pellets containing A144 + SF53 isolates); 2 (100 g of pellets containing A144 isolate); 3 (100 g of pellets containing SF53 isolate); 4 (Pellets without fungi). Feces were collected at 12, 24, 36, 48, 60 and 72 hours after pellet administration. The association of isolates, in vitro, was able to reduce L3 by 69.7%, compared with the control group; A144 isolate reduced L3 by 77.3%; and SF53 isolate reduced L3 by 84.3%. In vivo, the association of fungi A. musiformis and M. sinense showed predatory activity against L3 of cyathostomins with efficacy ranged from 63.2% to 83%. The association of the fungi A. musiformis and M. sinense both in vitro and in vivo proved to be efficient for controlling L3 of cyathostomins.

The aim was to detect correlations of IgG antibodies against N. caninum in serum and colostrum samples from ewes, through the IFAT, and to evaluate the presence of this immunoglobulin in the serum of newborn lambs after colostrum ingestion. Blood samples from 162 ewes that did not show any disease in the general physical examination and from their newborn lambs, not more than five days postpartum, along with 162 colostrum samples and 182 blood samples from the neonates, were analyzed. In total, 27.8% (45/162) of the mothers were positive for anti-N. caninum IgG, among which antibodies were detected in the colostrum in 46.7% (21/45). All the ewes with positive colostrum had reactive offspring. The kappa agreement for the correlation between the serological tests on the ewes and the colostrum results was 0.558. This correlation increased as the antibody titers of the mothers increased, and reached 1.000 from the titer of 1:400 from the mothers. Comparison of the antibody detection results between the offspring’s serum and colostrum showed a kappa agreement of 1.000. In conclusion, there was a good agreement regarding the detection of anti-N. caninum IgG between the colostrum samples and the lambs’ serum; the use of colostrum forms a noninvasive alternative for diagnosing N. caninum in sheep herds.

Abstract Bioverm® (Duddingtonia flagrans) is a fungal formulation indicated for controlling gastrointestinal nematodes in ruminants and horses, which has recently been authorized for commercialization in Brazil. The objective was to determine the efficiency of Bioverm® against larvae of gastrointestinal nematodes after passage through the gastrointestinal tract of cattle. Twelve animals were used, divided into two groups. In the treated group, a single dose of 1 g of Bioverm® per 10 kg of live weight (containing 105 chlamydospores of D. flagrans) was provided for each animal. Fecal samples were obtained from the animals in each group at 12, 24, 36, 48, 60 and 72 hours after administration. In assay A, 2 g of feces were added to Petri dishes containing 2% agar-water medium. In assay B, coprocultures were performed. In both assays, the peak of larval predation occurred within 48 hours after administration of Bioverm®. In assay A, a significant larval reduction (P < 0.05) was seen at 48 h (88.2%). In assay B, significant reductions (P < 0.05) were seen at 36 h (43.7%) and 48 h (82.3%). Bioverm® showed high predatory capacity after passage through the gastrointestinal tract of cattle and was effective for controlling gastrointestinal nematodes.

Sheep farming is of great importance in generating income to smallholders in semi-arid areas worldwide. Gastrointestinal helminths cause important damage to the animals with poor welfare conditions, impacting in local economies. The objective of this work was to evaluate the FAMACHA method (FMC) in a herd of 60 Santa Inês from Paraíba, Brazil, and to compare the data from previous production years. In order to determine anemic sheep, FMC chart was used in biweekly evaluations for 12 months by trained personnel. Blood and fecal samples were collected monthly to determine packed cell volume (PCV), and parasite fecal egg count (FEC), respectively. The animals were divided into: dry, pregnant and lactating ewes, lambs and rams. Haemonchus was the predominant helminth genus (94.7%) during the whole year. There were up to 97% of success on the interpretation of FMC with 90 to 100% efficacy in the treatment decision. Among the 1356 evaluations, in only 40 (3%) were required deworming, being 27 during the rainy season and 13 during the long dry season. A statistically significant correlation (P < 0.01) was observed between FEC and PCV for the dry, lactating, and pregnant categories. The data reports a 94% reduction in deworming, with a greater need for anthelmintic use in lambs and lactating ewes (P < 0.05). Our data validates the use of FMC in Santa Inês sheep, which is considered a local resilient breed in the semi-arid region of Brazil. We recommend that animals should be examined during the rainy season to prevent most of the clinical problems.

This study aimed to evaluate the use of a sodium alginate matrix-pelletized formulation of Duddingtonia flagrans for biological control of gastrointestinal nematodiasis in feedlot goats in the semiarid region of Northeastern Brazil. We used 20 Saanen female goats (age, 4 months; average weight, 12 kg) that did not receive anthelmintic treatment and had counting of eggs per gram of faeces (EPGs) ? 500. The animals were divided into two groups: in group 1 (D. flagrans group), each animal received 3 g of pellets (0.6 g of D. flagrans mycelium) per 10 kg of body weight, twice a week, over 4 months; and in group 2 (control group), each animal received 3 g of pellets without fungus per 10 kg of body weight, twice a week, over 4 months. Each group was maintained in a separate 15-m2 stall. Larval cultures and measurements of weight, EPG, and packed cell volume (PCV) were performed every 15 days. We observed low EPG levels in the D. flagrans group throughout the experimental period, with a significant difference (p < 0.05) on day 30 and from day 60, having, at the end of the experiment, average OPG values of only 150, reduction of 92.3% when compared to control group. Haemonchus sp. was the most prevalent helminth in all larval cultures. The D. flagrans group showed a mean weight gain of 8.8 kg at the end of the experiment (p < 0.05), while the control group showed a mean weight gain of 4.8 kg. The best PCV results (p < 0.05) were also observed in the D. flagrans group from day 30. Thus, the use of D. flagrans pellets in a sodium alginate matrix was effective in controlling gastrointestinal nematodiasis of feedlot goats in the semiarid region of Northeastern Brazil.

The objective of this study was to evaluate the in vivo predatory viability of the nematophagous fungus, Duddingtonia flagrans, after storage (36 months) and refrigeration (2-8 °C). This viability was evaluated using the infective larvae of gastrointestinal nematodes of sheep in the Northeastern semi-arid region of Brazil. Sixteen Santa Inês sheep with negative counting of eggs per gram of feces (EPG) were divided into four experimental groups, each group comprised of four animals. The pellets were administered at the dose of 3 g/10 kg of live weight (20% fungal micelyum), and a single administration was performed for each animal. Group I was administered pellets that had been stored for 36 months; Group II, freshly produced pellets; Group III, freshly produced pellets that did not contain fungi; and Group IV, pellets were not administered, and this was the control group. Feces were collected for 5 days, every 24 h for analysis. There was a significant decrease in the number of infective larvae of sheep nematodes that received D. flagrans pellets in a sodium alginate matrix, 82% was observed for Group I and 71% for Group II, compared to the control group. It is therefore concluded that the fungus, D. flagrans, pelleted in sodium alginate matrix after 36 months of storage at 2-8 °C, showed efficacy in reducing the number of infective larvae of gastrointestinal nematodes of sheep.

This study aimed to report the first occurrence of Leporacarus gibbus in a domestic rabbit (Oryctolagus cuniculus) from the state of Espírito Santo, Brazil. The two-year-old male rabbit came from a rural area and was admitted to the Veterinary Hospital Professor Ricardo Alexandre Hippler with intense pruritus throughout the body, severe hair loss, crustal lesions on the outer surface of the ears, and desquamation mainly in the dorsal region. A skin scraping and trichogram were performed. In the microscopic evaluation of the hairs obtained from the dorsal region, mites with morphology compatible with L. gibbus were observed. Cheyletiella parasitovorax and Psoropotes cuniculi were also detected in the same region. Treatment with ivermectin (0.4 mg/kg) was prescribed with three applications every 14 days, over a period of six weeks. This report presents the first description of the occurrence of L. gibbus in the state of Espírito Santo concomitant with poly-infestation of C. parasitovorax and P. cuniculi, and it is hoped that this will provide a research tool for future work in the region.

Abstract The objective was to evaluate the action of D. flagrans pellets in association with Levamisole Hydrochloride 5% for controlling sheep gastrointestinal nematodes in the northeastern Brazil. Three groups of six sheep each were formed: group 1 received 3 g of the pellets (0.6 g of D. flagrans mycelium) for each 10 kg b.w., twice a week for six months, and deworming with Levamisole Hydrochloride 5% when EPG ≥ 1500; group 2 received a dosage of Levamisole Hydrochloride 5% when EPG ≥ 1500; and group 3 received 3 g of pellets without fungi for each 10 kg b.w., twice a week for six months. EPG counts, larval cultures, packed cell volume (PCV) and weighing were performed every 15 days; monthly, samples of grass from each paddock were collected. The mean EPG of the groups began to statistically differ from day 30 (p < 0.05). Group 1 required less deworming with Levamisole Hydrochloride 5% and showed superiority of PCV values throughout the experiment (p < 0.05). There was a significant reduction (p < 0.05) in L3 recovery in the group 1 paddock from day 30 onwards. The use of D. flagrans pellets in association with Levamisole Hydrochloride 5% was effective for controlling gastrointestinal nematodes.

The objective of this study was to use chlamydospores of the fungus Pochonia chlamydosporia (isolates VC1 and VC4) against Toxocara canis eggs in a 15-day in vitro assay. One thousand T. canis eggs were placed in Petri dishes containing 2% water agar medium with different concentrations of chlamydospores (1,000, 10,000 or 100,000) of each fungal isolate of P. chlamydosporia (treated groups) and 1,000 eggs in Petri dishes without fungus (control group). Egg counts were performed to determine the ovicidal activity, which was classified as three effect levels: type 1, type 2 and type 3. Significant differences (P < 0.01) in egg destruction were found in comparison with the control group. The highest percentage of egg destruction was found in plates containing 100,000 chlamydospores (68.5% for VC1 and 70.5% for VC4). Chlamydospores of P. chlamydosporia were effective in destroying T. canis eggs and may contribute in the future towards combating the eggs of this parasite.O objetivo do trabalho foi utilizar clamidósporos do fungo Pochonia chlamydosporia (isolados VC1 e VC4) na destruição de ovos de Toxocara canis, num ensaio in vitro, realizado no intervalo de 15 dias. Em cada placa de Petri com ágar-água 2% foram vertidos 1.000 ovos de T. canis em 1.000, 10.000 ou 100.000 clamidósporos de cada isolado do fungo (grupos tratados). Foram realizadas as contagens para verificar a atividade ovicida, classificada em três níveis de efeito: tipo 1, tipo 2 e tipo 3. Os resultados demonstraram que houve diferença significativa (P < 0,01) na destruição dos ovos em relação aos ovos observados nas placas do grupo controle. O maior percentual de ovos destruídos foi observado nas placas contendo 100.000 clamidósporos (68,5% para VC1 e 70,5% para VC4). Clamidósporos do fungo P. chlamydosporia foram efetivos na destruição dos ovos de T. canis podendo contribuir no futuro para o combate aos ovos deste parasito.

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.

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 ágar–água 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.

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) e Monacrosporium 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 mesmo M. thaumasium no controle ambiental dos nematóides de bovinos naturalmente infectados.

The aim of this study was to evaluate the predatory activity of the fungus Duddingtonia flagrans (AC001) on infective larvae of Ancylostoma ceylanicum after gastrointestinal transit in hamsters. Twenty animals were used in the experiment, divided into two groups: a treated group (10 animals) and a control group (10 animals). In the group treated with D. flagrans, each animal received mycelium from the AC001 isolate, at an oral dose of 5 mg/25 g of live weight. To evaluate the predatory activity of the fungus, fecal samples were collected from the animals in both groups, at the times of 6, 8, 12, 24 and 36 hours after the treatment. Then, subsamples of 2 g of feces were placed in Petri dishes containing 2% water-agar (2% WA) culture medium and 1000 L3 of A. ceylanicum. Over the study period, the following percentage reductions were observed: 43.2% (6 hours), 30.8% (8 hours), 25.8% (12 hours), 30% (24 hours) and 11% (36 hours). The fungus D. flagrans presented predatory activity on the L3 of A. ceylanicum, after passing through the hamsters' gastrointestinal tract. It was therefore concluded that the fungus D. flagrans may be an alternative for biological control of the L3 of A. ceylanicum.O objetivo deste trabalho foi avaliar a atividade predatória do fungo Duddingtonia flagrans (AC001) sobre larvas infectantes de Ancylostoma ceylanicum após o trânsito gastrintestinal em hamsters. Foram utilizados vinte animais no experimento, divididos em dois grupos: um grupo tratado (10 animais) e um grupo controle (10 animais). No grupo tratado com D. flagrans, cada animal recebeu 5mg/25g de peso vivo de micélio do isolado AC001, por via oral. Para avaliar a atividade predatória do fungo, amostras fecais foram coletadas de ambos os grupos de animais nos horários de: 6, 8, 12, 24 e 36 após o tratamento. A seguir, 2g de fezes foram colocadas em placas de Petri contendo o meio de cultura ágar-água 2% (AA2%) e 1000 L3 de A. ceylanicum. Ao longo dos horários estudados os seguintes percentuais de redução foram observados: 43,2% (6 horas); 30,8% (8 horas); 25,8% (12 horas); 30% (24 horas) e 11% (36 horas). O fungo D. flagrans (AC001) apresentou atividade predatória sobre as L3 de A. ceylanicum após o trânsito pelo trato gastrintestinal de hamsters. Além disso, foi observada uma diferença significativa nos percentuais obtidos de cada horário em relação ao numero de L3 recuperadas (P < 0,01). Conclui-se, portanto, que o fungo D. flagrans pode ser uma alternativa de controle biológico das L3 de A. ceylanicum.

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.

The predatory capacity of three isolates of the nematodes predacious fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium and Arthrobotrys robusta on infective larvae of cyathostomes was evaluated in laboratorial conditions in medium water-agar 2% (WA 2%). The isolates AC001, NF34 and I31 had presented significant reduction (pKEY WORDS: Arthrobotrys robusta, Cyathostominae, equine, Duddingtonia flagrans, Monacrosporium thaumasium, Nematophagous fungi. A capacidade predatória de três isolados de fungos predadores de nematoides Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34) e Arthrobotrys robusta (I31) sobre larvas infectantes de ciatostomíneos foi avaliada em condições laboratoriais em ensaio experimental em meio ágar-água 2% (AA 2%). Os isolados AC001, NF34 e I31 apresentaram redução significativa (p PALAVRAS-CHAVES: Arthrobotrys robusta, ciatostomíneos, equinos, Duddingtonia flagrans, fungos nematófagos, Monacrosporium thaumasium.

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 (pCONCLUSIONS: The two isolates of fungi were efficient in the capture and destruction of A. vasorum L1.

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

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%. RESULTADOS: Ao final do experimento, os percentuais de redução das L1 de A. vasorum observados foram de: 53,5% (24h) e 71,3% (48h) CONCLUSÕES: O extrato bruto enzimático do fungo D. flagrans destruiu in vitro as L1, podendo ser utilizado como controle biológico desse nematóide.

INTRODUÇÃO: Strongyloides venezuelensis tem sido utilizado como um modelo para estudo da estrongiloidose humana. MÉTODOS: O objetivo deste trabalho foi comparar a capacidade predatória dos fungos nematófagos Duddingtonia flagrans (AC001), Arthrobotrys robusta (I-31) e Monacrosporium sinense (SF53) sobre larvas infectantes (L3) de Strongyloides venezuelensis em condições laboratoriais no meio ágar-água 2%. RESULTADOS: Ao final do experimento, os percentuais de redução de L3 de Strongyloides venezuelensis observados foram de: 93% (AC001); 77,2% (I-31) e 65,2% (SF53). CONCLUSÕES: Os fungos nematófagos foram capazes de capturar e destruir in vitro as L3, podendo ser utilizados como controladores biológicos de Strongyloides venezuelensis.

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% e em cultura de fezes. RESULTADOS: Observou-se um percentual de redução na eclosão dos ovos de Ancylostoma sp, de 76,8% 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.

INTRODUÇÃO: Strongyloides stercoralis é um nematoide que infecta grande parte da população mundial. MÉTODOS: O objetivo deste trabalho foi comparar a capacidade predatória dos fungos nematófagos Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34) e Arthrobotrys robusta (I-31) sobre larvas infectantes (L3) de Strongyloides stercoralis em condições laboratoriais no meio ágar-água 2%. RESULTADOS: Ao final do experimento, os percentuais de redução de L3 de Strongyloides stercoralis observados foram de: 83,7% (AC001); 75,5% (NF34) e 73,2% (I-31). CONCLUSÕES: Os fungos nematófagos foram capazes de capturar e destruir in vitro as L3, podendo ser utilizados como controladores biológicos de Strongyloides stercoralis.INTRODUCTION: Strongyloides stercoralis is a nematode that infects much of the population worldwide. METHODS: This study aimed to compare the ability of predatory nematophagous fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34) and Arthrobotrys robusta (I-31) on infective larvae (L3) of Strongyloides stercoralis in laboratory conditions on 2% water-agar. RESULTS: At the end of the experiment, the percentage reductions in Strongyloides stercoralis L3 were 83.7% (AC001), 75.5% (NF34) and 73.2% (I-31). CONCLUSIONS: The nematophagous fungi were able to capture and destroy the L3 in vitro and may be used as biological controls of Strongyloides stercoralis.

INTRODUÇÃO: Toxocara canis é um ascarídeo parasita do intestino delgado de cães, causador da larva migrans visceral em seres humanos. MÉTODOS: Com o objetivo de demonstrar a eficácia do fungo Pochonia chlamydosporia sobre ovos de Toxocara canis em condições laboratoriais, foi montado ensaio experimental em placas de Petri com ágar-água 2%. RESULTADOS: Houve atividade ovicida de 43,8% (pINTRODUCTION: Toxocara canis is an ascarid parasite of the small intestine of dogs that causes visceral larva migrans in humans. METHODS: With the aim of demonstrating the effectiveness of the fungus Pochonia chlamydosporia on Toxocara canis eggs under laboratory conditions, a trial was set up in Petri dishes with 2% agar-water. RESULTS: There was ovicidal activity of 43.8% (p < 0.01) in the treated group in relation to the control group over the periods studied. CONCLUSIONS: The results from the present study suggest that Pochonia chlamydosporia can potentially be used as an alternative biological control for embryonated Toxocara canis eggs.

Com o objetivo de demonstrar a eficácia do fungo Paecilomyces lilacinus sobre ovos de Taenia saginata em condições laboratoriais, foi montado ensaio em placas de Petri com agar - água 2%. Houve atividade ovicida (pWith the aim of demonstrating the effectiveness of the fungus Paecilomyces lilacinus on Taenia saginata eggs under laboratory conditions, a trial was set up in Petri dishes with water-agar 2%. There was ovicidal activity (p < 0.05) in relation to the control group on the tenth day of interaction and an internal colonization rate of 25.5% in the eggs.

A capacidade predatória de três isolados de fungos predadores de nematoides Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34) e Arthrobotrys robusta (I31) sobre larvas infectantes de ciatostomíneos foi avaliada em condições laboratoriais em ensaio experimental em meio ágar-água 2% (AA 2%). Os isolados AC001, NF34 e I31 apresentaram redução significativa (p

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.

The objective of this study was to use chlamydospores of the fungusPochonia chlamydosporia (isolates VC1 and VC4) against Toxocara canis eggs in a 15-day in vitro assay. One thousand T. canis eggs were placed in Petri dishes containing 2% water agar medium with different concentrations of chlamydospores (1,000, 10,000 or 100,000) of each fungal isolate of P. chlamydosporia (treated groups) and 1,000 eggs in Petri dishes without fungus (control group). Egg counts were performed to determine the ovicidal activity, which was classified as three effect levels: type 1, type 2 and type 3. Significant differences (P < 0.01) in egg destruction were found in comparison with the control group. The highest percentage of egg destruction was found in plates containing 100,000 chlamydospores (68.5% for VC1 and 70.5% for VC4). Chlamydospores of P. chlamydosporiawere effective in destroying T. canis eggs and may contribute in the future towards combating the eggs of this parasite.

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) andA. cladodes (CG719) were efficient at in vitro destruction of the L3 of L. douglassii.

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 fungusD. flagrans could be used for controlling cyathostomin L3, and suggested that at least one protease of approximately 38 kDa was present.

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

Abstract The purpose of this study was to evaluate the predatory activity of the nematode Butlerius spp. and fungal isolates of Duddingtonia flagrans, Clonostachys rosea, Arthrobotrys musiformis and Trichoderma esau against H. contortus infective larvae (L3) in grass pots. Forty-eight plastic gardening pots containing 140 g of sterile soil were used. Panicum spp. grass seeds (200 mg) were sown into each pot and individually watered with 10 mL of tap water. Twelve days after seeding, the pots were randomly divided into 6 groups (n=8). Two thousand H. contortus infective larvae (L3) were added to each group. Additionally, the following treatments were established: Group 1 – 2000 Butlerius spp. larvae; group 2 – A. musiformis (1x107 conidia); group 3 – T. esau (1x107 conidia); group 4 – C. rosea (1x107 conidia), group 5 – D. flagrans (1x107conidia) and Group 6 – no biological controller (control group). The larval population of H. contortus exposed to Butlerius spp. was reduced by 61.9%. Population reductions of 90.4, 66.7, 61.9 and 85.7% were recorded in the pots containing A. musiformis, T. esau, C. rosea and D. flagrans, respectively. The results of this study indicate that the predatory nematode Butlerius spp. and the assessed fungi display an important predatory activity can be considered suitable potential biological control agents.

Abstract The objectives of this study were to describe occurrences of Rhabditis spp. causing parasitic otitis in dairy cattle of Gir breed in the state of Espírito Santo, southeastern Brazil, and to evaluate the biological control of this nematode using the nematophagous fungi Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34). After nematode detection and collection, three groups were formed: two groups that were treated, respectively, with the fungal isolates; and a control group, without fungus. The treatments were as follows: (a) Petri dishes containing the culture medium 2% water agar (WA) + 250 nematodes + AC001; (b) Petri dishes containing 2% WA + 250 nematodes + NF34; and (c) Petri dishes containing only 2% WA + 250 nematodes. After seven days at 27 °C the treatments with fungi were able to capture and destroy the nematodes, with percentages of 82.0% (AC001) and 39.0% (NF34) in relation to the control group. The results demonstrate the occurrence of Rhabditis spp. after animals physical examination and that there was efficacy of the in vitro predatory activity of both fungal isolates. Thus, these results are important because they can assist in future in vivo control of this nematode in cattle.