Assessing the Microbial Quality of Shrimp (Xiphonaeus kroyeri) and Mussels (Perna perna) Illegally Sold in the Vitória Region, Brazil, and Investigating the Antimicrobial Resistance of Escherichia coli Isolates.

The consumption of seafood is crucial for food security, but poor hygiene along the food production chain can result in low microbiological quality, posing significant risks for public health and seafood quality. Thus, this study aimed to assess the microbiological quality and antimicrobial sensitivity of E. coli from 69 samples of illegally marketed shrimp and mussels in the Vitória Region, Brazil. These foods exhibited poor microbiological quality due to high counts of mesophilic, psychrotrophic, and enterobacteria microorganisms. While this issue is widespread in this area, shrimp samples displayed higher microbial counts compared to mussels, and fresh mussels had elevated counts of enterobacteria compared to frozen ones. Among the 10 E. coli isolates, none carried the genes blaCTX-M-1, blaCTX-M-2, blaCTX-M-3, blaCTX-M-15, mcr-1, mcr-2, mcr-3, mcr-4, and tet, associated with antibiotic resistance. Phenotypical resistance to tetracycline and fosfomycin was not observed in any isolate, while only 20% demonstrated resistance to ciprofloxacin. Regarding ampicillin and amoxicillin with clavulanic acid, 60% of isolates were resistant, 10% showed intermediate susceptibility, and 30% were sensitive. One isolate was considered simultaneously resistant to ß-lactams and quinolones, and none were conserved as ESBL producers. These findings highlight the inherent risks to local public health that arise from consuming improperly prepared seafood in this area.

Efficacy of Duddingtonia flagrans (Bioverm®) on the biological control of buffalo gastrointestinal nematodes.

We evaluated the efficacy of Bioverm®, a commercial product containing Duddingtonia flagrans, on the control of buffalo (Bubalus bubalis) gastrointestinal nematodes. We randomly divided 12 buffaloes into two groups of six animals. In the treated group, each animal received a Bioverm®`s single dose of 1g (105 chlamydospores of D. flagrans) to 10 kg of live weight; in the control group, each animal received 1g of corn bran for each 10 kg of live weight as a placebo. Fecal samples were individually collected from 12, 24, 36, 48, 60 and 72 h after treatments. To examine 1) viability of chlamydospores passed through the gastrointestinal tract, 2 g of faeces and 1000 infective larvae (L3) were added to Petri dishes with 2% water-agar, and 2) to examine larval predation by D. flagrans during fecal cultures, 2000 L3 were added. In the Petri dishes, were observed significant reductions (p < 0.01) in the treated group after 48 (56.7%) and 60 h (91.5%). In the fecal cultures, significant reductions (p < 0.01) occurred in the treated group from 36 h (75%), with larval reduction up to 72 h. High larval predation rate occurred 60 h after Bioverm® administration. Bioverm® maintained viability and predation capacity after passage through the buffalo's gastrointestinal tract, showing efficacy on gastrointestinal nematodes.

Association between Duddingtonia flagrans, dimethylsulfoxide and ivermectin for the control of Rhabditis spp. in cattle.

Cattle parasitic otitis caused by the nematode Rhabditis spp. is a serious health problem in Brazil, a situation which is confounded by lack of effective control measures. In vitro studies associating biological and chemical control as an alternative method showed promising results. The objective was to evaluate the combined use of Duddingtonia flagrans (AC001), 10% dimethylsulfoxide (DMSO), and 1.9% ivermectin for the in vivo control of Rhabditis spp., in naturally infected Gyr cattle. For this purpose, 48 animals, whose infection in both ears was diagnosed, were randomly assigned to 6 groups: group 1 (ivermectin 1.9%); group 2 (10% DMSO); group 3 (AC001); group 4 (ivermectin 1.9% + 10% DMSO w/v); group 5 (1.9% ivermectin + AC001 w/v); group 6 (10% DMSO + AC001 v/v). The treatments were performed in a single dose, in the right ears, with the left ears remaining untreated, as a control group. There was a significant reduction (p < 0.01) in the number of nematodes in the treated groups in relation to the control, with the following best efficacies: groups 1 and 2, 47% and 52.9%, respectively, 7 days after treatment; groups 3, 4, and 5, 47.8%, 48.6% and 36.7%, respectively, 14 days post-treatment; group 6, 38.4%, 21 days post-treatment. It was concluded that the combination of chemical compounds and D. flagrans in a single application was effective for the in vivo control of Rhabditis spp. in naturally infected cattle.

Lynxacarus radovskyi (Acari: Listrophoridae) in a domestic cat from Espírito Santo, Brazil / Lynxacarus radovskyi (Acari: Listrophoridae) em gato doméstico no Espírito Santo, Brasil

Lynxacarus radovskyi is of uncommon occurrence and there are few reports in the state of Espírito Santo, Southeastern Brazil. The purpose of the present note is to record the occurrence of L. radovskyi in a Persian domestic cat in Vila Velha, Espírito Santo, Brazil. The animal was asymptomatic, without evidence of dermatological signs. The present report can be informative, once the lack of knowledge in relation to the ectoparasite presence and potential of spread in the regional cat population is constant in the veterinary medicine practice,may cause underreporting of infestations.

Lynxacarus radovskyi é de ocorrência incomum e existem poucos relatos no estado do Espírito Santo, Sudeste do Brasil. O objetivo do presente relato foi reportar a ocorrência de L. radovskyi em felino doméstico da raça Persa em Vila Velha, Espírito Santo, Brasil. O animal apresentava-se assintomático à infestação, sem evidências de alterações dermatológicas. Este relato pode ser informativo, uma vez que a falta de conhecimento em relação à presença e potencial disseminativo deste ectoparasito é constante na prática da Medicina Veterinária, podendo causar subnotificações das infestações.

Lynxacarus radovskyi (Acari: Listrophoridae) in a domestic cat from Espírito Santo, Brazil / Lynxacarus radovskyi (Acari: Listrophoridae) em gato doméstico no Espírito Santo, Brasil

Lynxacarus radovskyi is of uncommon occurrence and there are few reports in the state of Espírito Santo, Southeastern Brazil. The purpose of the present note is to record the occurrence of L. radovskyi in a Persian domestic cat in Vila Velha, Espírito Santo, Brazil. The animal was asymptomatic, without evidence of dermatological signs. The present report can be informative, once the lack of knowledge in relation to the ectoparasite presence and potential of spread in the regional cat population is constant in the veterinary medicine practice,may cause underreporting of infestations.

Lynxacarus radovskyi é de ocorrência incomum e existem poucos relatos no estado do Espírito Santo, Sudeste do Brasil. O objetivo do presente relato foi reportar a ocorrência de L. radovskyi em felino doméstico da raça Persa em Vila Velha, Espírito Santo, Brasil. O animal apresentava-se assintomático à infestação, sem evidências de alterações dermatológicas. Este relato pode ser informativo, uma vez que a falta de conhecimento em relação à presença e potencial disseminativo deste ectoparasito é constante na prática da Medicina Veterinária, podendo causar subnotificações das infestações.

Germination capacity of the Pochonia chlamydosporia fungus after its passage through the gastrointestinal tract of domestic chickens (Gallus gallus domesticus).

This study evaluated the germination capacity of Pochonia chlamydosporia (VC4) fungus after its passage through the gastrointestinal tract of domestic chickens and its interaction with Ascaridia galli and Heterakis gallinarum eggs. Twenty-two domestic chickens were divided in two groups: control group (G1) received shredded corn substrate without VC4; and treatment group (G2) received a single dose of 29 g corn substrate containing 3.3 × 106 conidia/chlamydospores (VC4). Subsequently, chicken fecal samples were collected at intervals of 0, 6, 8, 10, 12, 18 and 24 h. Petri dishes from fecal samples of the treated group (G2) were subdivided (G2a and G2b), and then replicated in 2% agar-water medium for the microbiological test. After VC4 growth, approximately 200 eggs of A. galli (G2a) and H. gallinarum (G2b) were added to each subgroup to evaluation of ovicidal activity. There was fungal viability after passage through chicken gastrointestinal tract and egg predation of 59.9% and 43.2% for A. galli and H. gallinarum, respectively. The present work demonstrates the ability of the fungus P. chlamydosporia to survive after passing through the gastrointestinal tract of domestic chickens, an extreme environment (low pH, enzymes, microbiota and mechanical action), and still germinate after being excreted with feces.

Effect of silver nanoparticles (AgNP's) from Duddingtonia flagrans on cyathostomins larvae (subfamily: cyathostominae).

The in vitro effect of silver nanoparticles of the Duddingtonia flagrans filtrate enriched with chitin was evaluated on infective larvae of cyathostomins (L3). After biosynthesis, an assay was carried out with two experimental groups in microtubes, for a period of 24 h: G1 (AgNP's-D. flagrans (43.4 µg/mL) + 120 L3) and G2 (distilled water + 120 L3). At the end of this period, AgNP's-D. flagrans (G1) demonstrated an effect on L3 with a 43% reduction (p < 0.01) in relation to G2. Thus, the authors suggest new designs with AgNP's-D. flagrans for the control of cyathostomins.

Efficiency of the Bioverm ® (Duddingtonia flagrans) fungal formulation to control in vivo and in vitro of Haemonchus contortus and Strongyloides papillosus in sheep.

The objective of the present work was to evaluate the efficiency of Bioverm® fungal formulation (Duddingtonia flagrans-AC001) in controlling Haemonchus contortus and Strongyloides papillosus in sheep. In vitro predation tests were carried out in Petri dishes containing agar culture medium 2%. Four experimental groups were formed, with five replicates each: Group 1: 1 g of Bioverm ® and 1000 third-stage larvae (L3) of H. contortus; Group 2: 1000 L3 of H. contortus; Group 3: 1 g of Bioverm ® and 1000 L3 of S. papillosus; and Group 4: 1000 L3 of S. papillosus. In the in vivo tests, twelve 11-month-old sheep males positive for H. contortus were used. The animals were sorted in two groups (treatment and control), based on fecal egg counts (eggs per gram, EPG). Each group comprised six animals: treatment group-each animal received orally 100 g of Bioverm ® ; and control group-each animal received orally 100 g of rice. Subsequently, feces from these animals were collected at 12, 24, 36, 48, 60, 72, 84, and 96 h after Bioverm ® administration. In vitro results demonstrate that D. flagrans kept its predatory activity with 91.5% of mean reduction percentage of L3. After the passage test, Bioverm ® presented efficacy already after 12 h of its administration and kept similar results for 60 h. Bioverm® fungal formulation (D. flagrans-AC001) was efficient in reducing the population of H. contortus and S. papillosus under laboratory conditions in sheep feces. However, further studies are needed under natural conditions of ruminant grazing to prove the efficiency of this product.

Association between infestation by Lynxacarus radovskyi (Acari: Lystrophoridae) and the occurrence of Feline Eosinophilic Granuloma Complex.

Lynxacarus radovskyi are mites commonly found within domestic feline hair stems. The infested animal presents an opaque fur with a "salt and pepper" aspect. The contamination may occur by direct contact with other infected animals or by fomites, and the main diagnostic tool is the direct examination of the fur. The Feline Eosinophilic Granuloma Complex (FEGC) is a dermatological pattern of an immune answer against many types of injuries in felines, mainly during allergic reactions, and the treatment of choice are corticosteroids. The objective of this work was to document the linxcariosis cases at the Veterinary Hospital Professor Ricardo Alexandre Hippler-University of Vila Velha. Futhermore, the work also meant to research the association between the parasite contagion and the occurrence of FEGC injuries, which commonly occurred. Between April 2017 and August 2018, there were 8 reported cases of felines infested with L. radovskyi. The diagnoses were made with Acetate tape impression or by direct fur examination. Seven out of the eight cats presented with at least one characteristic injury of the FEGC. In all of them the chosen treatment was moxidectin or spot-on imidacloprid plus corticotherapy on the cats with FEGC, which were effective.

Combined use of ivermectin, dimethyl sulfoxide, mineral oil and nematophagous fungi to control Rhabditis spp.

Rhabditis spp., is a nematode known to cause otitis externa, an infection difficult to control, in cattle reared within tropical regions. The objective of this study was to assess the combined use of ivermectin 1%, dimethyl sulfoxide 1% and mineral oil 100% containing nematophagous fungi of both Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34) species to control in vitro Rhabditis spp. Thus, 12 experimental groups were designed with eight replicates each: G1 (nematodes + AC001); G2 (nematodes + NF34); G3 (nematodes + ivermectin 1%/positive control); G4 (nematodes + dimethyl sulfoxide 1%/positive control); G5 (nematodes + mineral oil 100%/positive control); G6 (nematodes + AC001 + ivermectin 1%); G7 (nematodes + NF34 + ivermectin 1%); G8 (nematodes + AC001 + mineral oil 100%); G9 (nematodes + NF34 + mineral oil 100%); G10 (nematodes + AC001 + dimethyl sulfoxide 1%); G11 (nematode + NF34 + dimethyl sulfoxide 1%); G12 (nematode + distilled water/negative control). The results demonstrated that all experimentally treated groups differed statistically (p < 0.01) from the control group. In the present study, the use of dimethyl sulfoxide 1% and mineral oil 100% in conjunction with conidia fungi portrayed noteworthy outcomes, which represents a future premise for the combined use of nematophagous fungi within these vehicles in both controlling Rhabditis spp.

Rhabditis spp., in the Espírito Santo, State of Brazil and evaluation of biological control.

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.

Nematicidal activity of silver nanoparticles from the fungus Duddingtonia flagrans.

BACKGROUND: Helminth parasites cause morbidity and mortality in both humans and animals. Most anthelmintic drugs used in the treatment of parasitic nematode infections act on target proteins or regulate the electrical activity of neurons and muscles. In this way, it can lead to paralysis, starvation, immune attack, and expulsion of the worm. However, current anthelmintics have some limitations that include a limited spectrum of activity across species and the threat of drug resistance, which highlights the need for new drugs for human and veterinary medicine. PURPOSE: Present study has been conducted to determine the anthelmintic activity of silver nanoparticles (AgNPs) synthesized from the extract of nematophagous fungus, Duddingtonia flagrans, on the infecting larvae of Ancylostoma caninum (L3). METHODS: The nanoparticles were characterized by visual, ultraviolet, Fourier-transform infrared spectroscopy, transmission electron microscopy (TEM) analysis, and X-ray diffraction. The in vitro study was based on experiments to inhibit the motility of infective larvae (L3), and the ultrastructural analysis of the nematode was performed by images obtained by TEM. RESULTS: The XRD studies revealed the crystalline nature of the nanoparticles, and FTIR results implied that AgNPs were successfully synthesized and capped with compounds present in the extract. The results showed that the green synthesis of AgNPs exhibited nematicidal activity, being the only ones capable of penetrating the cuticle of the larvae, causing changes in the tegmentum, and consequently, the death of the nematode. CONCLUSION: The extract of the fungus D. flagrans is able to synthesize AgNP and these have a nematicidal action.

Rhabditis spp, in the Espírito Santo, State of Brazil and evaluation of biological control / Rhabditis spp, no Espírito Santo, estado do Brasil e avaliação do controle biológico

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.

Resumo Os objetivos neste estudo foram descrever ocorrências do nematódeo Rhabditis spp., causando otite parasitária em bovinos leiteiros da raça Gir no estado do Espírito Santo, sudeste do Brasil, e avaliar o controle biológico desse nematódeo utilizando os fungos nematófagos Duddingtonia flagrans (AC001) e Monacrosporium thaumasium (NF34). Após a detecção e coleta dos nematódeos, três grupos foram formados: dois grupos que foram tratados com os isolados fúngicos, respectivamente; e um grupo controle, sem fungos. Os tratamentos foram os seguintes: (a) placas de Petri contendo o meio de cultura 2% ágar de água (WA) + 250 nematoides + AC001; (b) placas de Petri contendo 2% de WA + 250 nematoides + NF34; e (c) placas de contendo apenas 2% de nematódeos WA + 250. Após sete dias a 27 °C os tratamentos com fungos foram capazes de capturar e destruir os nematódeos, com porcentagens de 82,0% (AC001) e 39,0% (NF34) em relação ao grupo controle. Os resultados demonstram a ocorrência de Rhabditis spp., no Estado do Espírito Santo e a eficácia da atividade predatória in vitro dos isolados fúngicos utilizados. Assim, esses resultados são importantes, pois podem auxiliar no controle alternativo in vivo de Rhabditis spp. em bovinos com otite parasitária.

Leporacarus gibbus (Acari: listrophoridae) in a naturally-infested domestic rabbit: first report of its occurrence in the state of Espírito Santo, Brazil / Leporacarus gibbus (Acari: listrophoridae) em um coelho doméstico naturalmente infestado: primeiro relato de sua ocorrência no estado do Espírito Santo, Brasil

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.(AU)

Este trabalho teve como objetivo relatar a primeira ocorrência de Leporacarus gibbus em um coelho doméstico (Oryctolagus cuniculus) do estado do Espírito Santo, Brasil. O coelho macho de dois anos de idade veio de uma área rural e foi internado no Hospital Veterinário Professor Ricardo Alexandre Hippler com intenso prurido em todo o corpo, severa queda de cabelo, lesões crustais na superfície externa das orelhas e descamação principalmente na região dorsal. A raspagem da pele e o tricograma foram realizados. Na avaliação microscópica dos pêlos obtidos na região dorsal, observaram-se ácaros com morfologia compatível com L. gibbus. Cheyletiella parasitovorax e Psoropotes cuniculi também foram detectados na mesma região. O tratamento com ivermectina (0,4 mg / kg) foi prescrito com três aplicações a cada 14 dias, durante um período de seis semanas. Este relato apresenta a primeira descrição da ocorrência de L. gibbus no estado do Espírito Santo concomitante à poliinfestação de C. parasitovorax e P. cuniculi, e espera-se que esta seja uma ferramenta de pesquisa para futuros trabalhos na região.(AU)

Colonization and destruction of ants of the genus Camponotus sp. (Hymenoptera: Formicidae) in vitro by the fungus Pochonia chlamydosporia in the southeast region of Brazil.

The objective of this study was to evaluate, in vitro, the colonization and destruction of ants of the genus Camponotus sp. by the ovicidal fungus Pochonia chlamydosporia (VC4 isolate), in the southeast region of Brazil. The insects used in the experiment were worker ants of the genus Camponotus sp., collected periodically in the environment and immediately transported to the laboratory in test tubes. Then, VC4 growth was promoted in 2% chitin agar medium (2% WQ) to obtain a fungal solution containing conidia and/or chlamydospores. Two experimental groups were formed. Treated group consisted of Petri dishes containing 2% agar-water culture medium (2% WA) with nine live insects and 20 µL of fungal solution at the concentration of 15,000 conidia/chlamydospores. Control group consisted of Petri dishes containing 2% WA culture medium and nine live insects. The dishes in the treated and control groups were incubated in BOD at 25 ± 1 °C and 80 ± 10% relative humidity for 4 days. After 4 days, it was observed that the VC4 had grown, colonized, and caused the destruction of the ants. The fungus P. chlamydosporia was efficient at colonizing and destroying the urban ants collected on an experimental basis. Thus, it could open up new ways to reduce the use of chemical compounds in the future, decreasing health and environmental problems.

Coadministration of Nematophagous Fungi for Biological Control over Nematodes in Bovine in the South-Eastern Brazil.

This study compared the coadministration among the three nematode predatory fungi, Duddingtonia flagrans, Monacrosporium thaumasium, and Arthrobotrys robusta, in the biological control of cattle gastrointestinal nematodiasis in comparison with the use of the fungus D. flagrans alone. Five groups consisting of eight Girolando heifers were kept in paddocks of Brachiaria decumbens for six months. Each heifer received 1 g/10 kg of pellets containing the fungi (0.2 g of fungus/10 kg b.w.). Group 1 (G1) received pellets with D. flagrans and M. thaumasium in coadministration, G2 received D. flagrans and A. robusta, G3 received M. thaumasium, A. robusta, and D. flagrans, and G4 received the fungus D. flagrans alone. Group 5 (control) received pellets without fungi. The monthly mean of fecal egg count (FEC) of Groups 1, 2, 3, and 4 were 93.8, 85.3, 82.7, and 96.4% smaller than the mean of control group. The treatments with pellets containing D. flagrans or D. flagrans + M. thaumasium produced significantly better results than the D. flagrans + A. robusta or the combination of the three fungi. The associations which include A. robusta were less efficient in this study than D. flagrans alone or associated with M. thaumasium.

Duddingtonia flagrans formulated in rice bran in the control of Oesophagostomum spp. intestinal parasite of swine.

Three experimental assays with Duddingtonia flagrans (isolated AC001) were carried out. The growth of the genus Duddingtonia present in formulation of rice bran, its predatory capability on Oesophagostomum spp. infective larvae (L3) in petri dishes (assay 1), its action in faecal cultures with eggs of that parasite (assay 2) and isolate's capability of predation after passing through gastrointestinal tract of swine (assay 3) was evaluated. At assay 3, feces were collected at time intervals of 12, 24, 36, 48, and 60 h after feed animals with the formulation. Assays 1 and 2 showed a statistical difference (p < 0.01) by the F test when comparing the treated group with the control group. At the both assays, was observed in the treated group a reduction percentage of 74.18% and 88.38%, respectively. In assay 3, there was a statistical difference between the treated group and the control group at all collection times (p < 0.01). Regarding the collection periods, there was no statistical difference over time in the treatment group (p > 0.05). The results demonstrate that the fungal isolate AC001 formulated in rice bran can prey on L3 of Oesophagostomum spp., in vitro and after passing through the gastrointestinal tract, without loss of viability. This isolate may be an alternative in the control of Oesophagostomum spp. in swine.

Extracellular biosynthesis of silver nanoparticles using the cell-free filtrate of nematophagous fungus Duddingtonia flagrans.

The biosynthesis of metallic nanoparticles (NPs) using biological systems such as fungi has evolved to become an important area of nanobiotechnology. Herein, we report for the first time the extracellular synthesis of highly stable silver NPs (AgNPs) using the nematophagous fungus Duddingtonia flagrans (AC001). The fungal cell-free filtrate was analyzed by the Bradford method and 3,5-dinitrosalicylic acid assay and used to synthesize the AgNPs in the presence of a 1 mM AgNO3 solution. They have been characterized by UV-Vis spectroscopy, X-ray diffraction, transmission electron microscopy, dynamic light scattering, Zeta potential measurements, Fourier-transform infrared, and Raman spectroscopes. UV-Vis spectroscopy confirmed bioreduction, while X-ray diffractometry established the crystalline nature of the AgNPs. Dynamic light scattering and transmission electron microscopy images showed approximately 11, 38 nm monodisperse and quasispherical AgNPs. Zeta potential analysis was able to show a considerable stability of AgNPs. The N-H stretches in Fourier-transform infrared spectroscopy indicate the presence of protein molecules. The Raman bands suggest that chitinase was involved in the growth and stabilization of AgNPs, through the coating of the particles. Our results show that the NPs we synthesized have good stability, high yield, and monodispersion.

Nematophagous fungi combinations reduce free-living stages of sheep gastrointestinal nematodes in the field.

Gastrointestinal nematodes (GIN) can reduce or limit sheep production. Currently there is a clear deficiency in the action of drugs for the control of these parasites. Nematophagous fungi are natural enemies of GIN. Fungal combinations have potential for reducing GIN populations. The aim of this study was to evaluate the efficiency combinations of nematophagous fungi in sodium alginate matrix pellets for the biological control agents of gastrointestinal sheep nematode parasites in the field. The nematophagous fungi (0.2mg of fungus per kg of body weight), Arthrobotrys conoides, A. robusta, Duddingtonia flagrans, and Monacrosporium thaumasium were used. The treated groups were administered mycelium combinations in the following combinations: group 1 (D. flagrans+A. robusta); group 2 (M. thaumasium+A. conoides). The control group did not receive any fungal pellets. We used three groups with eight Santa Inês sheep each. Each animal was treated with approximately 1g of pellet per 10kg of live weight. During the experimental period, we evaluated: number of eggs per gram of feces (EPG), infective larvae (L3) per kg of dry matter, larvae recovered from coprocultures, packed cell volume, total plasma protein concentration of sheep, and environmental conditions. Group 2 EPG (M. thaumasium+A. conoides) differed from the control group in September and October. The number of L3/kg of dry matter recovered from animals of groups 1 and 2 at distances of 0-20 and 20-40cm from the fecal pats was lower than the control group. The packed cell volume and total plasma proteins of treated animals were similar to those of the control group. The combination of treatment groups (D. flagrans+A. robusta and M. thaumasium+A. conoides) reduced the number of L3/kg of pasture. Therefore, treatment of nematophagous fungal combinations have the potential to manage free-living stages of GIN in sheep.

Viability of Strongyloides venezuelensis eggs and larvae in vermiculite containing the fungus Duddingtonia flagrans.

Strongyloidiasis is the most clinically important disease among the infections caused by geohelminths, seeing that this parasite can cause autoinfection. The use of nematophagous fungi like Duddingtonia flagrans, that have predation action on eggs and infecciososas forms of helminths, emerges as an alternative method for environmental control. For this reason, analyzing the viability of larvae and eggs of Strongyloides venezuelensis and the action of Duddingtonia flagrans AC001 in vermiculite, as well as the action of the nematophagous fungi in different growth stages, is important to elaborate and define the best culture conditions that favor the activity of the fungus. Two different growth conditions were applied: both eggs and AC001 fungi were added at the same time to the vermiculite (assay A) and the addition of eggs after the growth of the AC001 fungi in the vermiculite (assay B). To recover the L3 larvae, the Baermann-Moraes method was applied, followed by the counting of L3 dead and alive. At last, it was observed that the vermiculite enriched with organic material is an adequate culture medium not only for the growth of the S. venezuelensis but also for the growth of the D. flagrans fungus, being therefore, a satisfactory culture medium for tests of viability and predatory action of this fungus. It was also observed that the activity of the AC001 fungus is greater when it is growing concomitantly with the eggs, in other words, when it is in the adaptation phase.