Novo: Integrated control of avian gastrointestinal parasites: optimizing their coprological diagnosis and biocontrol using predatory fungi. Inicial: Gastrointestinal Parasites of Domestic Birds and their Biological Control with Predatory Fungi

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Biocontrol of Avian Gastrointestinal Parasites Using Predatory Fungi: Current Status, Challenges, and Opportunities

Publication . Lozano, João; Almeida, Cristina; Oliveira, Manuela; Paz-Silva, Adolfo; Madeira de Carvalho, Luís

This review describes the current research status regarding the implementation of predatory fungi in the biological control approach of bird gastrointestinal (GI) parasitosis. The main GI parasites of Galliformes (e.g., broilers, layers, peacocks, pheasants) and Ratites (e.g., ostriches, emus, rheas) are addressed, as well as their impact on farms, zoos, and private collections. The main characteristics regarding biocontrol with predatory fungi are briefly described, such as their mode of action and efficacy against GI parasites of different animal hosts. The state of the art regarding the use of predatory fungi in birds is reviewed here by describing all associated articles already published in the main databases, techniques, and their main findings. Ovicidal fungi such as Pochonia chlamydosporia, Metarhizium spp. and Acremonium spp., and larvicidal fungi, namely Duddingtonia flagrans, Arthrobotrys spp. and Monacrosporium thaumasium, have shown promising predacious activity against ascarid eggs and nematode larvae from chickens and ostriches, both in vitro and in vivo, also revealing tolerance to the GI passage in chickens and maintenance of predacious capacity. Further studies are needed to understand the fungi–parasite–host gut microbiota interactions and target other avian GI parasitic species, such as nematodes, coccidia, cestodes, and trematodes.

2022-03-01Journal article

Open access

Integrated control of avian gastrointestinal parasites : optimizing their coprological diagnosis and biocontrol using predatory fungi

Publication . Lozano, João; Lozano, João Miguel Pestana; Carvalho, Luís Manuel Madeira de; Oliveira, Maria Manuela Castilho Monteiro de; Paz-Silva, Adolfo

In the last 30 years, several improvements were achieved in the control of animals’ gastrointestinal (GI) parasitism, including the development of more sensitive and precise coprological diagnosis techniques, like Mini-FLOTAC (MF), as well as the use of predatory fungi as a sustainable solution for parasite control. This project aimed to optimize the MF method for the diagnosis of avian GI parasitism, and to isolate predatory fungi and assess their potentialities in avian parasite control. First, the diagnosis of GI parasitism was performed in a poultry farm and three exotic bird collections, in Portugal mainland, using the MF technique. Eimeria spp. infections were identified in laying hens and peacocks, Capillaria spp., Trichostrongylus tenuis and Strongyloides pavonis in peacocks, as well as Libyostrongylus douglassii in ostriches and emus. Seven filamentous fungi were isolated from Galliformes feces, furtherly identified as Mucor circinelloides (n=6) and M. lusitanicus (n=1). All fungi were capable of destroying Eimeria spp. oocysts in vitro, with M. circinelloides isolate FR1 achieving the highest coccidicidal efficacy, 22%, after 14 days of exposure. All fungi were checked for potential in vitro susceptibilities to seven antiparasitic drugs (albendazole, fenbendazole, levamisole, ivermectin, lasalocid, amprolium and toltrazuril), and all were not susceptible to the tested antiparasitic drugs, independently of their concentrations. The characterization of the virulence profile of M. circinelloides FR1 revealed only the presence of genes coding for iron receptors (FOB1 and FOB2), iron permease (FTR1), ADP-ribosylation factors (ARF2, ARF6) and GTPase (CDC42), having the isolate only tested positive for lecithinase production. Oral administrations of spores of this fungal isolate were performed in laying hens and peacocks, which revealed that this fungus did not affect birds’ feces appearance and consistency (70 100% of normal feces), neither the relative abundances and alfa-diversities of their native gut bacteria and fungi. Finally, it was verified that the administration of this fungus spores to peacocks reduced significantly their Eimeria spp. fecal shedding up to 92%, after two months of trial. Results from this project point out the usefulness of MF in the coprological diagnosis of GI parasitism in birds, and provide the first report regarding the isolation of predatory fungi from bird feces. Finally, the safety and efficacy of using M. circinelloides FR1 in the integrated control of domestic and exotic birds GI parasitism was demonstrated

2025-02-07Doctoral thesis

Open access

First insights on the susceptibility of native coccidicidal fungi Mucor circinelloides and Mucor lusitanicus to different avian antiparasitic drugs

Publication . Lozano, João; Cunha, Eva; Madeira de Carvalho, Luís; Paz-Silva, Adolfo; Oliveira, Manuela

Background The combined application of predatory fungi and antiparasitic drugs is a sustainable approach for the integrated control of animal gastrointestinal (GI) parasites. However, literature addressing the possible interference of antiparasitic drugs on the performance of these fungi is still scarce. This research aimed to assess the in vitro susceptibility of six native coccidicidal fungi isolates of the species Mucor circinelloides and one Mucor lusitanicus isolate to several antiparasitic drugs commonly used to treat GI parasites’ infections in birds, namely anthelminthics such as Albendazole, Fenbendazole, Levamisole and Ivermectin, and anticoccidials such as Lasalocid, Amprolium and Toltrazuril (drug concentrations of 0.0078–4 µg/mL), using 96-well microplates filled with RPMI 1640 medium, and also on Sabouraud Agar (SA). Results This research revealed that the exposition of all Mucor isolates to the tested anthelminthic and anticoccidial drug concentrations did not inhibit their growth. Fungal growth was recorded in RPMI medium, after 48 h of drug exposure, as well as on SA medium after exposure to the maximum drug concentration. Conclusions Preliminary findings from this research suggest the potential compatibility of these Mucor isolates with antiparasitic drugs for the integrated control of avian intestinal parasites. However, further in vitro and in vivo studies are needed to confirm this hypothesis.

2024-02-23Journal article

Open access

Control of Strongyles in first-season grazing ewe lambs by integrating deworming and thrice-weekly administration of parasiticidal fungal spores

Publication . Voinot, Mathilde; Bonilla, Rodrigo; Sousa, Sérgio; Sanchís, Jaime; Canhão-Dias, Miguel; Delgado, José Romero; Lozano, João; Sánchez-Andrade, Rita; Arias, Maria Sol; Madeira de Carvalho, Luís

Parasiticidal fungi have been used in several in vivo experiments in livestock farms worldwide, constituting an effective tool for the biocontrol of gastrointestinal parasites in grazing animals. In the first year of study, two groups of eight first-season pasturing ewe lambs infected by strongyles were dewormed with albendazole, and then, the test group received an oral dose of 106 chlamydospores of Mucor circinelloides and 106 Duddingtonia flagrans individually and thrice a week from mid-September to May (FS1), while the control group remained without fungi (CT1). In the second year, two new groups of first-season grazing ewe lambs were treated with ivermectin and subjected to the same experimental design (FS2 and CT2, respectively). The anthelmintic efficacy was 96.6% (CT1), 95.6% (FS1), 96.1% (CT2), and 95.1% (FS2). The counts of strongyle egg output increased in the control groups (CT1 and CT2) throughout the study and reached numbers higher than 600 eggs per gram of feces (EPG), while in FS1 and FS2, they were <250 EPG. The values of red blood cell parameters registered for CT1 and CT2 were lower than those of the reference standards, while a significant increment was recorded in FS1 and FS2, and values within the physiological range were attained. It is concluded that integrating efficient anthelminthic deworming with rotational pasturing and the regular intake of chlamydospores of M. circinelloides and D. flagrans provides a helpful strategy for maintaining low levels of strongyle egg output in first-season grazing ewe lambs and improves their health status.

2021-10-17Journal article

Open access

Analyzing the safety of the parasiticide fungus Mucor circinelloides: first insights on its virulence profile and interactions with the avian gut microbial community

Publication . Lozano, João; Cunha, Eva; Almeida, Cristina; Nunes, Mónica; Dias, Ricardo; Vicente, Eduardo; Sebastião, Daniela; Henriques, Sérgio; Madeira de Carvalho, Luís; Paz-Silva, Adolfo; Oliveira, Manuela

Parasiticide fungi are considered an accurate, sustainable, and safe solution for the biocontrol of animal gastrointestinal (GI) parasites. This research provides an initial characterization of the virulence of the native parasiticide fungus Mucor circinelloides (FMV-FR1) and an assessment of its impact on birds’ gut microbes. The genome of this fungus was sequenced to identify the genes coding for virulence factors. Also, this fungus was checked for the phenotypic expression of proteinase, lecithinase, DNase, gelatinase, hemolysin, and biofilm production. Finally, an in vivo trial was developed based on feeding M. circinelloides spores to laying hens and peacocks three times a week. Bird feces were collected for 3 months, with total genomic DNA being extracted and subjected to long-read 16S and 25S-28S sequencing. Genes coding for an iron permease (FTR1), iron receptors (FOB1 and FOB2), ADP-ribosylation factors (ARFs) (ARF2 and ARF6), and a GTPase (CDC42) were identified in this M. circinelloides genome. Also, this fungus was positive only for lecithinase activity. The field trial revealed a fecal microbiome dominated by Firmicutes and Proteobacteria in laying hens, and Firmicutes and Bacteroidetes in peacocks, whereas the fecal mycobiome of both bird species was mainly composed of Ascomycetes and Basidiomycetes fungi. Bacterial and fungal alpha-diversities did not differ between sampling time points after M. circinelloides administrations (P = 0.62 and P = 0.15, respectively). Although findings from this research suggest the lack of virulence of this M. circinelloides parasiticide isolate, more complementary in vitro and in vivo research is needed to conclude about the safety of its administration to birds, aiming at controlling their GI parasites.

2024-05-02Journal article

Open access