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Evaluation of Fungal Parasitic Activity Under Field Conditions: A Soil Simulation Test
(MDPI, 2026-04-13) Magos Amado, Carlos Emiliano; Zubiria Ibarguren, Izaro; Hernández Malagón, José Ángel; Sanchís Polto, Jaime; Paz Silva, Adolfo; González Cerón, Fernando; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal
Several species of saprophytic filamentous fungi are able of disrupting the life cycle of certain soil-born parasites that are of veterinary and agronomy importance, offering a promising sustainable control alternative. This study consisted of designing an experimental model, using catnip (Nepeta cataria) trays to simulate a vegetated environment for evaluating the parasiticidal activity of Mucor circinelloides, Trichoderma atrobrunneum, and Duddingtonia flagrans. Fungal spores were added to treated trays before adding feces with protozoan (Eimeria spp.), and gastrointestinal nematodes (roundworms, strongyles), and untreatedcontrol water. No differences in plant growth or vigor, regardless of fungal presence, were observed, confirming the safety of these biological agents for vegetation. In the control trays, the viability of parasites ranged from 50% to 85%. In the treated trays, the viability of Eimeria and roundworms decreased by 40–100%, and the strongyle egg counts were reduced by 74% within 15 days. It is concluded that the vegetated tray model effectively simulates field conditions and provides a reliable platform for evaluating fungal efficacy against the free-living stages of parasites, offering a versatile tool for future research on soil-borne pathogens affecting animals and plants.
Nanoparticle-Based Approaches for Enhancing In Vitro Fertilization in Animal Reproduction
(MDPI, 2026-03-18) Gałęska, Elżbieta; Kowalczyk, Alicja; Samardžija, Marko; Gračner, Gregurić Gordana; Wrzecinska, Marcjanna; Araujo, Jose Pedro; Hernández Malagón, José Ángel; Camiña García, María Mercedes; Czerniawska-Piatkowska, Ewa; Dobrzanski, Zbigniew; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal
Nanotechnology, based on nanoparticles, has become an emerging interdisciplinary tool in reproductive biotechnology, offering innovative opportunities to improve fertilization efficiency and reproductive performance in farm animals. The purpose of this review is to provide an updated synthesis of current research on nanoparticle-based approaches that enhance in vitro fertilization outcomes and other assisted reproductive technologies. The focus is on the biological mechanisms, potential benefits, and limitations of nanoparticle use in animal reproduction. Nanoparticles—including gold, silver, zinc oxide, selenium, and magnetic iron oxide—exhibit distinctive physicochemical properties that enable targeted interactions with gametes and reproductive cells. When used in semen extenders or culture media, nanoparticles improve sperm motility, acrosome and membrane integrity, and reduce oxidative stress and apoptosis. These effects contribute to enhanced fertilization rates and higher embryo developmental competence. In addition, nanoparticles can function as carriers for hormones, antioxidants, and growth factors, stabilizing reagents essential for oocyte maturation, sperm capacitation, and early embryo culture. The review also discusses nanopurification (selectively isolating and removing particles) and nanosorting (separating or organizing nanoscale objects) techniques that allow for non-invasive selection of viable gametes, and fluorescence- and magnet-assisted sorting systems that increase precision in sperm sexing. The mechanical aspects of nanoparticle–cell interactions are analyzed, emphasizing the influence of particle size, dose, and surface modification on both biological efficacy and cytotoxicity. Safety, toxicological concerns, and regulatory frameworks—including International Organization for Standardization (ISO) standards and European Commission recommendations—are critically reviewed to highlight the need for harmonized biocompatibility criteria. Although nanoparticle use in animal reproduction remains largely experimental, accumulated evidence demonstrates its potential to improve reproductive efficiency and reduce economic losses. Integrating nanoparticle-based systems with existing reproduction platforms may represent a transformative step toward sustainable and precision-driven livestock breeding.
Testing a Sustainable Strategy Against Poultry Helminth Stages Developing in the Soil
(MDPI, 2025-11-15) León González, Jorge Alexander; Pérez Anzúrez, Gustavo; Abreu Ramos, Inês; Magos Amado, Carlos Emiliano; Boso Dafonte, David; Lozano, João; Hernández Malagón, José Ángel; Cazapal Monteiro, Cristiana Filipa; Bonilla, Rodrigo; Sanchís, Jaime; Paz Silva, Adolfo; Sánchez-Andrade Fernández, Rita; Carvalho, Luis Manuel Madeira de; Arias Vázquez, María Sol; Universidade de Santiago de Compostela. Facultade de Veterinaria; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal
Free-ranging hens are at risk of infection by parasites characterized by certain stages that develop in the soil until attaining the infective phase. To analyze the usefulness of a biological control strategy of helminths affecting pasturing hens, fecal samples containing eggs of the helminths Ascaridia galli and Capillaria spp. were collected and then homogenized with an electric mixer. A total of 64 small areas were established by placing wooden frames (15 × 40 × 30 cm) on the ground and then adding approximately 100 g of a fecal mixture (per area). Four batches of 16 areas were considered: G1, sprayed with 2 × 106 spores of the parasitophagous fungus Mucor circinelloides (day 0) at 0.5 L/m2 (=600 mL/area); G2, sprayed with spores twice (every two weeks); G3, sprayed four times (every week); and Control, sprayed weekly with water. After a four-week period, the egg viability reduced for ascarids and capillarids (26% and 27%, respectively) in the control group; 64% and 79% in G1; 71% and 82% in G2; and 79% and 80% in G3. It was concluded that spraying with fungal spores provides a very useful tool for preventing infection by helminths on free-range poultry.
Agar–Agar Gels Carrying Curative and Preventive Agents Against Helminths: An In Vitro Compatibility Evaluation
(MDPI, 2025-07-12) Zubiria Ibarguren, Izaro; Abreu Ramos, Inês; Boso Dafonte, David; Pérez Anzúrez, Gustavo; Cazapal Monteiro, Cristiana Filipa; Sánchez-Andrade Fernández, Rita; Arias Vázquez, María Sol; Paz Silva, Adolfo; Hernández Malagón, José Ángel; Camiña García, María Mercedes; Universidade de Santiago de Compostela. Facultade de Veterinaria; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Departamento de Fisioloxía
The global market size of animal parasiticides was valued at USD 12.9 billion in 2024. Animal deworming only results in temporary cures with little to no preventive effects; therefore, a strategy that combines animal deworming with prevention is essential in improving the control of helminths. The effectiveness of co-administrating curative and preventive agents and their compatibility were considered based on the parasitophagous fungus Mucor circinelloides, which was developed in edible agar–agar (red seaweed)-carrying dewormers. Accordingly, Petri dishes were prepared with either a biopolymer alone (control, G-C) or with the anthelmintic piperazine (550, 1102, 2210, and 5500 mg/plate) or levamisole (37.5, 75, 150, and 300 mg/plate) and were used to culture the fungus Mucor circinelloides. Strong fungal growth and high numbers of spores were observed in the presence of the anthelmintics. No differences were measured between the control plates and those containing parasiticide drugs. Similar mycelial growth patterns and sporogenesis rates were recorded for different amounts of each anthelmintic. In conclusion, this novel formulation based on biopolymers containing anthelmintics and enriched with the parasitophagous fungus represents a highly promising tool to consider for jointly deworming animals and minimizing the risks of helminth infection. Further studies are in progress to confirm these in vitro results.
Targeting the Risk of Diptera-Borne Zoonoses by a Sentinel Equestrian Centers Program
(MDPI, 2025-07-04) Cazapal Monteiro, Cristiana Filipa; Boso Dafonte, David; Abreu Ramos, Inês; Camiña García, María Mercedes; Sanchís Polto, Jaime Manuel; Paz Silva, Adolfo; Cardoso, Luis; Sánchez-Andrade Fernández, Rita; Arias Vázquez, María Sol; Hernández Malagón, José Ángel; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Departamento de Fisioloxía
Diptera-borne diseases pose a major threat to global health, and their distribution is constantly changing due to climate change, globalization, and environmental changes. To improve the knowledge of dipteran species and their distribution in equine facilities, CDC-UV and oviposition traps were placed, and the dipping technique was performed in 16 equestrian centers of Northwest (NW) Spain (Galicia and Castilla y León Autonomous Communities) between July and November 2023. A questionnaire was distributed among the horse owners to obtain additional information. Four genera of culicids, Culex (51.8%), Culiseta (38.6%), Anopheles (8.4%), and Aedes/Ochlerotatus (1.2%) were identified in the equestrian centers. Culex pipiens s.l. was the most prevalent and well-distributed species (93.8% of the centers), whereas Anopheles maculipennis s.l. and An. claviger/petragnani, the anopheline species, were the most frequent (37.5% and 31.2%, respectively). The Culiseta genus was found in approximately 81.2% of the equine facilities. All genera were collected at medium and high altitudes and in Csb (warm-summer Mediterranean climate) areas. Equestrian centers from NW Spain albeit a variety of culicids with high vectorial capacity, together with an ideal environment for their breeding, the presence of vectors and hosts (humans and animals). This potential problem for global health enhances the need for entomological surveillance.
Monitoring of questing tick species distribution in Galicia, north-western Spain, over a period of 5.5 years
(Elsevier, 2025) Vilá Pena, María; Abreu Ramos, Inês; Bautista García, Génesis; Íñiguez Pichel, Elvira; Cazapal Monteiro, Cristiana Filipa; Hernández Malagón, José Ángel; Sánchez-Andrade Fernández, Rita; Arias Vázquez, María Sol; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal
An active survey was performed by the Galician Vector Surveillance Network (ReGaViVec) to determine the distribution of questing tick species in the Autonomous Community of Galicia Galicia, north-western Spain. Monitoring of tick populations involved drag/flag sampling at 533 locations over a period of 5.5 years. The identification of tick species, sex, and stage was carried out according to morphological keys, and the results were analyzed considering three Köppen climate regions, i.e. Cfb (marine west coast climate), Csb (warm-summer Mediterranean climate), and Csa (hot-summer Mediterranean climate), season, environment (forest, rural, or urban), vegetation height (≤ 15 cm, 16–40 cm, and > 40 cm); and altitude (< 300 m, 301–500 m, 501–1000 m, and >1000 m). A total of 1378 ticks were collected at 260 locations: 62.92% in Csb, 24.38% in Cfb, and 12.70% in Csa. Of these, 2% were larvae, 45.2% were nymphs, and 52.8% were adults (58.3% females and 41.7% males). Six species were recorded, i.e. Ixodes ricinus (57.90%), Rhipicephalus sanguineus (sensu lato) (26.05%), Dermacentor reticulatus (10.95%), Dermacentor marginatus (2.10%), Haemaphysalis sp. (2.61%), and Rhipicephalus bursa (0.36%). A seasonal pattern was recorded, with the relative abundance of I. ricinus and R. sanguineus (s.l.) peaking in spring-summer, and that of Dermacentor spp. and Haemaphysalis sp. peaking in autumn-winter. Significant differences were demonstrated in the species abundance distribution according to climate region, season, environment, and altitude. The highest tick abundance was recorded in summer, in rural areas, and at altitudes of < 300 m. Because most of the tick species present in the environment of Galicia have vectorial competence for emerging tick-borne pathogens, it is important to maintain research and coordination of tick surveillance practices in the region.
First insights on the susceptibility of native coccidicidal fungi Mucor circinelloides and Mucor lusitanicus to different avian antiparasitic drugs
(Springer Nature, 2024-02-24) Lozano, Joao; Cunha, Eva; Carvalho, Luis M. Madeira de; Paz Silva, Adolfo; Oliveira, Manuela; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
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). 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. 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.
Determinants of the current and future distribution of the West Nile virus mosquito vector Culex pipiens in Spain
(Elsevier, 2020-09-15) Gangoso, L.; Aragonés, D.; Martínez-de la Puente, J.; Lucientes, J.; Delacour-Estrella, S.; Estrada Pena, R.; Montalvo, T.; Bueno-Mari, R.; Bravo-Barriga, D.; Frontera, E.; Marqués, E.; Ruiz-Arrondo, I.; Muñoz, A.; Oteo, J.A.; Miranda, M. A.; Barceló, C.; Arias Vázquez, María Sol; Silva-Torres, M. I.; Ferraguti, M.; Magallanes, S.; Muriel, J.; Marzal, A.; Aranda, C.; Ruiz, S.; González, M. A.; Morchon, R.; Gomez-Barroso, D.; Figuerola, J.; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
Changes in environmental conditions, whether related or not to human activities, are continuously modifying the geographic distribution of vectors, which in turn affects the dynamics and distribution of vector-borne infectious diseases. Determining the main ecological drivers of vector distribution and how predicted changes in these drivers may alter their future distributions is therefore of major importance. However, the drivers of vector populations are largely specific to each vector species and region. Here, we identify the most important human-activity-related and bioclimatic predictors affecting the current distribution and habitat suitability of the mosquito Culex pipiens and potential future changes in its distribution in Spain. We determined the niche of occurrence (NOO) of the species, which considers only those areas lying within the range of suitable environmental conditions using presence data. Although almost ubiquitous, the distribution of Cx. pipiens is mostly explained by elevation and the degree of urbanization but also, to a lesser extent, by mean temperatures during the wettest season and temperature seasonality. The combination of these predictors highlights the existence of a heterogeneous pattern of habitat suitability, with most suitable areas located in the southern and northeastern coastal areas of Spain, and unsuitable areas located at higher altitude and in colder regions. Future climatic predictions indicate a net decrease in distribution of up to 29.55%, probably due to warming and greater temperature oscillations. Despite these predicted changes in vector distribution, their effects on the incidence of infectious diseases are, however, difficult to forecast since different processes such as local adaptation to temperature, vector-pathogen interactions, and human-derived changes in landscape may play important roles in shaping the future dynamics of pathogen transmission.
Integrating the control of helminths in dairy cattle: Deworming, rotational grazing and nutritional pellets with parasiticide fungi
(Elsevier, 2020-03-17) Voinot, Mathilde; Cazapal Monteiro, Cristiana Filipa; Hernández Malagón, José Ángel; Palomero Salinero, Antonio Miguel; Arroyo, Fabián Leonardo; Sanchís, Jaime; Pedreira, Jose; Sánchez-Andrade Fernández, Rita; Paz Silva, Adolfo; Arias Vázquez, María Sol; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
Thirty-two Friesian cattle under a leaders/followers four-day rotation and passing eggs of trematodes and gastrointestinal nematodes (GIN) were studied in two trials for the integrated control of these helminths over two years. In the first trial, the effect of rotational pasturing was assessed on a group of leaders (milking cows, G-L1) and followers (dried-off cows and heifers, G-F1) supplemented daily with commercial nutritional pellets. In the second trial, leaders (G-L2) and followers (G-F2) were maintained under a rotational pasturing regime; the cows received daily commercial pelleted feed and heifers pellets manufactured with a blend of parasiticide fungi (3 x 10(5) chlamydospores of both Mucor circinelloides and Duddingtonia flagrans/kg pellet). Deworming via closantel and albendazole was performed in cows in each trial at the beginning of their drying periods, and fourteen days later, the fecal egg-count reductions (FECR) of Calicophoron daubneyi and GIN were from 94 to 100% (average 98 %), while the percentages of reduction of cattle shedding eggs (CPCR) were from 50 to 100% (average 77 % and 82 %, respectively). The heifers were dewormed one time only, at the beginning of each trial, and the values of FECR and CPCR were 100 % against C. daubneyi and 96 % and 83 %, respectively, against GIN. Over a period of 24 months, significantly higher numbers of helminth egg-output were observed in G-L1, with the lowest numbers in G-F2. C. daubneyi egg output was reduced by 5 % (G-L1) and 42 % (G-F1) at the end of trial 1 and by 83 % (G-L2) and 100 % (G-F2) at the end of trial 2; the numbers of GIN egg-output decreased by 13 % (G-L1) and 18 % (G-F1) at the end of trial 1, and by 72 % (G-L2) and 85 % (G-F2) at the end of trial 2. No adverse effects were detected in cattle taking pellets enriched with fungal spores (G-F2). It is concluded that long-term ingestion of spores of M. circinelloides and D. flagrans provides a valuable tool to improve the effect of rotational grazing and to lessen the risk of infection by C. daubneyi and GIN in dairy cattle, and accordingly, the performance of integrated control programs.
Feeding horses with industrially manufactured pellets with fungal spores to promote nematode integrated control
(Elsevier, 2016-10-15) Hernández Malagón, José Ángel; Arroyo, Fabián Leonardo; Suarez, José; Cazapal Monteiro, Cristiana Filipa; Romasanta, Ángel; Eugenia López-Arellano, María; Pedreira, Jose; Madeira de Carvalho, Luis Manuel; Sánchez-Andrade Fernández, Rita; Arias Vázquez, María Sol; Mendoza de Gives, Pedro; Paz Silva, Adolfo; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
The usefulness of pellets industrially manufactured with spores of parasiticide fungi as a contribution to integrated nematode control was assessed in grazing horses throughout sixteen months. Two groups of 7 Pura Raza Galega autochthonous horses (G-T and G-P) were dewormed pour-on (1 mg Ivermectin/kg bw) at the beginning of the trial, and other group (G-C) remained untreated. The G-P was provided daily with commercial pellets to which was added a mixture of fungal spores during the industrial manufacturing (2 × 106 spores of Mucor circinelloides and same dose of Duddingtonia flagrans/kg), and G-T and G-C received pellets without spores. The efficacy of the parasiticidal strategy was assessed by estimating the reduction in the faecal egg counts (FECR) and in the number of horses shedding eggs in the faeces (PHR), and also the egg reappearance periods (ERP). Blood analyses were performed to identify the changes in the red and white cell patterns. To ascertain if horses developed an IgG humoral response against the fungi, antigenic products collected from M. circinelloides and D. flagrans were exposed to the horse sera by using an ELISA. The faecal elimination of eggs of Parascaris equorum and strongyles ceased 2 weeks after treatment in G-T and G-P, thus the values of FECR and PHR were 100%. No P. equorum-eggs were detected later, and the strongyle egg reappearance period was 28 weeks in G-P, and 8 weeks in G-T. Strongyle egg-output values remained lower than 300 eggs per gram of faeces in the G-P, whereas numbers between 330 and 772 in G-C and G-T were recorded. Normal values for the erythrocytes, haemoglobin and haematocrit in horses consuming pellets with spores were recorded, and lower than normal in the other groups. Sensitization of horses to the fungal species was disproven. It is concluded that feeding horses with pellets industrially manufactured with fungal spores represents a very useful tool to implement an integrated control of helminths affecting horses. This strategy allows a decrease in their risk of infection, aids in reducing the frequency of anthelmintic treatment.
Horses are susceptible to natural, but resistant to experimental, infection with the liver fluke, Fasciola hepatica
(Elsevier, 2020-05) Aoife Quigley; Mary Sekiya; García-Campos, Adrián; Paz Silva, Adolfo; Alison Howell; Diana J. Williams; Grace Mulcahy; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
Fasciola hepatica is a common parasite of livestock in Ireland, causing significant economic losses and affecting animal welfare. A previous abattoir study of 200 horses led to an estimated 9.5 % prevalence of infection in horses slaughtered in Ireland. However, the epidemiology and pathogenic significance of this infection in this species is not well-described. The objectives of this study were to determine the susceptibility of horses to oral challenge infection with F. hepatica metacercariae, and to document the course of the infection along with serological and biochemical response. We attempted an experimental infection of horses (n = 10; 9 geldings and 1 mare) with F. hepatica. Four were given 1000 metacercariae, four 500 metacercariae and two were sham-infected. Blood and faecal samples were taken at intervals up to 18 weeks post-infection (wpi). ELISA assays were used to assess sero-conversion in the experimental horses and also in a panel of sera from horses of known fluke status. No flukes were recovered from any of the livers, and neither were any lesions that could be attributed to F. hepatica infection observed. Coproantigen ELISA was negative throughout for all horses. Three antibody detection ELISAs, useful in diagnosing fasciolosis in other species, had limitations as diagnostic aids as determined using a panel of sera from horses of known F. hepatica infection status. This study is limited by the relatively small number of animals included, and the relatively short duration of the study period. Failure to establish infection after oral challenge raises fundamental questions on the pathophysiology and epidemiology of equine fasciolosis.
The efficacy of predatory fungi on the control of gastrointestinal parasites in domestic and wild animals—A systematic review
(Elsevier, 2020-07) Canhao Dias, Miguel; Paz Silva, Adolfo; Carvalho, Luis Manuel Madeira de; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
Background: Gastrointestinal parasites like nematodes are associated with significant impacts on animal health, causing poor growth rates, diseases and even death. Traditional parasite control includes the use of anthelmintic drugs, albeit being associated with drug resistance and ecotoxicity. In the last decade, biological control of parasites using nematophagous or predatory fungi has been increasingly studied, although systematic evidence of its efficacy is still lacking. The aim of this work was to assess the evidence of efficacy of nematophagous fungi in the control of nematodes and other gastrointestinal parasites in different animal species. Methods: Using the PICO method (Population, Intervention, Comparison and Outcomes), we performed a systematic review on the subject to search for original papers published between January 2006 and October 2019, written in English, and indexed in PubMed/Medline. Medical Subject Headings (MeSH) terms were used in the syntax. Papers were selected for detailed review based on title and abstract. Inclusion and exclusion criteria were applied, and relevant data were collected from the remaining papers. Results: The literature search retrieved 616 papers. Eighty-nine were submitted to a detailed review. In the end, 53 papers were included in the analysis. The studies were very heterogeneous, using different fungi, doses, frequency of administration, duration of treatment, host animals, and target parasites. Considering the 53 papers, 44 studies (83 % of the interventions) showed efficacy, with only 9 studies (17 %) showing no significant differences when compared to control. Conclusion: With the increasing hazards of drug resistance and ecotoxicity, biological control with predatory fungi stands out as a good tool for future parasite management, whether as a complementary treatment or as an alternative to standard parasite control.
The capability of the fungus Mucor circinelloides to maintain parasiticidal activity after the industrial feed pelleting enhances the possibilities of biological control of livestock parasites
(Elsevier, 2016-01-01) Arroyo, Fabián Leonardo; Arias Vázquez, María Sol; Cazapal Monteiro, Cristiana Filipa; Hernández Malagón, José Ángel; Suárez, José; Miguélez, Silvia; Romasanta, Ángel; Sánchez-Andrade Fernández, Rita; Paz Silva, Adolfo; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
The ability of the spores of the ovicide fungus Mucor circinelloides to resist the industrial manufacturing of pelleted feed and retain their biological and parasitological activities has been tested. Firstly, survival of M. circinelloides spores at elevated temperatures was in vitro assayed. In a second assay, the spores of M. circinelloides were added in the mixing phase of the industrial pelleting of livestock (calves and horses) feed. The biological development (mycelium growth rates and sporogenesis) and the ovicidal activity on eggs of the parasites Fasciola hepatica and Parascaris equorum eggs were measured in plates. In the in vitro assay, a similar level of biological development in all the conditions except by heating the spores at 72 °C for 10 min were observed. Viability of F. hepatica eggs reduced to 55–60%, and 56–70% that of P. equorum eggs. After the addition of the spores to the meal previous to the pelletization phase, percentages of reduction of 54–58% viability F. hepatica eggs and 61–67% P. equorum eggs were recorded. It is concluded that the spores of M. circinelloides maintain their antagonistic effect against eggs of the parasites F. hepatica and P. equorum in industrially manufactured pellets, providing thus a very helpful tool to prevent infection by trematodes or ascarids among pasturing livestock.
Formulating fungal spores to prevent infection by trichostrongylids in a zoological park: Practical approaches to a persisting problem
(Elsevier, 2020-12-31) Palomero Salinero, Antonio Miguel; Cazapal Monteiro, Cristiana Filipa; Viña Pombo, Cándido; Hernández Malagón, José Ángel; Voinot Meissner, Mathilde; Vilá Pena, María; Silva Torres, María Isabel; Paz Silva, Adolfo; Sánchez-Andrade Fernández, Rita; Arias Vázquez, María Sol; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
Two different formulations containing a blend of spores of the filamentous fungi Mucor circinelloides and Duddingtonia flagrans have been assayed against trichostrongylids in wapitis captive in a grassy parcel from a zoological park and dewormed with fenbendazole. One approach (sprayed-on pellets) consisted of providing them, during 10 months, nutritional pellets sprayed the spores just before to be ingested, and the other (formulated pellets) involved the administration of pellets industrially manufactured with the spores for an identical period. Data collected on wapitis treated with fenbendazole without receiving spores during a 10-months interval were considered as controls. The effect of these strategies was evaluated through the analysis of feces collected directly from the soil, by estimating the reduction of the counts of eggs of trichostrongylids per gram of feces (EPG). The possibility of side effects in the digestive tract, respiratory apparatus or on the skin was assessed. The coprological analyses showed that, despite the successfulness of fenbendazole, the values of trichostrongylids egg-output increased four months later above 300 EPG. Through the administration of pellets sprayed spores, a significant EPG reduction of 69% (30-100) was recorded, and 71% (36-100) by giving the wapitis pellets industrially enriched with the spores. No unfavorable effects were observed among the wapitis taking the spores. It is concluded that ingestion of a blend of spores of M. circinelloides and D. flagrans every two days affords a beneficial long-term effect on controlling the infection by trichostrongylids in wild captive animals.
Isolation of saprophytic filamentous fungi from avian fecal samples and assessment of its predatory activity on coccidian oocysts
(Nature Research, 2023-06-29) Lozano, João; Louro, Mariana; Almeida, Cristina; Victório, Ana Cláudia; Melo, Pedro; Rodrigues, João Paulo; Oliveira, Manuela; Paz Silva, Adolfo; Madeira de Carvalho, Luís; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
Fungal strains used in the biocontrol of animal gastrointestinal parasites have been mainly isolated from pasture soil, decaying organic matter, and feces from herbivores and carnivores. However, their isolation from birds and assessment of predatory activity against avian GI parasites has been scarce thus far. This research aimed to isolate filamentous fungi from avian fecal samples and evaluate their predatory activity against coccidia. A pool of 58 fecal samples from chickens, laying hens, and peacocks, previously collected between July 2020-April 2021, were used for isolation of filamentous fungi and assessment of their in vitro predatory activity against coccidian oocysts, using Water-Agar medium and coprocultures. The Willis-flotation technique was also performed to obtain concentrated suspensions of oocysts. A total of seven Mucor isolates was obtained, being the only fungal taxa identified, and all presented lytic activity against coccidia. Isolates FR3, QP2 and SJ1 had significant coccidiostatic efficacies (inhibition of sporulation) higher than 70%, while isolates FR1, QP2 and QP1 had coccidicidal efficacies (destruction of the oocysts) of 22%, 14% and 8%, respectively, after 14 days of incubation, being a gradual and time-dependent process. To our knowledge, this is the first report regarding the isolation of native predatory fungi from avian feces and demonstration of their lytic activity against coccidia.
Assessing the efficacy of the ovicidal fungus Mucor circinelloides in reducing coccidia parasitism in peacocks
(Nature Research, 2024-05-31) Lozano, João; Almeida, Cristina; Vicente, Eduardo; Sebastião, Daniela; Palomero Salinero, Antonio Miguel; Cazapal Monteiro, Cristiana Filipa; Arias Vázquez, María Sol; Oliveira, Manuela; Carvalho, Luis Madeira de; Paz Silva, Adolfo; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
The biological control of gastrointestinal (GI) parasites using predatory fungi has been recently proposed as an accurate and sustainable approach in birds. The current study aimed to assess for the first time the efficacy of using the native ovicidal fungus Mucor circinelloides (FMV-FR1) in reducing coccidia parasitism in peacocks. For this purpose, an in vivo trial was designed in the resident peacock collection (n = 58 birds) of the São Jorge Castle, at Lisbon, Portugal. These animals presented an initial severe infection by coccidia of the genus Eimeria (20106 ± 8034 oocysts per gram of feces, OPG), and thus received commercial feed enriched with a M. circinelloides suspension (1.01 × 108 spores/kg feed), thrice-weekly. Fresh feces were collected every 15 days to calculate the coccidia shedding, using the Mini-FLOTAC technique. The same bird flock served simultaneously as control (t0 days) and test groups (t15–t90 days). The average Eimeria sp. shedding in peacocks decreased up to 92% following fungal administrations, with significant reduction efficacies of 78% (p = 0.004) and 92% (p = 0.012) after 45 and 60 days, respectively. Results from this study suggest that the administration of M. circinelloides spores to birds is an accurate solution to reduce their coccidia parasitism.
Biological control of soil transmitted helminths (STHs) in a zoological park by using saprophytic fungi
(Elsevier, 2018-05-31) Hernández Malagón, José Ángel; Cazapal Monteiro, Cristiana Filipa; Arroyo, Fabián Leonardo; Silva, María Isabel; Palomero Salinero, Antonio Miguel; Paz Silva, Adolfo; Sánchez-Andrade Fernández, Rita; Arias Vázquez, María Sol; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
Toxascaris leonina and Trichuris sp. are soil-transmitted helminths (STHs) infecting domestic and wild mammals. The antagonistic effect of the saprophytic filamentous fungi Mucor circinelloides and Verticillium sp. was examined on eggs of T. leonina passed in the feces of captive lynxes (Lynx lynx) kept in a zoological park. The activity of M. circinelloides and Trichoderma atrobrunneum was tested on eggs of Trichuris sp. shed by captive dromedaries (Camelus dromedarius). The parasiticide activity was assessed by measuring the ovistatic (delayed development) and ovicidal (non-viability) effects on eggs placed in Petri plates, and by spraying spores directly onto fecal samples. Based on the observation of that hyphae of M. circinelloides, Verticillium sp. and T. atrobrunneum adhered to the eggshells, penetrated and destroyed the inner embryo, an ovicidal type 3 effect was concluded. Development of eggs of T. leonina and Trichuris sp. in the feces was delayed in the presence of all fungi, and one third remained at the stage of zygote. A significant reduction of T. leonina viable eggs was recorded in the feces sprayed spores of M. circinelloides (58%) or Verticillium sp. (67%). Fifty percent of the eggs of Trichuris sp. became into non-viable by 30 days after the exposure to either M. circinelloides or T. atrobrunneum. It is concluded that distribution of the filamentous fungi M. circinelloides, Verticillium sp. and T. atrobrunneum constitutes a novel approach to conduct the biological control of the STHs (T. leonina and Trichuris sp.) affecting wild animals captive in a zoological park.
Soil fungi enable the control of gastrointestinal nematodes in wild bovidae captive in a zoological park: a 4-year trial
(Cambridge University Press, 2020-05-27) Palomero Salinero, Antonio Miguel; Cazapal Monteiro, Cristiana Filipa; Esther Valderrábano Cano; Paz Silva, Adolfo; Sánchez-Andrade Fernández, Rita; Arias Vázquez, María Sol; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal; Universidade de Santiago de Compostela. Facultade de Veterinaria
The control of gastrointestinal nematodes among ruminants maintained in zoological parks remains difficult due to infective stages develop in the soil. For the purpose to improve the possibilities of the control of gastrointestinal nematodes (genera Trichostrongylus, Nematodirus, Chabertia and Haemonchus) affecting wild captive bovidae ruminants belonging to the subfamilies Antilopinae, Caprinae, Bovinae and Reduncinae, commercial pelleted feed enriched with a blend of 10(4)-10(5) spores of both filamentous fungi Mucor circinelloides + Duddingtonia flagrans per kg meal was provided for a period of 3.5 years. All animals were dewormed at the beginning of the trial and also when exceeding a cut-off point of 300 eggs per gram of feces (EPG). The anthelmintic efficacy ranged between 96 and 100%. The need for repeating the administration of parasiticide treatment disappeared at the 24th month of study in the Antilopinae individuals, and at the 8th month in the Caprinae, Bovinae and Reduncinae. No side-effects were observed on the skin or in the digestive, respiratory or reproductive system. It was concluded that this strategy provides a sustainable tool for preventing the contamination of paddocks where captive ruminants are maintained, decreasing the risk of infection by gastrointestinal nematodes and consequently the need of frequent deworming.
Determination of exposure to Fasciola hepatica in horses from Uruguay using a recombinant-based ELISA
(CZECH ACADEMY AGRICULTURAL SCIENCES, 2015-09-30) Sanchís, J.; Suárez, J.; Hillyer, G.V.; Hernández Malagón, José Ángel; Solari, M.A; Cazapal Monteiro, Cristiana Filipa; Duque de Araujo, A.M.; Carvalho, Luís Manuel Madeira de; Paz Silva, Adolfo; Sánchez-Andrade Fernández, Rita; Arias Vázquez, María Sol; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal
The risk of exposure to Fasciola hepatica in horses from Uruguay was evaluated using ELISA and a recombinant surface protein (FhrAPS). Blood samples were collected from 368 horses from different districts. Detection of antibodies and the seasonal IgG pattern against the trematode was also established. The overall seroprevalence was 54% (ranging 23% November to 93% December). Significantly higher values were observed in the Anglo-Arab horses (86%) and in those older than xix year (63%). No sex- or origin-differences were observed. Two primary risk periods for infection in Uruguay have been identified, the end of spring and autumn. Exposure to the fluke should be taken into account among those horses feeding on pastures grazed by ruminants. Using FhrAPS-ELISA, helpful information concerning the risk of horses to the infection by trematode can be obtained. This test allows the opportunity to compare the results achieved in different laboratories by minimising specific regional effects.
A combined effort to avoid strongyle infection in horses in an oceanic climate region: rotational grazing and parasiticidal fungi
(BMC, 2018-12-28) Hernández Malagón, José Ángel; Sánchez-Andrade Fernández, Rita; Cazapal Monteiro, Cristiana Filipa; Arroyo, Fabián Leonardo; Sanchís, Jaime Manuel; Paz Silva, Adolfo; Arias Vázquez, María Sol; Universidade de Santiago de Compostela. Departamento de Patoloxía Animal
Background: An approach to preventing strongyle infection in horses was tested, comprising rotational pasturing and the administration of spores of two parasiticidal fungi, Mucor circinelloides and Duddingtonia flagrans. Methods: Twenty-two adult Spanish Sport Horses were dewormed with ivermectin (1 mg pour-on/kg body weight) and then randomly divided into three groups. G-1 was maintained with continuous grazing, and G-2 and G-3 were kept on a four-paddock rotation system. Commercial pelleted feed (2.5 kg/horse) was supplied to G-1 and G-2 twice a week; horses in G-3 received pellets containing 2 x 10(6) spores/kg of each fungus. Fecal samples were analyzed by the flotation method to estimate the reduction in the fecal egg counts (FECR), the percentage of horses shedding eggs (PHR), and the egg reappearance period (ERP). Results: Third-stage larvae were identified in fecal pats as Cyathostomum (sensu lato) types A, C and D, Gyalocephalus capitatus, Triodontophorus serratus, Poteriosthomum spp., Strongylus vulgaris and S. edentatus. Two weeks after treatment, the FECR values were 100% in G-1, 96% in G-2 and 99% in G-3; the PHR values were 100% in G-1, 75% in G-2 and 88% in G-3. A strongyle ERP of 6 weeks was observed in G-1, ERP of 10 weeks was observed in G-2, and ERP of 16 weeks was observed in G-3. The counts of eggs per gram of feces (EPG) were > 300 EPG in G-1 and G-2 but remained below 250 EPG in G-3 throughout the observation period of 12 months. Conclusions: These results suggest that horse strongyle infection could be decreased by combining rotational pasturing with feeding pellets containing the spores of parasiticidal fungi.

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