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

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.info:eu-repo/semantics/publishedVersio

Gastrointestinal parasitism in Miranda donkeys : epidemiology and selective control of strongyles infection in the Northeast of Portugal

Research Areas: Agriculture ; Veterinary Sciences ; ZoologyIn Portugal, equine parasitism in pasture animals is characterized by high parasitic burden and high helminthic biodiversity; both factors are potentially pathogenic for their hosts. The decrease in the number of donkeys over the last years in Portugal, their importance in rural lowland and mountain ecosystems and pastures and the scarce information regarding their parasitism led to this research, which aimed to evaluate the parasitological status of a Miranda donkey breed population, a native breed mainly located in the northeast of Portugal. This study provides better knowledge of their gastrointestinal parasitism, particularly strongyles, and the assessment of a targeted selective treatment (TST) as an alternative control approach of their parasitism. A longitudinal observational study was developed during a period of five years in a population of 62 Miranda donkeys. At first, strategic deworming of these animals was performed every semester, but this was progressively replaced by a TST approach according to the levels of Eggs per Gram (EPG). This new deworming strategy was conducted in association with a regular parasitological monitoring of the animals every three months, being dewormed with ivermectin when egg shedding was higher than 500 EPG. Over the study period, a decrease of the annual prevalence rate of infection by gastrointestinal strongyles was observed, from 35.5% to 19.4%, as well as a negative binomial distribution of parasitic strongyles in donkeys submitted to selective anthelminthic control. The prevalence rate of infection was higher in females (39.5%), in individuals under four years (46.7%) and in those presenting a lower body condition (40.8%). The egg output was higher in animals younger than four years (589.3 EPG) than in those older than 10 years (533.6 EPG) (p < 0.05). However, no differences were observed according to sex during the study period. Results from this study allowed to note the influence of swampy pastures and of the weather changes in the epidemiology of strongylosis in Miranda donkeys. Moreover, it was possible to establish the annual epidemiological curve of strongyle egg shedding, with June being the month with the highest EPG, December having the lowest EPG and March and September showing intermediate numbers. Overall, a lower biodiversity of gastrointestinal parasites was observed. Cyathostomum sensu lato was the most prevalent genus and Strongylus vulgaris was the most observed large strongyle of the Strongylidae family. Trichostrongylus axei and Parascaris sp. were other nematodes with a minor frequency. The higher prevalence of strongyles at the beginning of the study showed a progressive decrease throughout the research period, and also for parasite biodiversity. Therefore, a targeted selective treatment seems to be a rational anthelminthic control approach in Miranda donkey strongyle infection and in other gastrointestinal parasites, since it reduces the antiparasitic treatments, the parasite’ prevalence and the EPG level. However, a loss of parasite biodiversity was noted at the end of the study period, as Cyathostominae were the only isolated strongyles. This can be a challenging situation in the long run, taking the ability of these nematodes to adapt easily to any deworming program, meaning that fecal EPG monitoring should be kept as a rule to a rational parasite control program.info:eu-repo/semantics/publishedVersio

Diagnosis of coccidiosis by Eimeria spp. in free-range chickens using Mini-FLOTAC and McMaster techniques - preliminary results

Mini-FLOTAC is emerging as a more sensitive and accurate tool to identify gastrointestinal parasites in faecal samples from domestic animals, in comparison with the McMaster method. However, research regarding its specific application in poultry samples, particularly from free-range chickens, is scarce. The current research aimed to test the use of Mini-FLOTAC for the identification of Eimeria spp. in free-range chickens and compare its results with McMaster. For this study, 40 faecal samples were collected from free-range chickens in a poultry farm located in North-Western Lisbon (Portugal). Each sample was processed with McMaster and Mini-FLOTAC techniques for the detection and count of coccidian Eimeria spp. oocysts. The resulting OPG (oocysts per gram of faeces) data obtained by the two techniques were compared using the Wilcoxon Test and correlated with the Spearman Test, and Mini-FLOTAC’s relative sensitivity was assessed, using a significance level of p<0.05. The average OPG was higher with Mini-FLOTAC and doubled the one obtained using the McMaster method (2669.3 OPG and 1220 OPG, respectively), although these results were not significant. Mini-FLOTAC’s relative sensitivity obtained in this study reached 86% (70.5-95.3%, 95%CI), although this result was not statistically significant. However, correlation of OPG counts between Mini-FLOTAC and McMaster, was significant. These preliminary results suggest the potential interest in the use of Mini-FLOTAC for the diagnosis of coccidiosis by Eimeria spp. in poultry, based on its assessment in a free-range poultry production system.info:eu-repo/semantics/publishedVersio

Gastrointestinal parasites of free-range chickens : a worldwide issue

Gastrointestinal parasites with direct and/or indirect life cycles, namely Eimeria spp., Ascaridia galli, Heterakis gallinarum and Capillaria spp., are responsible for the most frequent and heavy economic losses in aviculture. The current review aimed to collect information about gastrointestinal parasites affecting poultry, as well as, research studies regarding parasitic diseases in free-range chicken production. From the references used in this review, 20% highlight research studies correlating prevalence of gastrointestinal parasites in free-range chickens and factors such as the production system, age and animal density, hygienic conditions and weather. Coccidiosis is responsible for the major economic losses in poultry farms. Diseases have different clinical signs and result in variable impacts on animals. Control programs include mainly vaccination, anti-coccidia and anthelminthic drugs, as well as, house cleaning and disinfection. This review concluded that coccidia and helminths are a global threat to free-range poultry production. More research in this area is necessary in order to understand the major factors influencing the prevalence of parasitic diseases in this type of poultry production. Also the search for new control strategies must be a priority, mainly using natural antiparasitic compounds and biological control approaches.info:eu-repo/semantics/publishedVersio

Long-term survey of Eimeria spp. prevalence and faecal shedding in a traditional Portuguese free-range broiler farm

In free-range broiler production, animals are highly exposed to gastrointestinal parasitism, namely by coccidia, due to their long contact with the outdoor environment. Also, the prevalence of gastrointestinal parasites in extensive poultry production, is frequently influenced by biotic and abiotic factors. The current national pioneer long-term survey aimed to assess the prevalence and faecal shedding of Eimeria spp. oocysts in a traditional Portuguese free-range broiler farm, on different production conditions. Between July 2018 and March 2019, a total of 350 faecal samples were collected from different groups of free-range broilers at the fattening phase, in a poultry farm located in North-western Lisbon district, Portugal. Quantitative and qualitative coprological techniques were performed, aiming to quantify the Eimeria spp. prevalence and faecal shedding level. Coccidia prevalence was higher in Summer and Autumn groups (72% and 80%, respectively), although faecal shedding levels were higher in the Autumn and Winter groups (1191 and 1562 Oocysts Per Gram, OPG). Animals showed clinical signs of coccidiosis during the survey, especially in Autumn’s 1st sampling, in which 42% of the faecal samples had blood. The lack of sanitary depopulations, length of the fattening phase (1 month), short pre-patent period of coccidia, exposure to different environmental conditions and irregular drug treatments with amprolium, were key factors responsible for the different parasitic scenarios observed during this survey.info:eu-repo/semantics/publishedVersio

Implementation of Mini-FLOTAC in routine diagnosis of Coccidia and Helminth infections in domestic and exotic birds

Mini-FLOTAC (MF) has recently been proposed for the fecal quantification of gastrointestinal (GI) parasites in birds due to its higher sensitivity and precision in comparison with the McMaster method. The current research aimed to test the use of MF in routine diagnosis of coccidia and helminth infections in several domestic and exotic bird collections in Portugal. Between July 2020 and April 2021, a total of 142 fecal samples from organic layers, peacocks and ratites were collected in four Portuguese bird collections and processed using MF and fecal cultures to identify and calculate GI parasite shedding and prevalence. The McMaster method was also used to compare the shedding levels obtained for both quantitative techniques. MF’s relative sensitivity and specificity were also assessed, using McMaster as the reference technique. The implementation of MF resulted in an average Eimeria spp. shedding higher in peacocks from bird collection 2 (502 OPG), followed by peacocks from collection 1 (107 OPG) and organic layers (24 OPG) and peacocks from collection 3 (9 OPG). Peacocks were also positive for Capillaria spp., Trichostrongylus tenuis and Strongyloides pavonis, whereas ostriches and emus were infected by L. douglassii. The MF protocol for exotic animals and the McMaster method did not differ significantly for each parasitic agent and bird species, and MF achieved relative sensitivities and specificities higher than 70% for Galliform Eimeria spp., peacock helminths and ratites’ L. douglassii infections. Higher L. douglassii EPG values were identified using the MF protocol for exotic species (2 g of feces/38 mL of sucrose solution), followed by McMaster 2/28, MF 5/45 and MF 2/18. The use of MF allowed for obtaining different intestinal parasitic populations in several bird species and locations, and MF 2/38 is globally proposed as the most suitable protocol for bird fecal samples as an alternative to the McMaster method in the diagnosis of avian intestinal parasitic infections.info:eu-repo/semantics/publishedVersio

Physico-chemical properties of mango pulp and cajarana blends

The mango and cajarana are pleasant-tasting fruits, yellow in color, have ascorbic acid with antioxidant properties that slow down oxidation and help the immune system to promote well-being. The production of blends can improve nutritional and sensory characteristics, combining macro and micronutrients that may be present in a higher concentration in one fruit and deficient in another, increasing the possibility of obtaining new flavors constituting a new product. The study had as objective the elaboration of blends of mango cv. espada and cajarana and to determine the physical-chemical characteristics and color parameters. Three formulations were prepared: F1 - 50% mango pulp and 50% cajarana pulp; F2 - 90% mango pulp and 10% cajarana pulp and F3 - 10% mango pulp and 90% cajarana pulp. The formulations were evaluated for physicochemical parameters of water activity, moisture content, ash, total soluble solids, pH, total titratable acidity, ascorbic acid, proteins, lipids, total sugars, reducers, non-reducers and color parameters. The data obtained were submitted to analysis of variance and comparison between means by the Tukey test. The blends caused significant changes in most of the parameters evaluated, with an increase in the moisture content, ash, total titratable acidity, proteins and lipids as the concentration of the cajarana pulp was increased. The increase in the proportion of mango pulp favored the achievement of lighter blends, with a predominance of yellowness. However, the development of blends from this study presents itself as a viable alternative for the use of fruit pulps, both in terms of nutrition, complementing the nutrient content through this mixture as a very promising product for the market.The mango and cajarana are pleasant-tasting fruits, yellow in color, have ascorbic acid with antioxidant properties that slow down oxidation and help the immune system to promote well-being. The production of blends can improve nutritional and sensory characteristics, combining macro and micronutrients that may be present in a higher concentration in one fruit and deficient in another, increasing the possibility of obtaining new flavors constituting a new product. The study had as objective the elaboration of blends of mango cv. espada and cajarana and to determine the physical-chemical characteristics and color parameters. Three formulations were prepared: F1 - 50% mango pulp and 50% cajarana pulp; F2 - 90% mango pulp and 10% cajarana pulp and F3 - 10% mango pulp and 90% cajarana pulp. The formulations were evaluated for physicochemical parameters of water activity, moisture content, ash, total soluble solids, pH, total titratable acidity, ascorbic acid, proteins, lipids, total sugars, reducers, non-reducers and color parameters. The data obtained were submitted to analysis of variance and comparison between means by the Tukey test. The blends caused significant changes in most of the parameters evaluated, with an increase in the moisture content, ash, total titratable acidity, proteins and lipids as the concentration of the cajarana pulp was increased. The increase in the proportion of mango pulp favored the achievement of lighter blends, with a predominance of yellowness. However, the development of blends from this study presents itself as a viable alternative for the use of fruit pulps, both in terms of nutrition, complementing the nutrient content through this mixture as a very promising product for the market

The Control of Zoonotic Soil-Transmitted Helminthoses Using Saprophytic Fungi

Soil-transmitted helminths (STHs) are parasites transmitted through contact with soil contaminated with their infective eggs/larvae. People are infected by exposure to human-specific species or animal species (zoonotic agents). Fecal samples containing eggs of Ascaris suum or Lemurostrongylus sp. were sprayed with spores of the soil saprophytic filamentous fungi Clonostachys rosea (CR) and Trichoderma atrobrunneum (TA). The antagonistic effect was assessed by estimating the viability of eggs and their developmental rate. Compared to the controls (unexposed to fungi), the viability of the eggs of A. suum was halved in CR and decreased by two thirds in TA, while the viability of the eggs of Lemurostrongylus sp. was reduced by one quarter and one third in CR and TA treatments, respectively. The Soil Contamination Index (SCI), defined as the viable eggs that attained the infective stage, reached the highest percentages for A. suum in the controls after four weeks (66%), with 21% in CL and 11% in TA. For Lemurostrongylus sp., the values were 80%, 49%, and 41% for control, CR and TA treatments, respectively. We concluded that spreading spores of C. rosea or T. atrobrunneum directly onto the feces of animal species represents a sustainable approach under a One Health context to potentially reduce the risk of zoonotic STHs in humansThis research was funded by the Research Projects CTM2015-65954-R and RYC-2016-21407 (Ministerio de Economía y Competitividad, Spain; FEDER), and ED431F 2018/03 (Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia, Spain). María Sol Arias Vázquez is recipient of a Ramón y Cajal (Ministerio de Economía y Competitividad, Spain) contractS

Mixed Production of Filamentous Fungal Spores for Preventing Soil-Transmitted Helminth Zoonoses: A Preliminary Analysis

Helminth zoonoses are parasitic infections shared by humans and animals, being the soil-transmitted helminths (STHs) mainly caused by roundworms (ascarids) and hookworms. This study was aimed to assess the individual and/or mixed production of two helminth-antagonistic fungi, one ovicide (Mucor circinelloides) and other predator (Duddingtonia flagrans). Fungi were grown both in Petri plates and in a submerged culture (composed by water, NaCl, Na2HPO4 · 12 H2O, and wheat (Triticum aestivum)). A Fasciola hepatica recombinant protein (FhrAPS) was incorporated to the cultures to improve fungal production. All the cultured plates showed fungal growth, without difference in the development of the fungi when grown alone or mixed. High counts of Mucor spores were produced in liquid media cultures, and no significant differences were achieved regarding single or mixed cultures, or the incorporation of the FhrAPS. A significantly higher production of Duddingtonia spores after the incorporation of the FhrAPS was observed. When analyzing the parasiticide efficacy of the fungal mixture, viability of T. canis eggs reduced to 51%, and the numbers of third stage cyathostomin larvae reduced to 4%. It is concluded, the capability of a fungal mixture containing an ovicide (Mucor) and a predator species (Duddingtonia) for growing together in a submerged medium containing the FhrAPS offers a very interesting tool for preventing STHs.This work was partly supported by the Research Projects 10MDS261023PR (Xunta de Galicia, Spain) and AGL2012-34355 (Ministerio de Economía y Competitividad, Spain; FEDER)S