Composition of the rumen microbiome and its association with methane yield in dairy cattle raised in tropical conditions.

BACKGROUND: Methane (CH4) emissions from rumen fermentation are a significant contributor to global warming. Cattle with high CH4 emissions tend to exhibit lower efficiency in milk and meat production, as CH4 production represents a loss of the gross energy ingested by the animal. The objective of this study was to investigate the taxonomic and functional composition of the rumen microbiome associated with methane yield phenotype in dairy cattle raised in tropical areas. METHODS AND RESULTS: Twenty-two Girolando (F1 Holstein x Gyr) heifers were classified based on their methane yield (g CH4 / kg dry matter intake (DMI)) as High CH4 yield and Low CH4 yield. Rumen contents were collected and analyzed using amplicon sequencing targeting the 16 and 18S rRNA genes. The diversity indexes showed no differences for the rumen microbiota associated with the high and low methane yield groups. However, the sparse partial least squares discriminant analysis (sPLS-DA) revealed different taxonomic profiles of prokaryotes related to High and Low CH4, but no difference was found for protozoa. The predicted functional profile of both prokaryotes and protozoa differed between High- and Low CH4 groups. CONCLUSIONS: Our results suggest differences in rumen microbial composition between CH4 yield groups, with specific microorganisms being strongly associated with the Low (e.g. Veillonellaceae_UCG - 001) and High (e.g., Entodinium) CH4 groups. Additionally, specific microbial functions were found to be differentially more abundant in the Low CH4 group, such as K19341, as opposed to the High CH4 group, where K05352 was more prevalent. This study reinforces that identifying the key functional niches within the rumen is vital to understanding the ecological interplay that drives methane production.

First record of Trypanosoma (Ornithotrypanum) infecting Neotropical birds.

Parasites play a pivotal role in ecosystem health, influencing human and zoonotic diseases, as well as biodiversity preservation. The genus Trypanosoma comprises approximately 500 species mostly found in wildlife animals. This study focuses on identifying trypanosomes found in the white-necked thrush (Turdus albicollis) and the yellow-legged thrush (Turdus flavipes) in the Neotropics. First, we demonstrate the utility of an 18S rDNA sequence-structure phylogeny as an alternative method for trypanosome classification, especially when gGAPDH sequences are unavailable. Subsequently, the sequence-structure phylogeny is employed to classify new trypanosome sequences discovered in wild birds, placing them within the Ornithotrypanum subgenus. This marks the first identification of Ornithotrypanum in Neotropical birds, contributing to the understanding of the distribution and ecological adaptation of avian trypanosomes. Beyond taxonomy, this study broadens our comprehension of the ecological implications of avian trypanosomes in the Neotropics, emphasizing the need for continued research in this field. These findings underscore the importance of alternative classification methods, which are essential to unravel the complex interactions between parasites, wildlife hosts, and their ecosystems.

Insights into the systematics of the family Ophryoscolecidae (Ciliophora, Entodiniomorphida).

The family Ophryoscolecidae currently comprises 225 species of trichostomatid ciliates, subdivided into three subfamilies (Entodiniinae, Diplodiniinae, and Ophryoscolecinae). The last taxonomic review of the family was performed 55 years ago, but recent morphological and molecular studies indicate the need for a profound review of the systematics of this taxon, since its current taxonomy is insufficient to organize the diversity of the group. Here, we briefly review the systematics of the family Ophryoscolecidae based on information recovered from the literature and new morphological and molecular data. We add four new 18S rDNA sequences of ophryoscolecids to molecular databases, which contributed to improving the comprehension of intrafamily relationships within this group. Finally, we discuss some systematic problems and suggest approaches to resolve such inconsistencies in the future.

Rediscovery of Plasmodium (Huffia) huffi (Apicomplexa, Haemosporida): a lost lineage from toucans.

Literature data on toucans haemosporidians are scarce and all reports come from investigations in Brazil. Muniz et al. (Rev Bras Malariol 3: 339-356, Muniz et al., Rev Bras Malariol 3:339-356, 1951) and Muniz and Soares (Rev Bras Malar 611-617, Muniz J, Soares R de RL (1954) Nota sôbre um parasita do gênero Plasmodium encontrado no Ramphastos toco Müller, 1776, "Tucano-Açu", e diferente do Plasmodium huffi: Plasmodium pinottii n. sp. Rev Bras Malar 611 - 617.) described two Plasmodium species, P. huffi and P. pinottii, in Ramphastos toco. Later, Manwell and Sessler (J Protozol 18: 570-574, Manwell and Sessler, Malaria Parasites of Toucans J Protozol 18:570-574, 1971) established a new subspecies, P. nucleophilum toucani. In the last review on avian haemosporidians, Valkiunas (Valkiunas, Avian malaria parasites and other haemosporidia, CRC Press, New York, 2005) highlighted that P. huffi was insufficiently characterized, considering it a lost lineage. Also, the original description of P. huffi was considered insufficiently clear, due to a possible co-infection of the toucan hosts with a Novyella-like species. Here, we redescribed the species Plasmodium (Huffia) huffi based on morphological and molecular data, which were found in two toucan species, Ramphastos toco and Pteroglossus aracari from Brazil. Morphological features of the specimens are markedly the same as the original description. In R. toco, we observe two individuals infected, one infected only with P. huffi and one co-infected with P. huffi and the Novyella-like species, as observed in the original description. Also, we observe one R. toco infected only with the Novyella-like species, identified by morphological and molecular data as Plasmodium (Novyella) nucleophilum nucleophilum. In this way, it was possible to redescribe Plasmodium huffi in detail, without the doubt characters observed in the original description. Moreover, by applying species delimitation algorithms to a large Plasmodium phylogeny, we were able to identify new possible hosts for P. huffi and extend its geographic distribution to include North America.

Haemoproteus syrnii (Haemosporida: Haemoproteidae) in owls from Brazil: morphological and molecular characterization, potential cryptic species, and exo-erythrocytic stages.

Haemoproteus syrnii is a haemosporidian parasite found in owls. Although morphological and molecular data on the species is available, its exo-erythrocytic development was never researched. In this study, we provide the morphological, morphometric, and molecular characterization of H. syrnii populations found in owl species from Minas Gerais, southeast Brazil. We also characterized the coalescent species delimitation based on the molecular and histopathology data. Samples from 54 owls from six different species were analyzed, generating 11 sequences of the cyt b gene, from which six were new sequences. The overall prevalence of infection was high (72.22%). The H. syrnii sequences were grouped into two well-supported independent clades, which included other Haemoproteus (Parahaemoproteus) species. This was supported by both the coalescent species delimitation analysis and by the genetic divergence between lineages of these distinct clades. There were small morphological and morphometric differences within the population presented in this study. However, when compared with other studies, the molecular analysis demonstrated considerable intraspecific variation and suggests potential cryptic species. The histopathological analysis revealed, for the first time, that lungs and skeletal muscle are exo-erythrocytic stage location of H. syrnii, and that the parasite is linked to the histopathological changes found in owls. This study brings new data from Haemoproteus species biology and host infection, and improves host-parasite relationship understanding under an owl conservation perspective.

Helmet-shaped Body of Entodiniomorphid Ciliates (Ciliophora, Entodiniomorphida), a Synapomorphy or a Homoplasy?

Triadinium was created to include Triadinium caudatum. Further, four other species were included, T. minimum, T. galea, T. elongatum, and T. magnum, all sharing a characteristic helmet-shaped body. Wolska and Grain argued that the inclusion of T. minimum and T. galea into Triadinium was done based on superficial morphological aspects, and established two new genera to accommodate these species: Circodinium and Gassovskiella. Although the phylogenetic relationships within Entodiniomorphida have been investigated by multiple authors, none of them discussed the evolutionary relationship of helmet-shaped entodiniomorphids. We performed molecular phylogenetics and revisited old literature digging for morphological data to explain our results. According to our analyses, the helmet-shaped body is homoplastic and may have evolved from at least three different entodiniomorphid ancestors. Circodinium minimum is phylogenetically related to members of Blepharocorythidae, T. caudatum emerged within Spirodiniidae and G. galea within Polydiniellidae. This phylogenetic hypothesis is partially supported by information on infraciliature and ultrastructure of C. minimum and T. caudatum. However, such morphological information is not available for polydiniellids. In order to shed some light into the evolution of the helmet-shaped ciliates, future works should focus to collect information on the infraciliature and the ultrastructure of Polydiniella mysorea and of other Triadinium species.

Molecular diversity and coalescent species delimitation of avian haemosporidian parasites in an endemic bird species of South America.

Haemoproteus spp. and Plasmodium spp. are blood parasites that occur in birds worldwide. Identifying the species within this group is complex, especially in wild birds that present low parasitemia when captured, making morphological identification very difficult. Thus, the use of alternative tools to identify species may be useful in the elucidation of the distribution of parasites that circulate in bird populations. The objectives of this study were to determine the prevalence and parasitemia of the genera Plasmodium and Haemoproteus in Tachyphonus coronatus in the Atlantic Forest, Brazil, and to evaluate the molecular diversity, geographic distribution, and specificity of these parasites based on coalescent species delimitation methods. Microscopic analysis, PCR, cyt b gene sequencing, phylogenetic analysis and coalescent species delimitation using single-locus algorithms were performed (Poisson tree process (PTP) and multi-rate Poisson tree process (MPTP) methods). The analyses were performed in 117 avian host individuals. The prevalence was 55.5% for Plasmodium and 1.7% for Haemoproteus, with a mean parasitemia of 0.06%. Twenty-five Plasmodium and two Haemoproteus lineages were recovered. The MPTP method recovered seven different evolutionarily significant units (ESUs) of Plasmodium and one of Haemoproteus, whereas PTP presented fourteen ESUs of Plasmodium and one of Haemoproteus. The MPTP was more consistent with current taxonomy, while PTP overestimated the number of lineages. These ESUs are widely distributed and have already been found in 22 orders of birds that, all together, inhabit every continent, except Antarctica. The computational methods of species delimitation proved to be effective in cases where the classification of Haemosporida based just on morphology is insufficient.

Morphology, molecular phylogeny, and taxonomy of trichodinids (Ciliophora, Mobilida) from Calanoid copepods.

Trichodinid ciliates were isolated from calanoid copepods and were characterized by morphological and molecular techniques. The trichodinids found in this study were morphologically similar to Trichodina acuta, but morphometric differences were observed between our specimens and T. acuta. The T. acuta specimens, isolated from fish in several geographical locations around the world, were slightly larger than specimens in our population. In fact, our morphological and morphometric data are more similar to Trichodina diaptomi populations, also found on copepods. Moreover, our phylogenetic analyses revealed that the 18S rRNA gene sequence from our samples emerged in a clade with two T. acuta sequences, infesting fish hosts in China. These three sequences showed 100% identity. Our data, along with a taxonomic review, suggest that the species T. diaptomi, described recently, is actually a synonym of T. acuta and that this species may be related to fish and copepod hosts. In conclusion, we highlight the urgent need to better investigate the polymorphism and host specificity within the Mobilida, as this data may represent important characters to better understand the evolution into this order.

Ciliates as model organisms for the ecotoxicological risk assessment of heavy metals: A meta-analysis.

Ciliates are key components of aquatic ecosystems, significantly contributing to the decomposition of organic matter and energy transfer to higher trophic levels. They are considered good biological indicators of chemical pollution and relatively sensitive to heavy metal contamination. In this study, we performed a meta-analysis of the available toxicity data of heavy metals and ciliates to assess: (1) the sensitivity of freshwater ciliates to different heavy metals, (2) the relative sensitivity of ciliates in comparison to the standard test species used in ecotoxicological risk assessment, and (3) the difference in sensitivity across ciliate taxa. Our study shows that the tolerance of ciliates to heavy metals varies notably, which is partly influenced by differences in methodological conditions across studies. Ciliates are, in general, sensitive to Mercury > Cadmium > Copper > Zinc > Lead > Chromium. Also, this study shows that most ciliates are more tolerant to heavy metal pollution than the standard test species used in ecotoxicological risk assessments, i.e., Raphidocelis subcapitata, Daphnia magna, and Onchornyncus mykiss. Threshold concentrations derived from toxicity data for these species is expected to confer sufficient protection for the vast majority of ciliate species. Our data analysis also shows that the most commonly tested ciliate species, Paramecium caudatum and Tetrahymena thermophila, are not necessarily the most sensitive ones to heavy metal pollution. Finally, this study stresses the importance of developing standard toxicity test protocols for ciliates, which could lead to a better comprehension of the toxicological impact of heavy metals and other contaminants to ciliate species.

Rhabdostylid Ciliates (Ciliophora, Peritrichia, Epistylididae) as Epibionts on Chironomid Larvae: Evidence of High Specificity and Association with Organic Pollution.

Although epibiont ciliates make up a significant part of the biomass in aquatic ecosystems and may cause perceptible alterations in the population dynamics of their hosts, studies on the extrinsic and intrinsic factors that control the abundance of these microorganisms are scarce in literature. In the present study, we investigated the colonization site and intensity of rhabdostylid epibiont upon chironomid larvae and assessed the influence of organic pollution and chironomid communities on the prevalence and abundance of epibiont ciliates at five sampling stations (two in rural areas and three in urban areas) along a neotropical urban stream over a period of 12 months. Among the 24 genera of chironomids found and the 31,976 larvae analyzed, 96.12% belonged to the Chironomus genus, of which 16.95% (5212) were colonized by Rhabdostyla aff. chironomi. The infestation intensity varied from one to 67 individuals per host with an average intensity of 4.86 (± 33.45). Ciliates were only found colonizing the chironomids' ventral tubules. The high number of chironomid larvae, high host- and site-specificity, low infestation intensity, and absence of apparent structural damage to hosts evidence an intimate relationship between epibiont and basibiont as well as a possibly long coevolutionary history. Both prevalence and abundance of epibiont ciliates were correlated to the pollution rate of the studied stream. There was an alteration in the composition and structure of the chironomid community along the sampling stations. Numeric dominance of Chironomus-tolerant chironomids and its direct correlation to infestation prevalence and to ciliates abundance highlights the predilection of both peritrich ciliates and Chironomus larvae for organically-enriched environments.

Phylogenetic Analyses Support Validity of Genus Eodinium (Ciliophora, Entodiniomorphida, Ophryoscolecidae).

The validity of genus Eodinium has been historically disputed due to morphological similarities with Diplodinium (absence of skeletal plates as well as adoral and dorsal ciliary zones at the same body level). To address this issue, the 18S rDNA of four Eodinium posterovesiculatum morphotypes and four Diplodinium anisacanthum morphotypes were sequenced and phylogenetically analyzed. The different inference methods suggest the existence of a last common ancestor of Eodinium and Ostracodinium that is not shared with Diplodinium, strongly supporting the validity of genus Eodinium. Since skeletal plates are present in all members of genus Ostracodinium, the most parsimonious is a secondary loss of skeletal plates in E. posterovesiculatum. This work represents a breakthrough in the taxonomy and phylogeny of the family Ophryoscolecidae indicating that the skeletal plates may not reflect evolutionary divergence within this group of ciliates as traditionally proposed.

Multidisciplinary re-description of Plasmodium (Novyella) paranucleophilum in Brazilian wild birds of the Atlantic Forest kept in captivity.

Haemosporidian blood parasites of the Plasmodium genus are the causative agents of avian malaria in many parts of the world. Despite the great diversity of Brazilian avifauna, few studies have been conducted to examine the haemosporidians of wild birds found in the Brazilian Atlantic Forest, especially those kept in captivity. This study aimed to re-examine and further characterize the South American avian parasite Plasmodium paranucleophilum using a multidisciplinary approach. Blood samples were collected from 68 captive birds representing 15 species found in the Atlantic Forest of southeastern Brazil. Morphometric and morphological characterization was performed, in addition to PCR and sequencing of the mitochondrial cytochrome b gene and subsequent phylogenetic analysis. The overall prevalence of P. paranucleophilum infection in the study was 13.23% (n = 9), with a mean parasitemia of 0.58%. We observed the highest parasitemia of 3.88% in Rupornis magnirostris. In our phylogenetic analysis, P. paranucleophilum and P lasmodium nucleophilum formed distinct, highly supported clades, with a mean genetic divergence of 2.48%. This study provides new morphological and molecular data, expanding our knowledge of the haemosporidians of wild birds in Brazil and highlighting the need for further investigation. The true depth of diversity in Brazilian avian haemosporidians remains largely unknown, and given the enormous variety of vectors and avian species, there may be many more species of these blood parasites yet to be described.

Methods for taxonomic studies of rumen ciliates (alveolata: ciliophora): a brief review.

This review presents the principal methods used in taxonomic studies of rumen ciliates: live observation, Lugol staining, fixation and staining with methyl-green formalin saline (MFS) solution, protargol staining, silver carbonate impregnation, scanning electron microscopy and molecular techniques. Mastering these techniques is essential for successful research on the taxonomy of rumen ciliates. No single technique reveals all of the characteristics required for a complete description of a rumen ciliate; therefore, it is necessary to combine the use of these techniques as appropriate to the rumen ciliate group under study. Tables are provided to summarize: 1) morphological methods more appropriate for revealing morphological structures of interest, 2) morphological methods indicated for each group of rumen ciliates, and 3) main primers used for PCR amplification of the 18S rDNA of rumen ciliates.

Redescription of the Little-known Ciliate Apoamphisiella vernalis (Stokes, 1887) Berger, 2006 (Ciliophora, Hypotricha) from the Atlantic Forest, Brazil.

Apoamphisiella Foissner, 1997 is a genus of hypotrichous ciliates reported to occur in various locations in the world, but that seems particularly widespread in the Americas. In this study, the first record of Apoamphisiella vernalis for Brazil is presented based on a population from the Atlantic Forest of the Minas Gerais state. A redescription of A. vernalis made from light and scanning electron microscopy observations unveils a unique combination of diagnostic features for this species, namely the presence of green cortical granules, one contractile vacuole lacking collecting canals, and the rear end of both left and right marginal cirral rows located at same level. Moreover, A. vernalis is found to be an omnivorous predator, feeding on arcelline testate amoebae, rotifers, and other organisms.

Feed efficiency and enteric methane emissions indices are inconsistent with the outcomes of the rumen microbiome composition.

The correlation between enteric methane emissions (eME) and feed efficiency (FE) in cattle is linked to the anaerobic fermentation of feedstuffs that occurs in the rumen. Several mathematical indices have been developed to predict feed efficiency and identify low methane emitters in herds. To investigate this, the current study aimed to evaluate the rumen microbial composition in the same group of animals ranked according to six different indices (three indices for FE and three for eME). Thirty-three heifers were ranked into three groups, each consisting of 11 animals, based on FE (feed conversion efficiency - FCE, residual weight gain - RG, and residual feed intake - RFI) and eME indices (production, yield, and intensity). Rumen fluids were collected using a stomach tube and analyzed using 16S rRNA and 18S rRNA, targeting rumen bacteria, archaea, and protozoa. The sequencing analysis revealed that the presence of unique microbial species in the rumen varies across animals ranked by the FE and eME indices. The High RG group harbored 17 unique prokaryotic taxa, while the High FCE group contained only seven. Significant differences existed in the microbial profiles of the animals based on the FE and eME indices. For instance, Raoultibacter was more abundant in the Intermediate RFI group but less so in the Intermediate RG and Intermediate FCE groups. The abundance of Entodinium was higher while Diplodinium was lower in the High FCE group, in contrast to the High RG and High RFI groups. Methanobrevibacter exhibited similar abundances across eME indices. However, the heifers did not demonstrate the same production, yield, and intensity of eME. The present findings underscore the importance of standardizing the FE and eME indices. This standardization is crucial for ensuring consistent and reliable assessments of the composition and function of the rumen microbiome across different herds.

Giants' cooperation: a draft genome of the giant ciliate Muniziella cunhai suggests its ecological role in the capybara's digestive metabolism.

Several hundred ciliate species live in animals' guts as a part of their microbiome. Among them, Muniziella cunhai (Trichostomatia, Pycnotrichidae), the largest described ciliate, is found exclusively associated with Hydrochoerus hydrochaeris (capybara), the largest known rodent reaching up to 90 kg. Here, we present the sequence, structural and functional annotation of this giant microeukaryote macronuclear genome and discuss its phylogenetic placement. The 85 Mb genome is highly AT rich (GC content 25.71 %) and encodes a total of 11 397 protein-coding genes, of which 2793 could have their functions predicted with automated functional assignments. Functional annotation showed that M. cunhai can digest recalcitrant structural carbohydrates, non-structural carbohydrates, and microbial cell walls, suggesting a role in diet metabolization and in microbial population control in the capybara's intestine. Moreover, the phylogenetic placement of M. cunhai provides insights on the origins of gigantism in the subclass Trichostomatia.

Insights on the taxonomy of Haemoproteus parasites infecting cracid birds.

The Haemosporida order is a well-supported clade of heteroxenous parasites transmitted by dipteran insects and frequently found parasitizing wild birds. These parasites have already been reported in all zoogeographic regions of the world, except for Antarctica. One of the potential hosts of haemosporidians is the Cracidae family, which includes approximately 50 species, 22 of which are present in Brazil, classified within nine genera. Data on haemosporidian infecting individuals of the Cracidae family is scarce, with only three Haemoproteus species being recorded in this group of birds. We found Haemoproteus spp. infection in all Penelope obscura bronzina analyzed. Among the parasites found, we observed two lineages of Haemoproteus (PENOBS02 and PENOBS03), which were characterized by morphological, molecular and phylogenetic approaches. The morphological data on cracid haemosporidian parasites, together with our phylogenetic results, allows discussions on the taxonomy of the Haemoproteus parasites that infect birds of the Cracidae family.

Taxonomic and predicted functional signatures reveal linkages between the rumen microbiota and feed efficiency in dairy cattle raised in tropical areas.

Ruminants digest plant biomass more efficiently than monogastric animals due to their symbiotic relationship with a complex microbiota residing in the rumen environment. What remains unclear is the relationship between the rumen microbial taxonomic and functional composition and feed efficiency (FE), especially in crossbred dairy cattle (Holstein x Gyr) raised under tropical conditions. In this study, we selected twenty-two F1 Holstein x Gyr heifers and grouped them according to their residual feed intake (RFI) ranking, high efficiency (HE) (n = 11) and low efficiency (LE) (n = 11), to investigate the effect of FE on the rumen microbial taxa and their functions. Rumen fluids were collected using a stomach tube apparatus and analyzed using amplicon sequencing targeting the 16S (bacteria and archaea) and 18S (protozoa) rRNA genes. Alpha-diversity and beta-diversity analysis revealed no significant difference in the rumen microbiota between the HE and LE animals. Multivariate analysis (sPLS-DA) showed a clear separation of two clusters in bacterial taxonomic profiles related to each FE group, but in archaeal and protozoal profiles, the clusters overlapped. The sPLS-DA also revealed a clear separation in functional profiles for bacteria, archaea, and protozoa between the HE and LE animals. Microbial taxa were differently related to HE (e.g., Howardella and Shuttleworthia) and LE animals (e.g., Eremoplastron and Methanobrevibacter), and predicted functions were significatively different for each FE group (e.g., K03395-signaling and cellular process was strongly related to HE animals, and K13643-genetic information processing was related to LE animals). This study demonstrates that differences in the rumen microbiome relative to FE ranking are not directly observed from diversity indices (Faith's Phylogenetic Diversity, Pielou's Evenness, Shannon's diversity, weighted UniFrac distance, Jaccard index, and Bray-Curtis dissimilarity), but from targeted identification of specific taxa and microbial functions characterizing each FE group. These results shed light on the role of rumen microbial taxonomic and functional profiles in crossbred Holstein × Gyr dairy cattle raised in tropical conditions, creating the possibility of using the microbial signature of the HE group as a biological tool for the development of biomarkers that improve FE in ruminants.

A checklist of species of the family Zoothamniidae (Ciliophora: Peritrichia), symbionts of crustaceans.

Eight genera are included in the family Zoothamniidae, and at the present the data of the epibiotic species on crustaceans is not updated. Thus, the main goal of the present work is to provide the checklist of zoothamniids worldwide reported as associated to several groups of crustaceans, including some data of their geographic distribution, and also to provide the complete list of species of this peritrich group. We obtained all available data of family Zoothamniidae and checked their taxonomic status. Then we provided the complete list of species including those reported as epibionts on crustacean hosts. We obtained 156 species included in eight genera of Zoothamniidae, being 85 species recorded as symbionts on 86 species of hosts. Genus Zoothamnium contained the higher number (56) of epibiotic species on crustaceans, and genus Pseudohaplocaulus has not been reported for any crustacean. The reviewed data highlights the concentration of records in Europe, and stands out the need to amplify sampling and studies in the Southern Hemisphere. The main groups of crustaceans infested by zoothamniids are the amphipods, isopods and decapods.

Hippoboscid flies (Diptera: Hippoboscidae) on birds of prey in the Atlantic Forest, Minas Gerais, Brazil.

Hippoboscid flies are potential ectoparasites of several avian orders, including birds of prey, a group formed by the orders Falconiformes, Strigiformes, Cathartiformes and Accipitriformes. In this study, we evaluated 155 birds of prey that were brought to the Wildlife Rehabilitation Center of Juiz de Fora, Minas Gerais, Brazil, between the years 2016 and 2019. Two species of hippoboscid flies (Ornithoctona erythrocephala and Icosta americana) were found in five species of birds of prey (Megascops choliba, Asio stygius, Athene cunicularia, Asio clamator and Caracara plancus). The average intensity found was 1.4 hippoboscids/bird and Megascops choliba (tropical screech-owl) was the most parasitized bird. This is the first record of parasitism of Asio stygius (stygian owl) by Ornithoctona erythrocephala in Brazil and of Athene cunicularia (burrowing owl) and Asio clamator (striped owl) by Icosta americana. Ornithoctona erythrocephala is recorded here for the first time infesting birds of prey in the state of Minas Gerais, Brazil.