Molecular identification and biological characterization of Eimeria columbarum from domestic pigeons (Columba livia domestica) in Guangdong, China.

Pigeon coccidiosis caused by Eimeria spp. is an important veterinary disease with a significant economic impact on the pigeon industry. Preventive measures for Eimeria columbarum in pigeons have been hampered by the lack of extensive genetic, morphological, and biological data on the oocysts. In this study, we examined the prevalence and identity of Eimeria spp. in domestic pigeons from seven cities in Guangdong Province, China. Data show that coccidiosis was prevalent in domestic pigeons in Guangdong Province, with an overall Eimeria spp. detection rate of 73.4%. Five Eimeria species were identified, including E. columbarum (73.4%), Eimeria kapotei (25.6%), Eimeria labbeana (19.6%), Eimeria duculai (19.6%), and Eimeria tropicalis (6.7%). We obtained single oocyst-derived lines of the dominant E. columbarum from fecal specimens. E. columbarum oocysts measured 20.06 ± 0.69 µm × 18.63 ± 1.03 µm, and sporocysts measured 10.29 ± 0.82 µm × 85.38 ± 0.46 µm. In infection experiment using obtained E. columbarum isolates, 60-day-old coccidia-free pigeons exhibited a prepatent period of 105 h and patent period of 9-10 days followed by severe diarrhea, depression, anorexia, and emaciation. Endogenous development of the parasite was observed mainly in the cytoplasm of epithelial cells in the duodenum, jejunum, ileum, and rectum. Two generations of meronts developed on days 3 and 4 after infection, respectively, while gamont and gamete developed on day 5 after infection. The morphological, genetic, and biological data are expected to be useful in elucidating the biological characterization of pigeon coccidiosis to develop measures against the treatment and containment of this disease.

Avian malaria in a feral-pet pigeon: a case report.

BACKGROUND: Avian malaria is caused by diverse parasite species of the genus Plasmodium, and it affects various bird species. The occurrence of this disease in some wild bird species is sparsely documented due to the scarce availability of samples. Hence the pathogenicity in some hosts is not completely known. In addition, feral birds may act as reservoirs bridging the transmission cycle from wild migratory birds to domestic and zoo-kept bird species. CASE PRESENTATION: An owner of pigeons adopted a feral pigeon (Columba livia forma domestica) and housed it together with his other pet-pigeons. The bird died unexpectedly a few weeks after a surgical procedure and necropsy revealed a severely anaemic carcass, with pale organs and hydropericardium. Histopathologic analysis revealed inflammatory infiltrates in the lung and liver, and monocytes and Kupffer cells contained haemozoin pigment indicative of phagocytosis of Plasmodium-infected erythrocytes. A high erythrocytic infection rate of 18% was evident in tissues and blood vessels in various organs. Furthermore, the thyroid had masses classified as thyroid carcinomas. Immunohistochemistry with anti- Plasmodium falciparum HSP70 antibody revealed positive signals in erythrocytes and intravascular leucocytes. Further microscopy analysis using a Hemacolor-stained impression smear revealed a high parasitaemia with an asynchronous infection showing all erythrocytic stages. Molecular diagnosis by PCR identified Plasmodium relictum, lineage GRW11 as the aetiological agent. The bird presented died most likely due to an acute infection as evidenced by the high blood parasitaemia, leading to major erythrocyte destruction. Further analyses of feral pigeons (n = 22) did not reveal any additional cases of Plasmodium infections. CONCLUSION: This study reports the first mortality associated with P. relictum lineage GRW11. The study supports previous studies, suggesting that Plasmodium infections are not frequent in pigeons. Host conditions like immunosuppression due to the tumour may have influenced the infection outcome in this fatal case. Use of anti-P. falciparum HSP70 antibody for detection of P. relictum antigens for immune assays in blood and tissue samples will be a useful tool for future studies.

Phylogenetic position of the pigeon mite, Ornithonyssus sylviarum, with amplification of its immunogenetic biomarkers in Egypt.

Ornithonyssus sylviarum (O. sylviarum) is an obligatory, blood-sucking ectoparasite widely distributed among poultry and other mammals, causing significant economic losses. This study represented the first report of molecular genotypic identification of O. sylviarum from pigeons, Columba livia domestica, in Egypt. PCR and sequencing of the 28S rRNA gene were conducted. The resulting mite sequences were subjected to BLAST analysis, revealing 90-100% similarity to O. sylviarum in all tested samples. The sequences were deposited in GenBank under the accession numbers PP049086 and PP033720. A phylogenetic tree was constructed to compare the obtained species with related species worldwide. Additionally, infected pigeons showed increased expression of IL-1, IL-10, IFN-γ, and TGF-ß3 genes and elevated serum levels of stress biomarkers. The increased level of these cytokines indicates there was a disturbance in the immune status of the infected host with parasite compared with control healthy ones. This increases the susceptibility to infection with other pathogens.

Molecular detection and characterization of Trichomonas gallinae isolated from ornamental birds in Tehran, Iran.

Trichomonas gallinae is a widespread protozoan parasite that primarily affects birds, causing a disease known as avian trichomonosis. The present study aimed to investigate the prevalence and genetic diversity of T. gallinae, a parasite causing avian trichomoniasis in feral pigeons, budgerigars, and finches in Tehran, Iran. The 5.8S ribosomal RNA locus, along with the internal transcribed spacer 2 (ITS2) region, has been extensively utilized for genotype identification and for determining inter- and intra-specific diversity. More recently, the Fe-hydrogenase (Fe-Hyd) gene has been suggested as an additional genetic marker to enhance the accuracy of strain subtyping discrimination. In the present study, a total of 12% (12/100) birds examined were infected with T. gallinae using microscopy and PCR methods. Infection was found in seven of 30 (23.3%) feral pigeons, three of 40 (7.5%) budgerigars, and two of 30 (6.66%) finches. Analysis of the ITS2 region of T. gallinae isolates revealed two highly similar sequences. The first sequence (GenBank: OQ689964-OQ689970) was found in five feral pigeons and two budgerigars, whereas the second sequence (GenBank: OQ689971-OQ689975) was identified in two feral pigeons, one budgerigar, and two finches. Phylogenetic analysis confirmed the presence of two distinct clusters (cluster I and cluster II) within the trichomonads based on the ITS2 region. However, further analysis using Fe-Hyd revealed greater diversity, with three subtypes identified (A1, A2, and C1). One isolate identified in the present study (GenBank accession number: OQ694508.1) belonged to subtype A1. Combining ITS2 and Fe-Hyd markers holds promise for a more comprehensive understanding of the population structure of T. gallinae and the potential role of ITS2 in host adaptation.

Innovative molecular and immunological approaches of heterophyiasis infecting some Egyptian marketed fishes.

Heterophyiasis is a highly endemic disease in the Nile Delta, Egypt, where people consume raw or undercooked Oreochromis niloticus and Mugil cephalus. Birds and rats play a crucial role in fish-borne zoonotic trematode transmission since they serve as natural and experimental hosts. This study aimed to update the epidemiological information, morphological description, molecular identification and gene expression of two distinct heterophyid metacercariae in Giza, Wadi Al-Rayan, and Lake Manzala, Egypt, whereas various heterophyid infections could be expected. The present Centrocestus formosanus, Heterophyes heterophyes, and Heterophyes nocens with accession numbers OR947651.1, OR947700.1, and OR947719.1, respectively, matched with those recorded in the GenBank. Findings of the current investigation indicated that various cytokines like IL-1ß, MHC-II, and TNF-α rapidly elevated in the infected pigeon's intestines. Additionally, the infection expanded due to the parasite's ejection from the host and the host's clinical affliction, which induced humoral immune responses. Interestingly, investigation of other trematode species is in extreme demand in terms of zoonoses. We suggest controlling snails, managing migratory birds, and examining and frying fishes to the point when the encysted metacercariae is destroyed.

Meningoencephalitis with malacia caused by Sarcocystis calchasi in a rock pigeon in Japan.

Sarcocystis spp. cause pigeon protozoan encephalitis, a neuronal disease. A female pigeon exhibiting torticollis had a necrotic area in the cerebral hemisphere surrounded by lesions with perivascular cuffing, gliosis, granulomatous foci, and meningitis. Non-necrotic lesions were also observed in the brainstem. Intact and degenerative schizonts were observed within the neuropils and neurons in the lesions. Deoxyribonucleic acid (DNA) was extracted from paraffin-embedded brain tissues and genetically analyzed after gel electrophoresis to determine Sarcocystis spp. using specific primer sets for 28S ribosomal ribonucleic acid and internal transcribed spacer region-1. DNA sequencing confirmed a significant homology with S. calchasi. This is the first report of meningoencephalitis with malacia caused by S. calchasi in a rock pigeon in Japan.

Selection of a suitable reference gene for gene-expression studies in Trichomonas gallinae under various biotic and abiotic stress conditions.

Trichomonas gallinae, a globally distributed protozoan parasite, significantly affects the pigeon-breeding industry. T. gallinae infection mainly causes yellow ulcerative nodules on the upper respiratory tract and crop mucosa of pigeons, impeding normal breathing and feeding and ultimately causing death. Real-time quantitative PCR (qPCR) is a crucial technique for gene-expression analysis in molecular biology. Reference-gene selection for normalization is critical for ensuring this technique's accuracy. However, no systematic screening or validation of T. gallinae reference genes has been reported. This study quantified the transcript levels of ten candidate reference genes in T. gallinae isolates with different genotypes and culture conditions using qPCR. Using the geNorm, NormFinder, and BestKeeper algorithms, we assessed these reference genes' stabilities and ranked them using RankAggreg analysis. The most stable reference gene was tubulin beta chain (TUBB), while the widely used reference genes TUBG and GAPDH demonstrated poor stability. Additionally, we evaluated these candidate reference genes' stabilities using the T. gallinae TgaAtg8 gene. On using TUBB as a reference gene, TgaAtg8's expression profiles in T. gallinae isolates with different genotypes remained relatively consistent under various culture conditions. Conversely, using ACTB as a reference gene distorted the data. These findings provide valuable reference-gene-selection guidance for functional gene research and gene-expression analysis in T. gallinae.

Finding a needle in a haystack: DNA Haemoproteus columbae enrichment using percoll density gradient and flow cytometry.

Isolation of genomic DNA of blood parasites in birds, reptiles, amphibians, and fishes is a challenging task, given that their red blood cells are nucleated; for that reason, parasite genomic DNA is only a fraction of the total extracted DNA, and it is challenging to obtain concentrated high-quality genetic material. Percoll Density Gradient (PDG) and flow cytometry are tools for separating and analyzing cell populations or even a single cell, and both represent potent approaches for isolating avian haemosporidians parasites. Our experimental design included several steps seeking to concentrate the parasite´s DNA. We used blood samples from a Rock pigeon infected with Haemoproteus columbae. After inducing parasite exflagellation and gametogenesis in vitro, we subjected the samples to a Percoll Density Gradient to separate the parasites from the rest of the blood cells. Following centrifugation, the layer containing extracellular parasites underwent a flow cytometry and cell sorting process, during which we selected two different subpopulations of cells for analysis. Based on qPCR analyses, we demonstrate parasite DNA enrichment in Percoll Density Gradient and flow cytometry samples; simultaneously, these samples showed the lowest concentration of Columba livia DNA. However, the concentration of parasite DNA was higher in the PDG than in the cell sorting sample. This study reports the concentration of the Haemoproteus parasite by flow cytometry without DNA-intercalating dyes, and this methodology can serve as a technique for DNA enrichment of blood parasites infecting nucleated red blood cells to improve techniques that allow obtaining complete genomes.

Prevalence and molecular characterization of Trichomonas gallinae from pigeons in Anhui, China.

Trichomonas gallinae, a protozoan parasite causing avian trichomonosis, exhibits a widespread global prevalence. It primarily affects the upper digestive tract of birds and has resulted in significant ecological problems worldwide. This study aimed to investigate the prevalence and genotypes of T. gallinae in Anhui Province, China. A total of 1612 oropharyngeal swab samples were collected from pigeon farms in Anhui Province to determine the prevalence of T. gallinae infection. The results revealed 565 (35.1%) positive samples of T. gallinae. Significant differences in infection rates were observed among different regions and age groups. Furthermore, the ITS1/5.8 S/ITS2 region was amplified, sequenced, and subjected to phylogenetic analysis. Genotypes A and B of T. gallinae were identified, and genotype B was the dominant genotype in Anhui Province. This is the first report on the prevalence and molecular characterization of T. gallinae in Anhui Province, China. Additionally, we integrated reports on the prevalence and genotype of T. gallinae in relevant provinces in China.

Distribution of phoretic mites and lice in Pseudolynchia canariensis living on pigeons and the relationship with seasonality, carrier sex, plumage coloration and age of definitive hosts.

Among the parasites, some groups that have a limited capacity for locomotion, such as mites and lice, the transmission is challenging to win. These ectoparasites disperse through direct contact between hosts or, in some cases, through phoresy. However, these processes are not well-documented in detail because they are difficult to observe and quantify. In the present study, the patterns of distribution of skin mites and phoretic lice on hippoboscid louse fly Pseudolynchia canariensis sampled from Columba livia were evaluated. The analyzed pigeons were juveniles and adults, with three distinct plumage colors: blue checker, spread, or wild type, and were caught over 24 months. A total of 1,381 hippoboscid flies were collected on 377 hosts. The plumage color did not influence the infestation patterns of louse flies on juvenile and adult pigeons, nor did it influence the infestation patterns of skin mites and phoretic lice on the hippoboscid flies. However, the environmental temperature was directly related to higher prevalence, mean infestation intensity, and phoretic species richness on P. canariensis during the hottest seasons. Furthermore, a higher abundance of phoretic mite eggs, including embryonated eggs, was observed in females of P. canariensis in all seasons.

MOLECULAR CHARACTERIZATION OF ORNITHOSTRONGYLUS QUADRIRADIATUS ISOLATED FROM A RACING PIGEON (COLUMBA LIVIA DOMESTICA).

An infestation of Ornithostrongylus quadriradiatus, has been found in a racing pigeon Columba livia domestica). In addition, this individual had a mild infection with Trichomonas spp. and Eimeria spp. The bird showed symptoms of increased appetite; unformed, dark- green feces; and lack of typical male behavior, but the general condition of this pigeon was good. The pigeon was euthanized because of a permanent inability to fly due to an injury to the wing. A postmortem examination revealed the presence of very small dark-pink twisted nematodes with the morphology of O. quadriradiatus in the small intestine, mainly the duodenum. Histopathology showed segmental inflammatory changes in the intestines caused by both blood-sucking nematodes and Eimeria spp. The 5.8S ribosomal ribonucleic acid polymerase chain reaction amplicons from deoxyribonucleic acid extracted from O. quadriradiatus had 82.40% identity with Australostrongylus victoriensis and 81.20% with Travassostrongylus spp. This is the first molecular characterization of O. quadriradiatus-a relatively rare pigeon parasite.

PHEROMONE COMMUNICATION IN FEATHER-FEEDING WING LICE (INSECTA: PHTHIRAPTERA).

Pheromone communication is central to the life history of insect parasites. Determining how pheromones affect parasite behavior can provide insights into host-parasite interactions and suggest novel avenues for parasite control. Lice infest thousands of bird and mammal species and feed on the host's feathers or blood. Despite the pervasiveness of lice in wild populations and the costs they exact on livestock and poultry industries, little is known about pheromone communication in this diverse group. Here, we test for pheromone communication in the wing lice (Columbicola columbae) of Rock Doves (Columba livia). Wing lice spend the majority of their lives on bird flight feathers where they hide from host preening by inserting their bodies between coarse feather barbs. To feed, wing lice must migrate to bird body regions where they consume the insulating barbs of contour feathers. We first show that wing lice readily form aggregations on flight feathers. Next, using a Y-tube olfactometer, we demonstrate that wing lice use pheromone communication to move toward groups of nearby conspecifics. This pheromone is likely an aggregation pheromone, as wing lice only produce the pheromone when placed on flight feathers. Finally, we found that when forced to choose between groups of male and female lice, male lice move toward male groups and females toward female groups, suggesting the use of multiple pheromones. Ongoing work aims to determine the chemical identity and function of these pheromones.

Parasite exchange and hybridisation at a wild-feral-domestic interface.

Interactions between wild, feral, and domestic animals are of economic and conservation significance. The pigeon Columba livia is a synanthropic species in a feral form, but it also includes the rare Rock Dove. Columba livia is an important player at the wild-domestic interface, acting as a carrier of avian diseases, and the feral form threatens Rock Doves with extinction via hybridisation. Despite its abundance, little is known about drivers of disease prevalence in C. livia, or how disease and hybridisation represent synergistic threats to Rock Doves. We focused on infection by the parasite Trichomonas, first collating prevalence estimates in domestic and free-living populations from relevant studies of C. livia. Second, we characterised variation in the diversity and prevalence of Trichomonas among three C. livia populations in the United Kingdom: a feral, a Rock Dove, and a feral-wild hybrid population. Across multiple continents, free-living pigeons had lower Trichomonas infection than captive conspecifics, but the effect was weak. Environmental factors which could impact Trichomonas infection status did not explain variation in infection among populations. Among the British populations, strain diversity varied, and there was lower parasite prevalence in Rock Doves than feral pigeons. Individual infection status was not explained by the available covariates, including hybrid score and site. The drivers of Trichomonas prevalence are unclear, perhaps due to idiosyncratic local-scale drivers. However, given the population-level variation in both infection prevalence and introgressive hybridisation, the potential combined effects could accelerate the extinction of the Rock Dove. Further study of the synergistic effects of multiple types of biotic interactions at the wild-feral-domestic interface is warranted, especially where vagile, globally distributed and superabundant animals are involved.

DOES PREENING BEHAVIOR REDUCE THE PREVALENCE OF AVIAN FEATHER LICE (PHTHIRAPTERA: ISCHNOCERA)?

Animals defend themselves against parasites in many ways. Defenses, such as physiological immune responses, are capable of clearing some infections. External parasites that do not feed on blood, however, are not controlled by the physiological immune system. Instead, ectoparasites like feather-feeding lice (Phthiraptera: Ischnocera) are primarily controlled by behavioral defenses such as preening. Here we test the hypothesis that birds able to preen are capable of clearing infestations of feather lice. We experimentally manipulated preening ability in a captive population of rock pigeons (Columba livia) that were infested with identical numbers of feather lice (Columbicola columbae or Campanulotes compar or both). We then monitored the feather louse infestations for 42 wk. Birds with impaired preening remained infested throughout the experiment; in contrast, the prevalence of lice on birds that could preen normally decreased by 50%. These data indicate that it is indeed possible for birds to clear themselves of feather lice, and perhaps other ectoparasites, by preening. We note, however, that captive birds spend more time preening than wild birds, and that they are less likely to be reinfested than wild birds. Thus, additional studies are necessary to determine under what circumstances wild birds can clear themselves of ectoparasites by preening.

Morphology and morphometry of Paratanaisia bragai () Freitas, 1959 (Digenea, Eucotylidae) through light and scanning electron microscopy.

Paratanaisia bragai is a digenetic trematode that reaches sexual maturity in the kidney collecting ducts of domestic and wild birds, while the snails Subulina octona and Leptinaria unilamellata serve as its intermediate hosts in Brazil. The present study analyzed the morphology and morphometry of P. bragai. Adult specimens of the parasite were collected from naturally infected Columba livia kidneys, fixed and prepared for observation via bright field and differential interference contrast light microscopy and scanning electron microscopy. The parasite has an elongated and flattened body, with a subterminal oral sucker located at the anterior end of the body, as observed by all techniques used. Staining the parasite with hematoxylin-eosin enabled observation of the pharynx, located posteriorly to the oral sucker, the vitelline glands, which are extra-cecal and extend anteriorly to the pre-ovarian region and later to the median region of the body, and intestinal caeca parallel to the vitelline glands. The presence and functionality of the acetabulum are controversial points in the literature, but it was observed in all specimens analyzed by scanning electron microscopy, with a major diameter of 38.36 ± 6.96 (28.77 - 45.39) and minor diameter of 31.59 ± 7.04 (21.75 - 38.16). Close to the acetabulum, scales were observed in the integument of the parasite. Scales with (1 - 5) blade divisions were identified. In the genital pore, it was possible to see the everted cirrus with rosette shape. The excretory pore (first morphometric record) is dorsal and subterminal, with major diameter of 12.27 ± 9.16 (5.79 - 18.75) and minor diameter of 3.95 ± 1.49 (2.89 - 5.00).

Pigeon mite: An underdiagnosed cause of papular urticaria.

BACKGROUND: Dermanyssus gallinae, known as bird mite, generally lives on nestlings' featherless skin. Humans are accidentally infected, and itchy dermatitis is induced when the mites are unable to use birds' blood. The diagnosis is difficult due to the very small size and rapid movement of the mites, which make them hard to spot. CASE PRESENTATION: A 14-year-old male and his mother were referred to the allergy clinic complaining of a 2-week generalized itchy cutaneous papular lesion, unresponsive to antihista-mines, with the feeling of an insect moving on the surface of the skin. Due to the history of recently hatched pigeons nesting on their balcony and finding very small bugs, diagnosed as D. gallinae, they were instructed to clean the pigeon's nest as the source of these parasites, which successfully solved the problem. CONCLUSION: Bird mite infestation should be considered in the differential diagnosis of recurrent pruritus and urticaria, refractory to conventional treatments. Physicians should be aware of this mite infestation in approach to any patient with papular urticaria.

Low parasite infestations in high densities: The paradox of woodpigeons in urban areas.

The type of habitat occupied by avian populations has a marked effect on the parasitises they host. The growth of cities and urban areas in recent decades has favoured some species of birds adapted to these types of habitats - urban exploiters - although the effects of urbanisation on the parasitism of wildlife are not always well known. This study compares the ectoparasites characteristic of two differentiated populations of woodpigeons, one located in a predominantly urban environment and the other in a rural one. Most of the species found were chewing lice, with Columbicola claviformis and Campanulotes bidentatus being dominant. Despite the higher density of the urban population, woodpigeon individuals were characterised by a lower abundance of chewing lice, as well as the presence of ectoparasites typical of feral pigeons such as Hohorstiella lata and the hippoboscids fly, Pseudolynchia canariensis. Similarly, birds with lower weights showed a higher parasitic load, which became more noticeable in urban woodpigeons. The lower ectoparasite load of urban hosts represents a health advantage compared with rural populations, which could be one of the causes of greater growth and reproductive success in urban populations of woodpigeons.

Evidence for strain-specific virulence of Trichomonas gallinae in African columbiformes.

Infection by parasites or pathogens can have marked physiological impacts on individuals. In birds, infection may affect moult and feather growth, which is an energetically demanding time in the annual cycle. Previous work has suggested a potential link between clinically visible Trichomonas gallinae infection and wing length in turtle doves Streptopelia turtur arriving on breeding grounds. First, T. gallinae infection was characterized in 149 columbids from 5 species, sampled on turtle dove wintering grounds in Senegal during the moulting period, testing whether infection by T. gallinae is linked to moult. Trichomonas gallinae prevalence was 100%, so rather than testing for differences between infected and uninfected birds, we tested for differences in moult progression between birds infected by different T. gallinae strains. Twelve strains of T. gallinae were characterized at the internal transcribed spacer 1 (ITS1)/5.8S/ITS2 region, of which 6 were newly identified within this study. In turtle doves only, evidence for differences in wing length by strain was found, with birds infected by strain Tcl-1 having wings nearly 6 mm longer than those infected with strain GEO. No evidence was found for an effect of strain identity within species on moult progression, but comparisons between infected and uninfected birds should be further investigated in species where prevalence is lower.

Detection and molecular characterization of Trichomonas gallinae recovered from domestic pigeons in Egypt.

Trichomonas gallinae is a protozoan parasite that causes canker in pigeons. Squabs (young pigeons) are frequently infected with T. gallinae and can die because of the infection, while adult pigeons can act as carriers showing no clinical signs. In the present study, 50 squabs, up to 1-month-old, were purchased from pigeon markets in different regions of the Giza governorate, Egypt. Direct wet mount preparations of the oral excretions of the squabs (mouth wash) and Giemsa staining revealed that 64% (32/50) were positive for T. gallinae. Experimental infection of ten squabs with 103 T. gallinae trophozoites/ml resulted in oral lesions on the mouth, tongue, and soft palate, with the presence of yellowish-white nodules (cheese-like) in the oral cavity on the sixth day post-infection in all squabs. A subset of five samples were cultured in modified Diamond's media, their DNA was extracted, and a portion of the ribosomal internal transcribed spacer region (ITS1/5.8S/ITS2) was amplified by polymerase chain reaction (PCR) followed by sequencing. Phylogenetic analysis of the five isolates revealed 64-91% homology with some reference isolates circulating in Egypt and related countries.

Trichomonas gallinae induces heterophil extracellular trap formation in pigeons.

Avian trichomonosis is a worldwide and cross-species epidemic, and the infection in pigeons is particularly severe. Although the disease causes a serious threat to poultry health resulting in significant economic losses, the relationship between Trichomonas gallinae (T. gallinae) and host innate immunity is still not clear. Extracellular traps (ETs) are an innate immunity response to parasitic infections. However, whether host cells can produce ETs after T. gallinae infection has not yet been reported. In the present study, the ability of T. gallinae to induce the production of heterophil extracellular traps (HETs) in pigeons was examined. T. gallinae-induced HETs were observed by scanning electron microscopy (SEM) and the main components of HETs were detected by fluorescence confocal microscopy. Changes in reactive oxygen species (ROS) and lactate dehydrogenase (LDH) were tested during the HETosis. A quantitative analysis of T. gallinae-induced HETs, the role of myeloperoxidase (MPO), store-operated Ca (2+) entry (SOCE), and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in T. gallinae-induced HET formation were conducted by inhibitor assays. The results showed that T. gallinae induced ET formation in pigeon heterophils. ETs consisted of a DNA skeleton, neutrophil elastase (NE), MPO, and Histone3 (H3). T. gallinae-induced HETs formation in a dose- and time-dependent process. The release of T. gallinae-induced HETs depends on MPO, SOCE, and NADPH oxidase. Furthermore, after T. gallinae stimulated pigeon heterophils, ROS production was significantly increased, while no significant differences in the LDH activity were observed.